A Study About The Biosynthesis Of Taxol In Cephalotaxus Mannii

Cephalotaxus mannii is an endangered, rare and national key protected plant. Its stems, leaves, seeds and bark contain a variety of secondary metabolites with anticancer activity.It’s potential commercial value has attract great attraction.This study is about the biosynthesis of Cephalotaxus mannii secondary metabolites, taxol in particular. On the basis of transcriptome sequencing,4genes involved in the biosynthesis of taxol were cloned from Cephalotaxus manni. After elicited by methyl jasmonate acid (MeJA), the gene expression and regulation characteristics were probed in Cephalotaxus manni. Meanwhile, the Cephalotaxus mannii cell suspension system was also established in this study. Elicitor treatment with MeJA can also effect secondary metabolites, such as taxol, biosynthesis in Cephalotaxus mannii suspension cell. The main results were as follows:(1) Two transcriptome of Cephalotaxus mannii leaves were sequenced. Combined results of Transcriptome Sequencing, Gene segments information, gene functional annotation, gene expression differences are analyzed respectively.(2) According to the BLAST results from transcriptome sequencing, genes fragment related to taxol biosynthesis, the taxane13-alpha-hydroxylase gene, taxadiene5-alpha hydroxylase gene, taxadiene synthase gene, CF943gene were cloned from Cephalataxus mannii by using the RACE techniques. And the4genes were analyzed in bioinformatics respectively.(3) According to the Real-Time PCR result, taxane13-alpha-hydroxylase gene expression level increased significantly, after stimulated by methyl jasmonate acid (MeJA).(4) The Cephalotaxus mannii cell suspension culture had been established. Meanwhile, the HPLC results showed:taxol content in suspension cell is high in lag phase,:taxol content in suspension cell is low in mitotic phase,; and it raise again during, plateau phase.(5) Methyl jasmonic acid(MeJA) treatment can promote the taxol biosynthesis in Cephalotaxus mannii suspension cell.