棉花黄萎病病原菌大丽轮枝菌的快速分子检测
Rapid molecular detection of Verticillium dahliae by PCR method

大丽轮枝菌Verticillium dahliae是引起棉花黄萎病的土传病害病原真菌。快速及时地检测出大丽轮枝菌，对棉花黄萎病的早期预警及后期防治具有重要意义。采用聚合酶链式反应（polymerase chain reaction，PCR）检测技术对已报道大丽轮枝菌的检测引物进行验证、筛选和改进，获得1对改进的特异性PCR引物VDS-F/VDS-R。在优化的反应体系与扩增条件下，能特异性地从大丽轮枝菌基因组扩增出l条约520 bp的产物条带，检测灵敏度达到10-2 ng/μL；利用该引物可特异性地从含有大丽轮枝菌的土壤及棉花植株组织中检测出病原菌；采用巢氏PCR法对人工病土的检测灵敏度达到了10个孢子/g土。表明本引物的PCR检测体系可用于棉花黄萎病的早期快速检测。
Verticillium dahliae, which can cause cotton Verticillium wilt, is a devastating soilborne fungus. In order to rapidly and accurately detect the fungus and timely prevent cotton Verticillium wilt, PCR primers for V. dahliae reported in the literatures were screened, verified and modified by using the PCR detection technology. A pair of modified V. dahliae specific PCR primers VDS-F/VDS-R was developed and only a single band of PCR product of 520 bp in length was obtained with V. dahliae genomic DNA as template under the optimized reaction system and amplification conditions. The detection sensitivity of the primers VDS-F/VDS-R was 10-2 ng/μL of the template DNA. The primers VDS-F/VDS-R could be used to specifically detect V. dahliae in soils or the diseased cotton tissues. The sensitivity of nested-PCR for detection of V. dahliae in soils was ten spores per gram soil. The results indicated that the modified PCR primers were very useful for early detection of cotton Verticillium wilt pathogen.