02944nas 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<p>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.</p>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-0