论文部分内容阅读
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 is the soil-borne disease-causing fungus causing cotton verticillium wilt. Rapid and timely detection of Verticillium dahliae is of great significance to the early warning and later prevention and treatment of cotton Verticillium wilt. One of the improved specific PCR primers VDS-F / VDS-R was obtained by polymerase chain reaction (PCR) detection technique to verify, screen and improve the detection primers of reported Verticillium dahliae. Under the optimized reaction conditions and amplification conditions, a band of about 520 bp of l product was amplified from the genome of V. dahliae specifically and the detection sensitivity was 10-2 ng / μL. Using this primer The pathogen could be detected specifically from the tissues of soil and cotton plants containing Verticillium dahliae. The detection sensitivity of artificial nematodes by nested PCR method was 10 spores / g soil. The PCR detection system of this primer can be used for the early rapid detection of cotton Verticillium wilt.