Cloning And Bioinformatics Analysis Of Caffeic Acid O-menthyl-transferase Gene From Broussonetia Papyrifera

Broussonetia papyrifera mainly distributed in Yellow River,Yangtze River and Pearl River Basin of China,which is a multi-functional species extensively applied in the industry of papermaking,feed and medicine etc.Caffeic acid-3-0-methl transferase(COMT)is an important methylase in plant lignin biosynthesis pathway,which plays an important role in biosynthesis of syringyl lignin monomer.To control expression of this gene can influence content of lignin and composition of monomer.Therefore,to research characteristics and mechanism of BpCOMT expression can provide reference for improvement on materials quality of forest.In this study,full-length sequence of B.papyrifera Caffeic acid 0-methyl transferase gene(BpCOMT)was cloned using degenerate-polymerase chain reaction(PCR)and rapid ampliication of cDNA ends(RACE)techniques.The cloned BpCOMT cDNA contains 1227 bp,which contains an open reading frame(ORF)of 1104 bp 65 bp of 5'-untranslated region(UTR)and 58 bp of 3'-UTR,encoding a protein of 367 amino acids with molecular weight of 40.04 kDa and theoretical isoelectric point of 5.51.The bioinformatics analysis showed BpCOMT belongs to methyltransferase(OMTs)family,and contains a conserved element specific to OMTs family.The phylogenetic analysis should that the BpCOMT is in the same sub differentiated group with COMTs from Castaneamollissima,Juglansregia,Betulapendula,and Betulaplatyphylla.Real time Q-PCR analysis showed that BpCOMT was stably expressed in all organs of B.papyrifera,while it is significantly expressed in roots than that in leaves and stems.The findings provided theoretical and experiment reference for further researching the regulation mechanism of BpCOMT in lignin monomer biosynthesis and genetic improvement of forest trees using the gene.