02417nas a2200265 4500008004100000022001400041245011200055210006900167260007300236300001200309490000900321520145600330653001901786653001901805653002501824653002201849653002501871100002001896700002801916700002001944700001801964700002401982700002102006856012402027 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. uhttp://library.ncl.res.in/content/vivo-and-vitro-effect-capsicum-annum-proteinase-inhibitors-helicoverpa-armigera-gut-003016nas a2200277 4500008004100000022001400041245013800055210006900193260007700262300001400339490000700353520197800360653001002338653003002348653003402378653001102412653002102423653002902444100001902473700002802492700002102520700002502541700002102566700002002587856013102607 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. uhttp://library.ncl.res.in/content/interaction-recombinant-canpis-helicoverpa-armigera-gut-proteases-reveals-their-processing-002474nas a2200253 4500008004100000022001400041245014800055210006900203260010600272300001200378490000700390520147200397653002001869653002501889653001101914653002901925653002401954100002801978700002202006700002102028700002102049700002002070856013002090 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. uhttp://library.ncl.res.in/content/identification-and-expression-profiling-helicoverpa-armigera-micrornas-and-their-possible-001936nas a2200205 4500008004100000245014900041210006900190260000800259300001200267490000700279520115700286100002301443700002401466700002301490700002801513700002201541700002001563700002101583856012601604 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. uhttp://library.ncl.res.in/content/improved-tolerance-against-helicoverpa-armigera-transgenic-tomato-over-expressing-multi02170nas a2200193 4500008004100000245015500041210006900196260000800265300001000273490000800283520141800291100002601709700002501735700002201760700002301782700002001805700002101825856013001846 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. uhttp://library.ncl.res.in/content/isolation-and-characterization-9-lipoxygenase-and-epoxide-hydrolase-2-genes-insight-lactone02196nas a2200229 4500008004100000245018300041210006900224260000800293300001400301490000700315520133000322100002801652700002001680700002201700700002001722700002301742700001301765700001801778700002101796700002001817856012901837 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. uhttp://library.ncl.res.in/content/integrated-transcriptomic-and-proteomic-analyses-suggest-participation-endogenous-protease02983nas a2200301 4500008004100000022001400041245008000055210006900135260000900204300000700213490000800220520207700228653001002305653001802315653001202333653001602345653001702361653002802378653001002406100001602416700002602432700002702458700002202485700002802507700002102535700002002556856010502576 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. uhttp://library.ncl.res.in/content/influence-domestication-specialized-metabolic-pathways-fruit-crops