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<p>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 &amp; 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.</p>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<p>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.</p>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-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<p>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.</p>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-0