根癌农杆菌介导遗传转化稻曲病菌

Agrobacterium tumefaciens-Mediated Transformation of the Pathogen of Ustilaginoidea virens

利用根癌农杆菌介导转化系统，通过潮霉素抗性筛选转化子，对稻曲病菌分生孢子进行了转化。农杆菌经乙酰丁香酮预先诱导，转化效率大约为390-450个转化子／10^6个孢子。PCR检测绿色荧光蛋白基因和潮霉素基因表达盒，结果显示被测转化子基因组中均成功整合了目的基因片段。同时，在488nm下这些转化子都具有荧光。表明利用根癌农杆菌可以成功转化稻曲病菌。

Conidia of Ustilaginoidea virens were transformed by a Agrobacteriurn tumefaciens-mediated transformation （AtMT） system using a binary plasmid vector pBIG2RHPH2-GFP-GUS including HPH gene. AtMT yielded stable hygromycin B-resistant clones （average 390-450 per 10^6 conidia）, after pre-culture strain C58C1 with acetosyringone （AS）. The presence of the fragment of EGFP gene and HPH gene cassette in the transformants were checked by PCR method, and all tested transformants showed EGFP fluorescence in mycelium under 488 nm wavelength. It suggested that Ustilaginoidea virens was successfully transformed by AtMT method.