GFP标记的马铃薯大丽轮枝菌生物学特性研究

The Study on Biological Characteristics of GFP Labeled Potato Verticillium dahliae

为了研究马铃薯黄萎病菌的侵染机制,将绿色荧光蛋白基因(GFP)通过农杆菌介导的遗传转化方法转入马铃薯黄萎病菌VD012中,经过潮霉素选择性培养基的筛选和分子鉴定,获得了47株有绿色荧光信号的阳性转化子。随机挑取8株阳性转化子,以野生型菌株为对照,对转化子的菌落形态、菌丝的生长速率、产孢量、粗毒素含量和致病力进行了研究。结果表明,8株阳性转化子中有3株微菌核产生的数量明显高于野生型,1株转化子微菌核产生量低于野生型菌株;各阳性转化子的生长速率和野生型菌株差异不显著;阳性转化子的产孢量与对照相比,有2株差异不显著。其余均有不同程度的降低。保湿培养8 h,所有阳性转化子的平均萌发率低于野生型菌株。相比对照,粗毒素的含量表现为升高趋势的转化子有7株,1株表现为下降趋势。致病力测定的结果表明,致病力增强的转化子有1株,2株转化子的致病力呈现降低的趋势。

In order to unravel the infection mechanism of V.dahliae to potato,a binary construct contained GFP gene was transferred via Agrobacterium tumefaciens-mediated system into V.dahliae isolate VD012.47 positive transformants were obtained by hygromycin B selection and molecular verification.Eight tansformants were randomly selected to characterize the colony morphology,growth rate,conidia production,toxin production and pathogenicity.The results showed that three transformants could produce more microsclerotia compared with the wild type strain;whereas,one transforment produced less microsclerotia.The significant difference on the growth rate of eight transformants.The conidia production ability of the transformants were decreased to certain degree.except two among them were not remarkable overall.The average germination rate of all the tested transformants was lower than that of the wild strain after 8 h germination.Regarding to the toxin production,seven transformants showed increased tendency,while only one decreased compared with the wild type.The pathogencity results showed that,compared with wild strain,only one transformant showed increased tendency,the left two strains showed decreased pattern.