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Gene Knockout Vector Construction and Function Assay of FvST12 in Fusarium verticillioides

ZHANG Yue-ping1, QU Hua-xiang2   

  1. 1、Longping College, Graduate School of Central South University, Changsha 410125;
    2、Agronomy College, Hunan Agricultural University, Changsha 410128
  • Received:2010-08-24 Online:2011-04-02 Published:2010-11-04

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Abstract: 【Objective】The objective of this study is to identify and analyze the STE12 homolog transcription factor gene in Fusarium verticillioides. 【Method】 The F. oxysporum Fost12 transcription factor was used as a target sequence to search its homolog by BlastP in F. verticillioides genome sequence. FVEG_05267.3, a predicted gene in F. verticillioides, was identified and it showed 98% similarity to Fost12. It was called FvST12 in this study. Based on the Double-Joint PCR, the knockout vector was constructed and null knockout mutants were generated through fungal protoplast transformation. The gene knockout mutants were confirmed by PCR and Southern blot. Furthermore, the function of the gene in F. verticillioides was systematically characterized 【Result】Compared with the wild type, Fvst12 mutant has normal growth rate, condiation, colony morphology, and stress responses (osmotic and oxidative stress) tested in this study. However, the mutant reduced the virulence on maize ear and stalk. 【Conclusion】The transcription factor FvST12 is an important virulence factor but it isn’t involved in the vegetative growth, conidiation, osmotic and oxidative stress responses in F. verticillioides.

Key words: Fusarium verticillioides , FvST12 , gene knockout , pathogenicity , function assay

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