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-002067nas a2200229   4500008004100000022001400041245009900055210006900154260005700223300001400280490000700294520120800301653003101509653003001540653001701570653002501587100002401612700002401636700002701660700002201687856012801709       2008                        eng d  a0959-399300aIsolation and heterologous expression of PHA synthesising genes from Bacillus thuringiensis R10 aIsolation and heterologous expression of PHA synthesising genes   a233 SPRING ST, NEW YORK, NY 10013 USAbSPRINGERcSEP  a1769-17740 v243 a<p>The polyhydroxyalkanoate biosynthesis gene locus from Bacillus thuringiensis R1 was isolated, cloned and analyzed at the molecular level. We found that a similar to 5 kb SacI-ClaI digested fragment of genomic DNA from B. thuringiensis R1 encoding the PHA synthesising genes, conferred PHA producing ability to E. coli. The fragment was sequenced and found to be of 4787 bp with five open reading frames. Sequence alignment with closely related species of Bacillus in the existing database revealed that the ORFs correspond to phaP, phaQ, phaR, phaB and phaC genes. However, E. coli harboring phaP, phaQ, phaR, phaB and phaC locus produced very low PHA. Furthermore, complementation of the locus with phaA from Ralstonia eutropha increased the PHA production in the recombinant E. coli from 3.0% to 24% of cell dry mass. The putative promoter regions and ribosome binding sites were identified for each of the gene. Conserved domains for PHA synthase and aceto-acetyl-coA reductase were also identified. We hence conclude that the PHA operon of Bacillus thuringiensis R1 consists of phaP, phaQ, phaR, phaB, phaC and complementation of the same with phaA is accountable for its high PHA production.</p>10aaceto-acetyl-coA reductase10aBacillus thuringiensis R110aPHA synthase10apolyhydroxyalkanoate1 aDesetty, Rohini, D.1 aMahajan, Vineet, S.1 aKhan, Bashir, Mohammad1 aRawal, Shuban, K.  uhttps://library.ncl.res.in/content/isolation-and-heterologous-expression-pha-synthesising-genes-bacillus-thuringiensis-r1-002593nas 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-002628nas a2200265   4500008004100000022001400041245019800055210006900253260007400322300001400396490000700410520155800417653003201975653002202007653002602029653002202055100002002077700001802097700001802115700002402133700003202157700002202189700002702211856012402238       2013                        eng d  a0301-485100aFunctional characterization, homology modeling and docking studies of beta-glucosidase responsible for bioactivation of cyanogenic hydroxynitrile glucosides from Leucaena leucocephala (subabul)0 aFunctional characterization homology modeling and docking studie  aVAN GODEWIJCKSTRAAT 30, 3311 GZ DORDRECHT, NETHERLANDSbSPRINGERcFEB  a1351-13630 v403 a<p>Glycosyl hydrolase family 1 beta-glucosidases are important enzymes that serve many diverse functions in plants including defense, whereby hydrolyzing the defensive compounds such as hydroxynitrile glucosides. A hydroxynitrile glucoside cleaving beta-glucosidase gene (Llbglu1) was isolated from Leucaena leucocephala, cloned into pET-28a (+) and expressed in E. coli BL21 (DE3) cells. The recombinant enzyme was purified by Ni-NTA affinity chromatography. The optimal temperature and pH for this beta-glucosidase were found to be 45 A degrees C and 4.8, respectively. The purified Llbglu1 enzyme hydrolyzed the synthetic glycosides, pNPGlucoside (pNPGlc) and pNPGalactoside (pNPGal). Also, the enzyme hydrolyzed amygdalin, a hydroxynitrile glycoside and a few of the tested flavonoid and isoflavonoid glucosides. The kinetic parameters K (m) and V (max) were found to be 38.59 mu M and 0.8237 mu M/mg/min for pNPGlc, whereas for pNPGal the values were observed as 1845 mu M and 0.1037 mu M/mg/min. In the present study, a three dimensional (3D) model of the Llbglu1 was built by MODELLER software to find out the substrate binding sites and the quality of the model was examined using the program PROCHEK. Docking studies indicated that conserved active site residues are Glu 199, Glu 413, His 153, Asn 198, Val 270, Asn 340, and Trp 462. Docking of rhodiocyanoside A with the modeled Llbglu1 resulted in a binding with free energy change (Delta G) of -5.52 kcal/mol on which basis rhodiocyanoside A could be considered as a potential substrate.</p>10aGlycosyl hydrolase family 110ahomology modeling10aLeucaena leucocephala10aMolecular docking1 aShaik, Noor, M.1 aMisra, Anurag1 aSingh, Somesh1 aFatangare, Amol, B.1 aRamakumar, Suryanarayanarao1 aRawal, Shuban, K.1 aKhan, Bashir, Mohammad  uhttps://library.ncl.res.in/content/functional-characterization-homology-modeling-and-docking-studies-beta-glucosidase-0