Analysis Of Differential Expressed Genes In The Suspension Cell Culture Of Tripterygium Wilfordii Hook. F. With Elictor Methyl Jasmonate Treated

As an excellent application and development prospects botanical pesticides,the market demand of Tripterygiun wilfordii Hook. f is gradually increasing. In order to provide the basis for taking advantage of modern biological technology regulation tripterygium wilfordii secondary metabolite biosynthesis of alkaloids, cell suspension culture of tripterygium wilfordii was taken as tested plant material in this study. cDNA amplified fragment length polymorphism analysis (cDNA-AFLP) was used to display transcripts whose expression is rapidly altered during the treatment of Methyl Jasmonate (MeJA) to cell suspension culture of tripterygium wilfordii. Two sixty and four primer combinations were used for selective amplification. The main results were as follows:1 The total alkaloids production was inhibited to some extent when MeJA was added into the medium. The total alkaloids production went to 624.13μg·g^-1 (DW)in the control. The alkaloids production in cell suspension culture with 200μM MeJA treated reached 478.76μgg^-1 (DW), lightly higher than the other concentrations, was still lower than control.2 Through optimization experiment steps including: restriction enzymes cut, ligation, pre- amplified and amplification, an optimized cDNA– AFLP system of differentially expressed genes in the suspension cell culture of tripterygiun wilfordii Hook. f with elictor MeJA was established.3 64 pairs of AFLP amplification primers were used to analyze the cDNA of the treated and control. Among the transcript derived fragments (TDFs), by further re-PCR amplification, cloning and sequencing,through blast analysis of the sequences we know that: seven fragments were found to be involved in the cells of the signal transduction, transcription regulation and energy metabolism in GenBank, however the other twelve fragments were found no significant homologous sequences in data base, probably they were new cDNA sequences specific to cell suspension culture of tripterygium wilfordii.4 By blast of these sequences in GenBank, Homologous analysis indicated that No.A2T4C4 fragment was acquainted identical to MYB-like protein gene(Maxident 100%). MYB protein as a kind of transcription factors is critical to reveal the mechanism of secondary metabolites transcriptional regulation. Presumably, MYB protein gene may be involved in the regulation of MeJA to tripterygium wilfordii cells suspension culture secondary metabolites biosynthesis. So, No.A2T4C4 fragment is important for further understanding of the function of MYB genes and the molecular mechanism of tripterygium wilfordii total alkaloids biosynthesis.