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以棉花黄萎病菌强致病力菌株Vd084、Vd082和弱致病力菌株Vd001为供试菌株,以耐病品种中棉所35和感病品种冀棉11为鉴别寄主,采用蛭石沙土无底纸钵定量蘸菌液体法,首先明确棉花黄萎病菌致病力与落叶间的关系,然后采用大丽轮枝菌落叶型菌株特异引物D-1/D-2和非落叶型菌株特异引物ND-1/ND-2对供试菌株进行分子检测。结果表明,1×107 mL-1的常规接种量下,3个菌株均能导致棉苗出现落叶;与弱致病力菌株相比,强致病力菌株出现落叶的时间早3d,且落叶株率高。在接种量为1×105~1×107 mL-1,强致病力菌株的落叶株率显著高于弱致病力菌株,当接种量达到1×108 mL-1时,强致病力菌株和弱致病力菌株间差异不明显。3个菌株在耐病品种上的落叶株率(28.8%)显著低于感病品种(37.7%)。分子检测表明,2个强致病力菌株Vd084和Vd082为落叶型菌株,弱致病力菌株Vd001为非落叶型菌株。结果显示,该2对引物检测出的非落叶型菌株Vd001在温室苗期人工接种条件下仍能导致棉苗落叶。可见,对于落叶型棉花黄萎病菌的检测还需要建立更完善的方法,尤其是模拟大田间条件的检测方法更为重要。
The virulent strain of Verticillium dahliae (Vd084, Vd082) and weak pathogenic strain (Vd001) were used as tested strains. The disease-resistant variety Zhongmiansuo 35 and susceptible variety Jiamian 11 were identified as hosts. Bowl quantitative dipped liquid method, first clear pathogenicity of Verticillium dahliae cotton and the relationship between the deciduous, and then using the leaves of Dallidogi deciduous strain specific primers D-1 / D-2 and non-deciduous strain-specific primers ND- 1 / ND-2 on the test strains for molecular testing. The results showed that all the three strains could cause defoliation of cotton seedlings under the conventional inoculum volume of 1 × 107 mL-1. Compared with the virulent virulent strains, the virulent strains had deciduous leaves as early as 3d, High rate. Inoculation amount of 1 × 105 ~ 1 × 107 mL-1, strong virulent strains of deciduous strains were significantly higher than the weak virulence strains, when the inoculum size reached 1 × 108 mL-1, strong virulence strains And weak virulence strains were not significantly different. The rate of deciduous plants (28.8%) in the three isolates was significantly lower than that of the susceptible cultivars (37.7%). Molecular tests showed that the two virulent strains Vd084 and Vd082 were deciduous and the weakly virulent strain Vd001 was non-deciduous. The results showed that the non-deciduous strain Vd001 detected by the two pairs of primers could still cause cottonseed leaves deciduous under artificial inoculation in greenhouse. Visible, for the detection of deciduous cotton Verticillium dahliae also need to establish a more complete method, in particular, to simulate the conditions between the field detection method is more important.