农杆菌介导的西瓜枯萎病菌遗传转化

为获得带GFP标记的西瓜枯萎病菌转化株,用于后期观察病原菌侵染过程,采用农杆菌介导的方法,对西瓜枯萎病菌1号生理小种进行了遗传转化。结果表明:共培养时间为36h,枯萎病菌孢子和农杆菌AGL1比例为1∶1时该菌株的遗传转化效率最高,可以达到117.33个转化子/107个孢子。转化株的孢子、菌丝体及萌发的孢子均能发出稳定而强的绿色荧光。转化株的致病力检测显示其致病力与转化前的野生菌株致病力无明显差异。结果表明本研究获得的带GFP标记的西瓜枯萎病菌转化株可用于观察病菌在西瓜根系的侵染过程。

Agrobacterium tumefaciens-mediated transformation of Fusarium oxysporum f.sp. niveum

The purpose of this study is to obtain GFP-tagged transformants of Fusarium oxysporum f. sp. niveum race 1, which can be used to study colonization of watermelon roots by the pathogen of Fusarium wilt. F.oxysporum f. sp. niveum race 1 was transformed with the pCH-sGFP transformation vector using Agrobacterium tumefaciens AGL1. The results indicated that the transformation efficiency was the highest at 117.33 transformants/107 conidia when the mix was co-cultivated for 36 h with an equal volume of conidial suspension and A.tumefaciens AGL1. The expression of green fluorescent protein in the hyphae, conidia and germinating spores of the transformants were stable with strong green fluorescent signal. Pathogenicity tests confirmed that the mean disease severity caused by the isolate FON-sGFP was not significantly different from that of the wild-type isolate FON1. FON1-sGFP might be a useful tool for visualizing colonization process of watermelon roots by F.oxysporum f. sp. niveum.