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以尖孢镰刀菌西瓜专化型(Fusarium oxysporum f.sp.niveum)FO-11-06菌株为受体,采用农杆菌介导的遗传转化方法,实现了农杆菌AGL1(含pATMT1)介导的西瓜枯萎病菌的转化,并研究了转化介质和pH值对转化效率的影响。结果表明:在25℃下,采用乙酰丁香酮(AS)浓度为200μmol/L、农杆菌OD600=0.15、尖孢镰刀菌分生孢子含量为1×106个/mL、pH值为5.1～5.8时可实现T-DNA的插入转化。当转化体系pH值为5.3时,转化效率最高,达到每106个分生孢子产生553个转化子。不同共转化介质对转化效率影响明显,玻璃纸和硝酸纤维素滤膜转化效率较高,每106个分生孢子获得551个和549个转化子,显著高于滤纸。通过转化,获得尖孢镰刀菌西瓜专化型转化子1 989个,对生长速度、菌落颜色和产孢量等性状进行分析,发现有46个性状特异的突变体。研究结果为进一步研究尖孢镰刀菌西瓜专化型的生长发育、致病机制和相关基因的功能奠定了基础。
Using Fusarium oxysporum f.sp. niveum FO-11-06 strain as the recipient, the Agrobacterium-mediated genetic transformation method was used to achieve the Agrobacterium tumefaciens AGL1 (containing pATMT1) -mediated Watermelon Fusarium oxysporum transformation, and the conversion media and pH on the conversion efficiency. The results showed that when the concentration of acetosyringone (AS) was 200μmol / L, the Agrobacterium OD600 was 0.15, the Fusarium oxysporum conidia content was 1 × 106 / mL and the pH value was 5.1-5.8 at 25 ℃, T-DNA can be inserted and transformed. When the pH value of the transformation system was 5.3, the transformation efficiency reached the highest, reaching 553 transformants per 106 conidia. Different co-transformation media had obvious effect on conversion efficiency, cellophane and nitrocellulose filter had higher transformation efficiency, 551 and 549 transformants per 106 conidia, which were significantly higher than that of filter paper. A total of 1 989 Fusarium oxysporum watermelon transformants were obtained by transformation. The growth rate, colony color and sporulation were analyzed, and 46 trait-specific mutants were found. The results laid the foundation for further study on the growth and development, pathogenic mechanism and related gene function of Fusarium oxysporum watermelon.