02875nas a2200349 4500008004100000022001400041245015400055210006900209260011000278300001200388490000600400520159700406653002302003653002502026653002902051653002402080100002002104700001702124700001802141700001802159700001802177700001702195700001802212700002402230700001802254700002102272700003002293700002702323700002202350700002402372856012902396 2008 eng d a0972-584900aDirect shoot organogenesis and plant regeneration from hypocotyl explants in selected genotypes of Leucaena leucocephala - a leguminous pulpwood tree0 aDirect shoot organogenesis and plant regeneration from hypocotyl aDR K S KRISHNAN MARG, PUSA CAMPUS, NEW DELHI 110 012, INDIAbNATL INST SCIENCE COMMUNICATION-NISCAIRcJUL a388-3930 v73 a

An efficient in vitro plant regeneration system in subabul (Leucaena leucocephala), a leguminous pulp wood tree species, was established. The induction of shoots was achieved from selected elite clones of subabul K-8, K-636 and also wild type on MS medium supplemented with 2% sucrose and different concentrations (0.88 to 24.6 mu M) of plant growth regulators (BA, Kn, 2iP & TDZ). The best medium for shoot regeneration was MS with 22.2 mu M BA (5 shoots per explant), followed by 22.7 mu M TDZ (4.6 shoots per explant). Addition of putriscine (9.3 mu M) to MS medium containing 22.2 mu M BA enhanced the number of multiple shoots to 7-8 but not the frequency of response. Shoot initials (measuring 1 cm) when separated and transferred on to MS medium containing 1.4 mu M GA(3) elongated to 2-5 cm in 15.20 d with 80% frequency. The per cent frequency of shoot differentiation was almost identical in the genotypes K-8 and K-636 but it differed significantly from the wild type. Leaf yellowing and abscission in all the genotypes was curtailed by supplementing the medium with 685 mu M glutamine or 540 mu M adenine. The excised shoots were transferred to root regeneration media containing 2.46 and 4.98 mu M IBA or 2.6 and 5.3 mu M NAA. Root regeneration was noticed with 100% frequency in all the three genotypes in presence of IBA or NAA. Plantlets were transferred successfully to the pots with 70% survival rate with no visible morphological variations. The protocol can be utilized for mass propagation and genetic transformation studies of this important pulpwood species.

10ahypocotyl explants10aLeucaena lucocephala10apulpwood leguminous tree10ashoot organogenesis1 aSirisha, V., L.1 aPrashant, S.1 aRanadheer, D.1 aRamprasad, P.1 aShaik, N., M.1 aArha, Manish1 aGupta, S., K.1 aSrivastava, Sarneer1 aYadav, A., K.1 aKulkarni, P., S.1 aAbhilash, Othalathara, U.1 aKhan, Bashir, Mohammad1 aRawal, Shuban, K.1 aKishor, P., B. Kavi uhttps://library.ncl.res.in/content/direct-shoot-organogenesis-and-plant-regeneration-hypocotyl-explants-selected-genotypes-003067nas a2200301 4500008004100000022001400041245019700055210006900252260005700321300001200378490000700390520193100397653004002328653001202368653001402380653002802394100003002422700001702452700001802469700002102487700002002508700001702528700002102545700002702566700001802593700002402611856013002635 2009 eng d a1610-294000aPhylogenetic analysis, homology modelling, molecular dynamics and docking studies of caffeoyl-CoA-O- methyl transferase (CCoAOMT 1 and 2) isoforms isolated from subabul (Leucaena leucocephala)0 aPhylogenetic analysis homology modelling molecular dynamics and  a233 SPRING ST, NEW YORK, NY 10013 USAbSPRINGERcFEB a203-2210 v153 a

Caffeoyl coenzyme A O-methyltransferase (CCoAOMT) is an important enzyme that participates in lignin biosynthesis especially in the formation of cell wall ferulic esters of plants. It plays a pivotal role in the methylation of the 3-hydroxyl group of caffeoyl CoA. Two cDNA clones that code CCoAOMT were isolated earlier from subabul and in the present study; 3D models of CCoAOMT1 and CCoAOMT2 enzymes were built using the MODELLER7v7 software to find out the substrate binding sites. These two proteins differed only in two amino acids and may have little or no functional redundancy. Refined models of the proteins were obtained after energy minimization and molecular dynamics in a solvated water layer. The models were further assessed by PROCHECK, WHATCHECK, Verify_3D and ERRAT programs and the results indicated that these models are reliable for further active site and docking analysis. The refined models showed that the two proteins have 9 and 10 alpha-helices, 6 and 7 beta-sheets respectively. The models were used for docking the substrates CoA, SAM, SAH, caffeoyl CoA, feruloyl CoA, 5-hydroxy feruloyl CoA and sinapyl CoA which showed that CoA and caffeoyl CoA are binding with high affinity with the enzymes in the presence and absence of SAM. It appears therefore that caffeoyl CoA is the substrate for both the isoenzymes. The results also indicated that CoA and caffeoyl CoA are binding with higher affinity to CCoAOMT2 than CCoAOMT1. Therefore, CCoAOMT2 conformation is thought to be the active form that exists in subabul. Docking studies indicated that conserved active site residues Met58, Thr60, Val63, Glu82, Gly84, Ser90, Asp160, Asp162, Thr169, Asn191 and Arg203 in CCoAOMT1 and CCoAOMT2 enzymes create the positive charge to balance the negatively charged caffeoyl CoA and play an important role in maintaining a functional conformation and are directly involved in donor-substrate binding.

10aCaffeoyl-CoA 3-O-methyl transferase10adocking10aModelling10aS-adenosyl homocysteine1 aPagadala, Nataraj, Sekhar1 aArha, Manish1 aReddy, P., S.1 aKumar, Ranadheer1 aSirisha, V., L.1 aPrashant, S.1 aReddy, Janardhan1 aKhan, Bashir, Mohammad1 aRawal, S., K.1 aKishor, P., B. Kavi uhttps://library.ncl.res.in/content/phylogenetic-analysis-homology-modelling-molecular-dynamics-and-docking-studies-caffeoyl-002397nas a2200241 4500008004100000022001400041245012000055210006900175260010900244300001200353490000800365520145200373653002301825653001801848653002901866100002001895700001701915700002401932700002801956700002401984700002002008856012702028 2010 eng d a0378-874100aCytotoxic activity of 9,10-dihydro-2,5-dimethoxyphenanthrene-1,7-diol from Eulophia nuda against human cancer cells0 aCytotoxic activity of 910dihydro25dimethoxyphenanthrene17diol fr aELSEVIER HOUSE, BROOKVALE PLAZA, EAST PARK SHANNON, CO, CLARE, 00000, IRELANDbELSEVIER IRELAND LTDcMAR a251-2530 v1283 a

Ethnopharmacological relevance: Eulophia nuda L. (Orchidaceae) is a medicinally important terrestrial orchid used for the treatment of tumours and various health problems by the local healers throughout the Western Ghats region in Maharashtra (India). Aim of the study: To isolate the active molecule from Eulophia nuda and to study its cytotoxic potential against human cancer cells. Materials and methods: The crude methanolic extract of Eulophia nuda tubers was fractionated by stepwise gradient of the solvents chloroform-methanol to isolate the pure compound. Isolated pure compound was assessed for its cytotoxic potential against human breast cancer cell lines, MCF-7 and MDA-MB-231 using MTT assay. Structure elucidation of the isolated active compound was carried out by extensive spectroscopic analysis including (1)H NMR, (13)C NMR, NOESY, COSY, LC-MS and IR. Results: The isolated active molecule was identified as phenanthrene derivative 9,10-dihydro-2,5-dimethoxyphenanthrene-1,7-diol. This compound showed good antiproliferative activity against human breast cancer cell lines MCF-7 (91%) and MDA-MB-231 (85%) at 1000 mu g/ml concentration. Conclusion: 9,10-Dihydro-2,5-dimethoxyphenanthrene-1,7-diol from Eulophia nuda tubers showed good growth suppressive effect against human cancer cell lines MCF-7 and MDA-MB-231 making it a potential biomolecule against human cancer. (C) 2009 Elsevier Ireland Ltd. All rights reserved.

10aCytotoxic activity10aEulophia nuda10aPhenanthrene derivatives1 aShriram, Varsha1 aKumar, Vinay1 aKishor, P., B. Kavi1 aSuryawanshi, Sharad, B.1 aUpadhyay, Ankur, K.1 aBhat, Manoj, K. uhttps://library.ncl.res.in/content/cytotoxic-activity-910-dihydro-25-dimethoxyphenanthrene-17-diol-eulophia-nuda-against-002593nas a2200265 4500008004100000022001400041245014100055210006900196260010300265300001200368490000700380520154900387653002801936653002401964653002601988653002402014100002302038700002502061700001702086700002702103700002202130700002402152700002702176856012402203 2011 eng d a0981-942800aExpression analysis of cinnamoyl-CoA reductase (CCR) gene in developing seedlings of Leucaena leucocephala: A pulp yielding tree species0 aExpression analysis of cinnamoylCoA reductase CCR gene in develo a23 RUE LINOIS, 75724 PARIS, FRANCEbELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIERcFEB a138-1450 v493 a

Removal of lignin is a major hurdle for obtaining good quality pulp. Leucaena leucocephala (subabul) is extensively used in paper industry in India; therefore, as a first step to generate transgenic plants with low lignin content, cDNA and genomic clones of CCR gene were isolated and characterized. The cDNA encoding CCR (EC 1.2.1.44) was designated as LI-CCR; the sequence analysis revealed an Open Reading Frame (ORF) of 1005 bp. Phylogenetic analysis showed that LI-CCR sequence is highly homologous to CCRs from other dicot plants. The 2992 bp genomic clone of Leucaena CCR consists of 5 exons and 4 introns. The haploid genome of L leucocephala contains two copies as revealed by DNA blot hybridization. LI-CCR gene was over-expressed in Escherichia coli, which showed a molecular mass of approximately 38 kDa. Protein blot analysis revealed that LI-CCR protein is expressed at higher levels in root and in stem, but undetectable in leaf tissues. Expression of CCR gene in Leucaena increased up to 15 d in case of roots and stem as revealed by QRT-PcR studies in 0-15 d old seedlings. ELISA based studies of extractable CCR protein corroborated with QRT-PCR data. CCR protein was immuno-cytolocalized around xylem tissue. Lignin estimation and expression studies of 5,10 and 15 d old stem and root suggest that CCR expression correlates with quantity of lignin produced, which makes it a good target for antisense down regulation for producing designer species for paper industry. (C) 2010 Elsevier Masson SAS. All rights reserved.

10aCinnamoyl-CoA reductase10aDeveloping seedling10aLeucaena leucocephala10aLignin biosynthesis1 aSrivastava, Sameer1 aGupta, Ranadheer, K.1 aArha, Manish1 aVishwakarma, Rishi, K.1 aRawal, Shuban, K.1 aKishor, P., B. Kavi1 aKhan, Bashir, Mohammad uhttps://library.ncl.res.in/content/expression-analysis-cinnamoyl-coa-reductase-ccr-gene-developing-seedlings-leucaena-002944nas a2200325 4500008004100000022001400041245017600055210006900231260007400300300001200374490000700386520175900393653003502152653002702187653003602214653001202250100002002262700001702282700002102299700001502320700001502335700001702350700002002367700001902387700001902406700002002425700002702445700002402472856012202496 2012 eng d a0167-690300aCloning, characterization and impact of up- and down-regulating subabul cinnamyl alcohol dehydrogenase (CAD) gene on plant growth and lignin profiles in transgenic tobacco0 aCloning characterization and impact of up and downregulating sub aVAN GODEWIJCKSTRAAT 30, 3311 GZ DORDRECHT, NETHERLANDSbSPRINGERcAPR a239-2530 v663 a

Both cDNA including 5'UTR and 3'UTR and genomic clones of cinnamyl alcohol dehydrogenase (CAD) were isolated and characterized from a pulp-yielding leguminous tree Leucaena leucocephala (LlCAD1). The deduced amino acid sequence shared high identity with orthologous sequences of Acacia mangium x Acacia auriculiformis (83%), Medicago sativa (83%), Nicotiana tabaccum (83%) and Aralia cordata (81%). Full length cDNA contained 78 bases of 5'UTR and 283 bases of 3'UTR, while the genomic clone contained 5 exons and 4 introns. Western blot analysis revealed elevated expression of LlCAD1 in seedling roots and shoots compared to leaves. Sense and antisense CAD tobacco transgenics showed increased and reduced CAD activity accompanied by a change in monomeric lignin composition. Histochemical staining of lignin in down-regulated plants suggested an increase in aldehyde units and a decrease in S/G ratio. Down-regulation of CAD resulted in accumulation of syringic, ferulic, p-coumaric and sinapic acids compared to untransformed controls. These observations were validated by anatomical studies of down-regulated transgenic stems which showed thin walled, elongated phloem and xylem fibres, accompanied by a reduction in the density of vessel elements and amount of secondary xylem when compared to untransformed plants. Furthermore, Klason lignin analysis of CAD antisense transgenics showed 7-32% reduced lignin and normal phenotype as compared to untransformed plants. Such a reduction was not noticed in up-regulated transgenics. These results demonstrate a unique opportunity to explore the significant role that down-regulation of CAD gene plays in reducing lignin content thereby offering potential benefits to the pulp and paper industry.

10aCinnamyl alcohol dehydrogenase10aLignin down-regulation10aSubabul (Leucaena leucocephala)10aTobacco1 aSirisha, V., L.1 aPrashant, S.1 aKumar, Ranadheer1 aPramod, S.1 aJalaja, N.1 aKumari, Hima1 aRao, Maheshwari1 aRao, Nageswara1 aMishra, Preeti1 aKarumanchi, Rao1 aKhan, Bashir, Mohammad1 aKishor, P., B. Kavi uhttps://library.ncl.res.in/content/cloning-characterization-and-impact-and-down-regulating-subabul-cinnamyl-alcohol-002422nas a2200301 4500008004100000022001400041245011200055210006900167260000800236300001100244490000800255520141300263653002501676653001501701653002201716653001501738653002701753653002101780100002401801700002701825700002401852700002001876700002701896700002401923700002401947700002301971856012601994 2024 eng d a0926-669000aUnlocking the genetic and biotechnological potential of Gloriosa superba to enhance its alkaloid production0 aUnlocking the genetic and biotechnological potential of Gloriosa cMAY a1181440 v2113 a

Gloriosa superba is a medicinally important plant that produces the pharmaceutically precious colchicine molecule. Colchicine is being used for treating gout, for enhancing the production of interleukin-8 (inhibits human pancreatic cancer) and many other human ailments. Since its seeds and rhizomes (tuberous roots) are the chief sources of colchicine, pharmaceutical industries exploit the natural sources, thus demolishing its habitat. Chemo- and genetic diversity among the G. superba populations exist, but has not been exploited profitably. Scope exists for generating mutants using the chemical mutagens and also via breeding methods to generate superior varieties for commercial cultivation. Innovative methods for breaking seed dormancy and superior colchicine extraction, besides biotechnological interventions are imperative to save the biodiversity and to produce colchicine on a large scale. Biorhizomes produced in vitro hold great promise for biomanufacturing colchicine in bioreactors commercially. Since the biosynthetic pathway of colchicine is now known, isolation of the pathway genes and their heterologous overexpression or the application of synthetic biology methods needs to be explored in future for industrial manufacturing of colchicine. Advances made in the recent past in these fields of colchicine production from G. superba are critically evaluated in this review.

10aBiosynthetic pathway10aColchicine10aGenetic diversity10aGlory lily10aMetabolite elicitation10aOmics approaches1 aMahajan, Yogesh, A.1 aShinde, Balkrishna, A.1 aShirke, Harshad, A.1 aGandra, Jawahar1 aSuravajhala, Prashanth1 aKishor, P., B. Kavi1 aKadoo, Narendra, Y.1 aNikam, Tukaram, D. uhttps://library.ncl.res.in/content/unlocking-genetic-and-biotechnological-potential-gloriosa-superba-enhance-its-alkaloid