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| Double-stranded RNA-mediated Gene Silencing and Over-expression of FmSTS in Polygonum multiflorum Thunb Hairy Roots |
| ZHU Kuan-peng1,2, ZHAO Shu-jin 1 |
1. General Hospital of Guangzhou Military Command, Guangzhou 510010,China;
2. School of Biological Science and Engineering, South China University of Technology, Guangzhou 510640,China |
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Abstract Objective: To construct a method to research the gene FM-STS’ function that the gene come by RACE amplified from the stilbene synthase gene’s consensus sequence in Polygonum multiflorum Thunb. Method: The over-expression vector pBIN-35S-STS-GFP was constructed from the blank plasmids pBIN-35S-GFP and the gene FM-STS sequence and also the interference expression vector pBIN-35S -forward-reverse-GFP was constructed from the blank plasmids and the forward/ reverse direction sequence on account of double stranded RNA.Sent the over-expression plasmid/the interference plasmid and the blank plasmid into wild-type Agrobacterium rhizogenes ATCC15834, then induced hairy roots and cultured them. In the end the hairy roots were analyzed by HPLC and Real-time PCR. Result: The PCR results showed that the gene rolB and gfp are both expressed in hairy roots, and the content of stilbene glucoside in blank group is 2.18mg/g,in the over-expression group is 4.67mg/g and in the interference group is 0.65mg/g, At the mRNA level to detect gene Fm-STS expression level, in the over-expression group is the highest, it’s 3.17 times in the blank group and 101.44 times in the interference group. Conclusion: The method binding both technique that double stranded RNA-mediated gene silencing and over-expression is made good use of researching the gene’ s function, and the gene FM-STS is the key enzyme gene in synthesis stilbene glucoside.
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Received: 27 April 2012
Published: 25 August 2012
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