Scientia Agricultura Sinica ›› 2011, Vol. 44 ›› Issue (8): 1603-1609 .doi: 10.3864/j.issn.0578-1752.2011.08.008

• PLANT PROTECTION • Previous Articles     Next Articles

Obtaining and analysis on silent transformants of Polyketide synthase gene(StPKS) in Setosphaeria turcica

  

  1. 1. 河北农业大学
    2. 河北,保定,河北农业大学毒素实验室
    3.
  • Received:2010-10-08 Revised:2010-12-02 Online:2011-04-15 Published:2011-04-15
  • Contact: CAO ZHI YAN

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Abstract: 【Objective】RNA interference technology was used to investigate the function of a polyketide synthase gene (StPKS) in Setosphaeria turcica.【Method】The silent vector, pSilent-StPKS1-2, was constructed by inserting StPKS fragments into plasmid pSilent-1 then trandformed into S. turcica 01-23 protoplasts through PEG. Transformants were selected on potato dextrose agar (PDA) containing hygromycin and the expression levels of StPKS was analysised by Semiquantitative RT-PCR. The hypae morphology of transformants and wide type was observed by microscope.【Results】The RNAi vector was effective to reduce the gene expression. Thirty hygromycin-resistant transformants were isolated and five melanin-deficient transformants were analysised by Semiquantitative RT-PCR. The results showed that StPKS mRNA was weakly expressed in white-color phenotypes. The transformants hypae morphology was anomalistic, including intumescent, anamorphic and ramose.【Conclusion】StPKS gene involved in the DHN-melanin biosynthesis and related to the mycelial morphology. The DHN-melanin biosynthesis was readuced when StPKS gene weakly expression.

Key words: Setosphaeria turcica, Polyketide synthase gene, DHN-melanin, RNAi

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