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Cloning and expression analysis of kenaf (Hibiscus cannabinus L.) major lignin and cellulose biosynthesis gene sequences and polymer quantification during plant development
Authors:Pasquale Chiaiese  Gianluca Ruotolo  Antonio Di MatteoAmalia De Santo Virzo  Anna De MarcoEdgardo Filippone
Institution:a Department of Soil, Plant, Environmental and Animal Production Sciences, School of Biotechnological Science, University of Naples Federico II, Via Università, 100, 80055 Portici (Naples), Italy
b Department of Soil, Plant, Environmental and Animal Production Sciences, School of Agriculture, University of Naples Federico II, Italy
c Department of Structural and Functional Biology, School of Mathematical, Physical and Natural Sciences, University of Naples Federico II, Italy
Abstract:In order to apply state-of-the-art molecular breeding techniques in fibre crop it is necessary to have a good knowledge of major polymer biosynthesis gene sequences and their expression pattern. Polymerase chain reaction was employed to isolate sequences of the major genes for lignin and cellulose biosynthesis in a kenaf cultivar. CeSA, 4cl, c4h, cad, and ccr gene primers were designed according to their conservative regions; partial sequences of lignin and cellulose biosynthesis genes were obtained. One actin II gene sequence was also isolated from the kenaf genome as a housekeeping gene to be employed in qPCR analysis. Expression levels of genes c4h, cad and CeSA in bark and core from plants harvested at three different growth stages were evaluated. Using qPCR analyses it was found that the expression levels of the two biosynthesis lignin genes in bark tissues increased during plant growth, while a negative trend was recorded in core tissues. In both bark and core, the quantity of lignin was positively correlated to plant growth while cellulose content decreased.
Keywords:4cl  4-coumarate:CoA ligase  c4 h  cinnamate 4-hydroxylase  cad  cinnamyl alcohol dehydrogenase  ccr  cinnamoyl-CoA oxidoreductase  CeSA  cellulose synthase
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