棉花植株内黄萎病菌致病类型的快速检测

目的]建立一种能从棉花植株中快速检测黄萎病菌致病类型的方法,为黄萎病菌致病类型的大面积普查打下基础.方法]利用Dneasy Plant Mini Kit试剂盒对落叶型标准菌株592和非落叶型标准菌株511及病圃中显症前后不同时期棉花植株DNA进行提取,然后用落叶型引物DB19/DB22、DB19/espdef01和非落叶型引物NDf/ NDr、InND2f/InND2r对所提DNA进行巢式PCR扩增.结果]使用落叶型引物只在592菌株和落叶型病圃的病株中检测到目标条带,使用非落叶型引物只在511菌株和非落叶型病圃的病株中检测到目标条带.结论]该试剂盒可以有效地提取棉花植株中的DNA,供试的两套引物对检测植株中棉花黄萎病菌致病类型具有高度特异性,该法可直接检测棉花植株中黄萎病菌的致病类型.

Rapid Detertion of Pathotype of Verticilliun dahliae in Cotton Plants

【Objective】This research was conducted to establish the method to rapidly detect to detcet pathotypes of Verticillium dahliae in cotton plants,which would serve as a basis of general investigation of V.dahliae in large area.【Method】DNA extracted from defoliating strain 592,nondefoliating strain 511,and cotton plants using Dneasy Plant Mini Kit was amplified with defoliating V.dahliae-specific primers DB19/DB22,DB19/espdef01 and nondefoliating V.dahliae-specific primers NDf/ NDr,InND2f/InND2r by nested PCR.【Result】Defoliating specific band only appeared in 592 strain and plants collected from defoliating disease garden when using DB19/DB22 and DB19/espdef01,nondefoliating specific band only appeared in 511 strain and plants collected from nondefoliating disease garden when using NDf/ NDr,InND2f/InND2r.【Conclusion】DNA can be extracted effectively from cotton plants by using Dneasy Plant Mini Kit,and the two sets of primers are higyly special in detection of defoliating and nondefoliating pathotypes of V.dahliae.This technique can be used to detect pathotypes of V.dahliae directly in cotton plants.