02058nas a2200229 4500008004100000022001400041245012000055210006900175260010600244300001200350490000700362520113700369653001301506653004101519653002801560653003201588100001901620700002001639700002401659700002101683856012401704 2005 eng d a0022-191000aCharacterization of two midgut proteinases of helicoverpa armigera and their interaction with proteinase inhibitors0 aCharacterization of two midgut proteinases of helicoverpa armige aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLANDbPERGAMON-ELSEVIER SCIENCE LTDcMAY a513-5220 v513 a

Two serine proteinases from the midgut of Helicoverpa armigera have been partially purified and characterized. One proteinase, HGP-1, was capable of hydrolyzing a synthetic substrate of elastase and was inhibited by elastatinal. The second proteinase, HGP-2, was inhibited by a trypsin inhibitor. Molecular weights of HGP-1 and HGP-2 were approximately 26.0 and 29.0 kDa, respectively. Both the proteinases exhibited alkaline pH optima in the range of 10-11. Furthermore, interaction of HGP-1 and HGP-2 with proteinase inhibitors (PIs) from host and non-host plants was studied. HGP-1 was not only insensitive to a PI from chickpea (host) but was also able to degrade it. The same PI from chickpea was able to inhibit over 50% activity of HGP-2. On the contrary, PIs from potato (non-host) showed strong inhibition of both, HGP-1 and HGP-2 and also demonstrated protection of chickpea seed proteins from digestion by both the HGPs. These results could provide important clues in designing strategies for sustainable use of plant PIs in developing insect-tolerant transgenic plants. (C) 2005 Elsevier Ltd. All rights reserved.

10aelastase10aHelicoverpa armigera gut proteinases10ainsect-resistant plants10aplant proteinase inhibitors1 aTelang, M., A.1 aGiri, Ashok, P.1 aSainani, Mohini, N.1 aGupta, Vidya, S. uhttps://library.ncl.res.in/content/characterization-two-midgut-proteinases-helicoverpa-armigera-and-their-interaction-002159nas a2200241 4500008004100000022001400041245006700055210006700122260010600189300001200295490000700307520131200314653001501626653002001641653002501661653002601686653001101712100002801723700002001751700002401771700002101795856010101816 2005 eng d a0965-174800aGene expression patterns of helicoverpa armigera gut proteases0 aGene expression patterns of helicoverpa armigera gut proteases aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLANDbPERGAMON-ELSEVIER SCIENCE LTDcAPR a355-3670 v353 a

Relative quantification of reported gut proteinase cDNAs from Helicoverpa armigera larvae fed on various host plants (cotton, chickpea, pigeonpea, tomato and okra), non-host plant Pis (winged bean, bitter gourd, ground nut, and capsicum) and during larval development has been carried out using semi-quantitative RT-PCR. Five trypsin-like and three chymotrypsin-like proteinases were categorized as insensitive or sensitive to most of the proteinase inhibitors (Pis) and insensitive/sensitive to specific Pis based on their expression analysis. These results were supported by amino acid sequence analysis, where diverged amino acids were observed in the regions, which are reported to be involved in typical trypsin trypsin inhibitor interactions and critical for proteinase inhibitor resistance. Among exopeptidases (five aminopeptidase and three carboxypeptidase), HaAmi4 and HaAmi5 of aminopeptidase and HaCar1 of carboxypeptidase exhibited considerable differential expression. Elastase and cathepsin B-like proteinases were expressed at negligible levels. The proteases identified in the study would be ideal candidates for further interactions studies with protease inhibitors to understand the structural reasons of protease inhibitor insensitivity (c) 2005 Elsevier Ltd. All rights reserved.

10aexpression10agut proteinases10aHelicoverpa armigera10aProteinase inhibitors10aRT-PCR1 aChougule, Nanasaheb, P.1 aGiri, Ashok, P.1 aSainani, Mohini, N.1 aGupta, Vidya, S. uhttps://library.ncl.res.in/content/gene-expression-patterns-helicoverpa-armigera-gut-proteases-002364nas a2200253 4500008004100000022001400041245022000055210006900275260010600344300001400450490000700464520128900471653002301760653002501783653002801808653002301836653001801859653002601877100001801903700002001921700002401941700002101965856012401986 2005 eng d a0031-942200aHigher accumulation of proteinase inhibitors in flowers than leaves and fruits as a possible basis for differential feeding preference of Helicoverpa armigera on tomato (Lycopersicon esculentum Mill, Cv. Dhanashree)0 aHigher accumulation of proteinase inhibitors in flowers than lea aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLANDbPERGAMON-ELSEVIER SCIENCE LTDcNOV a2659-26670 v663 a

Tomato (Lycopersicon esculentum, Mill; cultivar- Dhanashree) proteinase inhibitors (Pis) were tested for their trypsin inhibitory (TT) and Helicoverpa armigera gut proteinases inhibitory (HGPI) activity in different organs of the tomato plants. Analysis of TI and HGPI distribution in various parts of the plant showed that flowers accumulated about 300 and 1000 times higher levels of TI while 700 and 400 times higher levels of HGPI as compared to those in leaves and fruits, respectively. Field observation that H. armigera larvae infest leaves and fruits but not the flowers could be at least partially attributed to the protective role-played by the higher levels of Pis in the flower tissue. Tomato Pis inhibited about 50-80% HGP activity of H. armigera larvae feeding on various host plants including tomato, of larvae exposed to non-host plant Pis and of various larval instars. Tomato Pis were found to be highly stable to insect proteinases wherein incubation of inhibitor with HGP even for 3 h at optimum conditions did not affect inhibitory activity. Bioassay using H. armigera larvae fed on artificial diet containing tomato PIs revealed adverse effect on larval growth, pupae development, adult formation and fecundity. (c) 2005 Elsevier Ltd. All rights reserved.

10aaccumulation of PI10aHelicoverpa armigera10aLycopersicon esculentum10aMidgut proteinases10aPlant defense10aProteinase inhibitors1 aDamle, M., S.1 aGiri, Ashok, P.1 aSainani, Mohini, N.1 aGupta, Vidya, S. uhttps://library.ncl.res.in/content/higher-accumulation-proteinase-inhibitors-flowers-leaves-and-fruits-possible-basis-002418nas a2200265 4500008004100000022001400041245011200055210006900167260007300236300001200309490000900321520145600330653001901786653001901805653002501824653002201849653002501871100002001896700002801916700002001944700001801964700002401982700002102006856012502027 2005 eng d a0304-416500aIn vivo and in vitro effect of capsicum annum proteinase inhibitors on helicoverpa armigera gut proteinases0 aIn vivo and in vitro effect of capsicum annum proteinase inhibit aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcMAR a156-1670 v17223 a

Two proteinase inhibitors (PIs), CapA1 and CapA2, were purified from Capsicum annum Linn. Var. Phule Jyoti leaves and assessed for their in vitro and in vivo activity against Helicoverpa armigera gut proteinases (HGPs). Both the inhibitors exhibited molecular weights of about 12 kDa with inhibitory activity against bovine trypsin and chymotrypsin indicating presence of probable two-inhibitor repeats of PIN II family. CapA1 and CapA2 inhibited 60-80% HGP (azocaseinolytic) activity of fourth instar larvae feeding on various host plants while 45-65% inhibition of HGP activity of various instars (II to VI) larvae reared on artificial diet. The partial purification of HGP isoforms, their characterization with synthetic inhibitors and inhibition by C. annum PIs revealed that most of the trypsin-like activity (68-91%) of HGPs was sensitive to C. annum PIs while 39-85% chymotrypsin-like activity of HGPs was insensitive to these inhibitors. The feeding of C. annum leaf extracts and two purified PIs in various doses to H. armigera larvae for two successive generations through artificial diet demonstrated their potential in inhibiting larval growth and development, delay in pupation period and dramatic reduction in fecundity and fertility. This is the first report-demonstrating efficacy of C. annum PIs against insect gut proteinases as well as larval growth and development of H. armigera. (c) 2004 Elsevier B.V. All rights reserved.

10aCapsicum annum10agut proteinase10aHelicoverpa armigera10ainsect resistance10aProteinase inhibitor1 aTamhane, V., A.1 aChougule, Nanasaheb, P.1 aGiri, Ashok, P.1 aDixit, A., R.1 aSainani, Mohini, N.1 aGupta, Vidya, S. uhttps://library.ncl.res.in/content/vivo-and-vitro-effect-capsicum-annum-proteinase-inhibitors-helicoverpa-armigera-gut-002815nas a2200241 4500008004100000022001400041245014600055210006900201260009000270300001800360490000600378520190300384100001502287700001402302700001402316700001502330700002202345700001802367700002002385700001702405700002502422856012602447 2005 eng d a1475-285900aInactive enzymatic mutant proteins (phosphoglycerate mutase and enolase) as sugar binders for ribulose-1,5-bisphosphate regeneration reactors0 aInactive enzymatic mutant proteins phosphoglycerate mutase and e aMIDDLESEX HOUSE, 34-42 CLEVELAND ST, LONDON W1T 4LB, ENGLANDbBIOMED CENTRAL LTDcFEB aArticle No. 50 v43 a

Background: Carbon dioxide fixation bioprocess in reactors necessitates recycling of D- ribulose1,5-bisphosphate (RuBP) for continuous operation. A radically new close loop of RuBP regenerating reactor design has been proposed that will harbor enzyme-complexes instead of purified enzymes. These reactors will need binders enabling selective capture and release of sugar and intermediate metabolites enabling specific conversions during regeneration. In the current manuscript we describe properties of proteins that will act as potential binders in RuBP regeneration reactors. Results: We demonstrate specific binding of 3-phosphoglycerate (3PGA) and 3-phosphoglyceraldehyde ( 3PGAL) from sugar mixtures by inactive mutant of yeast enzymes phosphoglycerate mutase and enolase. The reversibility in binding with respect to pH and EDTA has also been shown. No chemical conversion of incubated sugars or sugar intermediate metabolites were found by the inactive enzymatic proteins. The dissociation constants for sugar metabolites are in the micromolar range, both proteins showed lower dissociation constant (Kd) for 3-phosphoglycerate ( 655 - 796 mu M) compared to 3-phosphoglyceraldehyde ( 822 - 966 mu M) indicating higher affinity for 3PGA. The proteins did not show binding to glucose, sucrose or fructose within the sensitivity limits of detection. Phosphoglycerate mutase showed slightly lower stability on repeated use than enolase mutants. Conclusions: The sugar and their intermediate metabolite binders may have a useful role in RuBP regeneration reactors. The reversibility of binding with respect to changes in physicochemical factors and stability when subjected to repeated changes in these conditions are expected to make the mutant proteins candidates for in-situ removal of sugar intermediate metabolites for forward driving of specific reactions in enzyme-complex reactors.

1 aDe, Dinesh1 aDutta, D.1 aKundu, M.1 aMahato, S.1 aSchiavone, M., T.1 aChaudhuri, S.1 aGiri, Ashok, P.1 aGupta, Vidya1 aBhattacharya, S., K. uhttps://library.ncl.res.in/content/inactive-enzymatic-mutant-proteins-phosphoglycerate-mutase-and-enolase-sugar-binders-102786nas a2200265 4500008004100000022001400041245014000055210006900195260007400264300001200338490000700350520181600357653001302173653002002186653001502206653002502221653001102246653002502257100001902282700002002301700002502321700002202346700002102368856013102389 2005 eng d a0167-441200aKunitz trypsin inhibitor from chickpea (Cicer arietinum L.) that exerts anti-metabolic effect on podborer (Helicoverpa armigera) larvae0 aKunitz trypsin inhibitor from chickpea Cicer arietinum L that ex aVAN GODEWIJCKSTRAAT 30, 3311 GZ DORDRECHT, NETHERLANDSbSPRINGERcFEB a359-3740 v573 a

Chickpea (Cicer arietinum L.) seeds contain Bowman-Birk proteinase inhibitors, which are ineffective against the digestive proteinases of larvae of the insect pest Helicoverpa armigera. We have identified and purified a low expressing proteinase inhibitor (PI), distinct from the Bowman-Birk Inhibitors and active against H. armigera gut proteinases (HGP), from chickpea seeds. N-terminal sequencing of this HGP inhibitor revealed a sequence similar to reported pea (Pisum sativum) and chickpea alpha-L-fucosidases and also homologous to legume Kunitz inhibitors. The identity was confirmed by matrix assisted laser desorption ionization - time of flight analysis of tryptic peptides and isolation of DNA sequence coding for the mature protein. Available sequence data showed that this protein forms a distinct phylogenetic cluster with Kunitz inhibitors from Glycine max, Medicago truncatula, P. sativum and Canavalia lineata. The isolated coding sequence was cloned into a yeast expression vector and produced as a recombinant protein in Pichia pastoris. alpha-L-fucosidase activity was not detectable in purified or recombinant protein, by solution assays. The recombinant protein did not inhibit chymotrypsin or subtilisin activity but did exhibit stoichiometric inhibition of trypsin, comparable to soybean Kunitz trypsin inhibitor. The recombinant protein exhibited higher inhibition of total HGP activity as compared to soybean kunitz inhibitor, even though it preferentially inhibited HGP-trypsins. H. armigera larvae fed on inhibitor-incorporated artificial diet showed significant reduction in average larval weight after 18 days of feeding demonstrating potent antimetabolic activity. The over-expression of this gene in chickpea could act as an endogenous source of resistance to H. armigera.

10aChickpea10aCicer arietinum10afucosidase10aHelicoverpa armigera10aKunitz10aProteinase inhibitor1 aSrinivasan, A.1 aGiri, Ashok, P.1 aHarsulkar, Abhay, M.1 aGatehouse, J., A.1 aGupta, Vidya, S. uhttps://library.ncl.res.in/content/kunitz-trypsin-inhibitor-chickpea-cicer-arietinum-l-exerts-anti-metabolic-effect-podborer-002480nas a2200253 4500008004100000022001400041245015100055210006900206260010600275300001400381490000700395520148200402653002001884653002501904653002001929653001501949653002501964100001901989700002802008700002002036700002202056700002102078856012702099 2005 eng d a0022-191000aPodborer (Helicoverpa armigera Hubn.) does not show specific adaptations in gut proteinases to dietary Cicer arietinum Kunitz proteinase inhibitor0 aPodborer Helicoverpa armigera Hubn does not show specific adapta aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLANDbPERGAMON-ELSEVIER SCIENCE LTDcNOV a1268-12760 v513 a

We investigated the response of Helicoverpa armigera larvae towards ingestion of Cicer arietinum Kunitz proteinase inhibitor (CaKPI), which caused antagonistic effects on developing H. armigera larvae. CaKPI-degrading proteinases were not detectable in either control or sensitized larvae. There were negligible increases in total proteinase activity, as well as in trypsin-like and chymotrypsin-like activities of H. armigera gut proteinases (HGPs). Decrease in sensitivity of HGPs to inhibition by CaKPI was not observed when the inhibitor was fed Suggesting that the insect had not shown a specific adaptive response to dietary CaKPI. Semi-quantitative reverse transcriptase polymerase chain reaction (Q RT-PCR) analysis showed a general up-regulation of proteases in larvae that ingested CaKPI and a specific regulation of individual transcripts was not observed. CaKPI had maximum inhibitory activity against HGP derived from fourth instar larvae. CaKPI was equally potent in inhibition of HGPs derived from larvae fed on different host plants, as well as various proteinase inhibitors (Pis) to which larval adaptation was previously reported. The lack of larval response to CaKPI was attributable to the atypical active site sequence and inhibitory activity of CaKPI and/or to the pre-adaptation of H. armigera larvae due to the constant exposure to basal levels of CaKPI in chickpea seeds or a chickpea seed-based diet. (c) 2005 Elsevier Ltd. All rights reserved.

10aCicer arietinum10aHelicoverpa armigera10ainsect response10aproteinase10aProteinase inhibitor1 aSrinivasan, A.1 aChougule, Nanasaheb, P.1 aGiri, Ashok, P.1 aGatehouse, J., A.1 aGupta, Vidya, S. uhttps://library.ncl.res.in/content/podborer-helicoverpa-armigera-hubn-does-not-show-specific-adaptations-gut-proteinases-002134nas a2200265 4500008004100000022001400041245013000055210006900185260007900254300001200333490000800345520113900353653002701492653001401519653001601533653002601549653003201575653001601607100002501623700002801648700002301676700002001699700002001719856012901739 2005 eng d a1096-495900aUnraveling biochemical properties of cockroach (Periplaneta americana) proteinases with a gel X-ray film contact print method0 aUnraveling biochemical properties of cockroach Periplaneta ameri a360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USAbELSEVIER SCIENCE INCcJUL a261-2660 v1413 a

Eleven proteinase activity bands were detected in American cockroach (Periplaneta americana) gut. These were partially purified and characterized using a gel X-ray film contact print method. Cockroach gut proteinases (CGPs) show activity over a broad range of pH with maximum activity between pH 6 and 10, and optimal activity at 50-70 degrees C. CGPs were partially purified by preparative gel electrophoresis and analyzed using synthetic substrates and inhibitors. Four of the proteases exhibited chymotrypsin-like (C1 to C4) activity and seven trypsin-like (T I to T7) activity. Accuracy of the gel X-ray film contact print method is confirmed by including bovine chymotrypsin in CGP analysis. Inhibition of CGPs with different plant proteinaceous proteinase inhibitors allowed identification of potential CGP inhibitors. Our results imply that presence of several CGP activity bands, and their stability and activity over a broad pH and temperature range might contribute to adaptation of P americana to extreme environmental conditions and the polyphagous nature of the species. (c) 2005 Elsevier Inc. All rights reserved.

10aactivity visualization10acockroach10agut enzymes10aPeriplaneta americana10aplant proteinase inhibitors10aProteinases1 aHivrale, Vandana, K.1 aChougule, Nanasaheb, P.1 aChhabda, Pavan, J.1 aGiri, Ashok, P.1 aKachole, M., S. uhttps://library.ncl.res.in/content/unraveling-biochemical-properties-cockroach-periplaneta-americana-proteinases-gel-x-ray-002480nas a2200277 4500008004100000022001400041245015400055210006900209260008900278300001200367490000700379520142000386653001401806653001301820653002001833653003501853653002301888653001801911100002501929700002501954700002001979700002401999700002802023700002102051856013002072 2006 eng d a0885-576500aDifferentially expressed gene transcripts in roots of resistant and susceptible chickpea plant (Cicer arietinum L.) upon Fusarium oxysporum infection0 aDifferentially expressed gene transcripts in roots of resistant  a24-28 OVAL RD, LONDON NW1 7DX, ENGLANDbACADEMIC PRESS LTD ELSEVIER SCIENCE LTDcAPR a176-1880 v683 a

Differentially expressed genes in chickpea, (Cicer arietinum L.) during root infection by Fusarium oxysportan f sp. ciceri Racel, were identified using cDNA-RAPD and cDNA-AFLP approaches. The former employed decamer primers on cDNA template and revealed nine differentially expressed transcripts in the resistant-infected chickpea variety. Among the 2000 transcript-derived fragments (TDFs) screened by cDNA-AFLP, 273 were differentially expressed in chickpea roots during Fusaritan infection. Only 13.65% of the TDFs were differentially regulated during pathogen challenge, while the other 86% were expressed non-differentially during the process of pathogen infection in chickpea roots. Nineteen TDFs, which expressed differentially in the resistant-infected chickpea variety were cloned and sequenced. Two of these TDFs were similar to transcription factors like WRKY proteins and 14-3-3 proteins, while three TDFs represented the NBS-LRR-type gene sequences. Two TDFs had sequence identity to genes known to have function in defense. The RAPID TDF CaFRi60 showed sequence identity to gamma-glutamyl-cysteine synthetase. Among the TDFs examined by cDNA-AFLP, 19 were confirmed by Reverse Northern blot to be differentially expressed. The data confirms the effectiveness of the cDNA-AFLP technique in detecting differentially expressed genes during pathogenesis. (c) 2006 Elsevier Ltd. All rights reserved.

10acDNA-AFLP10aChickpea10aCicer arietinum10adifferentially expressed genes10afusarium oxysporum10aTranscriptome1 aNimbalkar, Suhas, B.1 aHarsulkar, Abhay, M.1 aGiri, Ashok, P.1 aSainani, Mohini, N.1 aFranceschi, Vincent, R.1 aGupta, Vidya, S. uhttps://library.ncl.res.in/content/differentially-expressed-gene-transcripts-roots-resistant-and-susceptible-chickpea-plant-002961nas a2200217 4500008004100000022001400041245017900055210006900234260008000303300001400383490000800397520206000405100002002465700002202485700002602507700002202533700002002555700001702575700002102592856013002613 2006 eng d a0032-088900aMolecular interactions between the specialist herbivore Manduca sexta (Lepidoptera, Sphingidae) and its natural host Nicotiana attenuata. VII. changes in the plant's proteome0 aMolecular interactions between the specialist herbivore Manduca  a15501 MONONA DRIVE, ROCKVILLE, MD 20855 USAbAMER SOC PLANT BIOLOGISTScDEC a1621-16410 v1423 a

When Manduca sexta attacks Nicotiana attenuata, fatty acid-amino acid conjugates (FACs) in the larvae's oral secretions (OS) are introduced into feeding wounds. These FACs trigger a transcriptional response that is similar to the response induced by insect damage. Using two-dimensional gel electrophoresis, matrix-assisted laser desorption ionization-time of flight, and liquid chromatography-tandem mass spectrometry, we characterized the proteins in phenolic extracts and in a nuclear fraction of leaves elicited by larval attack, and/or in leaves wounded and treated with OS, FAC-free OS, and synthetic FACs. Phenolic extracts yielded approximately 600 protein spots, many of which were altered by elicitation, whereas nuclear protein fractions yielded approximately 100 spots, most of which were unchanged by elicitation. Reproducible elicitor-induced changes in 90 spots were characterized. In general, proteins that increased were involved in primary metabolism, defense, and transcriptional and translational regulation; those that decreased were involved in photosynthesis. Like the transcriptional defense responses, proteomic changes were strongly elicited by the FACs in OS. A semiquantitative reverse transcription-PCR approach based on peptide sequences was used to compare transcript and protein accumulation patterns for 17 candidate proteins. In six cases the patterns of elicited transcript accumulation were consistent with those of elicited protein accumulation. Functional analysis of one of the identified proteins involved in photosynthesis, RuBPCase activase, was accomplished by virus-induced gene silencing. Plants with decreased levels of RuBPCase activase protein had reduced photosynthetic rates and RuBPCase activity, and less biomass, responses consistent with those of herbivore-attacked plants. We conclude that the response of the plant's proteome to herbivore elicitation is complex, and integrated transcriptome-proteome-metabolome analysis is required to fully understand this ubiquitous ecological interaction.

1 aGiri, Ashok, P.1 aWuensche, Hendrik1 aMitra, Sirsha, Sribas1 aZavala, Jorge, A.1 aMuck, Alexander1 aSvatos, Ales1 aBaldwin, Ian, T. uhttps://library.ncl.res.in/content/molecular-interactions-between-specialist-herbivore-manduca-sexta-lepidoptera-sphingidae-002554nas a2200253 4500008004100000022001400041245014600055210006900201260007300270300001000343490000800353520157900361653002201940653001501962653002601977653002002003653003802023653001502061100002802076700002002104700002402124700002102148856013102169 2007 eng d a0378-111900aDiverse forms of pin-II family proteinase inhibitors from capsicum annuum adversely affect the growth and development of helicoverpa armigera0 aDiverse forms of pinII family proteinase inhibitors from capsicu aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcNOV a29-380 v4033 a

Novel forms of Pin-II type proteinase inhibitor (PIs) cDNAs (CanPIs) having three or four inhibitory repeat domains (IRD) were isolated from the developing green fruits of Capsicum annuum. Deduced amino acid (aa) sequences of the CanPIs showed up to 15% sequence divergence among each other or reported inhibitors (CanPI-1 AF039398, CanPI-2 AF221097). Amino acid sequence analysis of these CanPIs revealed that three IRD Pls have trypsin inhibitory sites, while four IRD CanPIs have both trypsin and chymotrypsin inhibitory sites. Four CanPIs, two having three IRD (CanPI-3 AY986465 and CanPI-5 DQ005912) and two having four IRD (CanPI-7 DQ005913 and CanPI-9 DQ005915), were cloned in Pichia pastoris to express recombinant CanPIs. Recombinant CanPIs inhibited 90% of bovine trypsin (TI), while chymotrypsin inhibition (0) varied with the number of chymotrypsin inhibitory sites in the CanPIs. Recombinant inhibitors inhibited over 70% of the gut proteinase activity of Helicoverpa armigera. H. armigera larvae fed on recombinant CanPIs individually incorporated into artificial diet, showed 35% mortality; in addition, weight gain in H. armigera larvae and pupae was severely reduced compared to controls. Of the four CanPIs, CanPI-7, which has two sites for TI and CI, was the only one to have a consistently antagonistic effect on H. armigera growth and development. We conclude that among the four recombinant Pls tested, CanPIs containing diverse IRDs are best suited for developing insect-resistant transgenic plants. (c) 2007 Elsevier B.V. All rights reserved.

10ainsect resistance10aIP repeats10aPI-insect interaction10aPichia pastoris10apotato type 2 inhibitors (Pot-II)10aSolanaceae1 aTamhane, Vaijayantl, A.1 aGiri, Ashok, P.1 aSainani, Mohini, N.1 aGupta, Vidya, S. uhttps://library.ncl.res.in/content/diverse-forms-pin-ii-family-proteinase-inhibitors-capsicum-annuum-adversely-affect-growth-001773nas a2200265 4500008004100000022001400041245009300055210006900148260008800217300001400305490000700319520078000326653002401106653003301130653002101163653003401184100002201218700002601240700002001266700002301286700002401309700002601333700002101359856012701380 2007 eng d a0011-389100aGenetic diversity analysis of mango cultivars using inter simple sequence repeat markers0 aGenetic diversity analysis of mango cultivars using inter simple aC V RAMAN AVENUE, PO BOX 8005, BANGALORE 560 080, INDIAbCURRENT SCIENCE ASSOCcOCT a1135-11410 v933 a

With 33 polymorphic Inter Simple Sequence Repeat (ISSR) markers, genetic diversity among 70 mango cultivars and a distant outgroup was analysed. Clustering was done using Dice and Jaccard coefficients with bootstrapping. Multivariate analysis was conducted using the Euclidean distances. Among the total 420 bands, 408 were polymorphic. Probability that any ISSR fragment is shared by two different cultivars was calculated to be 2.54 x 10(-1). Non-Indian mango cultivars were found genetically diverged from Indian mango germ pool. No separation was observed between North Indian and South Indian cultivars. Twelve different cultivar-specific bands were detected for six cultivars, which approved the effectiveness of ISSR markers in mango genetic diversity analysis.

10aDNA finger printing10ainter simple sequence repeat10aMangifera indica10aprinciple coordinate analysis1 aPandit, Sagar, S.1 aMitra, Sirsha, Sribas1 aGiri, Ashok, P.1 aPujari, Keshav, H.1 aPatil, Bhimarao, P.1 aJambhale, Narayan, D.1 aGupta, Vidya, S. uhttps://library.ncl.res.in/content/genetic-diversity-analysis-mango-cultivars-using-inter-simple-sequence-repeat-markers-002167nas a2200253 4500008004100000022001400041245016100055210006900216260010600285300001000391490000700401520116500408653002501573653001701598653001001615653002501625653001301650653002701663100002701690700002601717700002001743700002701763856012301790 2007 eng d a1226-923900aQuick method for isolating RNA from raw and ripe fleshy fruits as well as for co-isolating DNA and RNA from polysaccharide- and polyphenol-rich leaf tissues0 aQuick method for isolating RNA from raw and ripe fleshy fruits a aCATHOLIC UNIV KOREA, DEPT LIFE SCIENCES, PUCHON 420-743, SOUTH KOREAbBOTANICAL SOCIETY OF KOREAcFEB a60-640 v503 a

Qualitative and quantitative changes in its chemical composition make it difficult to use any single procedure for isolating good-quality RNA from fruits at various ripening stages. Although the CTAB method has eliminated some specific problems, e.g., low pH in raw fruit or high levels of polysaccharides, oligosaccharides and phenolics in raw and ripe fruits, the total time required is long and unsuitable for high throughput. Here, we successfully modified this CTAB protocol to isolate good-quality RNA from (i) fleshy fruits, especially raw and ripe mangos; (ii) the leaves of a succulent air plant; and (iii) an oligosaccharide-rich onion epidermis. This RNA proved useful for downstream transcriptomic applications, where RT-PCR followed by RACE yielded the complete open reading frame of the (mango) terpene synthase gene. We also extended the utility of this protocol to co-isolate good-quality genomic DNA from the supernatant that remained after RNA precipitation. This preparation was useful for the arbitrary primer multilocus amplification of genomic DNA as well as for single locus diversity marker amplifications of the ctDNA and mtDNA.

10aDNA-RNA co-isolation10afleshy fruit10aMango10aoligosaccharide-rich10aRipening10atranscriptomic studies1 aPandit, Sagar, Subhash1 aMitra, Sirsha, Sribas1 aGiri, Ashok, P.1 aGupta, Vidya, Shrikant uhttps://library.ncl.res.in/content/quick-method-isolating-rna-raw-and-ripe-fleshy-fruits-well-co-isolating-dna-and-rna02352nas a2200181 4500008004100000022001400041245015300055210006900208260008500277300001000362490000600372520157500378100002201953700002001975700002501995700002102020856012902041 2008 eng d a1932-620300aDigestive duet: midgut digestive proteinases of manduca sexta ingesting nicotiana attenuata with manipulated trypsin proteinase inhibitor expression0 aDigestive duet midgut digestive proteinases of manduca sexta ing a185 BERRY ST, STE 1300, SAN FRANCISCO, CA 94107 USAbPUBLIC LIBRARY SCIENCEcAPR ae20080 v33 a

Background: The defensive effect of endogenous trypsin proteinase inhibitors (NaTPIs) on the herbivore Manduca sexta was demonstrated by genetically altering NaTPI production in M. sexta's host plant, Nicotiana attenuata. To understand how this defense works, we studied the effects of NaTPI on M. sexta gut proteinase activity levels in different larval instars of caterpillars feeding freely on untransformed and transformed plants. Methodology/Principal Findings: Second and third instars larvae that fed on NaTPI-producing (WT) genotypes were lighter and had less gut proteinase activity compared to those that fed on genotypes with either little or no NaTPI activity. Unexpectedly, NaTPI activity in vitro assays not only inhibited the trypsin sensitive fraction of gut proteinase activity but also halved the NaTPI-insensitive fraction in third-instar larvae. Unable to degrade NaTPI, larvae apparently lacked the means to adapt to NaTPI in their diet. However, caterpillars recovered at least part of their gut proteinase activity when they were transferred from NaTPI-producing host plants to NaTPI-free host plants. In addition extracts of basal leaves inhibited more gut proteinase activity than did extracts of middle stem leaves with the same protein content. Conclusions/Significance: Although larvae can minimize the effects of high NaTPI levels by feeding on leaves with high protein and low NaTPI activity, the host plant's endogenous NaTPIs remain an effective defense against M. sexta, inhibiting gut proteinase and affecting larval performance.

1 aZavala, Jorge, A.1 aGiri, Ashok, P.1 aJongsma, Maarten, A.1 aBaldwin, Ian, T. uhttps://library.ncl.res.in/content/digestive-duet-midgut-digestive-proteinases-manduca-sexta-ingesting-nicotiana-attenuata-002339nas a2200313 4500008004100000022001400041245009400055210006900149260009800218300001400316490000700330520125300337653001101590653002301601653002101624653001901645653002301664653001001687100002201697700002201719700002501741700002001766700002301786700002501809700002201834700002501856700002101881856012301902 2009 eng d a0022-514200aChanges in volatile composition during fruit development and ripening of `Alphonso' mango0 aChanges in volatile composition during fruit development and rip aTHE ATRIUM, SOUTHERN GATE, CHICHESTER PO19 8SQ, W SUSSEX, ENGLANDbJOHN WILEY & SONS LTDcSEP a2071-20810 v893 a

BACKGROUND: Volatile blends of five developing and five ripening stages of mango (Mangifera indica L. cv. Alphonso) were investigated along with those of flowers and leaves. Raw and ripe fruits of cv. Sabia were also used for comparison. RESULTS: A total of 55 volatiles belonging to various chemical classes such as aldehydes, alcohols, mono- and sesquiterpene hydrocarbons, lactones and furanones were identified. In all Alphonso tissues monoterpenes quantitatively dominated, with 57-99% contribution; in particular, (Z)-ocimene was found in the highest amount. Ripeness was characterized by the de novo appearance of lactones and furanones in the blend of monoterpenes. Sabja was distinguished by the abundance of monoterpene hydrocarbons in the raw fruit, and that of sesquiterpene hydrocarbons and their derivatives in the ripe stage. CONCLUSION: Various stages of the Alphonso fruit during transition from flower to ripe fruit are characterized by unique volatile signatures that are distinguished from each other by the qualitative and quantitative appearance of different volatile compounds. Thus volatiles can be highly informative markers while studying the development and ripening of mango. (C) 2009 Society of Chemical Industry

10aDeogad10aGas chromatography10aMangifera indica10amango products10aripening indicator10aSabja1 aPandit, Sagar, S.1 aKulkarni, Ram, S.1 aChidley, Hemangi, G.1 aGiri, Ashok, P.1 aPujari, Keshav, H.1 aKoellner, Tobias, G.1 aDegenhardt, Joerg1 aGershenzon, Jonathan1 aGupta, Vidya, S. uhttps://library.ncl.res.in/content/changes-volatile-composition-during-fruit-development-and-ripening-alphonso-mango-002069nas a2200265 4500008004100000022001400041245006900055210006900124260009900193300001200292490000800304520117400312653001201486653001001498653002101508653002601529653001501555100002201570700002501592700002201617700002301639700002001662700002101682856010001703 2009 eng d a0308-814600aCultivar relationships in mango based on fruit volatile profiles0 aCultivar relationships in mango based on fruit volatile profiles aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLANDbELSEVIER SCI LTDcMAY a363-3720 v1143 a

Aroma volatiles of mango (Mangifera indica L.) determine consumer acceptability and influence selection, whilst breeding. To assess their variety, composition and possible impact on cultivar relationships, volatile blends of 22 Indian and five non-Indian cultivars were investigated using solvent extraction and gas chromatography. Totally 84 volatiles belonging to various chemical classes were detected. Based on the Cumulative Occurrence of members of these classes, cultivars were grouped as monoterpene or sesquiterpene dominant. a-Pinene, beta-myrcene and beta-caryophyllene were found in all 27 cultivars. For ordination, common compounds with high (relative) concentration provided quantitative characters, whereas the rare and lesser ones provided qualitative (binary) characters; non-Indian cultivars separated from Indian ones but displayed close relations within their groups. In conclusion, mango flavour is dominated qualitatively as well as quantitatively by terpene hydrocarbons; ancient selection of mango cultivars is hinted to be driven by different aroma characters in different parts of the world. (C) 2008 Elsevier Ltd. All rights reserved.

10aFlavour10aGC-MS10aMangifera indica10aMultivariate analysis10aOrdination1 aPandit, Sagar, S.1 aChidley, Hemangi, G.1 aKulkarni, Ram, S.1 aPujari, Keshav, H.1 aGiri, Ashok, P.1 aGupta, Vidya, S. uhttps://library.ncl.res.in/content/cultivar-relationships-mango-based-fruit-volatile-profiles-002751nas a2200277 4500008004100000022001400041245012500055210006900180260006400249300001200313490000800325520173500333653000902068653003702077653003202114653000902146653001302155653002102168653003002189653004702219100002002266700002002286700002002306700001702326856013002343 2009 eng d a0027-551400aIdentification of Indian pathogenic races of fusarium oxysporum f. sp ciceris with gene specific, ITS and random markers0 aIdentification of Indian pathogenic races of fusarium oxysporum  a810 E 10TH ST, LAWRENCE, KS 66044 USAbALLEN PRESS INCcAUG a484-4950 v1013 a

In this study we demonstrate the synergistic use of gene-specific markers, ITS-RFLP, ISSR and AFLP for distinguishing Indian F. oxysporum f. sp. ciceris races. We also report for the first time that F. oxysporum f. sp. ciceris race 3, a wilt pathogen of chickpea in India, is actually E proliferatum based on phylogenetic analysis with EF-1 alpha sequence data. E oxysporum f. sp. ciceris races 1, 2 and 4 were easily distinguished from ``race 3'' (F. proliferatum) by PCR amplification with oligonucleotides designed from conserved regions of Hop78 transposon (Hop 78), cutinase (Cut), desaturase (Dst). F oxysporum f. sp. ciceris race 4 was distinguished with the xylanase 3 (xyl3) gene by absence of amplification product only in this race. The Xyl3 amplified-DNA fragment isolated and sequenced from E oxysporum f. sp. ciceris race I was similar to the F-xylanase (Xyl3) gene of E oxysporum f. sp. lycopersici. A TELD motif, which is characteristic of the F-xylanases family, was detected within the deduced amino acid sequence of F. oxysporum f. sp. ciceris. Similarly the F oxysporum f. sp. ciceris Hop78 DNA fragment, which identified ``race 3'' (E proliferatum), was homologous to the Hop78 transposon of E oxysporum f. sp. melonis, including the 100 amino acid conserved domain and the characteristic CCHC motif. The internal transcribed spacer region-restriction fragment length polymorphism (ITS-RFLP) approach along with intersimple sequence repeat (ISSR) method also differentiated ``race 3'' (F. proliferatum). Races 1 and 2 were identified by unique AFLP patterns. Sequence characterization of race-specific AFLP products revealed significant homologies of these sequences with metabolically important genes.

10aAFLP10aF. oxysporum f. sp ciceris races10agene specific amplification10aISSR10aITS-RFLP10arace delineation10asequence characterization10atranslation elongation factor (EF-1 alpha)1 aGurjar, Gayatri1 aBarve, Maneesha1 aGiri, Ashok, P.1 aGupta, Vidya uhttps://library.ncl.res.in/content/identification-indian-pathogenic-races-fusarium-oxysporum-f-sp-ciceris-gene-specific-its-002551nas a2200253 4500008004100000022001400041245009100055210006900146260010600215300001200321490000700333520162700340653002301967653000901990653003201999653001902031653001602050100002402066700002202090700002802112700002102140700002002161856011602181 2009 eng d a0022-191000aResponses of midgut amylases of Helicoverpa armigera to feeding on various host plants0 aResponses of midgut amylases of Helicoverpa armigera to feeding  aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLANDbPERGAMON-ELSEVIER SCIENCE LTDcAUG a663-6700 v553 a

Midgut digestive amylases and proteinases of Helicoverpa armigera, a polyphagous and devastating insect pest of economic importance have been studied. We also identified the potential of a sorghum amylase inhibitor against H. armigera midgut amylase. Amylase activities were detected in all the larval instars, pupae, moths and eggs; early instars had lower amylase levels which steadily increased up to the sixth larval instar. Qualitative and quantitative differences in midgut amylases of H. amigera upon feeding on natural and artificial diets were evident. Natural diets were categorized as one or more members of legumes, vegetables, flowers and cereals belonging to different plant families. Amylase activity and isoform patterns varied depending on host plant and/or artificial diet. Artificial diet-fed H. armigera larvae had comparatively high amylase activity and several unique amylase isoforms. Correlation of amylase and proteinase activities of H. armigera with the protein and carbohydrate content of various diets suggested that H. armigera regulates the levels of these digestive enzymes in response to macromolecular composition of the diet. These adjustments in the digestive enzymes of H. armigera may be to obtain better nourishment from the diet and avoid toxicity due to nutritional imbalance. H. armigera, a generalist feeder experiences a great degree of nutritional heterogeneity in its diet. An investigation of the differences in enzyme levels in response to macronutrient balance and imbalance highlight their importance in insect nutrition. (C) 2009 Elsevier Ltd. All rights reserved.

10aAmylase inhibitors10aDiet10aHelicoverpa armigera larvae10aMidgut amylase10aProteinases1 aKotkar, Hemlata, M.1 aSarate, Priya, J.1 aTamhane, Vaijayanti, A.1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/responses-midgut-amylases-helicoverpa-armigera-feeding-various-host-plants-002542nas a2200253 4500008004100000022001400041245011400055210006900169260007300238300001000311490000800321520164500329653001001974653002001984653002002004653001102024653001802035653002502053100002802078700002002106700001702126700002102143856012402164 2009 eng d a0378-111900aSpatial and temporal expression patterns of diverse Pin-II proteinase inhibitor genes in Capsicum annuum Linn0 aSpatial and temporal expression patterns of diverse PinII protei aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcAUG a88-980 v4423 a

Pin-II type proteinase inhibitor (PI) genes were cloned from fruit and stem tissues of Capsicum annuum L var Phule Jyoti using primers designed from reported CanPI gene sequence (AF039398). In total, 21 novel CanPIs, members of the Pin-II PI family, were identified in the study, with three isoforms of 1-inhibitory repeat domain (IRD), eight isoforms of 2-IRD), three isoforms of 3-IRD, five isoforms of 4-IRD and two partial CanPI sequences. Most of the sequences showed variation (2 to 20%) in the deduced AA sequences which were pronounced close to the reactive site loop. Expression patterns of CanPIs in the fruit and stem tissues of mature C. annuum plants were shown to vary qualitatively and quantitatively using semi-quantitative RT-PCR expression analysis. In the fruit tissue. CanPIs with different IRDs (from 1 to 4) were expressed simultaneously. In stem tissue, 1- and 2-IRD CanPIs were strongly expressed along moderate expression of 3- and 4-IRD genes. Analysis of CanPI protein activity showed a range of active forms across the tissues. CanPI expression was differentially up-regulated upon wounding and insect attack. Although infestation by aphids (Myzus persicae) and lepidopteran pests (Spodoptera litura) specifically induced 4-IRD CanPIs, virus-infected leaves did not affect CanPI expression. Analysis of CanPI protein activity indicated that the up-regulation in CanPI expression was not always correlated with increase in PI activity. Our results demonstrated that CanPI expression is regulated spatially, temporally as well as qualitatively and quantitatively. (C) 2009 Elsevier B.V. All rights reserved.

10aCanPI10aCapsicum annuum10aEndogenous role10aPin-II10aPlant defense10aProteinase inhibitor1 aTamhane, Vaijayanti, A.1 aGiri, Ashok, P.1 aKumar, Pavan1 aGupta, Vidya, S. uhttps://library.ncl.res.in/content/spatial-and-temporal-expression-patterns-diverse-pin-ii-proteinase-inhibitor-genes-002302nas a2200265 4500008004100000022001400041245007900055210006900134260007300203300001000276490000800286520133100294653002501625653003001650653003701680653003501717653003201752100002301784700002001807700002401827700002101851700002301872700002801895856011301923 2009 eng d a0378-111900aWinged bean chymotrypsin inhibitors retard growth of Helicoverpla anmigera0 aWinged bean chymotrypsin inhibitors retard growth of Helicoverpl aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcFEB a80-850 v4313 a

Two putative Kunitz-type chymotrypsin inhibitor genes (WC12 and WC15) were isolated from winged bean (Psophocarpus tetragonolobus (L.) DC). While WC12 has previously been characterized, WC15 represents a new member of the WC1 family. WC15 was exclusively expressed in winged bean seeds. Theoretical translation of both the genes resulted into polypeptides of 207 amino acids with 86% sequence similarity. The polypeptide sequences contain four half-cysteine residues and a well-conserved Leu(65)-Ser(66) reactive site, typical for chymotrypsin inhibitors. WC15 and WC12 were expressed in Pichia pastoris and the recombinant proteins were assayed against various proteinases. Both the inhibitors strongly inhibited commercially available bovine chymotrypsin. More importantly, gut proteinases of Helicoverpa armigera larvae that damage many important crop plants, were inhibited by WC12 and WC15. In addition, both proteinase inhibitors demonstrated significant reduction of growth of H. armigera larvae after feeding on inhibitor incorporated artificial diets. The inhibitory effects of WC12 and WC15 on activity of proteinases and larval growth make these proteins and their genes promising candidates for enhancing plant defense against H. armigera using transgenic plants. (C) 2008 Elsevier B.V. All rights reserved.

10aAntimetabolic effect10aDigestive gut proteinases10aKunitz-type proteinase inhibitor10aOrgan-specific mRNA expression10aPsophocarpus tetragonolobus1 aTelang, Manasi, A.1 aGiri, Ashok, P.1 aPyati, Prashant, S.1 aGupta, Vidya, S.1 aTegeder, Mechthild1 aFranceschi, Vincent, R. uhttps://library.ncl.res.in/content/winged-bean-chymotrypsin-inhibitors-retard-growth-helicoverpla-anmigera-002812nas a2200265 4500008004100000022001400041245009300055210006900148260010300217300001200320490000700332520183900339653002102178653002002199653002302219653002702242100002202269700002202291700002002313700002502333700002202358700002502380700002102405856012002426 2010 eng d a0981-942800aExpression profiling of various genes during the fruit development and ripening of mango0 aExpression profiling of various genes during the fruit developme a23 RUE LINOIS, 75724 PARIS, FRANCEbELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIERcJUN a426-4330 v483 a

Mango (Mangifera indica L. cv. Alphonso) development and ripening are the programmed processes; conventional indices and volatile markers help to determine agronomically important stages of fruit life (fruit-setting, harvesting maturity and ripening climacteric). However, more and precise markers are required to understand this programming; apparently, fruit's transcriptome can be a good source of such markers. Therefore, we isolated 18 genes related to the physiology and biochemistry of the fruit and profiled their expression in developing and ripening fruits, flowers and leaves of mango using relative quantitation PCR. In most of the tissues, genes related to primary metabolism, abiotic stress, ethylene response and protein turnover showed high expression as compared to that of the genes related to flavor production. Metallothionin and/or ethylene-response transcription factor showed highest level of transcript abundance in all the tissues. Expressions of mono- and sesquiterpene synthases and 14-3-3 lowered during ripening; whereas, that of lipoxygenase, ethylene-response factor and ubiquitin-protein ligase increased during ripening. Based on these expression profiles, flower showed better positive correlation with developing and ripening fruits than leaf. Most of the genes showed their least expression on the second day of harvest, suggesting that harvesting signals significantly affect the fruit metabolism. Important stages in the fruit life were clearly indicated by the significant changes in the expression levels of various genes. These indications complemented those from the previous analyses of fruit development, ripening and volatile emission, revealing the harmony between physiological, biochemical and molecular activities of the fruit. (C) 2010 Elsevier Masson SAS. All rights reserved.

10aMangifera indica10aStress response10aTerpene metabolism10aTranscriptome analysis1 aPandit, Sagar, S.1 aKulkarni, Ram, S.1 aGiri, Ashok, P.1 aKoellner, Tobias, G.1 aDegenhardt, Joerg1 aGershenzon, Jonathan1 aGupta, Vidya, S. uhttps://library.ncl.res.in/content/expression-profiling-various-genes-during-fruit-development-and-ripening-mango-003017nas a2200277 4500008004100000022001400041245013800055210006900193260007700262300001400339490000700353520197800360653001002338653003002348653003402378653001102412653002102423653002902444100001902473700002802492700002102520700002502541700002102566700002002587856013202607 2010 eng d a1615-985300aInteraction of recombinant CanPIs with helicoverpa armigera gut proteases reveals their processing patterns, stability and efficiency0 aInteraction of recombinant CanPIs with helicoverpa armigera gut  aPO BOX 10 11 61, D-69451 WEINHEIM, GERMANYbWILEY-V C H VERLAG GMBHcJUL a2845-28570 v103 a

Six diverse representative Capsicum annuum (common name: hot pepper; Solanaceae) protease inhibitor genes, viz CanPI-5, -7, -13, -15, -19, and 22 comprising 1-4 inhibitory repeat domains (IRDs), were cloned and expressed in Pichia pastoris. The recombinant proteins were evaluated for their interactions with bovine trypsin, chymotrypsin, and Helicoverpa armigera gut proteases (HGP) using electrophoretic (native and denaturing) and mass spectrometric (MALDI-TOF-MS in combination with intensity fading assays) techniques. These techniques allow qualitative and semiquantitative analysis of multiple and processed IRDs of purified recombinant Capsicum annuum proteinase inhibitor (rCanPI) proteins. rCanPIs showed over 90% trypsin inhibition, varying chymotrypsin inhibition depending on the number of respective IRDs and over 60% inhibition of total HGP. rCanPI-15 that has only one IRD showed exceptionally low inhibition of these proteases. Interaction studies of rCanPIs with proteases using intensity fading-MALDI-TOF-MS revealed gradual processing of multi-IRD rCanPIs into single IRD forms by the action of HGP at the linker region, unlike their interactions with trypsin and chymotrypsin. Intensity fading-MALDI-TOF-MS assay showed that CanPI-13 and -15, possessing single IRD and expressed predominantly in stem tissue are degraded by HGP; indicating their function other than defense. In vitro and in vivo studies on rCanPI-5 and -7 showed maximum inhibition of HGP isoforms and their processed IRDs were also found to be stable in the presence of HGP. Even single amino acid variations in IRDs were found to change the HGP specificity like in the case of HGP-8 inhibited only by IRD-12. The presence of active PI in insect gut might be responsible for changed HGP profile. rCanPI-5 and -7 enhanced HGP-7, reduced HGP-4, -5, -10 expression and new protease isoforms were induced. These results signify isoform complexity in plant PIs and insect proteases.

10aCanPI10aH. armigera gut proteases10aIntensity fading MALDI-TOF-MS10aPin-II10aPlant proteomics10aPlant-insect interaction1 aMishra, Manasi1 aTamhane, Vaijayanti, A.1 aKhandelwal, Neha1 aKulkarni, Mahesh, J.1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/interaction-recombinant-canpis-helicoverpa-armigera-gut-proteases-reveals-their-processing-002324nas a2200253 4500008004100000022001400041245010900055210006900164260007100233300001400304490000700318520141500325653001501740653001601755653002001771653002501791653001501816100002301831700002401854700002101878700002101899700002001920856013001940 2011 eng d a1535-389300aAssimilatory potential of helicoverpa armigera reared on host (chickpea) and nonhost (cassia tora) diets0 aAssimilatory potential of helicoverpa armigera reared on host ch a1155 16TH ST, NW, WASHINGTON, DC 20036 USAbAMER CHEMICAL SOCcNOV a5128-51380 v103 a

Adaptation to plant allelochemicals is a crucial aspect of herbivore chemical ecology. To understand an insect ecology, we studied an effect of nonhost Cassia tora seed-based diet (Ct) on growth, development, and molecular responses in Helicoverpa armigera. We employed a comparative approach to investigate the proteomic differences in gut, hemolymph, and frass of H. armigera reared on a normal (chickpea seed-based, Cp) and Ct diet. In this study, a total of 46 proteins were identified by nano-LC-MS(E). Among them, 17 proteins were up-regulated and 29 proteins were down-regulated when larvae were exposed to the Ct diet. Database searches combined with GO analysis revealed that gut proteases engrossed in digestion, proteins crucial for immunity, adaptive responses to stress, and detoxification were down-regulated in the Ct fed larvae. Proteins identified in H. armigera hemolymph were found to be involved in defense mechanisms. Moreover, proteins found in frass of the Ct fed larvae were observed to participate in energy metabolism. Biochemical and quantitative real-time PCR analysis of selected candidate proteins showed differential gene expression patterns and corroborated with the proteomic data. Our results suggest that the Ct diet could alter expression of proteins related to digestion, absorption of nutrients, adaptation, defense mechanisms, and energy metabolism in H. armigera.

10aadaptation10aCassia tora10aGene expression10aHelicoverpa armigera10aproteomics1 aDawkar, Vishal, V.1 aChikate, Yojana, R.1 aGupta, Vidya, S.1 aSlade, Susan, E.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/assimilatory-potential-helicoverpa-armigera-reared-host-chickpea-and-nonhost-cassia-tora-002457nas a2200253 4500008004100000022001400041245017200055210006900227260006700296300001400363490000700377520145800384653002301842653002301865653001301888653002901901653002301930100002501953700002801978700002002006700002302026700003002049856012402079 2011 eng d a0022-514200aBiochemical characterisation of alpha-amylase inhibitors from achyranthes aspera and their interactions with digestive amylases of coleopteran and lepidopteran insects0 aBiochemical characterisation of alphaamylase inhibitors from ach a111 RIVER ST, HOBOKEN 07030-5774, NJ USAbWILEY-BLACKWELLcAUG a1773-17800 v913 a

BACKGROUND: Starchy seeds are an important food and a source of dietary ingredients in many countries. However, they suffer from extensive predation by bruchids (weevils) and other pests. alpha-Amylase inhibitors are attractive candidates for the control of seed weevils, as these insects are highly dependent on starch as an energy source. RESULTS: A proteinaceous alpha-amylase inhibitor from the seeds of Achyranthes aspera was identified, purified and characterised. In electrophoretic analysis, two prominent amylase inhibitor activity bands (AI1 and AI2) were detected. The inhibitor was purified 9.99-fold with 1206.95 total amylase inhibitor units mg(-1) protein. The molecular weight of the purified inhibitor was around 6 kDa. The isolated alpha-amylase inhibitor was found to be resistant to heat and proteolysis. Feeding analysis of Callosobruchus maculatus larvae on a diet containing seed powder of A. aspera revealed that survival of the larvae was severely affected, with the highest mortality rate occurring on the fifth day of feeding. The isolated inhibitor inhibited the majority of amylase isoforms of C. maculatus, Tribolium confusum and Helicoverpa armigera in electrophoretic analysis and solution assays. CONCLUSION: The information obtained in the present investigation could be useful for a genetic engineering approach that would make seeds resistant to storage pest infestations. (C) 2011 Society of Chemical Industry

10aAchyranthes aspera10aAmylase inhibitors10aamylases10aCallosobruchus maculatus10aTribolium confusum1 aHivrale, Vandana, K.1 aChougule, Nanasaheb, P.1 aGiri, Ashok, P.1 aChhabda, Pavan, J.1 aKachole, Andmanvendra, S. uhttps://library.ncl.res.in/content/biochemical-characterisation-alpha-amylase-inhibitors-achyranthes-aspera-and-their-002280nas a2200241 4500008004100000022001400041245010000055210006900155260007300224300000800297490000800305520142300313653001001736653002201746653002001768653002501788653001401813100002401827700002001851700002101871700002001892856012601912 2012 eng d a0378-111900aAmylase gene expression patterns in Helicoverpa armigera upon feeding on a range of host plants0 aAmylase gene expression patterns in Helicoverpa armigera upon fe aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcJUN a1-70 v5013 a

Expression of two amylase genes (HaAmy1 and HaAmy2) was studied in Helicoverpa armigera (Hubner; Lepidoptera: Noctuidae) feeding on different host plants and during larval development. Alignment of HaAmy1 and HaAmy2 with other insect amylases shows similarities with known Lepidopteran amylase transcripts. H. armigera amylase gene expression is influenced by the availability of reducing sugars, sucrose and starch content of host plants and further correlates to the pool of reducing sugars in the gut and haemolymph of larvae. HaAmy1 and HaAmy2 during larval development on two host plants viz., maize (cereal) and marigold (ornamental) showed their relative difference. Results support the view that when host plants differ in their macronutrients, relationships of enzymes and substrates are flexible. The present work highlights the distribution of HaAmy1 and HaAmy2 (i) during various stages of insect development (second, fourth and sixth instar, pupa, adult and egg), (ii) in various tissues viz., head, haemolymph, fat body, integument and whole larval body of H. armigera feeding on artificial diet and (iii) in three gut regions of larvae fed on various diets. Complexity in expression of amylase genes suggests existence of mechanisms involved to detect nutrient balance required for avoiding fitness costs and focus their importance in insect nutrition. (C) 2012 Elsevier B.V. All rights reserved.

10aDiets10aDigestive amylase10aGene expression10aHelicoverpa armigera10aNutrition1 aKotkar, Hemlata, M.1 aBhide, Amey, J.1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/amylase-gene-expression-patterns-helicoverpa-armigera-upon-feeding-range-host-plants-002341nas a2200265 4500008004100000022001400041245012500055210006900180260005800249300000700307490000700314520143700321653001301758653002301771653001201794653001401806100001901820700002001839700001901859700001901878700001401897700002101911700002001932856012301952 2012 eng d a1536-244200aDevelopmental and digestive flexibilities in the midgut of a polyphagous pest, the cotton bollworm, helicoverpa armigera0 aDevelopmental and digestive flexibilities in the midgut of a pol aLIBRARY C327, TUCSON, AZ 85721 USAbUNIV ARIZONAcMAR a420 v123 a

Developmental patterns and survival of the cotton bollworm, Helicoverpa armigera Hubner (Lepidoptera: Noctuidae), a polyphagous insect pest, have been studied with reference to the effect of diet on major gut digestive enzymes (amylases, proteases, and lipases). Significant correlations between nutritional quality of the diet and larval and pupal mass were observed when H. armigera larvae were fed on various host plants viz. legumes (chickpea and pigeonpea), vegetables (tomato and okra), flowers (rose and marigold), and cereals (sorghum and maize). Larvae fed on diets rich in proteins and/or carbohydrates (pigeonpea, chickpea, maize, and sorghum) showed higher larval mass and developed more rapidly than larvae fed on diets with low protein and carbohydrate content (rose, marigold, okra, and tomato). Low calorific value diets like rose and marigold resulted in higher mortality (25-35%) of H. armigera. Even with highly varying development efficiency and larval/pupal survival rates, H. armigera populations feeding on different diets completed their life cycles. Digestive enzymes of H. armigera displayed variable expression levels and were found to be regulated on the basis of macromolecular composition of the diet. Post-ingestive adaptations operating at the gut level, in the form of controlled release of digestive enzymes, might be a key factor contributing to the physiological plasticity in H. armigera.

10aamylases10alarval performance10aLipases10aProteases1 aSarate, P., J.1 aTamhane, V., A.1 aKotkar, H., M.1 aRatnakaran, N.1 aSusan, N.1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/developmental-and-digestive-flexibilities-midgut-polyphagous-pest-cotton-bollworm-101672nas a2200265 4500008004100000022001400041245010700055210006900162260007500231300001000306490000600316520072300322653001701045653002101062653002701083653001601110100001601126700002101142700002501163700002501188700002301213700002601236700002001262856012401282 2012 eng d a1862-834600aDynamic proteome in enigmatic preeclampsia: an account of molecular mechanisms and biomarker discovery0 aDynamic proteome in enigmatic preeclampsia an account of molecul aPOSTFACH 101161, 69451 WEINHEIM, GERMANYbWILEY-V C H VERLAG GMBHcJAN a79-900 v63 a

The coevolution of genomics and proteomics has led to advancements in the field of diagnosis and molecular mechanisms of disease. Proteomics is now stepping into the field of obstetrics, where early diagnosis of pregnancy complication such as preeclampsia (PE) is imperative. PE is a multifactorial disease characterized by hypertension with proteinuria, which is a leading cause of maternal and neonatal morbidity and mortality occurring in 5-7% of pregnancies worldwide. This review discusses the probable molecular mechanisms that lead to PE and summarizes the proteomics research carried out in understanding the pathogenicity of PE, and for identifying the candidate biomarker for diagnosis of the disease.

10aHypertension10aOxidative stress10aPregnancy complication10aProteinuria1 aMary, Sheon1 aPatil, Gouri, V.1 aKulkarni, Asmita, V.1 aKulkarni, Mahesh, J.1 aJoshi, Sadhana, R.1 aMehendale, Savita, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/dynamic-proteome-enigmatic-preeclampsia-account-molecular-mechanisms-and-biomarker-002359nas a2200265 4500008004100000022001400041245011200055210006900167260009500236300001200331490000800343520134200351653003101693653001701724653002801741653002301769653002601792653002301818100002801841700002501869700002101894700002001915700003101935856012701966 2012 eng d a0021-979700aEnhanced permeation, leaf retention, and plant protease inhibitor activity with bicontinuous microemulsions0 aEnhanced permeation leaf retention and plant protease inhibitor  a525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USAbACADEMIC PRESS INC ELSEVIER SCIENCEcOCT a177-1830 v3833 a

Bicontinuous microemulsions (BCMEs) have excellent solubulizing properties along with low interfacial tension and aqueous content that can be controlled. In this work, water soluble plant protease inhibitor (PI), well characterized for its activity against insect pests, was incorporated into a BCME system and explored for permeation on hydrophobic leaf surfaces and protease inhibition activity. The bicontinuous nature of the microemulsion containing water:2-propanol:1-butanol (55:35:10 w/w) was characterized using conductivity and self-diffusion coefficient measurements. The PI was soluble in the water-rich bicontinuous domains, stable in the microemulsions, and protease inhibition activity was retained for a prolonged duration. The microemulsions ensured greater wettability and a wider spread of the PI on hydrophobic leaf surfaces as revealed by contact angle measurements. Significantly, trypsin inhibition activity assays of the PI recovered from the leaves after delivery from the microemulsion indicated a significant increase in the PI retention on the leaf. This BCME enabled greater leaf permeation and retention of the PI can be attributed to a temporary disruption of the waxy leaf surface followed by self-repair without causing any long term damage to the plant. (C) 2012 Elsevier Inc. All rights reserved.

10aBicontinuous microemulsion10aBiopesticide10aMicroemulsion pesticide10aprotease inhibitor10aProtein encapsulation10aProtein permeation1 aTamhane, Vaijayanti, A.1 aDhaware, Deepika, G.1 aKhandelwal, Neha1 aGiri, Ashok, P.1 aPanchagnula, Venkateswarlu uhttps://library.ncl.res.in/content/enhanced-permeation-leaf-retention-and-plant-protease-inhibitor-activity-bicontinuous-001858nas a2200241 4500008004100000022001400041245006800055210006800123260009900191300001000290490000800300520103900308653001001347653002601357653001301383653001401396100002201410700002501432700002301457700002001480700002101500856009501521 2012 eng d a0308-814600aGeographic variation in the flavour volatiles of alphonso mango0 aGeographic variation in the flavour volatiles of alphonso mango aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLANDbELSEVIER SCI LTDcJAN a58-660 v1303 a

Alphonso, one of the most popular cultivars of mango in India is known to exhibit geographic variation in the flavour of ripe fruits. To get chemical insight into this difference, volatiles were studied in the ripening fruits of Alphonso mangoes from three cultivation locations in India. Ripe fruits from Deogad had lower content of mono- and sesquiterpenes and higher content of lactones and furanones as compared to the fruits from Dapoli: whereas fruits from Vengurle had average quantities of these chemicals in comparison with Deogad and Dapoli fruits. This variation was clearly reflected as separate clustering of the localities in the Principal Component Analysis. The localities were indistinguishable from each other in terms of raw fruit volatiles. This study exemplifies a case of phenotypic plasticity; since the plants chosen were clonally propagated, such geographic variation in the volatiles can be attributed to varied abiotic conditions at these three localities. (C) 2011 Elsevier Ltd. All rights reserved.

10aMango10aPhenotypic plasticity10aRipening10aVolatiles1 aKulkarni, Ram, S.1 aChidley, Hemangi, G.1 aPujari, Keshav, H.1 aGiri, Ashok, P.1 aGupta, Vidya, S. uhttps://library.ncl.res.in/content/geographic-variation-flavour-volatiles-alphonso-mango-002403nas a2200313 4500008004100000022001400041245010700055210006900162260007100231300001400302490000700316520127100323653004301594653001201637653001201649653003601661100002501697700002301722700002101745700002901766700002401795700002301819700002201842700002001864700002701884700002601911700002501937856012701962 2012 eng d a1535-389300aLow plasma albumin levels are associated with increased plasma protein glycation and HbA1c in diabetes0 aLow plasma albumin levels are associated with increased plasma p a1155 16TH ST, NW, WASHINGTON, DC 20036 USAbAMER CHEMICAL SOCcFEB a1391-13960 v113 a

Albumin is one of the most abundant plasma proteins and is heavily glycated in diabetes. In this study, we have addressed whether variation in the albumin levels influence glycation of plasma proteins and HbA1c. The study was performed in three systems: (1) streptozotocin (STZ)-induced diabetic mice plasma, (2) diabetic clinical plasma, and (3) in vitro glycated plasma. Diabetic mice and clinical plasma samples were categorized as diabetic high albumin plasma (DHAP) and diabetic low albumin plasma (DLAP) on the basis of their albumin levels. For the in vitro experiment, two albumin levels, high albumin plasma (HAP) and low albumin plasma (LAP), were created by differential depletion of plasma albumin. Protein glycation was studied by using a combination of two-dimensional electrophoresis (2DE), Western blotting, and LC-MSE. In both mice and clinical experiments, an increased plasma protein glycation was observed in DLAP than in DHAP. Additionally, plasma albumin levels were negatively correlated with HbA1c. The in vitro experiment with differential depletion of albumin mechanistically showed that the low albumin levels are associated with increased plasma protein glycation and that albumin competes for glycation with other plasma proteins.

10aadvanced glycation end products (AGEs)10aAlbumin10aglucose10apost translational modification1 aBhonsle, Hemangi, S.1 aKorwar, Arvind, M.1 aKote, Sachin, S.1 aGolegaonkar, Sandeep, B.1 aChougale, Ashok, D.1 aShaik, Mahemud, L.1 aDhande, Nitin, L.1 aGiri, Ashok, P.1 aShelgikar, Kishore, M.1 aBoppana, Ramanamurthy1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/low-plasma-albumin-levels-are-associated-increased-plasma-protein-glycation-and-hba1c-002528nas a2200277 4500008004100000022001400041245014300055210006900198260007100267300001400338490000700352520148300359653002501842653001601867653003201883653002601915653000801941653003701949100002501986700002502011700002102036700002402057700002002081700002102101856012802122 2012 eng d a1535-389300aProteome profiling of flax (linum usitatissimum) seed: characterization of functional metabolic pathways operating during seed development0 aProteome profiling of flax linum usitatissimum seed characteriza a1155 16TH ST, NW, WASHINGTON, DC 20036 USAbAMER CHEMICAL SOCcDEC a6264-62760 v113 a

Flax (Linum usitatissimum L.) seeds are an important source of food and feed due to the presence of various health promoting compounds, making it a nutritionally and economically important plant. An in-depth analysis of the proteome of developing flax seed is expected to provide significant information with respect to the regulation and accumulation of such storage compounds. Therefore, a proteomic analysis of seven seed developmental stages (4, 8, 12, 16, 22, 30, and 48 days after anthesis) in a flax variety, NL-97 was carried out using a combination of ID-SDS-PAGE and LC-MSE methods. A total 1716 proteins were identified and their functional annotation revealed that a majority of them were involved in primary metabolism, protein destination, storage and energy. Three carbon assimilatory pathways appeared to operate in flax seeds. Reverse transcription quantitative PCR of selected 19 genes was carried out to understand their roles during seed development. Besides storage proteins, methionine synthase, RuBisCO and S-adenosylmethionine synthetase were highly expressed transcripts, highlighting their importance in flax seed development. Further, the identified proteins were mapped onto developmental seed specific expressed sequence tag (EST) libraries of flax to obtain transcriptional evidence and 81% of them had detectable expression at the mRNA level. This study provides new insights into the complex seed developmental processes operating in flax.

10aalpha-linolenic acid10acarbon flux10aDigital expression analysis10aflax seed development10aMSE10asecoisolariciresinol diglucoside1 aBarvkar, Vitthal, T.1 aPardeshi, Varsha, C.1 aKale, Sandip, M.1 aKadoo, Narendra, Y.1 aGiri, Ashok, P.1 aGupta, Vidya, S. uhttps://library.ncl.res.in/content/proteome-profiling-flax-linum-usitatissimum-seed-characterization-functional-metabolic-002107nas a2200301 4500008004100000022001400041245009200055210006900147260007900216300001000295490000700305520106700312653000801379653001301387653002101400653001601421653003401437100002401471700002101495700002501516700002401541700002601565700002201591700002601613700002001639700002501659856012101684 2012 eng d a1073-608500aProteomic analysis of glycated proteins from streptozotocin-induced diabetic rat kidney0 aProteomic analysis of glycated proteins from streptozotocininduc a999 RIVERVIEW DRIVE SUITE 208, TOTOWA, NJ 07512 USAbHUMANA PRESS INCcJAN a28-380 v503 a

Glycation of proteins leading to formation of advanced glycation end products (AGEs) has been considered as one of the important causes of diabetic nephropathy. Therefore, in this study, glycated proteins were detected by anti-AGE antibodies from kidney of streptozotocin-induced diabetic rat showing nephropathic symptoms, by using two dimensional electrophoresis and western blot analysis. These glycated proteins were identified and characterized by using combination of peptide mass finger printing and tandem mass spectrometric approaches. Glycated proteins identified included proteins from metabolic pathways, oxidative stress, cell signaling, and transport. Several of the proteins modified by glycation were involved in glucose metabolism. The extent of glycation was higher in diabetes compared to control, in the glycated proteins that were common to both control and diabetic kidney. Two dimensional electrophoresis proteins profiling of glycated proteins suggest that four of the glycated proteins were significantly up regulated in diabetes.

10aAGE10aDiabetes10aGlucose toxicity10aNephropathy10aPost-translation modification1 aChougale, Ashok, D.1 aBhat, Shweta, P.1 aBhujbal, Swapnil, V.1 aZambare, Mandar, R.1 aPuntambekar, Shraddha1 aSomani, Rahul, S.1 aBoppana, Ramanamurthy1 aGiri, Ashok, P.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/proteomic-analysis-glycated-proteins-streptozotocin-induced-diabetic-rat-kidney-003296nas a2200277 4500008004100000022001400041245007100055210006900126260008100195300000800276490000700284520235500291653001002646653001402656653002002670653003802690653002902728100001902757700002202776700002802798700002502826700002102851700002102872700002002893856010502913 2012 eng d a1471-222900aStress inducible proteinase inhibitor diversity in capsicum annuum0 aStress inducible proteinase inhibitor diversity in capsicum annu a236 GRAYS INN RD, FLOOR 6, LONDON WC1X 8HL, ENGLANDbBIOMED CENTRAL LTDcNOV a2170 v123 a

Background: Wound-inducible Pin-II Proteinase inhibitors (PIs) are one of the important plant serine PIs which have been studied extensively for their structural and functional diversity and relevance in plant defense against insect pests. To explore the functional specialization of an array of Capsicum annuum (L.) proteinase inhibitor (CanPIs) genes, we studied their expression, processing and tissue-specific distribution under steady-state and induced conditions. Inductions were performed by subjecting C. annuum leaves to various treatments, namely aphid infestation or mechanical wounding followed by treatment with either oral secretion (OS) of Helicoverpa armigera or water. Results: The elicitation treatments regulated the accumulation of CanPIs corresponding to 4-, 3-, and 2-inhibitory repeat domains (IRDs). Fourty seven different CanPI genes composed of 28 unique IRDs were identified in total along with those reported earlier. The CanPI gene pool either from uninduced or induced leaves was dominated by 3-IRD PIs and trypsin inhibitory domains. Also a major contribution by 4-IRD CanPI genes possessing trypsin and chymotrypsin inhibitor domains was specifically revealed in wounded leaves treated with OS. Wounding displayed the highest number of unique CanPIs while wounding with OS treatment resulted in the high accumulation of specifically CanPI-4, -7 and -10. Characterization of the PI protein activity through two dimensional gel electrophoresis revealed tissue and induction specific patterns. Consistent with transcript abundance, wound plus OS or water treated C. annuum leaves exhibited significantly higher PI activity and isoform diversity contributed by 3- and 4-IRD CanPIs. CanPI accumulation and activity was weakly elicited by aphid infestation yet resulted in the higher expression of CanPI-26, -41 and -43. Conclusions: Plants can differentially perceive various kinds of insect attacks and respond appropriately through activating plant defenses including regulation of PIs at transcriptional and post-translational levels. Based on the differentially elicited CanPI accumulation patterns, it is intriguing to speculate that generating sequence diversity in the form of multi-IRD PIs is a part of elaborative plant defense strategy to obtain a diverse pool of functional units to confine insect attack.

10aCanPI10aHerbivory10aOral secretions10aPin-II type proteinase inhibitors10aPlant-insect interaction1 aMishra, Manasi1 aMahajan, Neha, S.1 aTamhane, Vaijayanti, A.1 aKulkarni, Mahesh, J.1 aBaldwin, Ian, T.1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/stress-inducible-proteinase-inhibitor-diversity-capsicum-annuum-002032nas a2200181 4500008004100000022001400041245013500055210006900190260009100259300001100350490000600361520125800367100002801625700002301653700002001676700002501696856012901721 2013 eng d a1932-620300aAlterations in the helicoverpa armigera midgut digestive physiology after ingestion of pigeon pea inducible leucine aminopeptidase0 aAlterations in the helicoverpa armigera midgut digestive physiol a1160 BATTERY STREET, STE 100, SAN FRANCISCO, CA 94111 USAbPUBLIC LIBRARY SCIENCEcSEP ae748890 v83 a

Jasmonate inducible plant leucine aminopeptidase (LAP) is proposed to serve as direct defense in the insect midgut. However, exact functions of inducible plant LAPs in the insect midgut remain to be estimated. In the present investigation, we report the direct defensive role of pigeon pea inducible LAP in the midgut of Helicoverpa armigera (Lepidoptera: Noctuidae) and responses of midgut soluble aminopeptidases and serine proteinases upon LAP ingestion. Larval growth and survival was significantly reduced on the diets supplemented with pigeon pea LAP. Aminopeptidase activities in larvae remain unaltered in presence or absence of inducible LAP in the diet. On the contrary, serine proteinase activities were significantly decreased in the larvae reared on pigeon pea LAP containing diet as compared to larvae fed on diet without LAP. Our data suggest that pigeon pea inducible LAP is responsible for the degradation of midgut serine proteinases upon ingestion. Reduction in the aminopeptidase activity with LpNA in the H. armigera larvae was compensated with an induction of aminopeptidase activity with ApNA. Our findings could be helpful to further dissect the roles of plant inducible LAPs in the direct plant defense against herbivory.

1 aLomate, Purushottam, R.1 aJadhav, Bhakti, R.1 aGiri, Ashok, P.1 aHivrale, Vandana, K. uhttps://library.ncl.res.in/content/alterations-helicoverpa-armigera-midgut-digestive-physiology-after-ingestion-pigeon-pea-001988nas a2200349 4500008004100000022001400041245009800055210006900153260008900222300001200311490000800323520087500331100001401206700001501220700001601235700002001251700001501271700002401286700002001310700001401330700002101344700001601365700001601381700001301397700001601410700001401426700001701440700001901457700002001476700001901496856012301515 2013 eng d a0036-807500aBiosynthesis of antinutritional alkaloids in solanaceous crops is mediated by clustered genes0 aBiosynthesis of antinutritional alkaloids in solanaceous crops i a1200 NEW YORK AVE, NW, WASHINGTON, DC 20005 USAbAMER ASSOC ADVANCEMENT SCIENCEcJUL a175-1790 v3413 a

Steroidal glycoalkaloids (SGAs) such as alpha-solanine found in solanaceous food plants-as, for example, potato-are antinutritional factors for humans. Comparative coexpression analysis between tomato and potato coupled with chemical profiling revealed an array of 10 genes that partake in SGA biosynthesis. We discovered that six of them exist as a cluster on chromosome 7, whereas an additional two are adjacent in a duplicated genomic region on chromosome 12. Following systematic functional analysis, we suggest a revised SGA biosynthetic pathway starting from cholesterol up to the tetrasaccharide moiety linked to the tomato SGA aglycone. Silencing GLYCOALKALOID METABOLISM 4 prevented accumulation of SGAs in potato tubers and tomato fruit. This may provide a means for removal of unsafe, antinutritional substances present in these widely used food crops.

1 aItkin, M.1 aHeinig, U.1 aTzfadia, O.1 aBhide, Amey, J.1 aShinde, B.1 aCardenas, Pablo, D.1 aBocobza, S., E.1 aUnger, T.1 aMalitsky, Sergey1 aFinkers, R.1 aTikunov, Y.1 aBovy, A.1 aChikate, Y.1 aSingh, P.1 aRogachev, I.1 aBeekwilder, J.1 aGiri, Ashok, P.1 aAharoni, Asaph uhttps://library.ncl.res.in/content/biosynthesis-antinutritional-alkaloids-solanaceous-crops-mediated-clustered-genes-002840nas a2200241 4500008004100000022001400041245009000055210006900145260008100214300000700295490000700302520195400309653001902263653002402282653002102306653002602327653002902353100002602382700002802408700002002436700002502456856011702481 2013 eng d a1471-209100aCharacterization of a chemostable serine alkaline protease from Periplaneta americana0 aCharacterization of a chemostable serine alkaline protease from  a236 GRAYS INN RD, FLOOR 6, LONDON WC1X 8HL, ENGLANDbBIOMED CENTRAL LTDcNOV a320 v143 a

Background: Proteases are important enzymes involved in numerous essential physiological processes and hold a strong potential for industrial applications. The proteolytic activity of insects' gut is endowed by many isoforms with diverse properties and specificities. Thus, insect proteases can act as a tool in industrial processes. Results: In the present study, purification and properties of a serine alkaline protease from Periplaneta americana and its potential application as an additive in various bio-formulations are reported. The enzyme was purified near to homogeneity by using acetone precipitation and Sephadex G-100 gel filtration chromatography. Enzyme activity was increased up to 4.2 fold after gel filtration chromatography. The purified enzyme appeared as single protein-band with a molecular mass of similar to 27.8 kDa in SDS-PAGE. The optimum pH and temperature for the proteolytic activity for purified protein were found around pH 8.0 and 60 degrees C respectively. Complete inhibition of the purified enzyme by phenylmethylsulfonyl fluoride confirmed that the protease was of serine-type. The purified enzyme revealed high stability and compatibility towards detergents, oxidizing, reducing, and bleaching agents. In addition, enzyme also showed stability towards organic solvents and commercial detergents. Conclusion: Several important properties of a serine protease from P. Americana were revealed. Moreover, insects can serve as excellent and alternative source of industrially important proteases with unique properties, which can be utilized as additives in detergents, stain removers and other bio-formulations. Properties of the P. americana protease accounted in the present investigation can be exploited further in various industrial processes. As an industrial prospective, identification of enzymes with varying essential properties from different insect species might be good approach and bioresource.

10aChemostability10aIndustrial catalyst10aInsect proteases10aPeriplaneta americana10aSerine alkaline protease1 aSanatan, Prashant, T.1 aLomate, Purushottam, R.1 aGiri, Ashok, P.1 aHivrale, Vandana, K. uhttps://library.ncl.res.in/content/characterization-chemostable-serine-alkaline-protease-periplaneta-americana-002972nas a2200313 4500008004100000022001400041245010400055210006900159260010300228300001200331490000700343520187800350653001302228653002102241653001002262653003502272653002102307653002202328100001802350700001802368700002502386700001902411700001802430700002502448700001902473700002002492700001702512856012902529 2013 eng d a0981-942800aCharacterization of three novel isoprenyl diphosphate synthases from the terpenoid rich mango fruit0 aCharacterization of three novel isoprenyl diphosphate synthases  a23 RUE LINOIS, 75724 PARIS, FRANCEbELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIERcOCT a121-1310 v713 a

Mango (cv. Alphonso) is popular due to its highly attractive, terpenoid-rich flavor. Although Alphonso is clonally propagated, its fruit-flavor composition varies when plants are grown in different geo-climatic zones. Isoprenyl diphosphate synthases catalyze important branch-point reactions in terpenoid biosynthesis, providing precursors for common terpenoids such as volatile terpenes, sterols and carotenoids. Two geranyl diphosphate synthases and a farnesyl diphosphate synthase were isolated from Alphonso fruits, cloned for recombinant expression and found to produce the respective products. Although, one of the geranyl diphosphate synthases showed high sequence similarity to the geranylgeranyl diphosphate synthases, it did not exhibit geranylgeranyl diphosphate synthesizing activity. When modeled, this geranyl diphosphate synthase and farnesyl diphosphate synthase structures were found to be homologous with the reference structures, having all the catalytic side chains appropriately oriented. The optimum temperature for both the geranyl diphosphate synthases was 40 degrees C and that for farnesyl diphosphate synthase was 25 degrees C. This finding correlated well with the dominance of monoterpenes in comparison to sesquiterpenes in the fruits of Alphonso mango in which the mesocarp temperature is higher during ripening than development. The absence of activity of these enzymes with the divalent metal ion other than Mg2+ indicated their adaptation to the Mg2+ rich mesocarp. The typical expression pattern of these genes through the ripening stages of fruits from different cultivation localities depicting the highest transcript levels of these genes in the stage preceding the maximum terpene accumulation indicated the involvement of these genes in the biosynthesis of volatile terpenes. (C) 2013 Elsevier Masson SAS. All rights reserved.

10aAlphonso10aFlavor variation10aFruit10aIsoprenyl diphosphate synthase10aMangifera indica10aTerpene volatiles1 aKulkarni, Ram1 aPandit, Sagar1 aChidley, Hemangi, G.1 aNagel, Raimund1 aSchmidt, Axel1 aGershenzon, Jonathan1 aPujari, Keshav1 aGiri, Ashok, P.1 aGupta, Vidya uhttps://library.ncl.res.in/content/characterization-three-novel-isoprenyl-diphosphate-synthases-terpenoid-rich-mango-fruit-002516nas a2200265 4500008004100000022001400041245012500055210006900180260007300249300001400322490000900336520153000345653001901875653002101894653003601915653001301951653002301964653003201987100002202019700001902041700001902060700002102079700002002100856013002120 2013 eng d a0304-416500aComplementation of intramolecular interactions for structural-functional stability of plant serine proteinase inhibitors0 aComplementation of intramolecular interactions for structuralfun aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcNOV a5087-50940 v18303 a

Background: Plant protease inhibitors (PIs) constitute a diverse group of proteins capable of inhibiting proteases. Among Pls, serine Pls (SPIs) display stability and conformational restrictions of the reactive site loop by virtue of their compact size, and by the presence of disulfide bonds, hydrogen bonds, and other weak interactions. Scope of review: The significance of various intramolecular interactions contributing to protein folding mechanism and their role in overall stability and activity of SPIs is discussed here. Furthermore, we have reviewed the effect of variation or manipulation of these interactions on the activity/stability of SPIs. Major conclusions: The selective gain or loss of disulfide bond(s) in SPIs can be associated with their functional differentiation, which is likely to be compensated by non-covalent interactions (hydrogen bonding or electrostatic interactions). Thus, these intramolecular interactions are collectively responsible for the functional activity of SPIs, through the maintenance of scaffold framework, conformational rigidity and shape complementarities of reactive site loop. General significance: Structural insight of these interactions will provide an in-depth understanding of kinetic and thermodynamic parameters involved in the folding and stability mechanisms of SPIs. These features can be explored for engineering canonical SPIs for optimizing their overall stability and functionality for various applications. (C) 2013 Elsevier B.V. All rights reserved.

10aDisulfide bond10ahydrogen bonding10aIntramolecular weak interaction10aProtease10aReactive site loop10aSerine proteinase inhibitor1 aJoshi, Rakesh, S.1 aMishra, Manasi1 aSuresh, C., G.1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/complementation-intramolecular-interactions-structural-functional-stability-plant-serine-002009nas a2200253 4500008004100000022001400041245007800055210006900133260006700202300001200269490000700281520116500288653001501453653001701468653002501485653001401510653001201524100002401536700002001560700002201580700002101602700002001623856011201643 2013 eng d a0962-107500aDifferential protease activity augments polyphagy in helicoverpa armigera0 aDifferential protease activity augments polyphagy in helicoverpa a111 RIVER ST, HOBOKEN 07030-5774, NJ USAbWILEY-BLACKWELLcJUN a258-2720 v223 a

Helicoverpa armigera (Lepidoptera: Noctuidae) and other polyphagous agricultural pests are extending their plant host range and emerging as serious agents in restraining crop productivity. Dynamic regulation, coupled with a diversity of digestive and detoxifying enzymes, play a crucial role in the adaptation of polyphagous insects. To investigate the functional intricacy of serine proteases in the development and polyphagy of H.armigera, we profiled the expression of eight trypsin-like and four chymotrypsin-like phylogenetically diverse mRNAs from different life stages of H.armigera reared on nutritionally distinct host plants. These analyses revealed diet- and stage-specific protease expression patterns. The trypsins expressed showed structural variations, which might result in differential substrate specificity and interaction with inhibitors. Protease profiles in the presence of inhibitors and their mass spectrometric analyses revealed insight into their differential activity. These findings emphasize the differential expression of serine proteases and their consequences for digestive physiology in promoting polyphagy in H.armigera.

10aadaptation10achymotrypsin10aHelicoverpa armigera10apolyphagy10aTrypsin1 aChikate, Yojana, R.1 aTamhane, V., A.1 aJoshi, Rakesh, S.1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/differential-protease-activity-augments-polyphagy-helicoverpa-armigera-001301nas a2200241 4500008004100000022001400041245005300055210005200108260007500160300001200235490000600247520057000253653001000823653001300833653001400846653000900860100002500869700002300894700001600917700002500933700002000958856008100978 2013 eng d a1862-834600aGlycated proteome: from reaction to intervention0 aGlycated proteome from reaction to intervention aPOSTFACH 101161, 69451 WEINHEIM, GERMANYbWILEY-V C H VERLAG GMBHcJAN a155-1700 v73 a

Glycation, a nonenzymatic reaction between reducing sugars and proteins, is a proteome wide phenomenon, predominantly observed in diabetes due to hyperglycemia. Glycated proteome of plasma, kidney, lens, and brain are implicated in the pathogenesis of various diseases, including diabetic complications, neurodegenerative diseases, cancer, and aging. This review discusses the strategies to characterize protein glycation, its functional implications in different diseases, and intervention strategies to protect the deleterious effects of protein glycation.

10aaging10aDiabetes10aGlycation10aRAGE1 aKulkarni, Mahesh, J.1 aKorwar, Arvind, M.1 aMary, Sheon1 aBhonsle, Hemangi, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/glycated-proteome-reaction-intervention-002593nas a2200277 4500008004100000022001400041245009700055210006900152260007100221300001400292490000700306520162800313653001601941653001701957653001601974653001601990653001502006653002602021100002302047700002402070700002802094700002602122700002102148700002002169856012602189 2013 eng d a1535-389300aMolecular insights into resistance mechanisms of lepidopteran insect pests against toxicants0 aMolecular insights into resistance mechanisms of lepidopteran in a1155 16TH ST, NW, WASHINGTON, DC 20036 USAbAMER CHEMICAL SOCcNOV a4727-47370 v123 a

Insect pests remain a major reason for crop loss worldwide despite extensive use of chemical insecticides. More than B 50% of all insecticides are organophosphates, followed by synthetic pyrethroids, organochlorines, carbamates, and biopesticides, and their continued use may have many environmental, agricultural, medical, and socioeconomic issues. Importantly, only a countable number of insects have acquired the status of crop pests, mostly due to monoculture of crop plants and polyphagous nature of the insects. We focus on A adaptations of Lepidopteran insects to phytochemicals and synthetic pesticides in native and modern agricultural systems. Because of heavy use of chemical insecticides, a strong selection pressure is imposed on insect populations, resulting in the emergence of resistance against candidate compound(s). Current knowledge suggests that insects generally implement a three-tier system to overcome the effect of toxic compounds at physiological, biochemical, and genetic levels. Furthermore, we have discussed whether the adaptation to phytochemicals provides an advantage to the insect while encountering synthetic insecticide molecules. Specific metabolic pathways employed by insects to convert deterrents into less toxic forms or their removal from the system are highlighted. Using the proteomics approach, insect proteins interacting with insecticides can be identified, and their modification in resistant insects can be characterized. Also, systems biology studies can offer useful cues to decipher the molecular networks participating in the metabolism of detrimental compounds.

10afield crops10ainsect-pests10ainsecticide10aLepidoptera10aproteomics10aresistance mechanisms1 aDawkar, Vishal, V.1 aChikate, Yojana, R.1 aLomate, Purushottam, R.1 aDholakia, Bhushan, B.1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/molecular-insights-resistance-mechanisms-lepidopteran-insect-pests-against-toxicants-002496nas a2200289 4500008004100000022001400041245011300055210006900168260009300237300000800330490000600338520148200344653001901826653002501845653001101870653002101881653001301902100001801915700002501933700002201958700001801980700001901998700002002017700002502037700001702062856012702079 2013 eng d a2193-180100aOxidoreductase from `Alphonso' mango catalyzing biosynthesis of furaneol and reduction of reactive carbonyls0 aOxidoreductase from Alphonso mango catalyzing biosynthesis of fu aGEWERBESTRASSE 11, CHAM, CH-6330, SWITZERLANDbSPRINGER INTERNATIONAL PUBLISHING AGcOCT a4940 v23 a

Two furanones, furaneol (4-hydroxy-2,5-dimethyl-3(2H)-furanone) and mesifuran (2,5-dimethyl-4-methoxy-3(2H)-furanone), are important constituents of flavor of the Alphonso cultivar of mango (Mangifera indica). To get insights into the biosynthesis of these furanones, we isolated an enone oxidoreductase gene from the Alphonso mango. It has high sequence similarity to an alkenal/one oxidoreductase from cucumber (79% identity) and enone oxidoreductases from tomato (73% identity) and strawberry (72% identity). The complete open reading frame was expressed in E. coli and the (his) 6-tagged recombinant protein was purified by affinity chromatography. The purified protein assayed with NADH as a reducing agent converted D-fructose-1,6-diphosphate into furaneol, the immediate precursor of mesifuran. The enzyme was also able to convert two highly reactive carbonyls, 3-buten-2-one and 1-penten-3-one, produced by lipid peroxidation in plants, into their saturated derivatives. Expression profiling in various ripening stages of Alphonso fruits depicted an expression maxima at 10 days after harvest stage, shortly before the appearance of the maximum amount of furanones (completely ripe stage, 15 days after harvest). Although no furanones were detected at the 0 day after harvest stage, significant expression of this gene was detected in the fruits at this stage. Overall, the results suggest that this oxidoreductase plays important roles in Alphonso mango fruits.

10aDetoxification10aEnone oxidoreductase10aFlavor10aMangifera indica10aRipening1 aKulkarni, Ram1 aChidley, Hemangi, G.1 aDeshpande, Ashish1 aSchmidt, Axel1 aPujari, Keshav1 aGiri, Ashok, P.1 aGershenzon, Jonathan1 aGupta, Vidya uhttps://library.ncl.res.in/content/oxidoreductase-alphonso-mango-catalyzing-biosynthesis-furaneol-and-reduction-reactive-002194nas a2200181 4500008004100000245013200041210006900173260000800242300001600250490000900266520149500275100002201770700001901792700002801811700002101839700002001860856013201880 2013 eng d00aPlasticity of protease gene expression in helicoverpa armigera upon exposure to multi-domain capsicum annuum protease inhibitor0 aPlasticity of protease gene expression in helicoverpa armigera u cMAR a3414–34200 v18303 a

Background A multi-domain Pin-II type protease inhibitor from Capsicum annuum (CanPI-7) is known to be effective against the insect pest, Helicoverpa armigera. The present study is an attempt to investigate the optimal dose of recombinant CanPI-7 (rCanPI-7) for effective antibiosis to H. armigera and further to characterize the responses of digestive proteases upon rCanPI-7 ingestion. Methods The gut protease activity was assessed biochemically and transcript accumulation pattern for selected trypsin and chymotrypsin genes was analyzed by quantitative Real-Time PCR. Results The growth retardation upon exposure to rCanPI-7 was more prominent in neonates as compared to third instar larvae. Influence of stage and dosage of rCanPI-7 was conspicuous on the expression and regulation of candidate trypsin and chymotrypsin genes in H. armigera. The transcript accumulation pattern correlated with the protease activity in rCanPI-7 exposed larvae. Conclusions We conclude that early exposure and specific dose of protease inhibitor are essential for effective antibiosis despite the large diversity and plasticity in the expression of protease genes in H. armigera. Moreover, it is also evident that the regulation and expression of H. armigera gut proteases are specific to the stage of PI exposure. General significance These results highlight the requirement of optimal PI concentration for effective growth retardation and for inhibiting the major gut proteases of H. armigera.

1 aMahajan, Neha, S.1 aMishra, Manasi1 aTamhane, Vaijayanti, A.1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/plasticity-protease-gene-expression-helicoverpa-armigera-upon-exposure-multi-domain-capsicum02124nas a2200217 4500008004100000022001400041245008100055210006900136260010300205300001200308490000700320520130500327100002301632700002401655700002201679700002001701700002701721700002501748700002501773856010801798 2013 eng d a1535-947600aProteomic analysis of protease resistant proteins in the diabetic rat kidney0 aProteomic analysis of protease resistant proteins in the diabeti a9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USAbAMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INCcJAN a228-2360 v123 a

Glycation induced protein aggregation has been implicated in the development of diabetic complications and neurodegenerative diseases. These aggregates are known to be resistant to proteolytic digestion. Here we report the identification of protease resistant proteins from the streptozotocin induced diabetic rat kidney, which included enzymes in glucose metabolism and stress response proteins. These protease resistant proteins were characterized to be advanced glycation end products modified and ubiquitinated by immunological and mass spectrometry analysis. Further, diabetic rat kidney exhibited significantly impaired proteasomal activity. The functional analysis of identified physiologically important enzymes showed that their activity was reduced in diabetic condition. Loss of functional activity of these proteins was compensated by enhanced gene expression. Aggregation prone regions were predicted by in silico analysis and compared with advanced glycation end products modification sites. These findings suggested that the accumulation of protein aggregates is an inevitable consequence of impaired proteasomal activity and protease resistance due to advanced glycation end products modification. Molecular & Cellular Proteomics 12: 10.1074/mcp.M112.020651, 228-236, 2013.

1 aBansode, Sneha, B.1 aChougale, Ashok, D.1 aJoshi, Rakesh, S.1 aGiri, Ashok, P.1 aBodhankar, Subhash, L.1 aHarsulkar, Abhay, M.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/proteomic-analysis-protease-resistant-proteins-diabetic-rat-kidney-002317nas a2200169 4500008004100000245015000041210006900191260000800260300001200268490000600280520165500286100001801941700002101959700002101980700002002001856012602021 2013 eng d00aResistome analysis of mycobacterium tuberculosis: identification of aminoglycoside 2’-nacetyltransferase (AAC) as co-target for drug desigining0 aResistome analysis of mycobacterium tuberculosis identification  cFEB a174-1810 v93 a

The emergence of multidrug resistant tuberculosis (MDRTB) highlights the urgent need to understand the mechanisms of resistance to the drugs and to develop a new arena of therapeutics to treat the disease. Ethambutol, isonazid, pyrazinamide, rifampicin are first line of drugs against TB, whereas aminoglycoside, polypeptides, fluoroquinolone, ethionamide are important second line of bactericidal drugs used to treat MDRTB, and resistance to one or both of these drugs are defining characteristic of extensively drug resistant TB. We retrieved 1,221 resistant genes from Antibiotic Resistance Gene Database (ARDB), which are responsible for resistance against first and second line antibiotics used in treatment of Mycobacterium tuberculosis infection. From network analysis of these resistance genes, 53 genes were found to be common. Phylogenetic analysis shows that more than 60% of these genes code for acetyltransferase. Acetyltransferases detoxify antibiotics by acetylation, this mechanism plays central role in antibiotic resistance. Seven acetyltransferase (AT-1 to AT-7) were selected from phylogenetic analysis. Structural alignment shows that these acetyltransferases share common ancestral core, which can be used as a template for structure based drug designing. From STRING analysis it is found that acetyltransferase interact with 10 different proteins and it shows that, all these interaction were specific to M. tuberculosis. These results have important implications in designing new therapeutic strategies with acetyltransferase as lead co-target to combat against MDR as well as Extreme drug resistant (XDR) tuberculosis.

1 aS., Joshi, R.1 aD., Jamdhade, M.1 aS., Sonawane, M.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/resistome-analysis-mycobacterium-tuberculosis-identification-aminoglycoside-2%E2%80%9902001nas a2200265 4500008004100000022001400041245010900055210006900164260009900233300001200332490000800344520106400352653001301416653001201429653001001441653002001451653001301471653001401484100002501498700002201523700002301545700002001568700002101588856012601609 2013 eng d a0308-814600aSpatial and temporal changes in the volatile profile of alphonso mango upon exogenous ethylene treatment0 aSpatial and temporal changes in the volatile profile of alphonso aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLANDbELSEVIER SCI LTDcJAN a585-5940 v1363 a

Alphonso is a highly favoured and exported mango cultivar among the vast mango germplasm of India. Being a climacteric fruit, ethylene plays an important role in ripening of mango. For deeper understanding of effect of pre-climacteric ethylene treatment on volatile profiles of Alphonso mango, 26 volatiles were tracked through six ripening stages of pulp and skin of ethylene-treated and control Alphonso fruits. The study revealed accelerated ripening in terms of early appearance of ripening-specific compounds, lactones and mesifuran, upon ethylene treatment. While the level of lactones remained unaffected, the mesifuran level vastly increased upon ethylene treatment. Skin showed high terpene content while pulp had higher amount of lactones compared to skin. This work points towards involvement of ethylene as a natural hormone in the biosynthesis of lactones and furanones in naturally ripened fruits; whereas, an increase in the terpene level during ripening appears to be independent of ethylene. (C) 2012 Elsevier Ltd. All rights reserved.

10aEthylene10aFlavour10aMango10aPre-climacteric10aRipening10aVolatiles1 aChidley, Hemangi, G.1 aKulkarni, Ram, S.1 aPujari, Keshav, H.1 aGiri, Ashok, P.1 aGupta, Vidya, S. uhttps://library.ncl.res.in/content/spatial-and-temporal-changes-volatile-profile-alphonso-mango-upon-exogenous-ethylene-002054nas a2200253 4500008004100000022001400041245009800055210006900153260009500222300001400317490000800331520110600339653001001445653002301455653002201478653003901500653002201539100001901561700002201580700002501602700002101627700002001648856013201668 2013 eng d a0006-291X00aStructural-functional insights of single and multi-domain capsicum annuum protease inhibitors0 aStructuralfunctional insights of single and multidomain capsicum a525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USAbACADEMIC PRESS INC ELSEVIER SCIENCEcJAN a1060-10650 v4303 a

Pin-II protease inhibitors (PIs) are the focus of research interest because of their large structural-functional diversity and relevance in plant defense. Two representative Capsicum annuum PI genes (CanPI-15 and -7) comprising one and four inhibitory repeat domains, respectively, were expressed and recombinant proteins were characterized. beta-Sheet and unordered structure was found predominant in CanPI-15 while -7 also displayed the signatures of polyproline fold, as revealed by circular dichroism studies. Inhibition kinetics against bovine typsin indicated three times higher potency of CanPI-7 (K-i similar to 57 mu M) than -15 (similar to 184 mu M). Activity and structural stability of these CanPIs were revealed under various conditions of pH, temperature and denaturing agent. Structure prediction, docking studies with proteases and mass spectroscopy revealed the organization of multiple reactive site loops of multi domain PIs in space as well as the steric hindrances imposed while binding to proteases due to their close proximity. (C) 2012 Elsevier Inc. All rights reserved.

10aCanPI10aCircular dichroism10aDisulfide bonding10aPotato type-II protease inhibitors10aProtein stability1 aMishra, Manasi1 aJoshi, Rakesh, S.1 aGaikwad, Sushama, M.1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/structural-functional-insights-single-and-multi-domain-capsicum-annuum-protease-inhibitors-002205nas a2200229 4500008004100000022001400041245014400055210006900199260010600268300001000374490000800384520127700392653002001669653002501689653002901714653001401743653002601757100002201783700002101805700002001826856012901846 2014 eng d a0031-942200aDifferential antibiosis against helicoverpa armigera exerted by distinct inhibitory repeat domains of capsicum annuum proteinase inhibitors0 aDifferential antibiosis against helicoverpa armigera exerted by  aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLANDbPERGAMON-ELSEVIER SCIENCE LTDcMAY a16-220 v1013 a

Plant defensive serine proteinase inhibitors (Pis) are known to have negative impact on digestive physiology of herbivore insects and thus have a crucial role in plant protection. Here, we have assessed the efficacy and specificity of three previously characterized inhibitory repeat domain (IRD) variants from Capsicum annuum PIs viz., IRD-7, -9 and -12 against gut proteinases from Helicoverpa armigera. Comparative study of in silico binding energy revealed that IRD-9 possesses higher affinity towards H. armigera serine proteinases as compared to IRD-7 and -12. H. armigera fed on artificial diet containing 5 TIU/g of recombinant IRD proteins exhibited differential effects on larval growth, survival rate and other nutritional parameters. Major digestive gut trypsin and chymotrypsin genes were down regulated in the IRD fed larvae, while few of them were up-regulated, this indicate alterations in insect digestive physiology. The results corroborated with proteinase activity assays and zymography. These findings suggest that the sequence variations among Pis reflect in their efficacy against proteinases in vitro and in vivo, which also could be used for developing tailor-made multi-domain inhibitor gene(s). (C) 2014 Elsevier Ltd. All rights reserved.

10aCapsicum annuum10aHelicoverpa armigera10aInhibitory repeat domain10aProteases10aProteinase inhibitors1 aJoshi, Rakesh, S.1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/differential-antibiosis-against-helicoverpa-armigera-exerted-distinct-inhibitory-repeat-002475nas a2200253 4500008004100000022001400041245014800055210006900203260010600272300001200378490000700390520147200397653002001869653002501889653001101914653002901925653002401954100002801978700002202006700002102028700002102049700002002070856013102090 2014 eng d a0965-174800aIdentification and expression profiling of helicoverpa armigera microRNAs and their possible role in the regulation of digestive protease genes0 aIdentification and expression profiling of helicoverpa armigera  aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLANDbPERGAMON-ELSEVIER SCIENCE LTDcNOV a129-1370 v543 a

The present investigation is an effort to determine the possible roles of microRNAs (miRNAs) in the regulation of protease gene expression in Helicoverpa armigera upon exposure to plant protease inhibitors (PIs). Using Illumina platform, deep sequencing of 12 small RNA libraries was performed from H. armigera larvae fed on artificial diet (AD) or recombinant Capsicum annuum PI-7 (rCanPI-7) incorporated diet, at various time intervals (0.5, 2, 6, 12, 24, and 48 h). Sequencing data were analyzed with miRDeep2 software; a total of 186 unique miRNAs were identified from all the 12 libraries, out of which 96 were conserved while 90 were novel. These miRNAs showed all the conserved characteristics of insect miRNAs. Homology analysis revealed that most of the identified miRNAs were insect-specific, and more than 50 miRNAs were Lepidoptera-specific. Several candidate miRNAs (conserved and novel) were differentially expressed in rCanPI-7 fed larvae as compared to the larvae fed on AD. H. armigera miRNAs were found to have target sites in several protease genes as well as in protease regulation related genes such as serine PI and immune reactive PI. As expected, negative correlation in the relative abundance miRNAs and their target mRNAs was evident from qualitative real time polymerase chain reaction analysis. The investigation revealed potential roles of miRNAs in H. armigera protease gene regulation. (C) 2014 Elsevier Ltd. All rights reserved.

10aDeep sequencing10aHelicoverpa armigera10amiRNAs10aProtease gene regulation10aProtease inhibitors1 aLomate, Purushottam, R.1 aMahajan, Neha, S.1 aKale, Sandip, M.1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/identification-and-expression-profiling-helicoverpa-armigera-micrornas-and-their-possible-001848nas a2200253 4500008004100000022001400041245005200055210005200107260007700159300001000236490000600246520106800252653001701320653002401337653001901361653002101380653000901401100001701410700001601427700002001443700002001463700002501483856008601508 2014 eng d a1862-834600aImmune response to chemically modified proteome0 aImmune response to chemically modified proteome aBOSCHSTRASSE 12, D-69469 WEINHEIM, GERMANYbWILEY-V C H VERLAG GMBHcFEB a19-340 v83 a

Both enzymatic and nonenzymatic PTMs of proteins involve chemical modifications. Some of these modifications are prerequisite for the normal functioning of cell, while other chemical modifications render the proteins as neo-self antigens, which are recognized as non-self leading to aberrant cellular and humoral immune responses. However, these modifications could be a secondary effect of autoimmune diseases, as in the case of type I diabetes, hyperglycemia leads to protein glycation. The enigma of chemical modifications and immune response is akin to the chick-and-egg paradox. Nevertheless, chemical modifications regulate immune response. In some of the well-known autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, and multiple sclerosis, chemically modified proteins act as autoantigens forming immune complexes. In some instances, chemical modifications are also involved in regulating immune response during pathogen infection. Further, the usefulness of proteomic analysis of immune complexes is briefly discussed.

10aAutoantibody10aAutoimmune diseases10aImmune complex10aImmunoproteomics10aPTMs1 aBhat, Shweta1 aMary, Sheon1 aBanarjee, Reema1 aGiri, Ashok, P.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/immune-response-chemically-modified-proteome-002477nas a2200205 4500008004100000022001400041245010900055210006900164260009100233300001200324490000600336520165100342100002001993700002902013700001802042700003702060700002502097700002002122856012902142 2014 eng d a1932-620300aInsecticidal potential of defense metabolites from ocimum kilimandscharicum against helicoverpa armigera0 aInsecticidal potential of defense metabolites from ocimum kilima a1160 BATTERY STREET, STE 100, SAN FRANCISCO, CA 94111 USAbPUBLIC LIBRARY SCIENCEcAUG ae1043770 v93 a

Genus Ocimum contains a reservoir of diverse secondary metabolites, which are known for their defense and medicinal value. However, the defense-related metabolites from this genus have not been studied in depth. To gain deeper insight into inducible defense metabolites, we examined the overall biochemical and metabolic changes in Ocimum kilimandscharicum that occurred in response to the feeding of Helicoverpa armigera larvae. Metabolic analysis revealed that the primary and secondary metabolism of local and systemic tissues in O. kilimandscharicum was severely affected following larval infestation. Moreover, levels of specific secondary metabolites like camphor, limonene and beta-caryophyllene (known to be involved in defense) significantly increased in leaves upon insect attack. Choice assays conducted by exposing H. armigera larvae on O. kilimandscharicum and tomato leaves, demonstrated that O. kilimandscharicum significantly deters larval feeding. Further, when larvae were fed on O. kilimandscharicum leaves, average body weight decreased and mortality the larvae increased. Larvae fed on artificial diet supplemented with O. kilimandscharicum leaf extract, camphor, limonene and beta-caryophyllene showed growth retardation, increased mortality rates and pupal deformities. Digestive enzymes of H. armigera - namely, amylase, protease and lipase- showed variable patterns after feeding on O. kilimandscharicum, which implies striving of the larvae to attain required nutrition for growth, development and metamorphosis. Evidently, selected metabolites from O. kilimandscharicum possess significant insecticidal activity.

1 aSingh, Priyanka1 aJayaramaiah, Ramesha, H.1 aSarate, Priya1 aThulasiram, Hirekodathakallu, V.1 aKulkarni, Mahesh, J.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/insecticidal-potential-defense-metabolites-ocimum-kilimandscharicum-against-helicoverpa-002993nas a2200289 4500008004100000022001400041245017700055210006900232260008600301300001000387490000700397520189300404653002002297653002502317653001402342653002502356100002202381700001902403700002802422700001902450700002402469700001902493700002002512700002102532700002002553856013002573 2014 eng d a0739-110200aRemarkable efficiency of a pin-II proteinase inhibitor sans two conserved disulfide bonds is due to enhanced flexibility and hydrogen bond density in the reactive site loop0 aRemarkable efficiency of a pinII proteinase inhibitor sans two c a325 CHESTNUT ST, SUITE 800, PHILADELPHIA, PA 19106 USAbTAYLOR & FRANCIS INCcJAN a13-260 v323 a

Capsicum annuum (L.) expresses diverse potato type II family proteinase inhibitors comprising of inhibitory repeat domain (IRD) as basic functional unit. Most IRDs contain eight conserved cysteines forming four disulfide bonds, which are indispensible for their stability and activity. We investigated the functional significance of evolutionary variations in IRDs and their role in mediating interaction between the inhibitor and cognate proteinase. Among the 18 IRDs encoded by C. annuum, IRD-7, -9, and -12 were selected for further characterization on the basis of variation in their reactive site loop, number of conserved cysteine residues, and higher theoretical G(bind) for interaction with Helicoverpa armigera trypsin. Moreover, inhibition kinetics showed that IRD-9, despite loss of some of the disulfide bonds, was a more potent proteinase inhibitor among the three selected IRDs. Molecular dynamic simulations revealed that serine residues in the place of cysteines at seventh and eighth positions of IRD-9 resulted in an increase in the density of intramolecular hydrogen bonds and reactive site loop flexibility. Results of the serine residues chemical modification also supported this observation and provided a possible explanation for the remarkable inhibitory potential of IRD-9. Furthermore, this natural variant among IRDs showed special attributes like stability to proteolysis and synergistic inhibitory effect on other IRDs. It is likely that IRDs have coevolved selective specialization of their structure and function as a response towards specific insect proteases they encountered. Understanding the molecular mechanism of pest protease-plant proteinaceous inhibitor interaction will help in developing effective pest control strategies.An animated interactive 3D complement (I3DC) is available in Proteopedia at http://proteopedia.org/w/Journal:JBSD:39

10aCapsicum annuum10aHelicoverpa armigera10aProteases10aProteinase inhibitor1 aJoshi, Rakesh, S.1 aMishra, Manasi1 aTamhane, Vaijayanti, A.1 aGhosh, Anirban1 aSonavane, Uddhavesh1 aSuresh, C., G.1 aJoshi, Rajendra1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/remarkable-efficiency-pin-ii-proteinase-inhibitor-sans-two-conserved-disulfide-bonds-due-002424nas a2200241 4500008004100000022001400041245014400055210006900199260009500268300001200363490000800375520148000383653001601863653003001879653002301909653001001932100002201942700002301964700002401987700002102011700002002032856013002052 2014 eng d a0006-291X00aResistance through inhibition: ectopic expression of serine protease inhibitor offers stress tolerance via delayed senescence in yeast cell0 aResistance through inhibition ectopic expression of serine prote a525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USAbACADEMIC PRESS INC ELSEVIER SCIENCEcSEP a361-3680 v4523 a

Protease inhibitors have been known to confer multiple stress tolerance in transgenic plants. We have assessed growth of yeast (Pichia pastoris GS115) strains expressing inhibitory repeat domains (PpIRD(+)) of previously characterized Capsicum annuum protease inhibitors under high salt, heavy metal and oxidative stress. PpIRD(+) strains exhibited multiple stress tolerance and showed differential molecular responses at transcriptional and translational level on exposure to stress inducing agents like heavy metal, high salt and H2O2. PpIRD(+) strains display significant reduction in metacaspase (Yca1) activity, the key enzyme in apoptosis, indicates the possibility of cross reactivity of IRDs (serine protease inhibitor) with cysteine proteases. PpIRD(+) and Saccharomyces cerevisiae knockout with Yca1 (Delta Yca1) strain showed similar growth characteristics under stress, which indicated the delayed senescence due to cellular metacaspase inhibition. Molecular docking study showed a close proximity of IRDs reactive site and the active site of metacaspase in the complex that signified their strong interactions. Maintenance of GAPDH activity, primary target of metacaspase, in PpIRD(+) strain evidenced the inhibition of metacaspase activity and survival of these cells under stress. This report demonstrates a potential molecular mechanism of protease inhibitor-based multiple stress tolerance in yeast strains. (C) 2014 Elsevier Inc. All rights reserved.

10aMetacaspase10aMultiple stress tolerance10aprotease inhibitor10aYeast1 aJoshi, Rakesh, S.1 aTanpure, Rahul, S.1 aSingh, Rajan, Kumar1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/resistance-through-inhibition-ectopic-expression-serine-protease-inhibitor-offers-stress-002056nas a2200229 4500008004100000022001400041245006500055210006500120260010300185300001200288490000700300520125600307653002001563653002101583653001301604100002201617700001901639700002801658700002101686700002001707856009901727 2014 eng d a0981-942800aStress inducible proteomic changes in capsicum annuum leaves0 aStress inducible proteomic changes in capsicum annuum leaves a23 RUE LINOIS, 75724 PARIS, FRANCEbELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIERcJAN a212-2170 v743 a

Herbivore attack induces defense responses in plants, activating several signaling cascades. As a result, molecules deterrent to the herbivores are produced and accumulated in plants. Expression of defense mechanism/traits requires reorganization of the plant metabolism, redirecting the resources otherwise meant for growth. In the present work, protein profile of Capsicum annuum leaves was examined after herbivore attack/induction. Majority of proteins identified as differentially accumulated, were having roles in redox metabolism and photosynthesis. For example, superoxide dismutase and NADP oxidoreductase were upregulated by 10- and 6-fold while carbonic anhydrase and fructose-1,6-bisphosphatase were downregulated by 9- and 4-fold, respectively. Also, superoxide dismutase, NADPH quinone oxidoreductase and NADP dependent isocitrate dehydrogenase transcripts showed a higher accumulation in induced leaf tissues at early time points. In general, proteins having role in defense and damage repair were upregulated while those involved in photosynthesis appeared downregulated. Thus metabolic reconfiguration to balance defense and tolerance was evident in the stress-induced leaves. (C) 2013 Elsevier Masson SAS. All rights reserved.

10aCapsicum annuum10aOxidative stress10aWounding1 aMahajan, Neha, S.1 aMishra, Manasi1 aTamhane, Vaijayanti, A.1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/stress-inducible-proteomic-changes-capsicum-annuum-leaves-002128nas a2200289 4500008004100000022001400041245013100055210006900186260007100255300001600326490000700342520109100349653001701440653002501457653002601482653002301508100002201531700001701553700001501570700002201585700001701607700002301624700002201647700002101669700002001690856012801710 2014 eng d a0021-856100aWay toward ``dietary pesticides'': molecular investigation of insecticidal action of caffeic acid against helicoverpa armigera0 aWay toward dietary pesticides molecular investigation of insecti a1155 16TH ST, NW, WASHINGTON, DC 20036 USAbAMER CHEMICAL SOCcNOV a10847-108540 v623 a

Bioprospecting of natural molecules is essential to overcome serious environmental issues and pesticide resistance in insects. Here we are reporting insights into insecticidal activity of a plant natural phenol. In silico and in vitro screening of multiple molecules supported by in vivo validations suggested that caffeic acid (CA) is a potent inhibitor of Helicoverpa armigera gut proteases. Protease activity and gene expression were altered in CA-fed larvae. The structure-activity relationship of CA highlighted that all the functional groups are crucial for inhibition of protease activity. Biophysical studies and molecular dynamic simulations revealed that sequential binding of multiple CA molecules induces conformational changes in the protease(s) and thus lead to a significant decline in their activity. CA treatment significantly inhibits the insect's detoxification enzymes, thus intensifying the insecticidal effect. Our findings suggest that CA can be implicated as a potent insecticidal molecule and explored for the development of effective dietary pesticides.

10aCaffeic acid10aHelicoverpa armigera10ainsecticidal activity10aprotease inhibitor1 aJoshi, Rakesh, S.1 aWagh, T., P.1 aSharma, N.1 aMulani, Fayaj, A.1 aSonavane, U.1 aThulasiram, H., V.1 aJoshi, Rakesh, S.1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/way-toward-dietary-pesticides-molecular-investigation-insecticidal-action-caffeic-acid-002746nas a2200265 4500008004100000022001400041245011600055210006900171260007300240300001400313490000900327520177500336653001902111653002302130653002202153653002002175653002502195100002002220700002402240700002302264700002102287700002602308700002002334856012602354 2015 eng d a0304-416500aBiochemical, structural and functional diversity between two digestive alpha-amylases from helicoverpa armigera0 aBiochemical structural and functional diversity between two dige aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcSEP a1719-17280 v18503 a

Background: Helicoverpa armigera (Lepidoptera) feeds on various plants using diverse digestive enzymes as one of the survival tool-kit. The aim of the present study was to understand biochemical properties of recombinant alpha-amylases of H. armigera viz., HaAmy1 and HaAmy2. Methods: The open reading frames of HaAmy1 and HaAmy2 were cloned in Pichia pastoris and expressed heterologously. Purified recombinant enzymes were characterized for their biochemical and biophysical attributes using established methods. Results: Sequence alignment and homology modeling showed that HaAmy1 and HaAmy2 were conserved in their amino acid sequences and structures. HaAmy1 and HaAmy2 showed optimum activity at 60 degrees C; however, they differed in their optimum pH. Furthermore, HaAmy2 showed higher affinity for starch and amylopectin whereas HaAmy1 had higher catalytic efficiency. HaAmy1 and HaAmy2 were inhibited to the same magnitude by a synthetic amylase inhibitor (acarbose) while wheat amylase inhibitor showed about 2-fold higher inhibition of HaAmy1 than HaAmy2 at pH 7 while 6-fold difference at pH 11. Interactions of HaAmy1 and HaAmy2 with wheat amylase inhibitor revealed 2:1 stoichiometric ratio and much more complex interaction with HaAmy1. Conclusions: The diversity of amylases in perspective of their biochemical and biophysical properties, and their differential interactions with amylase inhibitors signify the potential role of these enzymes in adaptation of H. armigera on diverse plant diets. General significance: Characterization of digestive enzymes of H. armigera provides the molecular basis for the polyphagous nature and thus could assist in designing future strategies for the insect control. (C) 2015 Elsevier B.V. All rights reserved.

10aalpha-amylases10aAmylase inhibitors10aDigestive enzymes10aEnzyme activity10aHelicoverpa armigera1 aBhide, Amey, J.1 aChannale, Sonal, M.1 aPatil, Sucheta, S.1 aGupta, Vidya, S.1 aRamasamy, Sureshkumar1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/biochemical-structural-and-functional-diversity-between-two-digestive-alpha-amylases-002373nas a2200265 4500008004100000022001400041245011300055210006900168260007300237300001000310490000800320520140200328653001601730653002401746653002101770653002501791653002301816100002101839700002401860700002501884700002501909700002401934700002001958856012901978 2015 eng d a0927-776500aBio-physical evaluation and in vivo delivery of plant proteinase inhibitor immobilized on silica nanospheres0 aBiophysical evaluation and in vivo delivery of plant proteinase  aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcJUN a84-920 v1303 a

Recombinant expression of Capsicum annuum proteinase inhibitors (CanPI-13) and its application via synthetic carrier for the crop protection is the prime objective of our study. Herein, we explored proteinase inhibitor peptide immobilization on silica based nanospheres and rods followed by its pH mediated release in vitro and in vivo. Initial studies suggested silica nanospheres to be a suitable candidate for peptide immobilization. Furthermore, the interactions were characterized biophysically to ascertain their conformational stability and biological activity. Interestingly, bioactive peptide loading at acidic pH on nanospheres was found to be 62% and showed 56% of peptide release at pH 10, simulating gut milieu of the target pest Helicoverpa armigera. Additionally, in vivo study demonstrated significant reduction in insect body mass (158 mg) as compared to the control insects (265 mg) on 8th day after feeding with CanPI-13 based silica nanospheres. The study confirms that peptide immobilized silica nanosphere is capable of affecting overall growth and development of the feeding insects, which is known to hamper fecundity and fertility of the insects. Our study illustrates the utility and development of peptide-nanocarrier based platform in delivering diverse biologically active complexes specific to gut pH of H. armigera. (C) 2015 Elsevier B.V. All rights reserved.

10aBiodelivery10aHelicovera armigera10aProtein `corona'10aProteinase inhibitor10aSilica nanospheres1 aKhandelwal, Neha1 aDoke, Dhananjay, S.1 aKhandare, Jayant, J.1 aJawale, Priyanka, V.1 aBiradar, Ankush, V.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/bio-physical-evaluation-and-vivo-delivery-plant-proteinase-inhibitor-immobilized-silica-001985nas a2200265 4500008004100000022001400041245006200055210006100117260010600178300001000284490000700294520110800301653001401409653001401423653003201437653001601469653001601485653001701501653002001518100002201538700002201560700002201582700002001604856009501624 2015 eng d a0965-174800aCathepsins of lepidopteran insects: aspects and prospects0 aCathepsins of lepidopteran insects aspects and prospects aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLANDbPERGAMON-ELSEVIER SCIENCE LTDcSEP a51-590 v643 a

Molecular understanding of lepidopteran physiology has revealed that proteases consist of one of the central regulatory/reacting system for insect growth and survival. Among the various proteases, cathepsins are the most crucial cellular proteases, which play vital roles during insect development. In the present review, we have discussed various aspects of the lepidopteran insect cathepsins, emphasizing their roles in processes like development, growth, metamorphosis, apoptosis and immunity. Cathepsins are categorized into different types on the basis of their sequence diversification, leading to variation in structure and catalytic function. Cathepsins exhibit tissue and stage specific expression pattern which is fine-tuned by a delicate balance of expression, compartmentalization, zymogen activation, inhibition by protein inhibitors and degradation. The indispensability of cathepsins as cellular proteases in the above mentioned processes proposes them as novel targets for designing effective and specific insect controlling strategies. (C) 2015 Elsevier Ltd. All rights reserved.

10aApoptosis10aCathepsin10aCysteine/aspartate protease10aDevelopment10aLepidoptera10aPest control10aStress response1 aSaikhedkar, Nidhi1 aSummanwar, Aarohi1 aJoshi, Rakesh, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/cathepsins-lepidopteran-insects-aspects-and-prospects-002633nas a2200265 4500008004100000022001400041245022300055210006900278260010300347300001400450490000700464520147500471100002301946700002901969700003901998700002902037700001702066700002502083700002702108700002002135700002302155700003202178700002502210856013202235 2015 eng d a1535-947600aDevelopment of diagnostic fragment ion library for glycated peptides of human serum albumin: targeted quantification in prediabetic, diabetic, and microalbuminuria plasma by parallel reaction monitoring, SWATH, and MSE0 aDevelopment of diagnostic fragment ion library for glycated pept a9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USAbAMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INCcAUG a2150-21590 v143 a

Human serum albumin is one of the most abundant plasma proteins that readily undergoes glycation, thus glycated albumin has been suggested as an additional marker for monitoring glycemic status. Hitherto, only Amadori-modified peptides of albumin were quantified. In this study, we report the construction of fragment ion library for Amadori-modified lysine (AML), N(epsilon)-(carboxymethyl) lysine (CML)-, and N(epsilon)-(carboxyethyl) lysine (CEL)-modified peptides of the corresponding synthetically modified albumin using high resolution accurate mass spectrometry (HR/AM). The glycated peptides were manually inspected and validated for their modification. Further, the fragment ion library was used for quantification of glycated peptides of albumin in the context of diabetes. Targeted Sequential Window Acquisition of all THeoretical Mass Spectra (SWATH) analysis in pooled plasma samples of control, prediabetes, diabetes, and microalbuminuria, has led to identification and quantification of 13 glycated peptides comprised of four AML, seven CML, and two CEL modifications, representing nine lysine sites of albumin. Five lysine sites namely K549, K438, K490, K88, and K375, were observed to be highly sensitive for glycation modification as their respective m/z showed maximum fold change and had both AML and CML modifications. Thus, peptides involving these lysine sites could be potential novel markers to assess the degree of glycation in diabetes.

1 aKorwar, Arvind, M.1 aVannuruswamy, Garikapati1 aJagadeeshaprasad, Mashanipalya, G.1 aJayaramaiah, Ramesha, H.1 aBhat, Shweta1 aRegin, Bhaskaran, S.1 aRamaswamy, Sureshkumar1 aGiri, Ashok, P.1 aMohan, Viswanathan1 aBalasubramanyam, Muthuswamy1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/development-diagnostic-fragment-ion-library-glycated-peptides-human-serum-albumin-targeted-003064nas a2200265 4500008004100000022001400041245009700055210006900152260005700221300001200278490000800290520213900298653002002437653001702457653001402474653002302488653002902511100001902540700002802559700002202587700002402609700002102633700002002654856012402674 2015 eng d a0032-093500aEcological turmoil in evolutionary dynamics of plant-insect interactions: defense to offence0 aEcological turmoil in evolutionary dynamics of plantinsect inter a233 SPRING ST, NEW YORK, NY 10013 USAbSPRINGERcOCT a761-7710 v2423 a

Available history manifests contemporary diversity that exists in plant-insect interactions. A radical thinking is necessary for developing strategies that can co-opt natural insect-plant mutualism, ecology and environmental safety for crop protection since current agricultural practices can reduce species richness and evenness. The global environmental changes, such as increased temperature, CO (2) and ozone levels, biological invasions, land-use change and habitat fragmentation together play a significant role in re-shaping the plant-insect multi-trophic interactions. Diverse natural products need to be studied and explored for their biological functions as insect pest control agents. In order to assure the success of an integrated pest management strategy, human activities need to be harmonized to minimize the global climate changes. Plant-insect interaction is one of the most primitive and co-evolved associations, often influenced by surrounding changes. In this review, we account the persistence and evolution of plant-insect interactions, with particular focus on the effect of climate change and human interference on these interactions. Plants and insects have been maintaining their existence through a mutual service-resource relationship while defending themselves. We provide a comprehensive catalog of various defense strategies employed by the plants and/or insects. Furthermore, several important factors such as accelerated diversification, imbalance in the mutualism, and chemical arms race between plants and insects as indirect consequences of human practices are highlighted. Inappropriate implementation of several modern agricultural practices has resulted in (i) endangered mutualisms, (ii) pest status and resistance in insects and (iii) ecological instability. Moreover, altered environmental conditions eventually triggered the resetting of plant-insect interactions. Hence, multitrophic approaches that can harmonize human activities and minimize their interference in native plant-insect interactions are needed to maintain natural balance between the existence of plants and insects.

10aClimatic change10aCo-evolution10aEcosystem10aHuman interference10aPlant-insect interaction1 aMishra, Manasi1 aLomate, Purushottam, R.1 aJoshi, Rakesh, S.1 aPunekar, Sachin, A.1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/ecological-turmoil-evolutionary-dynamics-plant-insect-interactions-defense-offence-002495nas a2200277 4500008004100000022001400041245012300055210006900178260010600247300001200353490000800365520149500373653001301868653001801881653001001899653001701909653001201926653001601938100002001954700002601974700002602000700002402026700002002050700002102070856012602091 2015 eng d a0031-942200aMetabolic profiling of chickpea-Fusarium interaction identifies differential modulation of disease resistance pathways0 aMetabolic profiling of chickpeaFusarium interaction identifies d aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLANDbPERGAMON-ELSEVIER SCIENCE LTDcAUG a120-1290 v1163 a

Chickpea is the third most widely grown legume in the world and mainly used as a vegetarian source of human dietary protein. Fusarium wilt, caused by Fusarium oxysporum f. sp. ciceri (Foc), is one of the major threats to global chickpea production. Host resistance is the best way to protect crops from diseases; however, in spite of using various approaches, the mechanism of Foc resistance in chickpea remains largely obscure. In the present study, non-targeted metabolic profiling at several time points of resistant and susceptible chickpea cultivars using high-resolution liquid chromatography-mass spectrometry was applied to better understand the mechanistic basis of wilt resistance or susceptibility. Multivariate analysis of the data (OPLS-DA) revealed discriminating metabolites in chickpea root tissue after Foc inoculation such as flavonoids, isoflavonoids, alkaloids, amino acids and sugars. Foc inoculated resistant plants had more flavonoids and isoflavonoids along with their malonyl conjugates. Many antifungal metabolites that were induced after Foc infection viz, aurantion-obstine beta-glucosides and querecitin were elevated in resistant cultivar. Overall, diverse genetic and biochemical mechanisms were operational in the resistant cultivar for Foc defense as compared to the susceptible plant. The resistant chickpea plants employed the above-mentioned metabolic pathways as potential defense strategy against Foc. (C) 2015 Elsevier Ltd. All rights reserved.

10aChickpea10aFusarium wilt10aLC-MS10aMetabolomics10aOPLS-DA10aPhytoalexin1 aKumar, Yashwant1 aDholakia, Bhushan, B.1 aPanigrahi, Priyabrata1 aKadoo, Narendra, Y.1 aGiri, Ashok, P.1 aGupta, Vidya, S. uhttps://library.ncl.res.in/content/metabolic-profiling-chickpea-fusarium-interaction-identifies-differential-modulation-002856nas a2200277 4500008004100000022001400041245009000055210006900145260005700214300001200271490000800283520193300291653001002224653001302234653002202247653001302269653002502282100002202307700002002329700002802349700002202377700001902399700002102418700002002439856011902459 2015 eng d a0032-093500aStructural features of diverse Pin-II proteinase inhibitor genes from capsicum annuum0 aStructural features of diverse PinII proteinase inhibitor genes  a233 SPRING ST, NEW YORK, NY 10013 USAbSPRINGERcFEB a319-3310 v2413 a

The proteinase inhibitor (PI) genes from Capsicum annuum were characterized with respect to their UTR, introns and promoter elements. The occurrence of PIs with circularly permuted domain organization was evident. Several potato inhibitor II (Pin-II) type proteinase inhibitor (PI) genes have been analyzed from Capsicum annuum (L.) with respect to their differential expression during plant defense response. However, complete gene characterization of any of these C. annuum PIs (CanPIs) has not been carried out so far. Complete gene architectures of a previously identified CanPI-7 (Beads-on-string, Type A) and a member of newly isolated Bracelet type B, CanPI-69 are reported in this study. The 5' UTR (untranslated region), 3'UTR, and intronic sequences of both the CanPI genes were obtained. The genomic sequence of CanPI-7 exhibited, exon 1 (49 base pair, bp) and exon 2 (740 bp) interrupted by a 294-bp long type I intron. We noted the occurrence of three multi-domain PIs (CanPI-69, 70, 71) with circularly permuted domain organization. CanPI-69 was found to possess exon 1 (49 bp), exon 2 (551 bp) and a 584-bp long type I intron. The upstream sequence analysis of CanPI-7 and CanPI-69 predicted various transcription factor-binding sites including TATA and CAAT boxes, hormone-responsive elements (ABRELATERD1, DOFCOREZM, ERELEE4), and a defense-responsive element (WRKY71OS). Binding of transcription factors such as zinc finger motif MADS-box and MYB to the promoter regions was confirmed using electrophoretic mobility shift assay followed by mass spectrometric identification. The 3' UTR analysis for 25 CanPI genes revealed unique/distinct 3' UTR sequence for each gene. Structures of three domain CanPIs of type A and B were predicted and further analyzed for their attributes. This investigation of CanPI gene architecture will enable the better understanding of the genetic elements present in CanPIs.

10aCanPI10aCapsicum10aGene architecture10aPromoter10aProteinase inhibitor1 aMahajan, Neha, S.1 aDewangan, Veena1 aLomate, Purushottam, R.1 aJoshi, Rakesh, S.1 aMishra, Manasi1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/structural-features-diverse-pin-ii-proteinase-inhibitor-genes-capsicum-annuum-002409nas a2200169 4500008004100000022001400041245013400055210006900189260000800258300001800266490000600284520176200290100002002052700002502072700002002097856012202117 2015 eng d a2046-206900aTowards comprehension of complex chemical evolution and diversification of terpene and phenylpropanoid pathways in Ocimum species0 aTowards comprehension of complex chemical evolution and diversif cNOV a106886-1069040 v53 a

Ocimum species present a wide array of diverse secondary metabolites possessing immense medicinal and economic value. The importance of this genus is undisputable and exemplified in the ancient science of Chinese and Indian (Ayurveda) traditional medicine. Unlike several other plant species of Artemisia, Salvia, Catharanthus, Taxus, Mentha, etc. that are largely exploited, detailed characterization and identification of important metabolites from Ocimum species remained unexplored. Till date, most of the analyzed Ocimum species are predominantly rich in either phenylpropanoids or terpenoids. Metabolite data suggests domination of a unique set of signature compounds in all species. However, molecular pathways leading to the production, accumulation and metabolism of these compounds are poorly understood. The past few years have witnessed an upsurge in our understanding of the complex and intricately woven secondary metabolic pathways. Such information is generated through systematic analysis and correlation of metabolite profiling with transcriptomics data sets from different Ocimum species. The present review is aimed at integrating our current knowledge to understand the active secondary metabolic pathways, the key players in flux regulation including external stimuli, differential gene expression, transcription factors, microRNAs, enzyme promiscuity, etc. Extensive analysis of available data identifies events that may have contributed to evolve Ocimum species rich with a specific set of metabolites, thus, shedding light on pathway diversification. We believe that a better understanding of the multi-level regulation of intermediates and metabolites will help us harness the inherent diversity of Ocimum species optimally.

1 aSingh, Priyanka1 aKalunke, Raviraj, M.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/towards-comprehension-complex-chemical-evolution-and-diversification-terpene-and-002290nas a2200205 4500008004100000245011900041210006900160260000800229300001200237490000800249520153200257100002101789700002401810700002701834700002201861700002401883700002501907700002001932856013201952 2016 eng d00aBudding trends in integrated pest management using advanced micro- and nano-materials: challenges and perspectives0 aBudding trends in integrated pest management using advanced micr cDEC a157-1690 v1843 aOne of the most vital supports to sustain human life on the planet earth is the agriculture system that has been constantly challenged in terms of yield. Crop losses due to insect pest attack even after excessive use of chemical pesticides, are major concerns for humanity and environment protection. By the virtue of unique properties possessed by micro and nano-structures, their implementation in Agri-biotechnology is largely anticipated. Hence, traditional pest management strategies are now forestalling the potential of micro and nanotechnology as an effective and viable approach to alleviate problems pertaining to pest control. These technological innovations hold promise to contribute enhanced productivity by providing novel agrochemical agents and delivery systems. Application of these systems engages to achieve: i) control release of agrochemicals, ii) site-targeted delivery of active ingredients to manage specific pests, iii) reduced pesticide use, iv) detection of chemical residues, v) pesticide degradation, vi) nucleic acid delivery and vii) to mitigate post-harvest damage. Applications of micro and nano-technology are still marginal owing to the perception of low economic returns, stringent regulatory issues involving safety assessment and public awareness over their uses. In this review, we highlight the potential application of micro and nano-materials with a major focus on effective pest management strategies including safe handling of pesticides. (C) 2016 Elsevier Ltd. All rights reserved.1 aKhandelwal, Neha1 aBarbole, Ranjit, S.1 aBanerjee, Shashwat, S.1 aChate, Govind, P.1 aBiradar, Ankush, V.1 aKhandare, Jayant, J.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/budding-trends-integrated-pest-management-using-advanced-micro-and-nano-materials-challenges02071nas a2200181 4500008004100000245008400041210006900125260000800194300001200202490000800214520144300222100002201665700002401687700002601711700002001737700002101757856011101778 2016 eng d00aCharacterization of roseomonas and nocardioides spp. for arsenic transformation0 aCharacterization of roseomonas and nocardioides spp for arsenic  cNOV a742-7500 v3183 aThe metalloid arsenic predominantly exists in the arsenite [As(III)] and arsenate [As(V)]. These two forms are respectively oxidized and reduced by microbial redox processes. This study was designed to bioprospect arsenic tolerating bacteria from Lonar lake and to characterize their arsenic redoxing ability. Screening of sixty-nine bacterial species isolated from Lonar lake led to identification of three arsenic oxidizing and seven arsenic-reducing species. Arsenite oxidizing isolate Roseomonas sp. L-159a being closely related to Roseomonas cervicalis ATCC 49957 oxidized 2 mM As(III) in 60 h. Gene expression of large and small subunits of arsenite oxidase respectively showed 15- and 17-fold higher expression. Another isolate Nocardioides sp. L-37a formed a clade with Nocardioides ghangwensis JC2055, exhibited normal growth with different carbon sources and pH ranges. It reduced 2 mM As(V) in 36 h and showed constitutive expression of arsenate reductase which increased over 4-fold upon As(V) exposure. Genetic markers related to arsenic transformation were identified and characterized from the two isolates. Moderate resistance against the arsenicals was exhibited by the two isolates in the range of 1-5 mM for As(III) and 1-200 mM for As(V). Altogether we provide multiple evidences to indicate that Roseomonas sp. and Nocardioides sp. exhibited arsenic transformation ability. (C) 2016 Elsevier B.V. All rights reserved.1 aBagade, Aditi, V.1 aBachate, Sachin, P.1 aDholakia, Bhushan, B.1 aGiri, Ashok, P.1 aKodam, Kisan, M. uhttps://library.ncl.res.in/content/characterization-roseomonas-and-nocardioides-spp-arsenic-transformation02696nas a2200301 4500008004100000022001400041245016000055210006900215260010600284300000900390490000700399520155800406653001301964653001801977653002801995653002902023653001502052653002402067100002402091700002002115700001902135700002002154700002202174700002302196700002602219700002002245856012902265 2016 eng d a0965-174800aCharacterization of two coleopteran alpha-amylases and molecular insights into their differential inhibition by synthetic alpha-amylase inhibitor, acarbose0 aCharacterization of two coleopteran alphaamylases and molecular  aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLANDbPERGAMON-ELSEVIER SCIENCE LTDcJUL a1-110 v743 a

Post-harvest insect infestation of stored grains makes them unfit for human consumption and leads to severe economic loss. Here, we report functional and structural characterization of two coleopteran alpha-amylases viz. Callosobruchus chinensis alpha-amylase (CcAmy) and Tribolium castaneum alpha-amylase (TcAmy) along with their interactions with proteinaceous and non-proteinaceous alpha-amylase inhibitors. Secondary structural alignment of CcAmy and TcAmy with other coleopteran alpha-amylases revealed conserved motifs, active sites, di-sulfide bonds and two point mutations at spatially conserved substrate or inhibitor binding sites. Homology modeling and molecular docking showed structural differences between these two enzymes. Both the enzymes had similar optimum pH values but differed in their optimum temperature. Overall, pattern of enzyme stabilities were similar under various temperature and pH conditions. Further, CcAmy and TcAmy differed in their substrate affinity and catalytic efficiency towards starch and amylopectin. HPLC analysis detected common amylolytic products like maltose and maltotriose while glucose and malto-tetrose were unique in CcAmy and TcAmy catalyzed reactions respectively. At very low concentrations, wheat alpha-amylase inhibitor was found to be superior over the acarbose as far as complete inhibition of amylolytic activities of CcAmy and TcAmy was concerned. Mechanism underlying differential amylolytic reaction inhibition by acarbose was discussed. (C) 2016 Elsevier Ltd. All rights reserved.

10aAcarbose10aalpha-amylase10aalpha-amylase inhibitor10aCallosobruchus chinensis10aColeoptera10aTribolium castaneum1 aChannale, Sonal, M.1 aBhide, Amey, J.1 aYadav, Yashpal1 aKashyap, Garima1 aPawar, Pankaj, K.1 aMaheshwari, V., L.1 aRamasamy, Sureshkumar1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/characterization-two-coleopteran-alpha-amylases-and-molecular-insights-their-differential02690nas a2200253 4500008004100000245013700041210006900178260000800247300001400255490000900269520176500278100001602043700002902059700002502088700002402113700002202137700002202159700002602181700002402207700002202231700003702253700002002290856012602310 2016 eng d00aComparative functional characterization of eugenol synthase from four different ocimum species: implications on eugenol accumulation0 aComparative functional characterization of eugenol synthase from cNOV a1539-15470 v18643 a

Isoprenoids and phenylpropanoids are the major secondary metabolite constituents in Ocimum genus. Though enzymes from phenylpropanoid pathway have been characterized from few plants, limited information exists on how they modulate levels of secondary metabolites. Here, we performed phenylpropanoid profiling in different tissues from five Ocimum species, which revealed significant variations in secondary metabolites including eugenol, eugenol methyl ether, estragole and methyl cinnamate levels. Expression analysis of eugenol synthase (EGS) gene showed higher transcript levels especially in young leaves and inflorescence; and were positively correlated with eugenol contents. Additionally, transcript levels of coniferyl alcohol acyl transferase, a key enzyme diverting pool of substrate to phenylpropanoids, were in accordance with their abundance in respective species. In particular, eugenol methyl transferase expression positively correlated with higher levels of eugenol methyl ether in Ocimum tenuiflorum. Further, EGSs were functionally characterized from four Ocimum species varying in their eugenol contents. Kinetic and expression analyses indicated, higher enzyme turnover and transcripts levels, in species accumulating more eugenol. Moreover, biochemical and bioinformatics studies demonstrated that coniferyl acetate was the preferred substrate over coumaryl acetate when used, individually or together, in the enzyme assay. Overall, this study revealed the preliminary evidence for varied accumulation of eugenol and its abundance over chavicol in these Ocimum species. Current findings could potentially provide novel insights for metabolic modulations in medicinal and aromatic plants. (C) 2016 Elsevier B.V. All rights reserved.

1 aAnand, Atul1 aJayaramaiah, Ramesha, H.1 aBeedkar, Supriya, D.1 aSingh, Priyanka, A.1 aJoshi, Rakesh, S.1 aMulani, Fayaj, A.1 aDholakia, Bhushan, B.1 aPunekar, Sachin, A.1 aGade, Wasudeo, N.1 aThulasiram, Hirekodathakallu, V.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/comparative-functional-characterization-eugenol-synthase-four-different-ocimum-species02747nas a2200253 4500008004100000022001400041245014700055210006900202260006700271300001000338490000700348520180200355653001702157653002502174653002702199653001502226653001802241100002302259700002402282700002102306700002102327700002002348856012502368 2016 eng d a1672-960900aExpression of proteins involved in digestion and detoxification are regulated in helicoverpa armigera to cope up with chlorpyrifos insecticide0 aExpression of proteins involved in digestion and detoxification  a111 RIVER ST, HOBOKEN 07030-5774, NJ USAbWILEY-BLACKWELLcFEB a68-770 v233 a

Helicoverpa armigera is a key pest in many vital crops, which is mainly controlled by chemical strategies. To manage this pest is becoming challenging due to its ability and evolution of resistance against insecticides. Further, its subsequent spread on nonhost plant is remarkable in recent times. Hence, decoding resistance mechanism against phytochemicals and synthetic insecticides is a major challenge. The present work describes that the digestion, defense and immunity related enzymes are associated with chlorpyrifos resistance in H. armigera. Proteomic analysis of H. armigera gut tissue upon feeding on chlorpyrifos containing diet (CH) and artificial diet (AD) using nano-liquid chromatography-mass spectrometry identified upregulated 23-proteins in CH fed larvae. Database searches combined with gene ontology analysis revealed that the identified gut proteins engrossed in digestion, proteins crucial for immunity, adaptive responses to stress, and detoxification. Biochemical and quantitative real-time polymerase chain reaction analysis of candidate proteins indicated that insects were struggling to get nutrients and energy in presence of CH, while at the same time endeavoring to metabolize chlorpyrifos. Moreover, we proposed a potential processing pathway of chlorpyrifos in H. armigera gut by examining the metabolites using gas chromatography-mass spectrometry. H. armigera exhibit a range of intriguing behavioral, morphological adaptations and resistance to insecticides by regulating expression of proteins involved in digestion and detoxification mechanisms to cope up with chlorpyrifos. In these contexts, as gut is a rich repository of biological information; profound analysis of gut tissues can give clues of detoxification and resistance mechanism in insects.

10achlorpyrifos10aHelicoverpa armigera10ainsecticide resistance10aproteomics10areal-time PCR1 aDawkar, Vishal, V.1 aChikate, Yojana, R.1 aMore, Tushar, H.1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/expression-proteins-involved-digestion-and-detoxification-are-regulated-helicoverpa-002545nas a2200301 4500008004100000022001400041245015600055210006900211260009500280300001200375490000800387520137900395653002301774653003301797653001101830653002101841653002501862100002901887700001601916700002501932700002601957700002401983700002502007700002202032700003702054700002002091856013202111 2016 eng d a0006-291X00aFunctional characterization and transient expression manipulation of a new sesquiterpene synthase involved in beta-caryophyllene accumulation in Ocimum0 aFunctional characterization and transient expression manipulatio a525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USAbACADEMIC PRESS INC ELSEVIER SCIENCEcAPR a265-2710 v4733 a

The genus Ocimum has a unique blend of diverse secondary metabolites, with major proportion of terpenoids including mono- and sesquiterpenes. Although, beta-Caryophyllene, bicyclic sesquiterpene, is one of the major terpene found in Ocimum species and known to possess several biological activities, not much is known about its biosynthesis in Ocimum. Here, we describe isolation and characterization of beta-caryophyllene synthase gene from Ocimum kilimandscharicum Gurke (OkBCS- GenBank accession no. KP226502). The open reading frame of 1629 bp encoded a protein of 542 amino acids with molecular mass of 63.6 kDa and pl value of 5.66. The deduced amino acid sequence revealed 50-70% similarity with known sesquiterpene synthases from angiosperms. Recombinant OkBCS converted farnesyl diphosphate to beta-caryophyllene as a major product (94%) and 6% alpha-humulene. Expression variation of OkBCS well corroborated with beta-caryophyllene levels in different tissues from five Ocimum species. OkBCS transcript revealed higher expression in leaves and flowers. Further, agro-infiltration based transient expression manipulation with OkBCS over-expression and silencing confirmed its role in beta-caryophyllene biosynthesis. These findings may potentially be further utilized to improve plant defense against insect pests. (C) 2016 Elsevier Inc. All rights reserved.

10abeta-Caryophyllene10aNatural product biosynthesis10aOcimum10aTerpene synthase10aTransient expression1 aJayaramaiah, Ramesha, H.1 aAnand, Atul1 aBeedkar, Supriya, D.1 aDholakia, Shushan, B.1 aPunekar, Sachin, A.1 aKalunke, Raviraj, M.1 aGade, Wasudeo, N.1 aThulasiram, Hirekodathakallu, V.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/functional-characterization-and-transient-expression-manipulation-new-sesquiterpene-synthase02652nas a2200373 4500008004100000022001400041245014200055210006900197260006700266300001400333490000700347520139300354653001301747653002301760653001701783653000801800653003101808653001501839100002001854700001701874700002601891700002601917700002001943700002301963700002501986700001602011700001302027700003402040700002402074700002002098700001602118700002102134856012302155 2016 eng d a1467-764400aFusarium oxysporum mediates systems metabolic reprogramming of chickpea roots as revealed by a combination of proteomics and metabolomics0 aFusarium oxysporum mediates systems metabolic reprogramming of c a111 RIVER ST, HOBOKEN 07030-5774, NJ USAbWILEY-BLACKWELLcJUL a1589-16030 v143 a

Molecular changes elicited by plants in response to fungal attack and how this affects plant-pathogen interaction, including susceptibility or resistance, remain elusive. We studied the dynamics in root metabolism during compatible and incompatible interactions between chickpea and Fusarium oxysporum f. sp. ciceri (Foc), using quantitative label-free proteomics and NMR-based metabolomics. Results demonstrated differential expression of proteins and metabolites upon Foc inoculations in the resistant plants compared with the susceptible ones. Additionally, expression analysis of candidate genes supported the proteomic and metabolic variations in the chickpea roots upon Foc inoculation. In particular, we found that the resistant plants revealed significant increase in the carbon and nitrogen metabolism; generation of reactive oxygen species (ROS), lignification and phytoalexins. The levels of some of the pathogenesis-related proteins were significantly higher upon Foc inoculation in the resistant plant. Interestingly, results also exhibited the crucial role of altered Yang cycle, which contributed in different methylation reactions and unfolded protein response in the chickpea roots against Foc. Overall, the observed modulations in the metabolic flux as outcome of several orchestrated molecular events are determinant of plant's role in chickpea-Foc interactions.

10aChickpea10afusarium oxysporum10aMetabolomics10aNMR10aplant-pathogen interaction10aproteomics1 aKumar, Yashwant1 aZhang, Limin1 aPanigrahi, Priyabrata1 aDholakia, Bhushan, B.1 aDewangan, Veena1 aChavan, Sachin, G.1 aKunjir, Shrikant, M.1 aWu, Xiangyu1 aLi, Ning1 aRajmohanan, Pattuparambil, R.1 aKadoo, Narendra, Y.1 aGiri, Ashok, P.1 aTang, Huiru1 aGupta, Vidya, S. uhttps://library.ncl.res.in/content/fusarium-oxysporum-mediates-systems-metabolic-reprogramming-chickpea-roots-revealed02264nas a2200301 4500008004100000022001400041245011700055210006900172260009000241300002600331490000600357520118200363100002401545700002701569700001901596700002701615700002001642700002001662700001401682700001601696700002001712700002101732700002001753700002001773700001801793700001901811856013201830 2016 eng d a2041-172300aGAME9 regulates the biosynthesis of steroidal alkaloids and upstream isoprenoids in the plant mevalonate pathway0 aGAME9 regulates the biosynthesis of steroidal alkaloids and upst aMACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLANDbNATURE PUBLISHING GROUPcFEB aArticle Number: 106540 v73 a

Steroidal glycoalkaloids (SGAs) are cholesterol-derived molecules produced by solanaceous species. They contribute to pathogen defence but are toxic to humans and considered as anti-nutritional compounds. Here we show that GLYCOALKALOID METABOLISM 9 (GAME9), an APETALA2/Ethylene Response Factor, related to regulators of alkaloid production in tobacco and Catharanthus roseus, controls SGA biosynthesis. GAME9 knockdown and overexpression in tomato and potato alters expression of SGAs and upstream mevalonate pathway genes including the cholesterol biosynthesis gene STEROL SIDE CHAIN REDUCTASE 2 (SSR2). Levels of SGAs, C24-alkylsterols and the upstream mevalonate and cholesterol pathways intermediates are modified in these plants. Delta(7)-STEROL-C5(6)-DESATURASE (C5-SD) in the hitherto unresolved cholesterol pathway is a direct target of GAME9. Transactivation and promoter-binding assays show that GAME9 exerts its activity either directly or cooperatively with the SlMYC2 transcription factor as in the case of the C5-SD gene promoter. Our findings provide insight into the regulation of SGA biosynthesis and means for manipulating these metabolites in crops.

1 aCardenas, Pablo, D.1 aSonawane, Prashant, D.1 aPollier, Jacob1 aBossche, Robin, Vanden1 aDewangan, Veena1 aWeithorn, Efrat1 aTal, Lior1 aMeir, Sagit1 aRogachev, Ilana1 aMalitsky, Sergey1 aGiri, Ashok, P.1 aGoossens, Alain1 aBurdman, Saul1 aAharoni, Asaph uhttps://library.ncl.res.in/content/game9-regulates-biosynthesis-steroidal-alkaloids-and-upstream-isoprenoids-plant-mevalonate-102412nas a2200277 4500008004100000022001400041245011500055210006900170260007900239300001200318490000700330520145300337653001301790653001501803653001201818653001001830653001401840653001301854100002501867700002201892700002601914700002301940700002001963700002101983856013002004 2016 eng d a1073-608500aMolecular cloning and characterization of O-methyltransferase from mango fruit (mangifera indica cv. alphonso)0 aMolecular cloning and characterization of Omethyltransferase fro a999 RIVERVIEW DRIVE SUITE 208, TOTOWA, NJ 07512 USAbHUMANA PRESS INCcMAY a340-3500 v583 a

Flavour of ripe Alphonso mango is invariably dominated by the de novo appearance of lactones and furanones during ripening. Of these, furanones comprising furaneol (4-hydroxy-2,5-dimethyl-3(2H)-furanone) and mesifuran (2,5-dimethyl-4-methoxy-3(2H)-furanone) are of particular importance due to their sweet, fruity caramel-like flavour characters and low odour detection thresholds. We isolated a 1056 bp complete open reading frame of a cDNA encoding S-adenosyl-l-methionine-dependent O-methyltransferase from Alphonso mango. The recombinantly expressed enzyme, MiOMTS showed substrate specificity towards furaneol and protocatechuic aldehyde synthesizing mesifuran and vanillin, respectively, in an in vitro assay reaction. A semi-quantitative PCR analysis showed fruit-specific expression of MiOMTS transcripts. Quantitative real-time PCR displayed ripening-related expression pattern of MiOMTS in both pulp and skin of Alphonso mango. Also, early and significantly enhanced accumulation of its transcripts was detected in pulp and skin of ethylene-treated fruits. Ripening-related and fruit-specific expression profile of MiOMTS and substrate specificity towards furaneol is a suggestive of its involvement in the synthesis of mesifuran in Alphonso mango. Moreover, a significant trigger in the expression of MiOMTS transcripts in ethylene-treated fruits point towards the transcriptional regulation of mesifuran biosynthesis by ethylene.

10aEthylene10aexpression10aFlavour10aMango10aMesifuran10aRipening1 aChidley, Hemangi, G.1 aOak, Pranjali, S.1 aDeshpande, Ashish, B.1 aPujari, Keshav, H.1 aGiri, Ashok, P.1 aGupta, Vidya, S. uhttps://library.ncl.res.in/content/molecular-cloning-and-characterization-o-methyltransferase-mango-fruit-mangifera-indica-cv02074nas a2200181 4500008004100000245017100041210006900212260000800281300001000289490000700299520135000306100001801656700002301674700002001697700002301717700002901740856012301769 2016 eng d00aParasitism by Chelonus blackburni (Hymenoptera) affects food consumption and development of Helicoverpa armigera (Lepidoptera) and cellular architecture of the midgut0 aParasitism by Chelonus blackburni Hymenoptera affects food consu cMAR a65-700 v193 aBiological control agents are vital components of an integrated pest management strategy, and this is frequently referred to as natural control. Natural enemies of insect pests include predators, parasitoids, and pathogens. Among them, a parasitoid, Chelonus blackburni (Cameron), was found to be the best biological control agent for the polyphagous pest, Helicoverpa armigera (Hubner). C. blackburni alters the feeding performance of H. annigera larvae upon parasitism and as a result severely affects growth and development. Moreover, it shortens the feeding period of H. armigera and increases mortality. Furthermore, total hemocyte count (THC) was significantly decreased in parasitized larvae than control. Parasitized H. armigera had 26% less number of blood cells compared to healthy larvae. Histological studies showed that the structure of midgut of H. annigera is drastically affected by C. blackburni leading to reduced food consumption, which ultimately led to larval death. The present study provides an insight to changes involved in H. annigera due to parasitism by C blackburni, a parasite that could be used as an effective biocontrol agent to manage H. armigera. (C) 2015 Korean Society of Applied Entomology, Taiwan Entomological Society and Malaysian Plant Protection Society. Published by Elsevier B.V. All rights reserved.1 aSanap, Yogita1 aDawkar, Vishal, V.1 aGiri, Ashok, P.1 aSen, Avalokiteswar1 aPandit, Radhakrishna, S. uhttps://library.ncl.res.in/content/parasitism-chelonus-blackburni-hymenoptera-affects-food-consumption-and-development02556nas a2200277 4500008004100000022001400041245012900055210006900184260009000253490000600343520147700349100002001826700002901846700002401875700002901899700002301928700001601951700002601967700002401993700002202017700002602039700002502065700003702090700002002127856013102147 2016 eng d a2045-232200aPotential dual role of eugenol in inhibiting advanced glycation end products in diabetes: proteomic and mechanistic insights0 aPotential dual role of eugenol in inhibiting advanced glycation  aMACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLANDbNATURE PUBLISHING GROUPcJAN0 v63 a

Medicinally important genus Ocimum harbors a vast pool of chemically diverse metabolites. Current study aims at identifying anti-diabetic candidate compounds from Ocimum species. Major metabolites in O. kilimandscharicum, O. tenuiflorum, O. gratissimum were purified, characterized and evaluated for anti-glycation activity. In vitro inhibition of advanced glycation end products (AGEs) by eugenol was found to be highest. Preliminary biophysical analysis and blind docking studies to understand eugenol-albumin interaction indicated eugenol to possess strong binding affinity for surface exposed lysines. However, binding of eugenol to bovine serum albumin (BSA) did not result in significant change in secondary structure of protein. In vivo diabetic mice model studies with eugenol showed reduction in blood glucose levels by 38% likely due to inhibition of alpha-glucosidase while insulin and glycated hemoglobin levels remain unchanged. Western blotting using anti-AGE antibody and mass spectrometry detected notably fewer AGE modified peptides upon eugenol treatment both in vivo and in vitro. Histopathological examination revealed comparatively lesser lesions in eugenol-treated mice. Thus, we propose eugenol has dual mode of action in combating diabetes; it lowers blood glucose by inhibiting a-glucosidase and prevents AGE formation by binding to epsilon-amine group on lysine, protecting it from glycation, offering potential use in diabetic management.

1 aSingh, Priyanka1 aJayaramaiah, Ramesha, H.1 aAgawane, Sachin, B.1 aVannuruswamy, Garikapati1 aKorwar, Arvind, M.1 aAnand, Atul1 aDhaygude, Vitthal, S.1 aShaikh, Mahemud, L.1 aJoshi, Rakesh, S.1 aBoppana, Ramanamurthy1 aKulkarni, Mahesh, J.1 aThulasiram, Hirekodathakallu, V.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/potential-dual-role-eugenol-inhibiting-advanced-glycation-end-products-diabetes-proteomic-002505nas a2200253 4500008004100000022001400041245010700055210006900162260010300231300001400334490000700348520151500355100001701870700003901887700002401926700002401950700002501974700002301999700002002022700003202042700002602074700002502100856012602125 2016 eng d a1535-947600aProteomic insight reveals elevated levels of albumin in circulating immunecomplexes in diabetic plasma0 aProteomic insight reveals elevated levels of albumin in circulat a9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USAbAMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INCcJUN a2011-20200 v153 a

A Hyperglycemic condition in diabetes promotes formation of advanced glycation end products, which are known to elicit immune response and form complexes with immunoglobulins called circulating immune complexes. To investigate the involvement of advanced glycation end product (AGE)-modified proteins in the elicitation of an immune response, circulating immune complexes were isolated and proteins associated were identified and characterized. Label-free-based mass spectrometric analysis of circulating immune complexes in clinical plasma of prediabetic, newly diagnosed diabetes, and diabetic microalbuminurea revealed elevated levels of serum albumin in the circulating immune complexes, which were also observed to be AGE modified. Further, to examine the role of glycation, circulating immune complexeswere analyzed in the streptozotocin-induced diabetic mice treated with or without aminoguanidine, a prototype glycation inhibitor. Mass spectrometric analysis of circulating immune complexes showed elevated levels of serum albumin in plasma from diabetic mice over that of control animals. Aminoguanidine-treated diabetic mice displayed decreased AGE modification of plasma albumin, accompanied by a reduced level of albumin in the circulating immune complexes. In addition, elevated levels of proinflammatory cytokines such as IL-1b, IL-2, and TNF-alpha were observed in diabetes, which were reduced with aminoguanidine treatment, suggesting the involvement of glycation in the immune response.

1 aBhat, Shweta1 aJagadeeshaprasad, Mashanipalya, G.1 aPatil, Yugendra, R.1 aShaikh, Mahemud, L.1 aRegin, Bhaskaran, S.1 aMohan, Viswanathan1 aGiri, Ashok, P.1 aBalasubramanyam, Muthuswamy1 aBoppana, Ramanamurthy1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/proteomic-insight-reveals-elevated-levels-albumin-circulating-immunecomplexes-diabetic02304nas a2200193 4500008004100000245009500041210006900136260000800205300001000213490000800223520161200231100002401843700002301867700002401890700002901914700002101943700002001964856012601984 2016 eng d00aRNAi of selected candidate genes interrupts growth and development of Helicoverpa armigera0 aRNAi of selected candidate genes interrupts growth and developme cOCT a44-510 v1333 aHelicoverpa armigera is one of the major crop pests and is less amenable to current pest control approaches. RNA interference (RNAi) is emerging as a potent arsenal for the insect pest control over current methods. Here, we examined the effect on growth and development in H. armigera by targeting various enzymes/proteins such as proteases like trypsins (HaTry2, 3, 4 and 6), chymotrypsin (HaChy4) and cysteine protease like cathepsin (HaCATHL); glutathione S-transferases (HaGST1a, 6 and 8); esterases (HaAce4, HaJHE); catalase (HaCAT); super-oxide-dismutase (HaCu/ZnSOD); fatty acid binding protein (HaFabp) and chitin deacetylase (HaCda5b) through dsRNA approach. Significant downregulation of cognate mRNA expression and reduced activity of trypsin and GST-like enzyme were evident upon feeding candidate dsRNAs to the larvae. Among these, the highest mortality was observed in HaAce4 dsRNA fed larvae followed by HaJHE; HaCAT; HaCuZnSOD; HaFabp and HaTry3 whereas remaining ones showed relatively lower mortality. Furthermore, the dsRNA fed larvae showed significant reduction in the larval mass and abnormalities at the different stages of H. armigera development compared to their control diets. For example, malformed larvae, pupae and moth at a dose of 60 pg/day were evident in high number of individual insects fed on dsRNA containing diets. Moreover, the growth and development of insects and moths were retarded in dsRNA fed larvae. These findings might provide potential new candidates for designing effective dsRNA as pesticide in crop protection. (C) 2016 Elsevier Inc. All rights reserved.1 aChikate, Yojana, R.1 aDawkar, Vishal, V.1 aBarbole, Ranjit, S.1 aTilak, Priyadarshini, V.1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/rnai-selected-candidate-genes-interrupts-growth-and-development-helicoverpa-armigera-001559nas a2200181 4500008004100000245007800041210006900119260000800188300001200196490000700208520094600215100001601161700002501177700002601202700002301228700002001251856010601271 2017 eng d00a Differential accumulation of vimentin fragments in preeclamptic placenta0 aDifferential accumulation of vimentin fragments in preeclamptic  cNOV a420-4250 v743 aPreeclampsia is a pregnancy complication that is the result of abnormal placentation because of inadequate trophoblast invasion into spiral arteries that prevent normal blood flow to the placenta. We report the alteration in vimentin protein proteolysis in placenta of normotensive and preeclamptic women, which is known to have a role in many physiological functions other than its major function in the structural integrity of the cell. Placental proteome from normotensive (n=25) and preeclamptic pregnancies (n=25) showed eight differentially accumulated protein spots of vimentin (proteolytic fragments) by two-dimensional electrophoresis. Immunoblots of normotensive and preeclamptic placenta revealed a difference in proteolytic processing of vimentin. In particular, lower molecular weight vimentin fragments of 32 and 20 kDa were 3.3 and 2.6-fold (p<0.0001) higher, respectively, in preeclampsia compared with normotensive placenta.1 aMary, Sheon1 aKulkarni, Mahesh, J.1 aMehendale, Savita, S.1 aJoshi, Sadhana, R.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/differential-accumulation-vimentin-fragments-preeclamptic-placenta02492nas a2200229 4500008004100000245017500041210006900216260000800285300001200293490000700305520162800312100002701940700002601967700002001993700002002013700001602033700002002049700001902069700002002088700002402108856013002132 2017 eng d00a Dynamic metabolic reprogramming of steroidal glycol-alkaloid and phenylpropanoid biosynthesis may impart early blight resistance in wild tomato (Solanum arcanum Peralta)0 aDynamic metabolic reprogramming of steroidal glycolalkaloid and  cNOV a411-4230 v953 aAlternaria solani severely affects tomato (Solanum lycopersicum L.) yield causing early blight (EB) disease in tropical environment. Wild relative, Solanum arcanum Peralta could be a potential source of EB resistance; however, its underlying molecular mechanism largely remains unexplored. Hence, non-targeted metabolomics was applied on resistant and susceptible S. arcanum accessions upon A. solani inoculation to unravel metabolic dynamics during different stages of disease progression. Total 2047 potential metabolite peaks (mass signals) were detected of which 681 and 684 metabolites revealed significant modulation and clear differentiation in resistant and susceptible accessions, respectively. Majority of the EB-triggered metabolic changes were active from steroidal glycol-alkaloid (SGA), lignin and flavonoid biosynthetic pathways. Further, biochemical and gene expression analyses of key enzymes from these pathways positively correlated with phenotypic variation in the S. arcanum accessions indicating their potential role in EB. Additionally, transcription factors regulating lignin biosynthesis were also up-regulated in resistant plants and electrophoretic mobility shift assay revealed sequence-specific binding of rSaWRKY1 with MYB20 promoter. Moreover, transcript accumulation of key genes from phenylpropanoid and SGA pathways along with WRKY and MYB in WRKY1 transgenic tomato lines supported above findings. Overall, this study highlights vital roles of SGAs as phytoalexins and phenylpropanoids along with lignin accumulation unrevealing possible mechanistic basis of EB resistance in wild tomato.1 aShinde, Balkrishna, A.1 aDholakia, Bhushan, B.1 aHussain, Khalid1 aPanda, Sayantan1 aMeir, Sagit1 aRogachev, Ilana1 aAharoni, Asaph1 aGiri, Ashok, P.1 aKamble, Avinash, C. uhttps://library.ncl.res.in/content/dynamic-metabolic-reprogramming-steroidal-glycol-alkaloid-and-phenylpropanoid-biosynthesis02353nas a2200193 4500008004100000245012300041210006900164260000800233300001400241490000900255520163300264100002501897700002601922700002201948700002301970700002001993700002102013856012502034 2017 eng d00aEffect of postharvest ethylene treatment on sugar content, glycosidase activity and its gene expression in mango fruit0 aEffect of postharvest ethylene treatment on sugar content glycos cMAR a1630-16390 v97 3 a

BACKGROUND: Ripening-associated softening is one of the important attributes that largely determines the shelf-life of mango (Mangifera indica Linn.) fruits. To reveal the effect of pre-climacteric ethylene treatment on ripening-related softening of Alphonso mango, ethylene treatment was given to mature, raw Alphonso fruits. Changes in the pool of reducing and non-reducing sugars, enzymatic activity of three glycosidases: ss-D-galactosidase, alpha-D-mannosidase and ss-D-glucosidase and their relative transcript abundance were analysed for control and ethylene treated fruits during ripening. RESULTS: Early activity of all the three glycosidases and accelerated accumulation of reducing and non-reducing sugars on ethylene treatment was evident. ss-D-Galactosidase showed the highest activity among three glycosidases in control fruits and marked increase in activity upon ethylene treatment. This was confirmed by the histochemical assay of its activity in control and ethylene treated ripe fruits. Relative transcript abundance revealed high transcript levels of ss-D-galactosidase in control fruits. Ethylene-treated fruits showed early and remarkable increase in the ss-D-galactosidase transcriptswhile alpha-D-mannosidase transcript variants displayed early accumulation. CONCLUSION: Thefindings suggest reduction in the shelf-life of Alphonso mango upon pre-climacteric ethylene treatment, a significant role of ss-D-galactosidase and alpha-D-mannosidase in the ripening related softening of Alphonso fruits and transcriptional regulation of their expression by ethylene. (C) 2016 Society of Chemical Industry

1 aChidley, Hemangi, G.1 aDeshpande, Ashish, B.1 aOak, Pranjali, S.1 aPujari, Keshav, H.1 aGiri, Ashok, P.1 aGupta, Vidya, S. uhttps://library.ncl.res.in/content/effect-postharvest-ethylene-treatment-sugar-content-glycosidase-activity-and-its-gene02629nas a2200229 4500008004100000245012800041210006900169260000800238300001200246490001000258520179900268100002002067700002402087700001902111700002602130700002202156700002302178700002102201700002602222700002002248856013102268 2017 eng d00aGenomic and functional characterization of coleopteran insect-specific alpha-amylase inhibitor gene from amaranthus species0 aGenomic and functional characterization of coleopteran insectspe cJUN a319-3320 v 94 3 aThe smallest 32 amino acid alpha-amylase inhibitor from Amaranthus hypochondriacus (AAI) is reported. The complete gene of pre-protein (AhAI) encoding a 26 amino acid (aa) signal peptide followed by the 43 aa region and the previously identified 32 aa peptide was cloned successfully. Three cysteine residues and one disulfide bond conserved within known alpha-amylase inhibitors were present in AhAI. Identical genomic and open reading frame was found to be present in close relatives of A. hypochondriacus namely Amaranthus paniculatus, Achyranthes aspera and Celosia argentea. Interestingly, the 3'UTR of AhAI varied in these species. The highest expression of AhAI was observed in A. hypochondriacus inflorescence; however, it was not detected in the seed. We hypothesized that the inhibitor expressed in leaves and inflorescence might be transported to the seeds. Sub-cellular localization studies clearly indicated the involvement of AhAI signal peptide in extracellular secretion. Full length rAhAI showed differential inhibition against alpha-amylases from human, insects, fungi and bacteria. Particularly, alpha-amylases from Helicoverpa armigera (Lepidoptera) were not inhibited by AhAI while Tribolium castaneum and Callosobruchus chinensis (Coleoptera) alpha-amylases were completely inhibited. Molecular docking of AhAI revealed tighter interactions with active site residues of T. castaneum alpha-amylase compared to C. chinensis alpha-amylase, which could be the rationale behind the disparity in their IC50. Normal growth, development and adult emergence of C. chinensis were hampered after feeding on rAhAI. Altogether, the ability of AhAI to affect the growth of C. chinensis demonstrated its potential as an efficient bio-control agent, especially against stored grain pests.1 aBhide, Amey, J.1 aChannale, Sonal, M.1 aYadav, Yashpal1 aBhattacharjee, Kabita1 aPawar, Pankaj, K.1 aMaheshwari, V., L.1 aGupta, Vidya, S.1 aRamasamy, Sureshkumar1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/genomic-and-functional-characterization-coleopteran-insect-specific-alpha-amylase-inhibitor01948nas a2200217 4500008004100000245014600041210006900187260000800256300001400264490000700278520115500285100001801440700002101458700002001479700002701499700002301526700001501549700002101564700002101585856012401606 2017 eng d00aGlucosylation of smoke-derived volatiles in grapevine (vitis vinifera) is catalyzed by a promiscuous resveratrol/guaiacol glucosyltransferase0 aGlucosylation of smokederived volatiles in grapevine vitis vinif cJUL a5681-56890 v653 aVinification of grapes (Vitis vinifera) exposed to forest fire smoke can yield unpalatable wine due to the presence of taint compounds from smoke and the release of smoke derived volatiles from their respective glycosides during the fermentation process or in-mouth during consumption. To identify glycosyltransferases (GTs) involved in the formation of glycosidically bound smoke-derived volatiles we performed gene expression analysis of candidate GTs in different grapevine tissues. Second, substrates derived from bushfire smoke or naturally occurring in grapes were screened with the candidate recombinant GTs. A resveratrol GT (UGT72B27) gene, highly expressed in grapevine leaves and berries was identified to be responsible for the production of the phenolic glucosides. UGT72B27 converted the stilbene trans-resveratrol mainly to the 3-O-glucoside. Kinetic analyses yielded specificity constants (k(cat)/K-m) of 114, 17, 9, 8, and 2 mM(-1) s(-1) for guaiacol, trans-resveratrol, syringol, methylsyringol, and methylguaiacol, respectively. This knowledge will help to design strategies for managing the risk of producing smoke-affected wines.1 aHaratl, Katja1 aHuang, Fong-Chin1 aGiri, Ashok, P.1 aFranz-Oberdorf, Katrin1 aFrotscher, Johanna1 aShao, Yang1 aHoffmann, Thomas1 aSchwab, Wilfried uhttps://library.ncl.res.in/content/glucosylation-smoke-derived-volatiles-grapevine-vitis-vinifera-catalyzed-promiscuous02383nas a2200205 4500008004100000245017400041210006900215260000800284300001400292490000700306520157700313100002301890700002301913700002001936700002601956700002001982700002602002700002202028856012702050 2017 eng d00aGlycoprotein alpha-amylase inhibitor from Withania somnifera differentially inhibits various alpha-amylases and affects the growth and development of Tribolium castaneum0 aGlycoprotein alphaamylase inhibitor from Withania somnifera diff cJUL a1382-13900 v733 a

BACKGROUND: Identification and characterisation of plant defensive molecules enrich our resources to design crop protection strategies. In particular, plant-derived proteinaceous inhibitor(s) of insect digestive enzymes appear to be a safe, sustainable and attractive option. RESULTS: A glycoprotein having non-competitive alpha-amylase inhibitory activity with a molecular weight of 8.3 kDa was isolated and purified from seeds of Withania somnifera alpha-amylase inhibitor (WSAI). Its mass spectrometry analysis revealed 59% sequence coverage with Wrightide II-type alpha-amylase inhibitor from Wrightia religiosa. A dose-dependent inhibition of alpha-amylases from Aspergillus oryzae, Bacillus subtilis, Helicoverpa armigera and Tribolium castaneumwas recorded. Interestingly, WSAI did not inhibit human salivary alpha-amylase significantly. When adults of T. castaneum were fed with WSAI (1.6mg g(-1)), decrease inconsumption, growthandefficiency of conversion of ingested foodwas evident, along withover fourfold increases in feedingdeterrence index. Adecline inlarval residual alpha-amylase activity after feedingofWSAI resulted ina reduction in longevity of T. castaneum. CONCLUSION: The study reflects the significance of WSAI in affecting the overall growth and development of T. castaneum. Pre-and post-harvest pest resistive capability makes WSAI a potential candidate for insect pest management. Further, the effectiveness of this inhibitor could be explored either in formulations or through a transgenic approach. (C) 2016 Society of Chemical Industry

1 aKasar, Sainath, S.1 aMarathe, Kiran, R.1 aBhide, Amey, J.1 aHerwade, Abhijeet, P.1 aGiri, Ashok, P.1 aMaheshwari, Vijay, L.1 aPawar, Pankaj, K. uhttps://library.ncl.res.in/content/glycoprotein-alpha-amylase-inhibitor-withania-somnifera-differentially-inhibits-various01937nas a2200205 4500008004100000245014900041210006900190260000800259300001200267490000700279520115700286100002301443700002401466700002301490700002801513700002201541700002001563700002101583856012701604 2017 eng d00aImproved tolerance against helicoverpa armigera in transgenic tomato over-expressing multi-domain proteinase inhibitor gene from capsicum annuum0 aImproved tolerance against helicoverpa armigera in transgenic to cJUL a597-6040 v233 aPlant proteinase inhibitors (PIs) are plant defense proteins and considered as potential candidates for engineering plant resistances against herbivores. Capsicum annuum proteinase inhibitor (CanPI7) is a multi-domain potato type II inhibitor (Pin-II) containing four inhibitory repeat domains (IRD), which target major classes of digestive enzymes in the gut of Helicoverpa armigera larvae. Stable integration and expression of the transgene in T1 transgenic generation, were confirmed by established molecular techniques. Protein extract of transgenic tomato lines showed increased inhibitory activity against H. armigera gut proteinases, supporting those domains of CanPI7 protein to be effective and active. When T1 generation plants were analyzed, they exhibited antibiosis effect against first instar larvae of H. armigera. Further, larvae fed on transgenic tomato leaves showed delayed growth relative to larvae fed on control plants, but did not change mortality rates significantly. Thus, better crop protection can be achieved in transgenic tomato by overexpression of multi-domain proteinase inhibitor CanPI7 gene against H. armigera larvae.1 aTanpure, Rahul, S.1 aBarbole, Ranjit, S.1 aDawkar, Vishal, V.1 aWaichal, Yashashree, A.1 aJoshi, Rakesh, S.1 aGiri, Ashok, P.1 aGupta, Vidya, S. uhttps://library.ncl.res.in/content/improved-tolerance-against-helicoverpa-armigera-transgenic-tomato-over-expressing-multi02171nas a2200193 4500008004100000245015500041210006900196260000800265300001000273490000800283520141800291100002601709700002501735700002201760700002301782700002001805700002101825856013101846 2017 eng d00aIsolation and characterization of 9-lipoxygenase and epoxide hydrolase 2 genes: insight into lactone biosynthesis in mango fruit (Mangifera indica L.)0 aIsolation and characterization of 9lipoxygenase and epoxide hydr cJUN a65-750 v1383 aUniqueness and diversity of mango flavour across various cultivars are well known. Among various flavour metabolites lactones form an important class of aroma volatiles in certain mango varieties due to their ripening specific appearance and lower odour detection threshold. In spite of their biological and biochemical importance, lactone biosynthetic pathway in plants remains elusive. Present study encompasses quantitative real-time analysis of 9-lipoxygenase (Mi9LOX), epoxide hydrolase 2 (MiEH2), peroxygenase, hydroperoxide lyase and acyl-CoA-oxidase genes during various developmental and ripening stages in fruit of Alphonso, Paid and Kent cultivars with high, low and no lactone content and explains their variable lactone content. Study also covers isolation, recombinant protein characterization and transient over-expression of Mi9LOX and MiEH2 genes in mango fruits. Recombinant Mi9LOX utilized linoleic and linolenic acids, while MiEH2 utilized aromatic and fatty acid epoxides as their respective substrates depicting their role in fatty acid metabolism. Significant increase in concentration of delta-valerolactone and gamma-decalactone upon Mi9LOX over-expression and that of delta-valerolactone, gamma-hexalactone and delta-hexalactone upon MiEH2 over-expression further suggested probable involvement of these genes in lactone biosynthesis in mango. (C) 2017 Elsevier Ltd. All rights reserved.1 aDeshpande, Ashish, B.1 aChidley, Hemangi, G.1 aOak, Pranjali, S.1 aPujari, Keshav, H.1 aGiri, Ashok, P.1 aGupta, Vidya, S. uhttps://library.ncl.res.in/content/isolation-and-characterization-9-lipoxygenase-and-epoxide-hydrolase-2-genes-insight-lactone02108nas a2200193 4500008004100000245012100041210006900162260000800231300001400239490000700253520138400260100001601644700002501660700002201685700003101707700002601738700002001764856013001784 2017 eng d00aPlacental proteomics provides insights into pathophysiology of pre-eclampsia and predicts possible markers in plasma0 aPlacental proteomics provides insights into pathophysiology of p cFEB a1050-10600 v163 a

Pre-eclampsia is a hypertensive disorder characterized by the new onset of hypertension >140/90 mmHg and proteinuria after the 20th week of gestation. The disorder is multi-factorial and originates with abnormal placentation. Comparison of the placental proteome of normotensive (n=25) and pre-eclamptic (n=25) patients by gel-free proteomic techniques, identified a total of 2145 proteins in the placenta of which 180 were differentially expressed (>1.3 fold, p<0.05). Gene ontology enrichment analysis of biological process suggested that the differentially expressed proteins belonged to various physiological processes such as angiogenesis, apoptosis, oxidative stress, hypoxia, placental development, which are implicated in the pathophysiology of pre-eclampsia. Some of the differentially expressed proteins were monitored in the plasma by multiple reaction monitoring (MRM) analysis, which showed an increase in Apolipoprotein AI and AII in gestational week 26-30th (2-fold, p<0.01) while haptoglobin and hemopexin decreased in gestational week 26-30th and 40th week/at delivery (1.8 fold, p<0.01) in pre-eclamptic patients. This study provides a proteomic insight into the pathophysiology of pre-eclampsia. Identified candidate proteins can be evaluated further for the development of potential biomarkers associated with pre-eclampsia pathogenesis.

1 aMary, Sheon1 aKulkarni, Mahesh, J.1 aMalakar, Dipankar1 aJoshi, Sadhana, Ramchandra1 aMehendale, Savita, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/placental-proteomics-provides-insights-pathophysiology-pre-eclampsia-and-predicts-possible02303nas a2200241 4500008004100000022001400041245010400055210006900159260000800228300001200236490000700248520148400255653002401739653001601763653001501779653003501794653001601829100002301845700002001868700002101888700002401909856012801933 2017 eng d a0141-813000aStructure-function relationship of a bio-pesticidal trypsin/chymotrypsin inhibitor from winged bean0 aStructurefunction relationship of a biopesticidal trypsinchymotr cMAR a532-5370 v963 a

Protease inhibitors are essential bio-molecules that serve as a model system for the study of protein structure and protease-protease inhibitor interaction. We here report a bi-functional serine protease inhibitor from winged bean (WBCTI) that completely retains its inhibitory property against trypsin and chymotrypsin even after heating at 70 degrees C. Detailed circular dichroism and fluorescence studies at different temperatures, 30-90 degrees C, have been performed to understand the reason behind thermal stability of the protein. On the basis of our results it appears that WBCTI maintains its canonical structure up to 70 degrees C. Above that the heat induced conformational change becomes irreversible which causes aggregation followed by precipitation of the protein. Moreover, the activity and stability of the secondary structure are found to decrease drastically in presence of dithiothreitol indicating that the protein acquires additional stability for the occurrence of two disulfide bonds. In addition to the structural characterization, an important property of WBCTI against the polyphagous pest Helicoverpa armigera has been explored in present study. WBCTI has showed reasonable inhibition of the mid-gut proteases of H. armigera. In artificial feeding trial through addition of WBCTI in diet resulted in significant growth retardation, delayed pupae formation and higher mortality of H. armigera larvae. (C) 2016 Elsevier B.V. All rights reserved.

10aDisulfide reduction10aHeat stable10aPesticidal10aTrypsin/Chymotrypsin inhibitor10aWinged bean1 aBanerjee, Sayanika1 aGiri, Ashok, P.1 aGupta, Vidya, S.1 aDutta, Samir, Kumar uhttps://library.ncl.res.in/content/structure-function-relationship-bio-pesticidal-trypsinchymotrypsin-inhibitor-winged-bean02190nas a2200217 4500008004100000245016800041210006900209260000800278490000600286520134900292100002601641700002401667700001601691700002501707700002201732700002401754700002301778700002001801700002101821856013001842 2017 eng d00aTranscriptional transitions in alphonso mango (Mangifera indica L.) during fruit development and ripening explain its distinct aroma and shelf life characteristics0 aTranscriptional transitions in alphonso mango Mangifera indica L cAUG0 v73 aAlphonso is known as the "King of mangos" due to its unique flavor, attractive color, low fiber pulp and long shelf life. We analyzed the transcriptome of Alphonso mango through Illumina sequencing from seven stages of fruit development and ripening as well as flower. Total transcriptome data from these stages ranged between 65 and 143 Mb. Importantly, 20,755 unique transcripts were annotated and 4,611 were assigned enzyme commission numbers, which encoded 142 biological pathways. These included ethylene and flavor related secondary metabolite biosynthesis pathways, as well as those involved in metabolism of starch, sucrose, amino acids and fatty acids. Differential regulation ( p-value <= 0.05) of thousands of transcripts was evident in various stages of fruit development and ripening. Novel transcripts for biosynthesis of mono-terpenes, sesqui-terpenes, di-terpenes, lactones and furanones involved in flavor formation were identified. Large number of transcripts encoding cell wall modifying enzymes was found to be steady in their expression, while few were differentially regulated through these stages. Novel 79 transcripts of inhibitors of cell wall modifying enzymes were simultaneously detected throughout Alphonso fruit development and ripening, suggesting controlled activity of these enzymes involved in fruit softening.1 aDeshpande, Ashish, B.1 aAnamika, Krishanpal1 aJha, Vineet1 aChidley, Hemangi, G.1 aOak, Pranjali, S.1 aKadoo, Narendra, Y.1 aPujari, Keshav, H.1 aGiri, Ashok, P.1 aGupta, Vidya, S. uhttps://library.ncl.res.in/content/transcriptional-transitions-alphonso-mango-mangifera-indica-l-during-fruit-development-and01944nas a2200181 4500008004100000245011200041210006900153260000800222300002200230490000700252520126400259100001601523700002501539700002601564700002301590700002001613856012901633 2017 eng d00aTubulointerstitial nephritis antigen-like 1 protein is downregulated in the placenta of pre-eclamptic women0 aTubulointerstitial nephritis antigenlike 1 protein is downregula cMAR aArticle Number: 80 v143 aBackground: Tubulointerstitial nephritis antigen-like 1 protein (TINAGL1), is a matricellular protein, known to play role in cell adhesion and cell receptor interaction. Research related to TINAGL1 is limited to cell culture and animal models. Demonstration of TINAGL1 as a positive regulator of angiogenesis and its expression in the decidua of postimplantation mouse uterus, prompted us to validate its expression in human placenta during impaired angiogenesis in pre-eclamptic condition. Methods: Placental tissue from normotensive (n = 25) and pre-eclamptic (n = 25) pregnancies were used to study the differentially expressed proteins by two-dimensional gel electrophoresis and TINAGL1 protein was validated with Western blotting. Results: A total of 55 protein spots were differentially expressed (fold change > 1.5, p < 0.05), of which 27 were upregulated and 28 were downregulated in the pre-eclamptic placenta. TINAGL1 was found to be downregulated in pre-eclamptic compared to normotensive pregnant women. Conclusion: This is the first study reporting TINAGL1 to be present in human placenta and differentially expressed in pre-eclamptic condition. The functional role of TINAGL1 in association to human pregnancy needs to be explored further.1 aMary, Sheon1 aKulkarni, Mahesh, J.1 aMehendale, Savita, S.1 aJoshi, Sadhana, R.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/tubulointerstitial-nephritis-antigen-1-protein-downregulated-placenta-pre-eclamptic-women02197nas a2200229 4500008004100000245018300041210006900224260000800293300001400301490000700315520133000322100002801652700002001680700002201700700002001722700002301742700001301765700001801778700002101796700002001817856013001837 2018 eng d00aIntegrated transcriptomic and proteomic analyses suggest the participation of endogenous protease inhibitors in the regulation of protease gene expression in helicoverpa armigera0 aIntegrated transcriptomic and proteomic analyses suggest the par cJUL a1324-13360 v173 aInsects adapt to plant protease inhibitors (PIs) present in their diet by differentially regulating multiple digestive proteases. However, mechanisms regulating protease gene expression in insects are largely enigmatic. Ingestion of multi-domain recombinant Capsicum annuum protease inhibitor-7 (CanPI-7) arrests growth and development of Helicoverpa armigera (Lepidoptera: Noctuidae). Using de novo RNA sequencing and proteomic analysis, we examined the response of H. armigera larvae fed on recombinant CanPI-7 at different time intervals. Here, we present evidence supporting a dynamic transition in H. armigera protease expression on CanPI-7 feeding with general down-regulation of protease genes at early time points (0.5 to 6 h) and significant up-regulation of specific trypsin, chymotrypsin and aminopeptidase genes at later time points (12 to 48 h). Further, coexpression of H. armigera endogenous PIs with several digestive protease genes were apparent. In addition to the differential expression of endogenous H. armigera PIs, we also observed a distinct novel isoform of endogenous PI in CanPI-7 fed H. armigera larvae. Based on present and earlier studies, we propose potential mechanism of protease regulation in H. armigera and subsequent adaptation strategy to cope with anti-nutritional components of plants.1 aLomate, Purushottam, R.1 aDewangan, Veena1 aMahajan, Neha, S.1 aKumar, Yashwant1 aKulkarni, Abhijeet1 aWang, Li1 aSaxena, Smita1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/integrated-transcriptomic-and-proteomic-analyses-suggest-participation-endogenous-protease02649nas a2200169 4500008004100000022001500041245009100056210006900147260000800216300001400224490000800238520204500246100002502291700002502316700002002341856011802361 2018 eng d a 0032-093500a Structural, functional and evolutionary diversity of 4-coumarate-CoA ligase in plants0 aStructural functional and evolutionary diversity of 4coumarateCo cNOV a1063-10780 v2483 aMain conclusions The 4-coumarate-CoA ligases (4CL) contribute in channelizing flux of different phenylpropanoid biosynthetic pathways. Expression of 4CL is optimized at developmental stages and in response to environmental triggers such as biotic and abiotic stresses. The enzyme is valuable in metabolic pathway engineering for curcuminoids, resveratrol, biofuel production and nutritional improvement. Vigorous analysis of regulation at functional and expression level is obligatory to attain efficient commercial production of candidate metabolites using 4CL. Phenylpropanoid pathway provides precursors for numerous secondary metabolites in plants. In this pathway, 4-coumarate-CoA ligase (EC 6.2.1.12, 4CL) is the main branch point enzyme which generates activated thioesters. Being the last enzyme of three shared common steps in general phenylpropanoid pathway, it contributes to channelize precursors for different phenylpropanoids. In plants, 4CL enzymes are present in multiple isoforms and encoded by small gene family. It belongs to adenylate-forming enzyme family and catalyzes the reaction that converts hydroxy or methoxy cinnamic acid derivatives to corresponding thioesters. These thioesters are further utilized for biosynthesis of phenylpropanoids, which are known for having numerous nutritional and medicinal applications. In addition, the 4CL enzymes have been characterized from various plants for their role in plant physiology or in biotic and abiotic stresses. Furthermore, specific isoforms are differentially regulated upon exposure to diverse stimuli leading to flux diversion toward the particular metabolite biosynthesis. Evolutionary studies showed that 4CL separately evolved after monocot and dicot segregation. Here, we provide a comprehensive review on 4CL, which includes evolution, function, gene/protein structure, role in metabolite biosynthesis and cellular partition, and their regulation. Based on the available data, we have explored the scope for pathway engineering by utilizing 4CL enzymes.1 aLavhale, Santosh, G.1 aKalunke, Raviraj, M.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/structural-functional-and-evolutionary-diversity-4-coumarate-coa-ligase-plants02335nas a2200253 4500008004100000022001200041245015200053210006900205260000900274300001000283490000700293520141700300653002501717653001301742653002301755653003401778100002601812700002201838700002401860700002401884700002301908700002001931856013001951 2018 eng d a965174800aTripeptides derived from reactive centre loop of potato type II protease inhibitors preferentially inhibit midgut proteases of Helicoverpa armigera0 aTripeptides derived from reactive centre loop of potato type II  cAPR  a17-250 v953 a

Potato type II protease inhibitors (Pin-II PIs) impede the growth of lepidopteran insects by inhibiting serine protease-like enzymes in the larval gut. The three amino acid reactive centre loop (RCL) of these proteinaceous inhibitors is crucial for protease binding and is conserved across the Pin-II family. However, the molecular mechanism and inhibitory potential of the RCL tripeptides in isolation of the native protein has remained elusive. In this study, six peptides corresponding to the RCLs of the predominant Pin-II PIs were identified, synthesized and evaluated for in vitro and in vivo inhibitory activity against serine proteases of the polyphagous insect, Helicoverpa armigera. RCL peptides with sequences PRN, PRY and TRE were found to be potent inhibitors that adversely affected the growth and development of H. armigera. The binding mechanism and differential affinity of the RCL peptides with serine proteases was delineated by crystal structures of complexes of the RCL peptides with trypsin. Residues P1 and P2 of the inhibitors play a crucial role in the interaction and specificity of these inhibitors. Important features of RCL peptides like higher inhibition of insect proteases, enhanced efficacy at alkaline gut pH, longer retention and high stability in insect gut make them suitable molecules for the development of sustainable pest management strategies for crop protection.

10aHelicoverpa armigera10aProtease10aprotease inhibitor10aReactive centre loop peptides1 aSaikhedkar, Nidhi, S.1 aJoshi, Rakesh, S.1 aBhoite, Ashwini, S.1 aMohandasan, Radhika1 aYadav, Amit, Kumar1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/tripeptides-derived-reactive-centre-loop-potato-type-ii-protease-inhibitors-preferentially02610nas a2200277 4500008004100000022001400041245012100055210006900176260000800245300001400253490000700267520170200274653002201976653001701998653002002015653001102035653001002046653000902056100002702065700002602092700002002118700001902138700002002157700002402177856013102201 2018 eng d a1467-764400aWRKY1 acts as a key component improving resistance against Alternaria solani in wild tomato, Solanum arcanum Peralta0 aWRKY1 acts as a key component improving resistance against Alter cAUG a1502-15130 v163 a

Early blight (EB), caused by Alternaria solani, is a major threat to global tomato production. In comparison with cultivated tomato (Solanum lycopersicum), a wild relative, S.arcanum exhibits strong resistance against EB. However, molecular cascades operating during EB resistance in wild or cultivated tomato plants are largely obscure. Here, we provide novel insight into spatio-temporal molecular events in S.arcanum against A.solani. Transcriptome and co-expression analysis presented 33-WRKYs as promising candidates of which 12 SaWRKYs displayed differential expression patterns in resistant and susceptible accessions during EB disease progression. Among these, SaWRKY1 exhibited induced expression with significant modulation in xyloglucan endotrans hydrolase 5 (XTH5) and MYB2 expressions that correlated with the disease phenotypes. Electro-mobility shift assay confirmed physical interaction of recombinant SaWRKY1 to SaXTH5 and SaMYB2 promoters. Comparative WRKY1 promoter analysis between resistant and susceptible plants revealed the presence of crucial motifs for defence mechanism exclusively in resistant accession. Additionally, many defence-related genes displayed significant expression variations in both the accessions. Further, WRKY1 overexpressing transgenic plants exhibited higher levels of EB resistance while RNAi silencing lines had increased susceptibility to A.solani with altered expression of XTH5 and MYB2. Overall, these findings demonstrate the positive influence of WRKY1 in improving EB resistance in wild tomato and this could be further utilized as a potential target through genetic engineering to augment protection against A.solani in crop plants.

10aAlternaria solani10aearly blight10aSolanum arcanum10aTomato10aWRKY110aXTH51 aShinde, Balkrishna, A.1 aDholakia, Bhushan, B.1 aHussain, Khalid1 aAharoni, Asaph1 aGiri, Ashok, P.1 aKamble, Avinash, C. uhttps://library.ncl.res.in/content/wrky1-acts-key-component-improving-resistance-against-alternaria-solani-wild-tomato-solanum03076nas a2200265 4500008004100000022001400041245011900055210006900174260000800243300001400251490000600265520217000271100002302441700002202464700002402486700002002510700002002530700001902550700002202569700002102591700003702612700001802649700002002667856012302687 2019 eng d a2470-134300aAzadirachtin-A from azadirachta indica impacts multiple biological targets in cotton bollworm helicoverpa armigera0 aAzadirachtinA from azadirachta indica impacts multiple biologica cMAY a9531-95410 v43 a

Azadirachtin-A (AzaA) from the Indian neem tree (Azadirachta indica) has insecticidal properties; however, its molecular mechanism remains elusive. The ``targeted and nontargeted proteomic profiling'', metabolomics, matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) imaging, gene expression, and in silico analysis provided clues about its action on Helicoverpa armigera. Fourth instar H. armigera larvae fed on AzaA-based diet (AzaD) suffered from significant mortality, growth retardation, reduced larval mass, complications in molting, and prolonged development. Furthermore, death of AzaD-fed larvae was observed with various phenotypes like bursting, blackening, and half-molting. Liquid chromatography-mass spectrometry (LC-MS) data showed limited catabolic processing of ingested AzaA and dramatic alternations of primary metabolism in H. armigera. MALDI-TOF imaging indicated the presence of AzaA in midgut of H. armigera. In the gut, out of 79 proteins identified, 34 were upregulated, which were related to digestion, immunity, energy production, and apoptosis mechanism. On the other hand, 45 proteins were downregulated, including those from carbohydrate metabolism, lipid metabolism, and energy transfer. In the hemolymph, 21 upregulated proteins were reported to be involved in immunity, RNA processing, and mRNA-directed protein synthesis, while 7 downregulated proteins were implicated in energy transfer, hydrolysis, lipid metabolism, defense mechanisms, and amino acid storage-related functions. Subsequently, six target proteins were identified using labeled AzaA that interacted with whole insect proteins. In silico analysis suggests that AzaA could be efficiently accommodated in the hydrophobic pocket of juvenile hormone esterase and showed strong interaction with active site residues, indicating plausible targets of AzaA in H. armigera. Quantitative polymerase chain reaction analysis suggested differential gene expression patterns and partly corroborated the proteomic results. Overall, data suggest that AzaA generally targets more than one protein in H. armigera and hence could be a potent biopesticide.

1 aDawkar, Vishal, V.1 aBarage, Sagar, H.1 aBarbole, Ranjit, S.1 aFatangare, Amol1 aGrimalt, Susana1 aHaldar, Saikat1 aHeckel, David, G.1 aGupta, Vidya, S.1 aThulasiram, Hirekodathakallu, V.1 aSvatos, Ales.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/azadirachtin-azadirachta-indica-impacts-multiple-biological-targets-cotton-bollworm01924nas a2200265 4500008004100000022001400041245018700055210006900242260000800311520091700319653001801236653002001254653002501274653003301299653001801332653003001350100001901380700002201399700002201421700002001443700002001463700002201483700002601505856012701531 2019 eng d a0739-110200aNMR structure and dynamics of inhibitory repeat domain variant 12, a plant protease inhibitor from Capsicum annuum, and its structural relationship to other plant protease inhibitors0 aNMR structure and dynamics of inhibitory repeat domain variant 1 cAPR3 a

Although several plant protease inhibitors have been structurally characterized using X-ray crystallography, very few have been studied using NMR techniques. Here, we report an NMR study of the solution structure and dynamics of an inhibitory repeat domain (IRD) variant 12 from the wound-inducible Pin-II type proteinase inhibitor from Capsicum annuum. IRD variant 12 (IRD12) showed strong anti-metabolic activity against the Lepidopteran insect pest, Helicoverpa armigera. The NMR-derived three-dimensional structure of IRD12 reveals a three-stranded anti-parallel beta-sheet rigidly held together by four disulfide bridges and shows structural homology with known IRDs. It is interesting to note that the IRD12 structure containing similar to 75% unstructured part still shows substantial amount of rigidity of N-H bond vectors with respect to its molecular motion. Communicated by Ramaswamy H. Sarma

10aBt transgenic10aCapsicum annuum10aHelicoverpa armigera10ainhibitory repeating domains10aNMR structure10aProtease inhibitors (PIs)1 aGartia, Janeka1 aAnangi, Raveendra1 aJoshi, Rakesh, S.1 aGiri, Ashok, P.1 aKing, Glenn, F.1 aBarnwal, Ravi, P.1 aChary, Kandala, V. R. uhttps://library.ncl.res.in/content/nmr-structure-and-dynamics-inhibitory-repeat-domain-variant-12-plant-protease-inhibitor02795nas a2200265 4500008004100000022001400041245014500055210006900200260000800269300001400277490000900291520187200300653001902172653001602191653001702207653002302224653001802247100002702265700002202292700002002314700002002334700002402354700002002378856013102398 2019 eng d a0304-416500aPhyto-inspired cyclic peptides derived from plant Pin-II type protease inhibitor reactive center loops for crop protection from insect pests0 aPhytoinspired cyclic peptides derived from plant PinII type prot cAUG a1254-12620 v18633 a

Background: Natural defence of plants against insect pests involves protease inhibitors (PIs) that interfere with insect digestive proteases. Pin-II type plant PIs are wound inducible upon insect damage and possess multiple inhibitory repeat domains that can inhibit trypsin and chymotrypsin-like proteases in the insect midgut. Yet, their agricultural ex-vivo application is limited due to large molecular size and environmental instability, which could be overcome by small peptides. Methods: Bicyclic peptides were designed by grafting Pin-II PIs derived reactive center loop (RCL) on synthetic tris(bromomethyl)benzene scaffold. In vitro binding with trypsin-like proteases was evaluated by biochemical and biophysical assays, followed by molecular dynamics simulations. In vivo effects on two major lepidopteran insect pests, Helicoverpa armigera and Spodoptera litura were studied upon feeding with peptide treated leaves. Affinity based pull down assays were used to identify target proteins in insect gut. Results: Bicyclic RCLs showed ten-fold enhanced protease inhibition compared to their linear counterparts. They exhibited feeding deterrence and growth reduction of lepidopteran insects. Bicyclic peptides predominantly interact with midgut serine proteases. Possible binding modes involve simultaneous interaction with the active site and specificity-determining residues of insect gut trypsin. Conclusion: Bicyclic peptides are potent inhibitors of serine proteases in the insect midgut. They cause feeding aversion and larval growth retardation. Bi-domain cyclic peptides interact with two sites on trypsin, leading to enhanced efficacy over linear RCL peptides. General significance: Bicyclic peptides mimic natural PIs by inhibiting insect proteases leading to growth reduction, thus, could be used as pest control molecules in agriculture.

10aCyclic peptide10aLepidoptera10aPest control10aprotease inhibitor10aReactive loop1 aSailthedkar, Nidhi, S.1 aJoshi, Rakesh, S.1 aYadav, Amit, K.1 aSeal, Shubhendu1 aFernandes, Moneesha1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/phyto-inspired-cyclic-peptides-derived-plant-pin-ii-type-protease-inhibitor-reactive-center02292nas a2200217 4500008004100000022001400041245014700055210006900202260000900271300001200280490000800292520149700300653001501797653001801812653002701830100002601857700002001883700002501903700002001928856012601948 2019 eng d a0141-813000aPotent chitin-hydrolyzing enzyme from Myrothecium verrucaria affects growth and development of Helicoverpa armigera and plant fungal pathogens0 aPotent chitinhydrolyzing enzyme from Myrothecium verrucaria affe cDEC  a517-5280 v1413 a

Chitin, a crucial structural and functional component of insects and fungi, serves as a target for pest management by utilizing novel chitinases. Here, we report the biocontrol potential of recombinant Myrothecium verrucaria endochitinase (rMvEChi) against insect pest and fungal pathogens. A complete ORF of MvEChi (1185 bp) was cloned and heterologously expressed in Escherichia coli. Structure based sequence alignment of MvEChi revealed the presence of conserved domains SXGG and DXXDXDXE specific for GH-18 family, involved in substrate binding and catalysis, respectively. rMvEChi (46.6 kDa) showed optimum pH and temperature as 7.0 and 30 degrees C, respectively. Furthermore, rMvEChi remained stable within the pH range of 6.0 to 8.0 and up to 40 degrees C. rMvEChi exhibited k(cat)/K-m values of 129.83 x 10(3) [(g/L)(-1) s(-1)] towards 4MU chitotrioside. Hydrolysis of chitooligosaccharides with various degrees of polymerization (DP) using rMvEChi indicated the release of DP2 as main end product with order of reaction as DP6 > DP5 > DP4 > DP3. Bioassay of rMvEChi against Helicoverpa armigera displayed potent anti-feedant activity and induced mortality. In vitro antifungal activity against plant pathogenic fungi (Ustilago maydis and Bipolaris sorokiniana) exhibited significant inhibition of mycelium growth. These results suggest that MvEChi has significant potential in enzyme-based pest and pathogen management. (C) 2019 Elsevier B.V. All rights reserved.

10aBiocontrol10aendochitinase10aMyrothecium verrucaria1 aVidhate, Ravindra, P.1 aBhide, Amey, J.1 aGaikwad, Sushama, M.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/potent-chitin-hydrolyzing-enzyme-myrothecium-verrucaria-affects-growth-and-development02608nas a2200313 4500008004100000022001400041245010700055210006900162260000800231300001000239490000700249520155900256653002801815653002501843653001101868653002101879653001301900653001801913100001601931700002901947700002501976700002602001700002502027700002402052700002202076700003702098700002002135856013902155 2019 eng d a0971-589400aTerpene profiling, transcriptome analysis and characterization of cis–terpineol synthase from Ocimum0 aTerpene profiling transcriptome analysis and characterization of cJAN a47-570 v253 a

Ocimum species produces a varied mix of different metabolites that imparts immense medicinal properties. To explore this chemo-diversity, we initially carried out metabolite profiling of different tissues of five Ocimum species and identified the major terpenes. This analysis broadly classified these five Ocimum species into two distinct chemotypes namely, phenylpropanoid-rich and terpene-rich. In particular, -caryophyllene, myrcene, limonene, camphor, borneol and selinene were major terpenes present in these Ocimum species. Subsequently, transcriptomic analysis of pooled RNA samples from different tissues of Ocimum gratissimum, O. tenuiflorum and O. kilimandscharicum identified 38 unique transcripts of terpene synthase (TPS) gene family. Full-length gene cloning, followed by sequencing and phylogenetic analysis of three TPS transcripts were carried out along with their expression in various tissues. Terpenoid metabolite and expression profiling of candidate TPS genes in various tissues of Ocimum species revealed spatial variances. Further, putative TPS contig 19414 (TPS1) was selected to corroborate its role in terpene biosynthesis. Agrobacterium-mediated transient over-expression assay of TPS1 in the leaves of O. kilimandscharicum and subsequent metabolic and gene expression analyses indicated it as a cis–terpineol synthase. Overall, present study provided deeper understanding of terpene diversity in Ocimum species and might help in the enhancement of their terpene content through advanced biotechnological approaches.

10aAgro-infiltration assay10aMetabolite profiling10aOcimum10aTerpene synthase10aTerpenes10aTranscriptome1 aAnand, Atul1 aJayaramaiah, Ramesha, H.1 aBeedkar, Supriya, D.1 aDholakia, Bhushan, B.1 aLavhale, Santosh, G.1 aPunekar, Sachin, A.1 aGade, Wasudeo, N.1 aThulasiram, Hirekodathakallu, V.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/terpene-profiling-transcriptome-analysis-and-characterization-cis%E2%80%93terpineol-synthase-ocimum02560nas a2200313 4500008004100000022001400041245010100055210006900156260000800225300001100233490000800244520153800252653002601790653001201816653001201828653003301840653001401873653002801887653001701915653002901932653002001961100002001981700002502001700001902026700001802045700003302063700002002096856013002116 2020 eng d a0031-942200aBiosynthesis and tissue-specific partitioning of camphor and eugenol in Ocimum kilimandscharicum0 aBiosynthesis and tissuespecific partitioning of camphor and euge cSEP a1124510 v1773 a

In Ocimum kilimandscharicum, the relative volatile composition of camphor in leaves was as high as 55%, while that of eugenol in roots was 57%. These metabolites were differentially partitioned between the aerial and root tissues. Global metabolomics revealed tissue-specific biochemical specialization, evident by the differential distribution of 2588 putative metabolites across nine tissues. Next-generation sequencing analysis indicated differential expression of 51 phenylpropanoid and 55 terpenoid pathway genes in aerial and root tissues. By integrating metabolomics with transcriptomics, the camphor biosynthesis pathway in O. kilimandscharicum was elucidated. In planta bioassays revealed the role of geranyl diphosphate synthase (gpps) and borneol dehydrogenase (bdh) in camphor biosynthesis. Further, the partitioning of camphor was attributed to tissue-specific gene expression of both the pathway entry point (gpps) and terminal (bdh) enzyme. Unlike camphor, eugenol accumulated more in roots; however, absence of the eugenol synthase gene in roots indicated long distance transport from aerial tissues. In silico co-expression analysis indicated the potential involvement of ATP-binding cassette, multidrug and toxic compound extrusion, and sugar transporters in eugenol transport. Similar partitioning was evident across five other Ocimum species. Overall, our work indicates that metabolite partitioning maybe a finely regulated process, which may have implications on plant growth, development, and defense.

10aBorneol dehydrogenase10aCamphor10aEugenol10aGeranyl diphosphate synthase10aLamiaceae10aMetabolite partitioning10aMetabolomics10aOcimum kilimandscharicum10aTranscriptomics1 aSingh, Priyanka1 aKalunke, Raviraj, M.1 aShukla, Anurag1 aTzfadia, Oren1 aThulasiram, Hirekodathakallu1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/biosynthesis-and-tissue-specific-partitioning-camphor-and-eugenol-ocimum-kilimandscharicum02661nas a2200241 4500008004100000022001400041245022700055210006900282260000900351520169200360653002002052653001002072653002202082653002602104653001902130653002602149100001802175700002602193700002902219700002402248700002002272856012702292 2020 eng d a0721-759500aCombinatorial approach through in vitro regeneration and phytochemical profiling of ceropegia media (huber) ans.: a potential way forward in the conservation of an endangered medicinal plant from the western ghats in India0 aCombinatorial approach through in vitro regeneration and phytoch cJUN 3 a

Ceropegia mediais an endemic and endangered plant as its propagation through seeds is unreliable due to low germination, slow growth and seedling decay under natural conditions. Also, tubers of this plant are edible serving as carbohydrate source with medicinal values leading to severe population decline in the natural habitat. To provide a sustainable solution, an efficient in vitro propagation protocol along with phytochemical profiling was developed forC. media. Callus cultures were induced from seedling and wild leaf tissues using the most effective Murashige and Skoog's (MS) medium with 2,4-dichlorophenoxyacetic acid (2,4-D; 2 mu M) and sucrose (3%). Somatic embryos were acquired on MS medium with 1 mu M 6-Benzylaminopurine (BAP) and 1 mu M 2,4-D. Conversion into plantlets was attained only from tissue culture-derived seedling leaf (TCDSL) explant. Further, in vitro tuberization was achieved from TCDSL callus with BAP and Naphthalene acetic acid (NAA). AgNO(3)as an elicitor had a positive effect on both fresh and dry weights of callus. Successful acclimatization (58%) was attained after two months resulting in normal phenotype in pots. Further, metabolite profiles of ten different tissues from wild and in vitro plants were compared. Total 82 compounds comprising alkaloids, fatty acids, fatty acid ester, steroids, terpenes and hydrocarbons were identified. Overall, results suggested enhanced production of selected metabolites with in vitro propagation and AgNO3, alleviating the problem of unavailability of planting materials. Thus, the current study might offer potential ways for the conservation of such RED enlisted species asC. media. [GRAHICS]

10aCeropegia media10aGC-MS10aMicrotuberization10aSecondary metabolites10aSilver nitrate10asomatic embryogenesis1 aPandey, Meena1 aDholakia, Bhushan, B.1 aJayaramaiah, Ramesha, H.1 aPunekar, Sachin, A.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/combinatorial-approach-through-vitro-regeneration-and-phytochemical-profiling-ceropegia02365nas a2200301 4500008004100000022001400041245011000055210006900165260000800234300001200242490000700254520141200261653001301673653001001686653001601696653002301712653002101735653001501756653001701771100001601788700002601804700002501830700002101855700002401876700002001900700002001920856012301940 2020 eng d a1073-608500aDifferential modulation in metabolites revealed with the improvement in the shelf-life of alphonso fruits0 aDifferential modulation in metabolites revealed with the improve cOCT a508-5200 v623 a

Globally farmers have difficulty in extending the shelf-life of the tropical fruits due to their perishable nature. The present study aimed to assess the effect of hexanal nano-formulation treatment (NFT) on the shelf-life of Alphonso mango. Further, volatilomics was performed to explore the molecular basis of such effect. Untreated and treated fruits were sampled starting from 5th to 21st day after NFT at an interval of 4 days. Moderate changes in visual and digital colour parameters were evident from the intact and dissected fruits of NFT set compared to untreated fruits. Biochemical assays affirmed the phenotypic differences with significant changes in the colour imparting compounds like carotenoids and anthocyanins among them. Further, gas chromatography-mass spectrometry analysis revealed significant qualitative and quantitative variations in the different classes of compounds like lactones, furanones, esters, aldehydes and alcohols. Some of the key metabolites showed differential modulations among the NFT and untreated fruit sets indicating their potential role in various processes, which ultimately might have resulted in delayed ripening of the mango. Overall, this study has demonstrated the beneficial effect of hexanal and identified important metabolites with the enhanced shelf-life in Alphonso that could be useful for farmers and mango-based food/flavour industries.

10aAlphonso10aGC-MS10aMetabolites10aNano-biotechnology10aNano-formulation10aShelf-life10aVolatilomics1 aDar, Saleem1 aDholakia, Bhushan, B.1 aShanmugam, Haripriya1 aGupta, Vidya, S.1 aSubramanian, K., S.1 aSubramanian, J.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/differential-modulation-metabolites-revealed-improvement-shelf-life-alphonso-fruits02932nas a2200313 4500008004100000022001400041245013200055210006900187260000800256520189100264653001602155653001302171653001102184653000902195653003302204653002302237653000902260653002102269100002202290700002502312700002302337700001902360700002602379700002702405700001702432700002002449700002502469856012402494 2020 eng d a0739-110200aDiscovery of potential multi-target-directed ligands by targeting host-specific SARS-CoV-2 structurally conserved main protease0 aDiscovery of potential multitargetdirected ligands by targeting  cMAY3 a

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has resulted in the current COVID-19 pandemic. Worldwide this disease has infected over 2.5 million individuals with a mortality rate ranging from 5 to 10%. There are several efforts going on in the drug discovery to control the SARS-CoV-2 viral infection. The main protease (M-Pro) plays a critical role in viral replication and maturation, thus can serve as the primary drug target. To understand the structural evolution of M-Pro, we have performed phylogenetic and Sequence Similarity Network analysis, that depicted divergence of Coronaviridae M-Pro in five clusters specific to viral hosts. This clustering was corroborated with the comparison of M-Pro structures. Furthermore, it has been observed that backbone and binding site conformations are conserved despite variation in some of the residues. These attributes can be exploited to repurpose available viral protease inhibitors against SARS-CoV-2 M-Pro. In agreement with this, we performed screening of similar to 7100 molecules including active ingredients present in the Ayurvedic anti-tussive medicines, anti-viral phytochemicals and synthetic anti-virals against SARS-CoV-2 M-Pro as the primary target. We identified several natural molecules like delta-viniferin, myricitrin, taiwanhomoflavone A, lactucopicrin 15-oxalate, nympholide A, afzelin, biorobin, hesperidin and phyllaemblicin B that strongly binds to SARS-CoV-2 M-Pro. Intrestingly, these molecules also showed strong binding with other potential targets of SARS-CoV-2 infection like viral receptor human angiotensin-converting enzyme 2 (hACE-2) and RNA dependent RNA polymerase (RdRp). We anticipate that our approach for identification of multi-target-directed ligand will provide new avenues for drug discovery against SARS-CoV-2 infection. Communicated by Ramaswamy H. Sarma

10aCoronavirus10aCOVID-1910ahACE-210aMPro10amulti-target-directed ligand10aprotease inhibitor10aRdRp10aSARS-CoV-2 virus1 aJoshi, Rakesh, S.1 aJagdale, Shounak, S.1 aBansode, Sneha, B.1 aShankar, Shiva1 aTellis, Meenakshi, B.1 aPandya, Vaibhav, Kumar1 aChugh, Anita1 aGiri, Ashok, P.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/discovery-potential-multi-target-directed-ligands-targeting-host-specific-sars-cov-202440nas a2200277 4500008004100000022001400041245013600055210006900191260000800260300001100268490000800279520148000287653001401767653001801781653003401799653002501833653001701858653002501875653001101900100002201911700002901933700002401962700002601986700002002012856013002032 2020 eng d a0926-669000aGeneration of novelties in the genus Ocimum as a result of natural hybridization: a morphological, genetical and chemical appraisal0 aGeneration of novelties in the genus Ocimum as a result of natur cNOV a1128590 v1563 a

The genus Ocimum is a boutique of a diverse set of specialized metabolites such as terpenoids and phenylpropanoids. Each Ocimum species and its cultivars represent a characteristic chemical profile. The present study explored the two interspecific Ocimum hybrids originating through a serendipitous natural cross between O. kilimandscharicum and O. basilicum. These two novel Ocimum hybrids exhibited intermediate morphological features of two parental species. Inter simple sequence repeats (ISSR) analysis and DNA barcoding with the plastid non-coding trnH-psbA intergenic spacer region reaffirmed unambiguous parental identification and differentiation of these natural hybrids from other available Ocimum species. Consequently, gas chromatographymass spectrometry-based metabolite profiling of two hybrids identified them as specific chemotypes with the presence of a unique blend of specialized metabolites from the parental species, which are either rich in terpenes or phenylpropanoids. Additionally, expression analysis of key genes from terpenoid and phenylpropanoid pathways corroborated with differential metabolite accumulation. Thus, these two Ocimum hybrids represented the novel chemotypes, which could be useful in commercial cultivation to produce novel essential oil and bioactive constituents. Further, the resulting metabolite diversity could have potential industrial applications in the areas of healthcare, bioremediation, and crop protection.

10aChemotype10aDNA barcoding10aInter Simple Sequence Repeats10aInterspecific hybrid10aO. basilicum10aO. kilimandscharicum10aOcimum1 aGurav, Tanuja, P.1 aJayaramaiah, Ramesha, H.1 aPunekar, Sachin, A.1 aDholakia, Bhushan, B.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/generation-novelties-genus-ocimum-result-natural-hybridization-morphological-genetical-and02789nas a2200301 4500008004100000022001400041245015300055210006900208260000800277300000700285490000800292520177100300653002102071653001402092653001302106653001002119653001502129653001802144100002302162700002702185700002302212700002402235700002602259700002902285700002902314700002002343856012402363 2020 eng d a0032-093500aHigh-throughput metabolomic and transcriptomic analyses vet the potential route of cerpegin biosynthesis in two varieties of Ceropegia bulbosa Roxb.0 aHighthroughput metabolomic and transcriptomic analyses vet the p cJAN a280 v2513 a

Ceropegia bulbosa is an important medicinal plant, used in the treatment of various ailments including diarrhea, dysentery, and syphilis. This is primarily attributed to the presence of pharmaceutically active secondary metabolites, especially cerpegin. As this plant belongs to an endemic threatened category, genomic resources are not available hampering exploration on the molecular basis of cerpegin accumulation till now. Therefore, we undertook high-throughput metabolomic and transcriptomic analyses using different tissues from two varieties namely, C. bulbosa var. bulbosa and C. bulbosa var. lushii. Metabolomic analysis revealed spatial and differential accumulation of various metabolites. We chemically synthesized and characterized the cerpegin and its derivatives by liquid chromatography tandem-mass spectrometry (LC-MS/MS). Importantly, these comparisons suggested the presence of cerpegin and 5-allyl cerpegin in all C. bulbosa tissues. Further, de novo transcriptome analysis indicated the presence of significant transcripts for secondary metabolic pathways through the Kyoto encyclopedia of genes and genomes database. Tissue-specific profiling of transcripts and metabolites showed a significant correlation, suggesting the intricate mechanism of cerpegin biosynthesis. The expression of potential candidate genes from the proposed cerpegin biosynthetic pathway was further validated by qRT-PCR and NanoString nCounter. Overall, our findings propose a potential route of cerpegin biosynthesis. Identified transcripts and metabolites have built a foundation as new molecular resources that could facilitate future research on biosynthesis, regulation, and engineering of cerpegin or other important metabolites in such non-model plants.

10a5-Allyl cerpegin10aCeropegia10aCerpegin10aLC-MS10ametabolite10aTranscriptome1 aGharat, Sachin, A.1 aShinde, Balkrishna, A.1 aMule, Ravindra, D.1 aPunekar, Sachin, A.1 aDholakia, Bhushan, B.1 aJayaramaiah, Ramesha, H.1 aRamaswamy, Gopalakrishna1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/high-throughput-metabolomic-and-transcriptomic-analyses-vet-potential-route-cerpegin02635nas a2200229 4500008004100000022001400041245012200055210006900177260000800246300001100254490000900265520184800274653001602122653001802138653002802156653001102184653002702195100002202222700002202244700002002266856011902286 2020 eng d a0304-416500aMolecular determinant for specificity: differential interaction of alpha-amylases with their proteinaceous inhibitors0 aMolecular determinant for specificity differential interaction o cDEC a1297030 v18643 a

Background: alpha-Amylase inhibitors (alpha-AIs) belong to the discrete classes, and exhibited differential specificities against alpha-amylases from various sources. Several alpha-amylases and their complexes with inhibitors at the molecular level have been studied in detail. Interestingly, some alpha-AIs depict specific and selective interactions amid different insect alpha-amylases. Scope of review: There are studies to understand evolutionary variability and functional differentiation of insect alpha-amylases and their cognate inhibitors. We have examined sequence, structural, and interaction diversity between various alpha-amylases and alpha-AIs. Based on these analyses, we are providing a potential basis for the functional differentiation among certain insect a-amylases concerning mammalian counterparts and their interactions with different proteinaceous alpha-AIs. Major conclusions: Insect alpha-amylases have conserved domain architecture with differences in length, number of disulfide bonds, and secondary structure. Furthermore, few of them exhibit variable characteristics like chloride dependent activity, the presence of N-terminal glutamine residue to protect against proteolytic degradation, and loop variations near the enzyme active site. Conformation of alpha-AI protein could be an essential factor for their specificity and binding affinities towards target alpha-amylase(s). Furthermore, variation into the enzyme binding pocket residues might contribute to differential interactions with inhibitors. General significance: Molecular insights in the interactions between insect alpha-amylases and plant alpha-AI will provide the details of mechanisms assisting the inhibitor specificity. Furthermore, this information will help to design potent and effective alpha-AIs against specific alpha-amylase.

10aActive site10aalpha-amylase10aalpha-amylase inhibitor10aInsect10amolecular interactions1 aRane, Ashwini, S.1 aJoshi, Rakesh, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/molecular-determinant-specificity-differential-interaction-alpha-amylases-their02489nas a2200241 4500008004100000022001400041245010400055210006900159260000800228300001400236490000800250520167700258653001801935653002801953653001801981653001501999653001702014100002202031700002102053700002202074700002002096856013102116 2020 eng d a0141-813000aMolecular investigation of coleopteran specific alpha-amylase inhibitors from amaranthaceae members0 aMolecular investigation of coleopteran specific alphaamylase inh cNOV a1444-14500 v1633 a

alpha-Amylase inhibitors (alpha-AIs) target alpha-amylases and interfere with the carbohydrate digestion of insects. Among different classes of alpha-AIs, a knottin-type inhibitor from Amaranthus hypochondriacus (AhAI) was found to be specific against coleopteran storage pests. In this report, we have characterized three previously unidentified knottin-type alpha-AIs from various Amaranthaceae plants namely, Amaranthus hypochondriacus (AhAI2), Alternanthera sessilis (AsAI) and Chenopodium quinoa (CqAI). They contain a signal peptide, pro-peptide, and mature peptide. The mature peptides of the new alpha-AIs shared 68 to 78% identity with AhAI and have highly variable pro-peptide regions. Along with the cystine-knot fold, they showed conservation of reactive site residues. All recombinant alpha-AIs were successfully expressed in their active form and native state using an oxidative cytoplasmic environment. Inhibition studies against various amylases revealed that these inhibitors showed selective inhibition of coleopteran recombinant insect alpha-amylases viz., Tribolium castaneum, and Callosobruchus chinensis. Tribolium castaneum alpha-amylase inhibition potency was highest for AhAI2 (Ki similar to 15 mu M) followed by AsAI (Ki similar to 43 mu M) and CqAI (Ki similar to 61 mu M). Interaction analysis of these inhibitors illustrated that the reactive site of inhibitors make several non-covalent interactions with the substrate-binding pocket of coleopteran alpha-amylases. The selectivity of these inhibitors against coleopteran a-amylases highlights their potential in storage grain pest control. (C) 2020 Elsevier B.V. All rights reserved.

10aalpha-amylase10aalpha-amylase inhibitor10aAmaranthaceae10aColeoptera10aKnottin-type1 aRane, Ashwini, S.1 aVenkatesh, Varun1 aJoshi, Rakesh, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/molecular-investigation-coleopteran-specific-alpha-amylase-inhibitors-amaranthaceae-members02341nas a2200289 4500008004100000022001400041245008700055210006900142260000900211300001400220490000800234520143200242653002101674653001101695653001801706653003301724653001501757100002201772700002001794700002501814700001801839700001801857700002101875700002301896700001701919856011501936 2021 eng d a0011-389100aAntiviral drugs prioritization for COVID-19 management based on rational selection0 aAntiviral drugs prioritization for COVID19 management based on r cMAY  a1464-14700 v1203 a

The SARS-CoV-2 infection has resulted in COVID-19 pandemic worldwide. It has infected around 0.1 billion individuals and caused 2 million fatalities across the globe till mid-January 2021. Drug repurposing has been utilized as the most preferred therapeutic intervention for COVID-19 mitigation due to its necessity and feasibility. To prioritize therapeutic regime against COVID-19, we used 61 antiviral drugs and their combinations. Selected molecules were subjected to virtual screening against: (i) human angiotensin-converting enzyme 2 receptor binding domain (hACE-2) which serves as an anchor for virus attachment and entry, (ii) SARS-CoV-2 RNA dependent RNA polymerase (RdRp) responsible for viral RNA replication, and (iii) SARS-CoV-2 main protease (M-Pro) needed for viral polyprotein slab proteolytic processing. Based on docking score, pharmacodynamic and pharmacokinetic parameters, combinations of Daclatasvir, Elbasvir, Indinavir, Ledipasvir, Paritaprevir and Rilpivirine were analysed further. Our analysis suggested Sofosbuvir in combination with Ledipasvir and Daclatasvir as potential therapeutic agents for SARS-CoV-2. The combined score suggests that these combinations have superior anti-SARS-CoV-2 potential than Remdesivir and other investigational drugs. The present work provides a rationale-based approach to select drugs with possible anti-SARS-CoV-2 activity for further clinical evaluation.

10aDrug repurposing10ahACE-210amain protease10aRNA dependent RNA polymerase10aSARS-CoV-21 aJoshi, Rakesh, S.1 aGiri, Ashok, P.1 aKulkarni, Mahesh, J.1 aGupta, Mahesh1 aVerma, Savita1 aChaudhry, Dhruva1 aDeshmukh, Narendra1 aChugh, Anita uhttps://library.ncl.res.in/content/antiviral-drugs-prioritization-covid-19-management-based-rational-selection02228nas a2200229 4500008004100000022001400041245007100055210006900126260000800195300001100203490000800214520154200222653001001764653001101774653001701785653001701802653001001819100002901829700002101858700002001879856009901899 2021 eng d a0168-945200aAuxin: an emerging regulator of tuber and storage root development0 aAuxin an emerging regulator of tuber and storage root developmen cMAY a1108540 v3063 a

Many tuber and storage root crops owing to their high nutritional values offer high potential to overcome food security issues. The lack of information regarding molecular mechanisms that govern belowground storage organ development (except a tuber crop, potato) has limited the application of biotechnological strategies for improving storage crop yield. Phytohormones like gibberellin and cytokinin are known to play a crucial role in governing potato tuber development. Another phytohormone, auxin has been shown to induce tuber initiation and growth, and its crosstalk with gibberellin and strigolactone in a belowground modified stem (stolon) contributes to the overall potato tuber yield. In this review, we describe the crucial role of auxin biology in development of potato tubers. Considering the emerging reports from commercially important storage root crops (sweet potato, cassava, carrot, sugar beet and radish), we propose the function of auxin and related gene regulatory network in storage root development. The pattern of auxin content of stolon during various stages of potato tuber formation appears to be consistent with its level in various developmental stages of storage roots. We have also put-forward the potential of three-way interaction between auxin, strigolactone and mycorrhizal fungi in tuber and storage root development. Overall, we propose that auxin gene regulatory network and its crosstalk with other phytohormones in stolons/roots could govern belowground tuber and storage root development.

10aAuxin10aPotato10aStorage root10aSweet potato10aTuber1 aKondhare, Kirtikumar, R.1 aPatil, Aruna, B.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/auxin-emerging-regulator-tuber-and-storage-root-development02694nas a2200229 4500008004100000245008200041210006900123260000800192300000900200490000700209520197100216653001302187653002402200653001902224653001202243653001102255653001702266100002102283700002902304700002002333856011102353 2021 eng d00aDiversity in chemical structures and biological properties of plant alkaloids0 aDiversity in chemical structures and biological properties of pl cJUN a33740 v263 aPhytochemicals belonging to the group of alkaloids are signature specialized metabolites endowed with countless biological activities. Plants are armored with these naturally produced nitrogenous compounds to combat numerous challenging environmental stress conditions. Traditional and modern healthcare systems have harnessed the potential of these organic compounds for the treatment of many ailments. Various chemical entities (functional groups) attached to the central moiety are responsible for their diverse range of biological properties. The development of the characterization of these plant metabolites and the enzymes involved in their biosynthesis is of an utmost priority to deliver enhanced advantages in terms of biological properties and productivity. Further, the incorporation of whole/partial metabolic pathways in the heterologous system and/or the overexpression of biosynthetic steps in homologous systems have both become alternative and lucrative methods over chemical synthesis in recent times. Moreover, in-depth research on alkaloid biosynthetic pathways has revealed numerous chemical modifications that occur during alkaloidal conversions. These chemical reactions involve glycosylation, acylation, reduction, oxidation, and methylation steps, and they are usually responsible for conferring the biological activities possessed by alkaloids. In this review, we aim to discuss the alkaloidal group of plant specialized metabolites and their brief classification covering major categories. We also emphasize the diversity in the basic structures of plant alkaloids arising through enzymatically catalyzed structural modifications in certain plant species, as well as their emerging diverse biological activities. The role of alkaloids in plant defense and their mechanisms of action are also briefly discussed. Moreover, the commercial utilization of plant alkaloids in the marketplace displaying various applications has been enumerated.10aalkaloid10aBiological activity10aclassification10adefense10aEnzyme10amodification1 aBhambhani, Sweta1 aKondhare, Kirtikumar, R.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/diversity-chemical-structures-and-biological-properties-plant-alkaloids01973nas a2200157 4500008004100000022001400041245007500055210006900130260000800199490000700207520144800214100002101662700001301683700002001696856009901716 2021 eng d a1664-462X00aEffectors of root-knot nematodes: an arsenal for successful parasitism0 aEffectors of rootknot nematodes an arsenal for successful parasi cDEC0 v123 aRoot-knot nematodes (RKNs) are notorious plant-parasitic nematodes first recorded in 1855 in cucumber plants. They are microscopic, obligate endoparasites that cause severe losses in agriculture and horticulture. They evade plant immunity, hijack the plant cell cycle, and metabolism to modify healthy cells into giant cells (GCs) – RKN feeding sites. RKNs secrete various effector molecules which suppress the plant defence and tamper with plant cellular and molecular biology. These effectors originate mainly from sub-ventral and dorsal oesophageal glands. Recently, a few non-oesophageal gland secreted effectors have been discovered. Effectors are essential for the entry of RKNs in plants, subsequently formation and maintenance of the GCs during the parasitism. In the past two decades, advanced genomic and post-genomic techniques identified many effectors, out of which only a few are well characterized. In this review, we provide molecular and functional details of RKN effectors secreted during parasitism. We list the known effectors and pinpoint their molecular functions. Moreover, we attempt to provide a comprehensive insight into RKN effectors concerning their implications on overall plant and nematode biology. Since effectors are the primary and prime molecular weapons of RKNs to invade the plant, it is imperative to understand their intriguing and complex functions to design counter-strategies against RKN infection.1 aJagdale, Shounak1 aRao, Uma1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/effectors-root-knot-nematodes-arsenal-successful-parasitism02270nas a2200241 4500008004100000022001400041245012300055210006900178260000900247300001200256490000800268520144100276653002701717653001501744653001101759653002901770653002101799100002501820700002201845700002001867700002001887856012101907 2021 eng d a0141-813000aFunctional insights into two Ocimum kilimandscharicum 4-coumarate-CoA ligases involved in phenylpropanoid biosynthesis0 aFunctional insights into two Ocimum kilimandscharicum 4coumarate cJUN  a202-2100 v1813 a

Plant 4-coumarate-CoA ligase (4CL) catalyzes the ligation of CoA to cinnamic acid and its derivatives. Activated CoA esters are utilized for the biosynthesis of phenolic metabolites and lignin that play essential function in plants. Here, we characterize the diversity of Ocimum kilimandscharicum 4CLs (Ok4CLs). Phylogenetic analysis suggest that Ok4CLs could be grouped into three classes, class I -enzymes mostly involved in lignin biosynthesis, class II -non-structural phenylpropanoid biosynthesis and class III -yet to be characterized for specific role(s). We selected two Ok4CLs namely Ok4CL7 and Ok4CL15 for further characterization. Gene expression analysis sug-gested that Ok4CL7 is highly expressed in leaf trichomes, whereas Ok4CL15 is abundant in the roots. The recom-binant Ok4CL7 and Ok4CL15 had optimal enzyme activities at 40 degrees C in pH 8 and 7, respectively. Ok4CL7 showed substrate preference towards p-coumaric acid, ferulic acid and caffeic acid. While, Ok4CL15 preferredp-coumaric acid, ferulic acid and sinapic acid. Feruloyl adenylate showed higher number of contacts and lowers binding en-ergy with Ok4CL7 and 15 compared to cinnamoyl adenylate. Based on root-specific expression and preference for sinapic acid, Ok4CL15 might be involved in lignin biosynthesis. Further exploration is needed to unravel the role of diverse Ok4CLs in O. kilimandscharicum. (c) 2021 Elsevier B.V. All rights reserved.

10a4-Coumarate-CoA ligase10aFlavonoids10alignin10aOcimum kilimandscharicum10aPhenylpropanoids1 aLavhale, Santosh, G.1 aJoshi, Rakesh, S.1 aKumar, Yashwant1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/functional-insights-two-ocimum-kilimandscharicum-4-coumarate-coa-ligases-involved02984nas a2200301 4500008004100000022001400041245008000055210006900135260000900204300000700213490000800220520207700228653001002305653001802315653001202333653001602345653001702361653002802378653001002406100001602416700002602432700002702458700002202485700002802507700002102535700002002556856010602576 2021 eng d a0032-093500aInfluence of domestication on specialized metabolic pathways in fruit crops0 aInfluence of domestication on specialized metabolic pathways in  cFEB  a610 v2533 a

Main conclusionDuring the process of plant domestication, the selection and traditional breeding for desired characters such as flavor, juiciness and nutritional value of fruits, probably have resulted in gain or loss of specialized metabolites contributing to these traits. Their appearance in fruits is likely due to the acquisition of novel and specialized metabolic pathways and their regulation, driven by systematic molecular evolutionary events facilitated by traditional breeding.AbstractPlants change their armory of specialized metabolism to adapt and survive in diverse ecosystems. This may occur through molecular evolutionary events, such as single nucleotide polymorphism, gene duplication and transposition, leading to convergent or divergent evolution of biosynthetic pathways producing such specialized metabolites. Breeding and selection for improved specific and desired traits (fruit size, color, taste, flavor, etc.) in fruit crops through conventional breeding approaches may further alter content and profile of specialized metabolites. Biosynthetic routes of these metabolites have been studied in various plants. Here, we explore the influence of plant domestication and breeding processes on the selection of biosynthetic pathways of favorable specialized metabolites in fruit crops. An orderly clustered arrangement of genes associated with their production is observed in many fruit crops. We further analyzed selection-based acquisition of specialized metabolic pathways comparing first the metabolic profiles and genes involved in their biosynthesis, followed by the genomic organization of such genes between wild and domesticated horticultural crops. Domestication of crop plants favored the acquisition and retention of metabolic pathways that enhanced the fruit value while eliminated those which produced toxic or unfavorable metabolites. Interestingly, unintentional reorganization of complex metabolic pathways by selection and traditional breeding processes has endowed us with flavorful, juicy and nutritionally rich fruits.

10aAroma10aDomestication10aFlavour10aFruit crops10aGene cluster10aSpecialized metabolites10aTaste1 aDar, Saleem1 aDholakia, Bhushan, B.1 aKulkarni, Abhijeet, P.1 aOak, Pranjali, S.1 aShanmugam, Dhanasekaran1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/influence-domestication-specialized-metabolic-pathways-fruit-crops02705nas a2200241 4500008004100000022001400041245008400055210006900139260000900208300000800217490000700225520190800232653001502140653001802155653001302173653002902186653003502215653003002250100002502280700002602305700002002331856011202351 2021 eng d a1471-222900aPINIR: a comprehensive information resource for Pin-II type protease inhibitors0 aPINIR a comprehensive information resource for PinII type protea cJUN  a2670 v213 aBackground: Serine protease inhibitors belonging to the Potato type-II Inhibitor family Protease Inhibitors (Pin-II type PIs) are essential plant defense molecules. They are characterized by multiple inhibitory repeat domains, conserved disulfide bond pattern, and a tripeptide reactive center loop. These features of Pin-II type PIs make them potential molecules for protein engineering and designing inhibitors for agricultural and therapeutic applications. However, the diversity in these PIs remains unexplored due to the lack of annotated protein sequences and their functional attributes in the available databases. Results: We have developed a database, PINIR (Pin-II type PIs Information Resource), by systematic collection and manual annotation of 415 Pin-II type PI protein sequences. For each PI, the number and position for signature sequences are specified: 695 domains, 75 linkers, 63 reactive center loops, and 10 disulfide bond patterns are identified and mapped. Database analysis revealed novel subcategories of PIs, species-correlated occurrence of inhibitory domains, reactive center loops, and disulfide bond patterns. By analyzing linker regions, we predict that alternative processing at linker regions could generate PI variants in the Solanaceae family. Conclusion: PINIR (https://pinir.ncl.res.in) provides a web interface for browsing and analyzing the protein sequences of Pin-II type PIs. Information about signature sequences, spatio-temporal expression, biochemical properties, gene sequences, and literature references are provided. Analysis of PINIR depicts conserved species-specific features of Pin-II type PI protein sequences. Diversity in the sequence of inhibitory domains and reactive loops directs potential applications to engineer Pin-II type PIs. The PINIR database will serve as a comprehensive information resource for further research into Pin-II type PIs.10aannotation10aData analysis10aDatabase10aKnowledge representation10aPin-II type protease inhibitor10aProtein sequence analysis1 aYadav, Nikhilesh, K.1 aSaikhedkar, Nidhi, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/pinir-comprehensive-information-resource-pin-ii-type-protease-inhibitors02884nas a2200409 4500008004100000022001400041245012200055210006900177260000800246300001400254490000800268520160500276653001201881653002701893653003601920653002501956653001101981100002701992700001902019700002002038700002002058700001902078700002102097700002302118700001602141700001702157700001602174700001602190700002002206700002602226700002202252700002102274700002002295700001702315700001902332856012302351 2022 eng d a0028-646X00a2-oxoglutarate-dependent dioxygenases drive expansion of steroidal alkaloid structural diversity in the genus Solanum0 a2oxoglutaratedependent dioxygenases drive expansion of steroidal cMAY a1394-14100 v2343 a

Solanum steroidal glycoalkaloids (SGAs) are renowned defence metabolites exhibiting spectacular structural diversity. Genes and enzymes generating the SGA precursor pathway, SGA scaffold and glycosylated forms have been largely identified. Yet, the majority of downstream metabolic steps creating the vast repertoire of SGAs remain untapped. Here, we discovered that members of the 2-OXOGLUTARATE-DEPENDENT DIOXYGENASE (2-ODD) family play a prominent role in SGA metabolism, carrying out three distinct backbone-modifying oxidative steps in addition to the three formerly reported pathway reactions. The GLYCOALKALOID METABOLISM34 (GAME34) enzyme catalyses the conversion of core SGAs to habrochaitosides in wild tomato S. habrochaites. Cultivated tomato plants overexpressing GAME34 ectopically accumulate habrochaitosides. These habrochaitoside enriched plants extracts potently inhibit Puccinia spp. spore germination, a significant Solanaceae crops fungal pathogen. Another 2-ODD enzyme, GAME33, acts as a desaturase (via hydroxylation and E/F ring rearrangement) forming unique, yet unreported SGAs. Conversion of bitter alpha-tomatine to ripe fruit, nonbitter SGAs (e.g. esculeoside A) requires two hydroxylations; while the known GAME31 2-ODD enzyme catalyses hydroxytomatine formation, we find that GAME40 catalyses the penultimate step in the pathway and generates acetoxy-hydroxytomatine towards esculeosides accumulation. Our results highlight the significant contribution of 2-ODD enzymes to the remarkable structural diversity found in plant steroidal specialized metabolism.

10aSolanum10aspecialized metabolism10asteroidal glycoalkaloids (SGAs)10astructural diversity10aTomato1 aSonawane, Prashant, D.1 aJozawiak, Adam1 aBarbole, Ranjit1 aPanda, Sayantan1 aAbebie, Bekele1 aKazachkova, Yana1 aGharat, Sachin, A.1 aRamot, Ofir1 aUnger, Tamar1 aWizler, Guy1 aMeir, Sagit1 aRogachev, Ilana1 aDoron-Faigenboim, Adi1 aPetreikov, Marina1 aSchaffer, Arthur1 aGiri, Ashok, P.1 aScherf, Tali1 aAharoni, Asaph uhttps://library.ncl.res.in/content/2-oxoglutarate-dependent-dioxygenases-drive-expansion-steroidal-alkaloid-structural02377nas a2200241 4500008004100000022001400041245012000055210006900175260000800244300001000252490000700262520155300269653000901822653002501831653001101856653001701867653001901884653003401903100002101937700002901958700002001987856012802007 2022 eng d a1568-776700aAdvanced genome editing strategies for manipulation of plant specialized metabolites pertaining to biofortification0 aAdvanced genome editing strategies for manipulation of plant spe cFEB a81-990 v213 a

Emerging trends in molecular biology have progressed the knowledge of plant specialized metabolites with respect to diversity in structure, function and biosynthetic pathways. Being powerful genome-editing tools, Zinc Finger Nuclease, Transcription Activator-Like Effector Nuclease, and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) systems have found potential application in genome and epigenome engineering. CRISPR/Cas9 is being used for studying the functions of multiple genes and gene families in plants. Our analysis suggests that although a rapid progress has occurred for utilization of CRISPR/Cas9 tool in crop improvement, limited studies are available for its application in manipulation of gene clusters of useful specialized metabolites in plants. In this review, after describing briefly about the recent advancements in genome editing techniques, we have further discussed their applicability in the modulation of metabolite production and biofortification of food crops. We have also emphasized the importance of CRISPR/Cas9-based targeted deletion of larger chromosomal fragments or gene clusters towards value addition of crop plants. The current policies for CRISPR/Cas9-edited crop plants in different countries and their acceptability in market place is also discussed. We propose that advanced genome editing techniques, including a multiplex CRISPR/Cas9 system could serve as a versatile tool for rewiring of metabolite gene clusters and improving the levels of useful metabolites in plants.

10aCas910aChromosomal fragment10aCRISPR10aGene cluster10aGenome editing10aPlant specialized metabolites1 aBhambhani, Sweta1 aKondhare, Kirtikumar, R.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/advanced-genome-editing-strategies-manipulation-plant-specialized-metabolites-pertaining02181nas a2200253 4500008004100000022001400041245012300055210006900178260000800247300001100255490000800266520133300274653001301607653001301620653002201633653001901655653001801674653001501692100002301707700002001730700002701750700002001777856013001797 2022 eng d a0031-942200aCharacterized constituents of insect herbivore oral secretions and their influence on the regulation of plant defenses0 aCharacterized constituents of insect herbivore oral secretions a cJAN a1130080 v1933 aFor more than 350 million years, there have been ongoing dynamic interactions between plants and insects. In several cases, insects cause-specific feeding damage with ensuing herbivore-associated molecular patterns that invoke characteristic defense responses. During feeding on plant tissue, insects release oral secretions (OSs) containing a repertoire of molecules affecting plant defense (effectors). Some of these OS components might elicit a defense response to combat insect attacks (elicitors), while some might curb the plant defenses (suppressors). Few reports suggest that the synthesis and function of OS components might depend on the host plant and associated microorganisms. We review these intricate plant-insect interactions, during which there is a continuous exchange of molecules between plants and feeding insects along with the associated microorganisms. We further provide a list of commonly identified inducible plant produced defensive molecules released upon insect attack as well as in response to OS treatments of the plants. Thus, we describe how plants specialized and defense-related metabolism is modulated at innumerable phases by OS during plant-insect interactions. A molecular understanding of these complex interactions will provide a means to design eco-friendly crop protection strategies.10aEffector10aElicitor10aHerbivore insects10aOral secretion10aPlant defense10aSuppressor1 aKallure, Gopal, S.1 aKumari, Archana1 aShinde, Balkrishna, A.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/characterized-constituents-insect-herbivore-oral-secretions-and-their-influence-regulation02347nas a2200265 4500008004100000022001400041245014700055210006900202260000800271300001100279490000800290520144700298653002101745653001201766653001601778653001301794653001901807653001401826100002301840700002701863700002501890700002001915700002001935856012601955 2022 eng d a0168-945200aDietary influence on modulation of Helicoverpa armigera oral secretion composition leading to differential regulation of tomato plant defense.0 aDietary influence on modulation of Helicoverpa armigera oral sec cJAN a1111200 v3143 a

Little is known about how different plant-based diets influence the insect herbivores' oral secretion (OS) composition and eventually the plant defense responses. We analyzed the OS composition of the generalist Lepidopteran insect, Helicoverpa armigera feeding on the host plant tomato (OSH), non-host plant capsicum (OSNH), and artificial diet (OSAD) using Liquid Chromatography-Quadrupole Time of Flight Mass Spectrometry. Higher numbers and levels of alkaloids and terpenoids were observed in OSH and OSNH, respectively while OSAD was rich in phospholipids. Interestingly, treatment of H. armigera OSAD, OSH and OSNH on wounded tomato leaves showed differential expression of (i) genes involved in JA and SA biosynthesis and their responsive genes, and (ii) biosynthetic pathway genes of chlorogenic acid (CGA) and trehalose, which exhibited increased accumulation along with several other plant defensive metabolites. Specifically, high levels of CGA were detected after OSH and OSNH treatments in tomato leaves. There was higher expression of the genes involved in phenyl-propanoid biosynthesis, which may lead to the increased accumulation of CGA and related metabolites. In the insect bioassay, CGA significantly inhibited H. armigera larval growth. Our results underline the differential accumulation of plant and insect OS metabolites and identified potential plant metabolite(s) affecting insect growth and development.

10aChlorogenic acid10adefense10aMetabolites10aNon-host10aOral secretion10atrehalose1 aKallure, Gopal, S.1 aShinde, Balkrishna, A.1 aBarvkar, Vitthal, T.1 aKumari, Archana1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/dietary-influence-modulation-helicoverpa-armigera-oral-secretion-composition-leading-002246nas a2200265 4500008004100000022001400041245007900055210006900134260000800203300001200211490000700223520141000230653002101640653000701661653002701668653001601695653002201711100002301733700002601756700002501782700002201807700002001829700002401849856010701873 2022 eng d a0022-284400aFunctional diversity of the lepidopteran ATP-binding cassette transporters0 aFunctional diversity of the lepidopteran ATPbinding cassette tra cAUG a258-2700 v903 a

The ATP-binding cassette (ABC) transporter gene family is ubiquitous in the living world. ABC proteins bind and hydrolyze ATP to transport a myriad of molecules across various lipid-containing membrane systems. They have been studied well in plants for transport of a variety of compounds and particularly, in vertebrates due to their direct involvement in resistance mechanisms against several toxic molecules/metabolites. ABC transporters in insects are found within large multigene families involved in the efflux of chemical insecticides and toxic/undesired metabolites originating from food and endogenous metabolism. This review deals with ABC transporter subfamilies of few agronomically important Lepidopteran pests. The transcriptional dynamics and regulation of ABC transporters during insect development emphasizes their functional diversity against insecticides, Cry toxins, and plant specialized metabolites. To generate insights about molecular function and physiological roles of ABCs, functional and structural characterization is necessary. Also, expansion and divergence of ABC transporter gene subfamilies in Lepidopteran insects needs more systematic investigation. We anticipate that newer methods of insect control in agriculture can benefit from an understanding of ABC transporter interactions with a vast range of natural specialized molecules and synthetic compounds.

10aABC transporters10aBt10ainsecticide resistance10aLepidoptera10aPlant metabolites1 aBarve, Pranoti, R.1 aTellis, Meenakshi, B.1 aBarvkar, Vitthal, T.1 aJoshi, Rakesh, S.1 aGiri, Ashok, P.1 aKotkar, Hemlata, M. uhttps://library.ncl.res.in/content/functional-diversity-lepidopteran-atp-binding-cassette-transporters02527nas a2200241 4500008004100000022001400041245014400055210006900199260000800268300001200276490000700288520167700295653001901972653001401991653001802005653002602023653001102049653002802060100002202088700002602110700002002136856012902156 2022 eng d a1568-776700aGlance at the chemodiversity of Ocimum species: Trends, implications, and strategies for the quality and yield improvement of essential oil0 aGlance at the chemodiversity of Ocimum species Trends implicatio cJUN a879-9130 v213 a

Ocimum species represent commercially important medicinal and aromatic plants. The essential oil biosynthesized by Ocimum species is enriched with specialized metabolites specifically, terpenoids and phenylpropanoids. Interestingly, various Ocimum species are known to exhibit diverse chemical profiles, and this chemical diversity has been at the center of many studies to identify commercially important chemotypes. Here, we present various chemotypes from the Ocimum species and emphasize trends, implications, and strategies for the quality and yield improvement of essential oil. Globally, many Ocimum species have been analyzed for their essential oil composition in over 50 countries. Asia represents the highest number of chemotypes, followed by Africa, South America, and Europe. Ocimum basilicum L. has been the most widespread and well-studied species, followed by O. gratissimum L., O. tenuiflorum L., O. canum Sims, O. americanum and O. kilimandscharicum Gurke. Moreover, various molecular reasons, benefits, adverse health effects and mechanisms behind this vast chemodiversity have been discussed. Different strategies of plant breeding, metabolic engineering, transgenic, and tissue-culture, along with anatomical modifications, are surveyed to enhance specific chemotypic profiles and essential oil yield in numerous Ocimum species. Consequently, chemical characterization of the essential oil obtained from Ocimum species has become indispensable for its proper utilization. The present chemodiversity knowledge from Ocimum species will help to exploit various applications in the industrial, agriculture, biopharmaceutical, and food sectors.

10aChemodiversity10aChemotype10aEssential oil10aMetabolic engineering10aOcimum10aSpecialized metabolites1 aGurav, Tanuja, P.1 aDholakia, Bhushan, B.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/glance-chemodiversity-ocimum-species-trends-implications-and-strategies-quality-and-yield02685nas a2200229 4500008004100000022001400041245016600055210006900221260000800290300001600298490000600314520183500320100001802155700002402173700002002197700002302217700002002240700002002260700002002280700002502300856013002325 2022 eng d a2470-134300aInvestigation of the captopril-insulin interaction by mass spectrometry and computational approaches reveals that captopril induces structural changes in insulin0 aInvestigation of the captoprilinsulin interaction by mass spectr cJUL a23115-231260 v73 a

ABSTRACT: Post-translational modifications remarkably regulate proteins' biological function. Small molecules such as reactive thiols, metabolites, and drugs may covalently modify the proteins and cause structural changes. This study reports the covalent modification and noncovalent interaction of insulin and captopril, an FDA-approved antihypertensive drug, through mass spectrometric and computation-based approaches. Mass spectrometric analysis shows that captopril modifies intact insulin, reduces it into its ``A'' and ``B'' chains, and covalently modifies them by forming adducts. Since captopril has a reactive thiol group, it might reduce the insulin dimer or modify it by reacting with cysteine residues. This was proven with dithiothreitol treatment, which reduced the abundance of captopril adducts of insulin A and B chains and intact Insulin. Liquid chromatography tandem mass spectrometric analysis identified the modification of a total of four cysteine residues, two in each of the A and B chains of insulin. These modifications were identified to be Cys6 and Cys7 of the A chain and Cys7 and Cys19 of the B chain. Mass spectrometric analysis indicated that captopril may simultaneously modify the cysteine residues of intact insulin or its subunits A and B chains. Biophysical studies involving light scattering and thioflavin T assay suggested that the binding of captopril to the protein leads to the formation of aggregates. Docking and molecular dynamics studies provided insights into the noncovalent interactions and associated structural changes in insulin. This work is a maiden attempt to understand the detailed molecular interactions between captopril and insulin. These findings suggest that further investigations are required to understand the long-term effect of drugs like captopril.

1 aGhosh, Amrita1 aPawar, Aiswarya, B.1 aChirmade, Tejas1 aJathar, Swaraj, M.1 aBhambure, Rahul1 aSengupta, Durba1 aGiri, Ashok, P.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/investigation-captopril-insulin-interaction-mass-spectrometry-and-computational-approaches02802nas a2200445 4500008004100000022001400041245013400055210006900189260000800258300001400266490000800280520142600288653001901714653001201733653002101745653001101766653001901777653003601796653001101832100002001843700001801863700002701881700002301908700002101931700001901952700003101971700002602002700002002028700002002048700001802068700001602086700001602102700002002118700002202138700001702160700001802177700002002195700001902215856012202234 2022 eng d a0028-646X00aSteroidal alkaloids defence metabolism and plant growth are modulated by the joint action of gibberellin and jasmonate signalling0 aSteroidal alkaloids defence metabolism and plant growth are modu cFEB a1220-12370 v2333 aSteroidal glycoalkaloids (SGAs) are protective metabolites constitutively produced by Solanaceae species. Genes and enzymes generating the vast structural diversity of SGAs have been largely identified. Yet, mechanisms of hormone pathways coordinating defence (jasmonate; JA) and growth (gibberellin; GA) controlling SGAs metabolism remain unclear. We used tomato to decipher the hormonal regulation of SGAs metabolism during growth vs defence tradeoff. This was performed by genetic and biochemical characterisation of different JA and GA pathways components, coupled with in vitro experiments to elucidate the crosstalk between these hormone pathways mediating SGAs metabolism. We discovered that reduced active JA results in decreased SGA production, while low levels of GA or its receptor led to elevated SGA accumulation. We showed that MYC1 and MYC2 transcription factors mediate the JA/GA crosstalk by transcriptional activation of SGA biosynthesis and GA catabolism genes. Furthermore, MYC1 and MYC2 transcriptionally regulate the GA signalling suppressor DELLA that by itself interferes in JA-mediated SGA control by modulating MYC activity through protein-protein interaction. Chemical and fungal pathogen treatments reinforced the concept of JA/GA crosstalk during SGA metabolism. These findings revealed the mechanism of JA/GA interplay in SGA biosynthesis to balance the cost of chemical defence with growth.10aalpha-tomatine10adefence10agibberellin (GA)10agrowth10ajasmonate (JA)10asteroidal glycoalkaloids (SGAs)10aTomato1 aPanda, Sayantan1 aJozwiak, Adam1 aSonawane, Prashant, D.1 aSzymanski, Jedrzej1 aKazachkova, Yana1 aVainer, Andrii1 aKilambi, Himabindu, Vasuki1 aAlmekias-Siegl, Efrat1 aDikaya, Varvara1 aBocobza, Samuel1 aShohat, Hagai1 aMeir, Sagit1 aWizler, Guy1 aGiri, Ashok, P.1 aSchuurink, Robert1 aWeiss, David1 aYasuor, Hagai1 aKamble, Avinash1 aAharoni, Asaph uhttps://library.ncl.res.in/content/steroidal-alkaloids-defence-metabolism-and-plant-growth-are-modulated-joint-action02358nas a2200253 4500008004100000245011500041210006900156260000800225300000900233490000700242520149500249100002701744700002301771700001801794700002001812700002001832700002601852700001601878700002001894700002401914700002001938700001901958856012701977 2023 eng d00aBAHD-type acyltransferase concludes the biosynthetic pathway of non-bitter glycoalkaloids in ripe tomato fruit0 aBAHDtype acyltransferase concludes the biosynthetic pathway of n cJUL a45400 v143 a

During tomato fruit ripening, bitter and toxic steroidal glycoalkaloids (SGAs) are converted to nonbitter and less toxic forms, but proposed acylating enzyme in pathway remain unknown. Here, authors report BAHD-type acyltransferase that catalyze acylation step in biosynthesis of non-bitter SGAs in tomato. Tomato is the highest value fruit and vegetable crop worldwide, yet produces & alpha;-tomatine, a renowned toxic and bitter-tasting anti-nutritional steroidal glycoalkaloid (SGA) involved in plant defense. A suite of modifications during tomato fruit maturation and ripening converts & alpha;-tomatine to the non-bitter and less toxic Esculeoside A. This important metabolic shift prevents bitterness and toxicity in ripe tomato fruit. While the enzymes catalyzing glycosylation and hydroxylation reactions in the Esculeoside A pathway have been resolved, the proposed acetylating step remains, to date, elusive. Here, we discovered that GAME36 (GLYCOALKALOID METABOLISM36), a BAHD-type acyltransferase catalyzes SGA-acetylation in cultivated and wild tomatoes. This finding completes the elucidation of the core Esculeoside A biosynthetic pathway in ripe tomato, allowing reconstitution of Esculeoside A production in heterologous microbial and plant hosts. The involvement of GAME36 in bitter SGA detoxification pathway points to a key role in the evolution of sweet-tasting tomato as well as in the domestication and breeding of modern cultivated tomato fruit.

1 aSonawane, Prashant, D.1 aGharat, Sachin, A.1 aJozwiak, Adam1 aBarbole, Ranjit1 aHeinicke, Sarah1 aAlmekias-Siegl, Efrat1 aMeir, Sagit1 aRogachev, Ilana1 aO'Connor, Sarah, E.1 aGiri, Ashok, P.1 aAharoni, Asaph uhttps://library.ncl.res.in/content/bahd-type-acyltransferase-concludes-biosynthetic-pathway-non-bitter-glycoalkaloids-ripe02323nas a2200265 4500008004100000022001400041245013000055210006900185260000800254300001200262490000700274520144300281653001201724653001301736653002401749653001801773653001501791653001201806653001701818100002201835700002601857700002201883700002001905856013201925 2023 eng d a1572-388700aDomain shuffling and site-saturation mutagenesis for the enhanced inhibitory potential of amaranthaceae α-amylase inhibitors0 aDomain shuffling and sitesaturation mutagenesis for the enhanced cOCT a519-5320 v423 a

Amaranthaceae a-amylase inhibitors (AAIs) are knottin-type proteins with selective inhibitory potential against coleopteran a-amylases. Their small size and remarkable stability make them exciting molecules for protein engineering to achieve superior selectivity and efficacy. In this report, we have designed a set of AAI pro- and mature peptides chimeras. Based on in silico analysis, stable AAI chimeras having a stronger affinity with target amylases were selected for characterization. In vitro studies validated that chimera of the propeptide from Chenopodium quinoa a-AI and mature peptide from Beta vulgaris a-AI possess 3, 7.6, and 4.26 fold higher inhibition potential than parental counterparts. Importantly, recombinant AAI chimera retained specificity towards target coleopteran a-amylases. In addition, to improve the inhibitory potential of AAI, we performed in silico site-saturation mutagenesis. Computational analysis followed by experimental data showed that substituting Asparagine at the 6th position with Methionine had a remarkable increase in the specific inhibition potential of Amaranthus hypochondriacus a-AI. These results provide structural-functional insights into the vitality of AAI propeptide and a potential hotspot for mutagenesis to enhance the AAI activity. Our investigation will be a toolkit for AAI's optimization and functional differentiation for future biotechnological applications.

10a& alpha10a-Amylase10a-Amylase inhibitors10aAmaranthaceae10aColeoptera10aPeptide10aPest control1 aRane, Ashwini, S.1 aNair, Vineetkumar, S.1 aJoshi, Rakesh, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/domain-shuffling-and-site-saturation-mutagenesis-enhanced-inhibitory-potential-amaranthaceae01935nas a2200253 4500008004100000022001400041245008900055210006900144260000800213300001000221490000700231520111400238653001501352653001501367653002101382653002701403653002301430653001701453100002601470700002301496700002401519700002001543856011801563 2023 eng d a0095-362800aGenomic determinants of entomopathogenic fungi and their involvement in pathogenesis0 aGenomic determinants of entomopathogenic fungi and their involve cJAN a49-600 v853 a

Entomopathogenic fungi offer an effective and eco-friendly alternative to curb insect populations in biocontrol strategy. The evolutionary history of selected entomopathogenic fungi indicates their ancestral relationship with plant endophytes. During this host shifting, entomopathogenic fungi must have acquired multiple mechanisms, including a combination of various biomolecules that make them distinguishable from other fungi. In this review, we focus on understanding various biochemical and molecular mechanisms involved in entomopathogenesis. In particular, we attempt to explain the indispensable role of enlarged gene families of various virulent factors, viz. chitinases, proteases, lipases, specialized metabolites, and cytochrome P450, in entomopathogenesis. Our analysis suggests that entomopathogenic fungi recruit a different set of gene products during the progression of pathogenesis. Knowledge of these bio-molecular interactions between fungi and insect hosts will allow researchers to execute pointed efforts towards the development of improved entomopathogenic fungal strains.

10aBiocontrol10aChitinases10aEndophytic fungi10aEntomopathogenic fungi10aHydrolytic enzymes10aInsect pests1 aVidhate, Ravindra, P.1 aDawkar, Vishal, V.1 aPunekar, Sachin, A.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/genomic-determinants-entomopathogenic-fungi-and-their-involvement-pathogenesis02238nas a2200205 4500008004100000022001400041245012800055210006900183260000900252300001600261490000600277520148500283100002101768700002001789700002701809700002301836700002201859700002001881856013101901 2024 eng d a2470-134300aAdsorption based downstream processing approach for penicillin V from a Penicillium chrysogenum BIONCL I22 culture filtrate0 aAdsorption based downstream processing approach for penicillin V cJUN  a25859-258690 v93 a

Penicillin V (phenoxy methyl penicillin) is highly sought after among natural penicillins because of its exceptional acid stability and effectiveness against common skin and respiratory infections. Given its wide-ranging therapeutic uses, there is a need to establish a greener method for its maximum recovery to reduce the carbon footprint. Here, we have identified and validated optimized operational conditions for resin-based penicillin V recovery. It was observed that Amberlite XAD4 had the highest penicillin V hydrophobic adsorption capacity among the other screened resins. Kinetic and isothermal studies using linear and nonlinear regression analysis showed that the adsorption process well fitted with pseudo-second-order kinetics (R-2 = 0.9816) and the Freundlich adsorption isotherm model (R-2 = 0.9871). Adsorption equilibrium was attained within 4 h, while maximum adsorption was observed at 3 mg/mL penicillin V concentration. Furthermore, the optimized extraction protocol was compared with the conventional butyl acetate-based downstream processing. Under optimum conditions resin-based penicillin V recovery was 2-fold higher as compared to the solvent extraction method and the resin could be reused for over six cycles without compromising the yield. These findings signify substantial progress toward the development of an environmentally sustainable approach for penicillin V recovery and a potentially viable method for extractive fermentation.

1 aMishal, Bela, H.1 aDas, Sancharini1 aMahajan, Vaishnavi, N.1 aDharne, Mahesh, S.1 aJoshi, Rakesh, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/adsorption-based-downstream-processing-approach-penicillin-v-penicillium-chrysogenum-bioncl02322nas a2200253 4500008004100000022001400041245014300055210006900198260000800267490000800275520140100283653001701684653003701701653001401738653002501752653001901777100002301796700003201819700002101851700002501872700002201897700002001919856012901939 2024 eng d a0965-174800aAminoacylase efficiently hydrolyses fatty acid amino acid conjugates of Helicoverpa armigera potentially to increase the pool of glutamine0 aAminoacylase efficiently hydrolyses fatty acid amino acid conjug cFEB0 v1653 a

One of the most prevalent bioactive molecules present in the oral secretion (OS) of lepidopteran insects is fatty acid amino acid conjugates (FACs). Insect dietary components have influence on the synthesis and retaining the pool of FACs in the OS. We noted differential and diet-specific accumulation of FACs in the OS of Helicoverpa armigera by using Liquid Chromatography-Quadrupole Time of Flight Mass Spectrometry. Interestingly, we identified FACs hydrolyzing enzyme aminoacylase (HaACY) in the OS of H. armigera through proteomic analysis. Next, we have cloned, expressed, and purified active recombinant HaACY in the bacterial system. Recombinant HaACY hydrolyzes all the six identified FACs in the OS of H. armigera larvae fed on host and non-host plants and releases respective fatty acid and glutamine. In these six FACs, fatty acid moieties vary while amino acid glutamine was common. Glutamine obtained upon hydrolysis of FACs by HaACY might serve as an amino acid pool for insect growth and development. To understand the substrate choices of HaACY, we chemically synthesized, purified, and characterized all the six FACs. Interestingly, rHaACY also shows hydrolysis of synthetic FACs into respective fatty acid and glutamine. Our results underline the importance of diet on accumulation of FACs and role of aminoacylase(s) in regulating the level of FACs and glutamine.

10aAminoacylase10aFatty acid amino acid conjugates10aGlutamine10aHelicoverpa armigera10aOral secretion1 aKallure, Gopal, S.1 aSahoo, Shubhranshu, Shekhar1 aKale, Rutuja, S.1 aBarvkar, Vitthal, T.1 aKontham, Ravindar1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/aminoacylase-efficiently-hydrolyses-fatty-acid-amino-acid-conjugates-helicoverpa-armigera02343nas a2200217 4500008004100000022001400041245012900055210006900184260000800253520157400261653001001835653001101845653001001856653002001866653002401886653002301910100002001933700002201953700002001975856013001995 2024 eng d a1568-776700aAroma Alchemy: uridine diphosphate-dependent glycosyltransferases mediated regulation of fruit aroma and flavor biosynthesis0 aAroma Alchemy uridine diphosphatedependent glycosyltransferases  cDEC3 a

The aroma compounds contribute to fruits flavor, taste, and nutritional value. These compounds include various chemical classes such as terpenoids, lactones, ketones, esters, acids, alcohols, and their derivatives. Uridine diphosphate-dependent glycosyltransferases (UGTs) modify these compounds by covalently adding one or multiple sugar molecules. This glycosylation process converts volatile, unstable, and hydrophobic aroma compounds into hydrophilic, stable, and slow-releasing reservoirs of fruit flavor. The diversity and spatio-temporal expression patterns of UGTs play a crucial role in forming a wide range of glycosylated aroma compounds. This review focuses on aroma-related compounds in both free and glycosylated-bound forms found in commercial vital fruits. We discuss various fruit-specific UGTs and their role in the glycosylation of aroma compounds. Based on structural and functional information on UGTs, we have assessed sugar donor specificity and the residues responsible for the same. Moreover, phylogenetic analysis of characterized UGTs provides insights into their substrate preferences. We also surveyed the expression dynamics of UGTs during fruit ripening, as the switching between aglycon and glycosylated-bound forms of aroma compounds significantly impacts fruit quality. Potential applications of UGTs in the food and fragrance industry have been discussed. The information reviewed could be beneficial for developing novel methods for flavor manipulation of commercially important glycosides derived from natural resources.

10aAroma10aFlavor10aFruit10aGlycoconjugates10aGlycosyltransferase10aVolatile compounds1 aVare, Tejas, B.1 aJoshi, Rakesh, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/aroma-alchemy-uridine-diphosphate-dependent-glycosyltransferases-mediated-regulation-fruit01182nas a2200169 4500008004100000245014600041210006900187260000900256490000700265520050900272100002400781700002000805700001800825700002000843700002200863856012700885 2024 eng d00aArticle chitinase inhibition induces transcriptional dysregulation altering ecdysteroid-mediated control of spodoptera frugiperda development0 aArticle chitinase inhibition induces transcriptional dysregulati cMAR 0 v273 a

Chitinases and ecdysteroid hormones are vital for insect development. Crosstalk between chitin and ecdysteroid metabolism regulation is enigmatic. Here, we examined chitinase inhibition effect on Spodopsilencing and overexpression resulted in ecdysone receptor deregulation. Transcription factors, like Neverland, and other ecdysteroid biosynthesis genes might lead to their upregulation in berberine-fed chitinase activity's impact on ecdysone biosynthesis and its transcriptional crosstalk.

1 aBarbole, Ranjit, S.1 aSharma, Shivani1 aPatil, Yogita1 aGiri, Ashok, P.1 aJoshi, Rakesh, S. uhttps://library.ncl.res.in/content/article-chitinase-inhibition-induces-transcriptional-dysregulation-altering-ecdysteroid02460nas a2200241 4500008004100000022001400041245014600055210006900201260000900270300001100279490000800290520157900298653001901877653002901896653002301925653003201948653001401980653002701994100002402021700002202045700002002067856013102087 2024 eng d a0006-291X00aEngineering inhibitory repeat domains of Pin-II type proteinase inhibitors indicate their high structural-functional tolerance to mutagenesis0 aEngineering inhibitory repeat domains of PinII type proteinase i cNOV  a1508080 v7353 a

Plant proteinase inhibitors (PIs) are critical in defending against biotic stress. Most PIs contain an inhibitory repeat domain (IRD), which serves as the functional component, displaying a high degree of sequence and structural conservation. In this study, we examined the structural and functional resilience of IRDs using a combination of computational modeling and experimental validation. We have taken an evolution-based approach to enhance the PIs effectiveness of two previously identified Capsicum annuum IRDs, IRD4 and IRD10. Through in silico site-saturation mutagenesis of IRD4 and IRD10, we identified key sites associated with enhanced PI activity for targeted mutagenesis. Binding energy predictions for a mutant IRD library, tested against target proteases, suggested that positions R11 and N32 in IRD4 and N32 and H33 in IRD10 were promising candidates for further modification to improve inhibitory potential. Subsequent experimental validation revealed that the mutant proteins IRD4_R11K and IRD4_N32S exhibited stronger chymotrypsin inhibition than the wild-type (WT) IRD4. Similarly, the mutants IRD10_N32S and IRD10_H33 N demonstrated improved trypsin inhibition relative to the WT IRD10. These findings indicate that engineered IRD variants can tolerate structural changes while maintaining or enhancing their inhibitory activity against target proteases. Overall, this study demonstrates the potential of engineering PIs to increase their structural and functional resilience, offering new opportunities for biotechnological applications.

10aCapsicum annum10aInhibitory repeat domain10amolecular modeling10aplant proteinase inhibitors10aProteases10aSaturation mutagenesis1 aBarbole, Ranjit, S.1 aJoshi, Rakesh, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/engineering-inhibitory-repeat-domains-pin-ii-type-proteinase-inhibitors-indicate-their-high02333nas a2200289 4500008004100000022001400041245007800055210006900133260000800202300001100210490000800221520142300229653001401652653002401666653002401690653002201714653003301736653002201769100001801791700001601809700002401825700002201849700002001871700002501891700002001916856010701936 2024 eng d a0141-813000aMulti-dimensional impact of captopril modification on human serum albumin0 aMultidimensional impact of captopril modification on human serum cAUG a1332890 v2743 a

Captopril is a thiol drug, widely used for the management of hypertension and cardiovascular diseases. Reactive thiols are found to covalently modify the cysteines of plasma proteins and affect their structure and function. Human serum albumin (HSA) is prone to undergo modification by various low molecular weight compounds, including drugs. Cysteine34 (Cys34) in HSA has a free thiol group with antioxidant properties, considered to be the most redox-sensitive amino acid in plasma. Through mass-spectrometric analysis, we demonstrate for the first time that captopril forms a disulfide adduct at Cys34 residue and increases the protease susceptibility of HSA to trypsin. As evidenced by our biophysical and electron microscopy studies, HSA undergoes structural alteration, aggregation and morphological changes when treated with different captopril concentrations. Molecular dynamics studies further revealed the regions of secondary structural changes in HSA due to disulfide adduct formation by captopril at Cys34. It also elucidated the residues involved in the noncovalent interactions with captopril. It is envisaged that structural change in HSA may influence the efficacy of drug delivery as well as its own biological function. These findings may thus provide significant insights into the field of pharmacology intriguing further investigation into the effects of long-term captopril treatment.

10aCaptopril10aElectron microscopy10aHuman serum albumin10aMass spectrometry10aMolecular dynamic simulation10aSpectrophotometry1 aGhosh, Amrita1 aJani, Vinod1 aSonavane, Uddhavesh1 aNaphade, Anvi, N.1 aJoshi, Rajendra1 aKulkarni, Mahesh, J.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/multi-dimensional-impact-captopril-modification-human-serum-albumin03081nas a2200325 4500008004100000022001400041245012700055210006900182260000800251300001200259490000800271520196900279653002702248653004002275653002202315653002002337653001502357653001702372653002902389653002902418653001302447100002502460700002902485700003202514700002502546700002102571700002202592700002002614856012102634 2024 eng d a0960-741200aOcimum kilimandscharicum 4CL11 negatively regulates adventitious root development via accumulation of flavonoid glycosides0 aOcimum kilimandscharicum 4CL11 negatively regulates adventitious cJUL a176-1960 v1193 a

4-Coumarate-CoA Ligase (4CL) is an important enzyme in the phenylpropanoid biosynthesis pathway. Multiple 4CLs are identified in Ocimum species; however, their in planta functions remain enigmatic. In this study, we independently overexpressed three Ok4CL isoforms from Ocimum kilimandscharicum (Ok4CL7, -11, and -15) in Nicotiana benthamiana. Interestingly, Ok4CL11 overexpression (OE) caused a rootless or reduced root growth phenotype, whereas overexpression of Ok4CL15 produced normal adventitious root (AR) growth. Ok4CL11 overexpression in N. benthamiana resulted in upregulation of genes involved in flavonoid biosynthesis and associated glycosyltransferases accompanied by accumulation of specific flavonoid-glycosides (kaempferol-3-rhamnoside, kaempferol-3,7-O-bis-alpha-l-rhamnoside [K3,7R], and quercetin-3-O-rutinoside) that possibly reduced auxin levels in plants, and such effects were not seen for Ok4CL7 and -15. Docking analysis suggested that auxin transporters (PINs/LAXs) have higher binding affinity to these specific flavonoid-glycosides, and thus could disrupt auxin transport/signaling, which cumulatively resulted in a rootless phenotype. Reduced auxin levels, increased K3,7R in the middle and basal stem sections, and grafting experiments (intra and inter-species) indicated a disruption of auxin transport by K3,7R and its negative effect on AR development. Supplementation of flavonoids and the specific glycosides accumulated by Ok4CL11-OE to the wild-type N. benthamiana explants delayed the AR emergence and also inhibited AR growth. While overexpression of all three Ok4CLs increased lignin accumulation, flavonoids, and their specific glycosides were accumulated only in Ok4CL11-OE lines. In summary, our study reveals unique indirect function of Ok4CL11 to increase specific flavonoids and their glycosides, which are negative regulators of root growth, likely involved in inhibition of auxin transport and signaling.

10a4-Coumarate-CoA ligase10a7-O-bis-alpha-l-rhamnoside10aadventitious root10aauxin transport10aFlavonoids10akaempferol-310aOcimum kilimandscharicum10aquercetin-3-O-rutinoside10arootless1 aLavhale, Santosh, G.1 aKondhare, Kirtikumar, R.1 aSinthadurai, Veenothini, S.1 aBarvkar, Vitthal, T.1 aKale, Rutuja, S.1 aJoshi, Rakesh, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/ocimum-kilimandscharicum-4cl11-negatively-regulates-adventitious-root-development02451nas a2200253 4500008004100000022001400041245014700055210006900202260000900271300000800280490000700288520156100295653002801856653002701884653001001911653002501921653001701946100001801963700002601981700002102007700002402028700002002052856012502072 2025 eng d a2190-572X00aComprehensive tissue specific analysis of antioxidant potential and metabolite profiling of a vulnerable therapeutic herb Ceropegia sahyadrica0 aComprehensive tissue specific analysis of antioxidant potential  cNOV  a4340 v153 a

This study investigated tissue-culture of an endangered medicinal herb, Ceropegia sahyadrica, along with tissue-specific variation in antioxidant activity and GC-MS based phytochemical profiling using wild and in vitro explants. In vitro leaf explants exhibited callus induction rate of 100% in Murashige and Skoog media containing 6-benzylaminopurine (4 mu M) and 2,4-dichlorophenoxyacetic acid (2 mu M) with 5% sucrose, whereas in vitro root explant achieved 66.7% with the combination of 2 mu M 6-benzylaminopurine and 1 mu M 2,4-dichlorophenoxyacetic acid in Murashige and Skoog medium. Microtuberization was most pronounced in in vitro root explants with a maximum response of 73.3% in 1 mu M indole-3-acetic acid, 2 mu M 6-benzylaminopurine and 5% sucrose. Scanning electron microscopy imaging revealed distinct cellular organization among tissues. Further, antioxidant potential assessed through 1,1-diphenyl-2-picrylhydrazyl radical scavenging assays and estimated total phenolic content showed metabolic variation and commonality between in vitro and wild tissues. Additionally, phytochemical profiling by gas chromatography-mass spectrometry identified 201 compounds, including alkaloids, fatty acid methyl esters, flavonoids, phytosterols and other bioactive molecules with alterations in metabolite profiles between in vitro and wild tissues. These findings demonstrated that explant origin and culture condition might significantly influence metabolic responses for conservation and ensures a reliable source of bioactive compound.

10aCeropegia- Conservation10aEndemic medicinal herb10aGC-MS10aMetabolite profiling10aTuberization1 aPandey, Meena1 aDholakia, Bhushan, B.1 aSaraf, Purva, S.1 aPunekar, Sachin, A.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/comprehensive-tissue-specific-analysis-antioxidant-potential-and-metabolite-profiling03688nas a2200145 4500008004100000245010300041210006900144260000800213300001100221490000700232520314100239100001703380700002003397856012503417 2025 eng d00aDeciphering the role of metabolites and phytohormones in plant resilience to drought and herbivory0 aDeciphering the role of metabolites and phytohormones in plant r cMAR a1007370 v153 a

Climate change is expected to result in increased variability in precipitation and more frequent outbreaks of insect pests. Thus, it is important to understand how plant-environment interactions are affected by both abiotic and biotic stresses. Water is essential for plant growth, development and interactions with other organisms, including insects. This review synthesizes current studies on the impact of drought and herbivore defense mechanisms and associated metabolic changes in plants. Severe drought can enhance plant tolerance to herbivores by promoting escape strategies whereas mild or intermittent drought may benefit insects by increasing nutrient availability. We discuss how plants adjust their metabolism to mitigate the effects of combined stresses. We further highlight the role of hormonal signaling pathways, such as abscisic acidjasmonic acid, salicylic acid and ethylene in coordinating plant responses. Research on metabolic changes accompanying hormonal crosstalk involved in managing multiple stresses is still emerging. The available evidence suggests that the outcome of drought and herbivory varies depending on factors such as stress intensity, duration, plant-herbivore species, and insect-feeding guilds. We propose open questions and anticipate further advances in molecular understanding of plant resilience to combined stresses such as drought and herbivory in the near future.

1 aSahu, Monika1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/deciphering-role-metabolites-and-phytohormones-plant-resilience-drought-and-herbivory02816nas a2200265 4500008004100000022001400041245008900055210006900144260000800213300001100221490000700232520198500239653002002224653001202244653002002256653001502276653002402291653001302315653001802328100002302346700002502369700002002394700002402414856011202438 2025 eng d a1744-117X00aHigh levels of sinigrin trigger synthesis of fatty acids in Plutella xylostella (L.)0 aHigh levels of sinigrin trigger synthesis of fatty acids in Plut cJUN a1014240 v543 a

Diamondback moth (Lepidoptera: Plutellidae; Plutella xylostella L.) is a specialist insect of the Brassicaceae family, damaging economically important crops, such as cabbage and cauliflower. Glucosinolates, also known as `mustard oil bombs' are present in all Brassicaceae members, of which sinigrin (allyl-glucosinolate or 2-propenylglucosinolate) is a major aliphatic compound. During herbivory, glucosinolates are converted to toxic isothiocyanates that deter insect pests. P. xylostella possesses glucosinolate sulfatases that desulfate them. Such a conversion renders them unfit for degradation to toxic products. Changes in the larval performance prompted us for RNA sequencing to understand probable adaptation mechanism under sinigrin stress. Differentially expressed genes were found to be related to larval cuticle proteins. Further, gene ontology and KEGG (Kyoto Encyclopedia of Genes and Genomes) analyses depict genes belonging to the categories, integral component of membrane, cellular processes and those involved in biosynthesis of fatty acids. Upregulation of cuticular genes viz. larval cuticle protein-17 (LCP-17), cuticular protein-19 (2CP-19) and ATP binding cassette transporter C7 (ABCC7), ABCC16 was validated by qRT-PCR. Liquid chromatography quadrupole time of flight mass spectrometry analysis of whole larvae feeding on sinigrin and their separated cuticle, depicted abundance of fatty acids. Changes in the topography of the larval cuticle were evident by scanning electron microscopy. Expression of PxABCH1 was corroborated to its role in the transport of cuticular lipids. Notably, molecular docking of PxABCH1 with cuticular fatty acids showed favorable binding interactions. To summarize, integrated transcriptomic and metabolomic analyses suggest that in response to a diet containing a high dose of sinigrin, P. xylostella re-programs metabolic pathways related to fatty acid biosynthesis that directly influence insect development.

10aABC transporter10aCuticle10aHost metabolite10aMetabolome10aPlutella xylostella10aSinigrin10aTranscriptome1 aBarve, Pranoti, R.1 aBarvkar, Vitthal, T.1 aGiri, Ashok, P.1 aKotkar, Hemlata, M. uhttps://library.ncl.res.in/content/high-levels-sinigrin-trigger-synthesis-fatty-acids-plutella-xylostella-l02001nas a2200253 4500008004100000022001400041245012600055210006900181260000800250300001100258490000800269520115500277653001401432653001201446653001601458653001601474653001401490653002601504100002401530700002201554700002901576700002001605856012201625 2025 eng d a0141-813000aMYB proteins: Versatile regulators of plant development, stress responses, and secondary metabolite biosynthetic pathways0 aMYB proteins Versatile regulators of plant development stress re cFEB a1385880 v2883 a

MYB proteins are ubiquitous in nature, regulating key aspects of plant growth and development. Although MYB proteins are known for regulating genes involved in secondary metabolite biosynthesis, particularly phenylpropanoids, their roles in terpenoid, glucosinolate, and alkaloid biosynthesis remain less understood. This review explores the structural and functional differences between activator and repressor MYB proteins along with their roles in plant growth, development, stress responses, and secondary metabolite production. MYB proteins serve as central hubs in protein-protein interaction networks that regulate expression of numerous genes involved in the adaptation of plants to varying environmental conditions. Thus, we also highlight key interacting partners of MYB proteins and their roles in these adaptation mechanisms. We further discuss the mechanisms regulating MYB proteins, including autoregulation, epigenetics, and post-transcriptional and post-translational modifications. Overall, we propose MYB proteins as versatile regulators for improving plant traits, stress responses, and secondary metabolite production.

10aActivator10adefense10aDevelopment10aMYB protein10aRepressor10aSecondary metabolites1 aBhatt, Preshita, A.1 aGurav, Tanuja, P.1 aKondhare, Kirtikumar, R.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/myb-proteins-versatile-regulators-plant-development-stress-responses-and-secondary02666nas a2200241 4500008004100000022001400041245009700055210006900152260000800221300001100229490000800240520185700248653002502105653002002130653002002150653002602170653001602196100002302212700002802235700002002263700001902283856012202302 2025 eng d a0960-741200aNavigating the challenges of engineering composite specialized metabolite pathways in plants0 aNavigating the challenges of engineering composite specialized m cMAR ae701000 v1213 a

Plants are a valuable source of diverse specialized metabolites with numerous applications. However, these compounds are often produced in limited quantities, particularly under unfavorable ecological conditions. To achieve sufficient levels of target metabolites, alternative strategies such as pathway engineering in heterologous systems like microbes (e.g., bacteria and fungi) or cell-free systems can be employed. Another approach is plant engineering, which aims to either enhance the native production in the original plant or reconstruct the target pathway in a model plant system. Although increasing metabolite production in the native plant is a promising strategy, these source plants are often exotic and pose significant challenges for genetic manipulation. Effective pathway engineering requires comprehensive prior knowledge of the genes and enzymes involved, as well as the precursor, intermediate, branching, and final metabolites. Thus, a thorough elucidation of the biosynthetic pathway is closely linked to successful metabolic engineering in host or model systems. In this review, we highlight recent advances in strategies for biosynthetic pathway elucidation and metabolic engineering. We focus on efforts to engineer complex, multi-step pathways that require the expression of at least eight genes for transient and three genes for stable transformation. Reports on the engineering of complex pathways in stably transformed plants remain relatively scarce. We discuss the major hurdles in pathway elucidation and strategies for overcoming them, followed by an overview of achievements, challenges, and solutions in pathway reconstitution through metabolic engineering. Recent advances including computer-based predictions offer valuable platforms for the sustainable production of specialized metabolites in plants.

10aBiosynthetic pathway10aEnzyme activity10aGene expression10aMetabolic engineering10aMetabolites1 aGharat, Sachin, A.1 aTamhane, Vaijayanti, A.1 aGiri, Ashok, P.1 aAharoni, Asaph uhttps://library.ncl.res.in/content/navigating-challenges-engineering-composite-specialized-metabolite-pathways-plants02356nas a2200265 4500008004100000022001400041245014500055210007000200260000800270300001100278490000800289520139600297653002901693653002701722653002201749653002401771653002601795100002701821700002301848700002001871700002201891700002001913700002201933856013501955 2025 eng d a0141-813000aNovel bifunctional inhibitor of protease and α-amylase from Clitorea ternatea restricts the growth and development in Spodoptera frugiperda0 aNovel bifunctional inhibitor of protease and αamylase from Clito cMAY a1411800 v3053 a

An inhibitor molecule capable of inhibiting a wide range of digestive enzymes without affecting endogenous enzymes is always desirable. We report characterization of CteTAI (M.W. 14 kDa), a bifunctional inhibitor (BFI) protein from the seeds of Clitoria ternatea capable of inhibiting trypsin and alpha-amylase. It retains trypsin inhibition activity up to 60 degrees C and alpha-amylase inhibition up to 40 degrees C. Trypsin inhibition is stable across pH 1-12, while alpha-amylase inhibition is stable between pH 3-7. CteTAI is a noncompetitive inhibitor of trypsin and an uncompetitive inhibitor of alpha-amylase. It selectively inhibits proteases and alpha-amylases from various sources, without affecting alpha-amylase from human saliva and Bacillus spp. Proteomic analysis identified CteTAI as a bifunctional inhibitor exhibiting 41 % similarity to a bifunctional inhibitor from Sesbania bispinosa. Feeding Spodoptera frugiperda larvae with CteTAI-infused diet impaired energy metabolism, resulting in undernourished larvae and malformed adults incapable of flight and mating. Key nutritional indices (RGR, RCR, %ECI, %FDI) were severely reduced, indicating that CteTAI disrupts growth and development by inhibiting multiple protease and alpha-amylase isoforms. Biochemical characterization of newly identified CteTAI suggests its potential application in crop protection.

10aalpha-Amylase inhibitors10aBifunctional inhibitor10aClitoria ternatea10aProtease inhibitors10aSpodoptera frugiperda1 aDongare, Pratiksha, M.1 aMadage, Varsha, A.1 aDeshpande, Neha1 aJoshi, Rakesh, S.1 aGiri, Ashok, P.1 aPawar, Pankaj, K. uhttps://library.ncl.res.in/content/novel-bifunctional-inhibitor-protease-and-%CE%B1-amylase-clitorea-ternatea-restricts-growth-and02916nas a2200313 4500008004100000022001400041245012200055210006900177260000800246300001100254490000900265520183400274653002602108653002602134653003202160653003002192653002002222653003502242100002202277700001902299700002502318700002002343700001802363700002502381700002002406700002202426700002402448856013002472 2026 eng d a0022-286000aNovel prodrug-inspired eugenol derivatives with enhanced bioavailability, anti-diabetic and anti-glycation efficacies0 aNovel prodruginspired eugenol derivatives with enhanced bioavail cAPR a1451750 v13573 a

In this study, we present the design, synthesis, and evaluation of novel eugenol analogs aimed to overcome its limited bioavailability due to insolubility in aqueous media. Thus, we re-engineered eugenol using prodruginspired structural modifications to improve pharmacokinetic properties. First, we structurally modified eugenol and synthesized its natural amino acid conjugates as esters and carbamates. These were prepared in NBoc protected, free amine, and HCl salt forms. These modifications are expected to improve the polarity and solubility of eugenol congeners in biological systems. They can also release the parent eugenol through enzymatic hydrolysis, enhancing its therapeutic potential. Next, we comprehensively screened for these derivatives through in silico studies followed by in vitro and in vivo assays. These include DPPH radical scavenging (IC50 range: 37.7 to 103.7 mu M), inhibition of (i) alpha-amylase (IC50 23.1 to 67.3 mu M), (ii) alpha-glucosidase (IC50 43.6 to 50.4 mu M), (iii) glycation (IC50 31.9 to 110.3 mu M) along with pharmacokinetic profiling and toxicity assessments. These experiments collectively demonstrated improved activity of eugenol analogs for several important parameters. Specifically, six analogs-epoxy eugenol (39), hydroxy eugenol (43), aspartate eugenol (26), isoleucinate eugenol (24), glutamate eugenol (37), and glutamate-salt eugenol (27) exhibited superior bioavailability, absorption, and distribution over to the parent compound eugenol. These analogs were found to be non-toxic and safe for oral administration. Overall, the study establishes a mechanistic and rational framework for modifying eugenol to overcome its inherent biopharmaceutical limitations, positioning them as promising candidates for treating diabetes and glycation-related conditions.

10aAmino acid conjugates10aAntidiabetic activity10aBioavailability enhancement10aDrug design and synthesis10aEugenol analogs10aIn silico and in vitro studies1 aKataria, Priyanka1 aVairale, Shiva1 aMote, Chandrashekhar1 aJoshi, Kaustubh1 aJoshi, Rakesh1 aKulkarni, Mahesh, J.1 aGiri, Ashok, P.1 aKontham, Ravindar1 aAgawane, Sachin, B. uhttps://library.ncl.res.in/content/novel-prodrug-inspired-eugenol-derivatives-enhanced-bioavailability-anti-diabetic-and-anti02660nas a2200205 4500008004100000022001400041245017000055210006900225260000800294300001600302490000700318520187100325100002002196700002302216700002002239700002802259700002002287700002002307856012702327 2026 eng d a2470-134300aPlant protease inhibitors induced oxidative and nutritional stress in helicoverpa armigera unveils multifaceted survival strategies: a lipidomic and metabolomic view0 aPlant protease inhibitors induced oxidative and nutritional stre cMAR a18664-186820 v113 a

Plant protease inhibitors impede insect growth and development by targeting digestive proteases. In response, insects adapt by modulating protease expression; however, their survival may not rely solely on digestive plasticity and likely involves broader physiological adjustments. To investigate this, we performed an integrated lipidomic and metabolomic analysis of Helicoverpa armigera larvae fed on the recombinant Capsicum annuum protease inhibitor (rCanPI-7), a four-domain inhibitor with potent activity against insect trypsins and chymotrypsins. The results revealed extensive metabolic reprogramming in H. armigera under rCanPI-7-induced stress, prominently affecting amino acid, glutathione, and pyrimidine metabolic pathways with distinct shifts in alanine, arginine, histidine, and branched-chain amino acid metabolism. Suppression of glycolysis and the tricarboxylic acid cycle indicated reduced primary energy metabolism, while oxidative stress was evident from glutathione depletion, lipid peroxidation, and ceramide accumulation, which are hallmarks of mitochondrial dysfunction. To offset this metabolic imbalance, H. armigera mobilized triglycerides and amino acids as alternative energy sources. Reinforcing antioxidant defenses, membrane remodeling, and activating apoptotic and neuromodulatory pathways plausibly aided in restoring cellular homeostasis. Collectively, these findings reveal a tightly coordinated physiological tug of war in H. armigera, where the detrimental effects of rCanPI-7-induced oxidative and nutritional stress were countered through compensatory metabolic, structural, and signaling adjustments. This study provides the first lipidomic insight into this pest species, offering a deeper understanding of its biochemical resilience and potential metabolic vulnerabilities for targeted biopesticide strategies.

1 aDewangan, Veena1 aGupta, Sonu, Kumar1 aDeshpande, Neha1 aLomate, Purushottam, R.1 aKumar, Yashwant1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/plant-protease-inhibitors-induced-oxidative-and-nutritional-stress-helicoverpa-armigera02499nas a2200229 4500008004100000022001400041245003300055210003300088260000800121300001100129490000800140520189500148653001002043653001402053653002402067653002602091653001302117100002902130700002502159700002002184856006502204 2026 eng d a0168-945200aRootless survivors in plants0 aRootless survivors in plants cMAR a1129510 v3643 a

The root system provides anchorage, uptakes of nutrients and water, and forms different associations within soil environments that govern plant fitness, crop performance, and yield. Auxin controls almost all aspects of root development. Both shoot- and root-derived auxins contribute to formation of polar auxin transport, which is crucial for establishing and maintaining normal root architecture. The coordinated activities of auxin influx and efflux carriers establish necessary polar auxin transport. A variety of natural metabolites and synthetic compounds are shown to interfere with auxin metabolism, transporters and signaling pathways having a negative impact on root growth. In this review, we highlight the reports demonstrating the observance of rootless phenotypes in plants and associated molecular mechanisms. Rootless phenotypes can be produced under in vitro culture conditions by modulation of phytohormone combinations (especially auxin and cytokinin), and supplementation of naturally-occurring flavonoids and their glycosides or synthetic auxin transport inhibitors (1-Nnaphthylphthalamic acid and 2,3,5-triiodobenzoic acid) or under in vivo conditions by modulation of several genes directly or indirectly associated with auxin biology. Further, we describe the crosstalk of naturallyoccurring flavonoids (e.g. kaempferol, quercetin), their glycosides, and other metabolites (e.g. azelaic acid, cis-cinnamic acid) with auxin transporters, their mobile nature, and influence on root development. Moreover, we provide evolutionary perspective on the auxin and flavonoid pathways and their possible roles in naturally rootless plants. We also emphasize the importance of rootless or reduced root growth phenotypes in modern agriculture, and the pressing needs to utilize naturally occurring auxin transport inhibitors for industrial and research applications.

10aAuxin10aFlavonoid10aFlavonoid-glycoside10aPolar auxin transport10arootless1 aKondhare, Kirtikumar, R.1 aLavhale, Santosh, G.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/rootless-survivors-plants