【摘要】 由大丽轮枝菌（Verticillium dahliae）引起的黄栌（Cotinus coggygria）枯萎病可造成黄栌大量死亡,严重威胁北京香山公园的红叶景观。建立快速、高灵敏度的黄栌枯萎病菌（V.dahliae）检测技术对于黄栌枯萎病的防治具有重要意义。基于大丽轮枝菌特异性引物P1∕P2扩增出的ITS序列设计3对巢式PCR引物,并根据其可扩增性和产物的特异性进行筛选。结果表明:以引物P1∕P2为第一轮扩增引物,以PN1∕PN2为第二轮扩增引物进行巢式PCR扩增,可检测出的DNA最低浓度为2.325×10^-4ng∕μL,其灵敏度是普通PCR的100倍;研究建立的巢式PCR检测体系可为黄栌枯萎病的快速检测和早期诊断提供有效的方法。更多还原

【Abstract】 Verticillium wilt of Cotinus coggygria,caused by Verticillium dahliae,is a soil-borne disease with high mortality that pose a serious threat to the red leaf landscape in Beijing Fragrant Hills Park.For long incubation period,the disease is difficult to be diagnosed at the early stage by exsiting methods.So,it is urgent to establish a rapid and more accurate detection method for V.dahliae in soil and plants.Based on the DNA sequences of fragments amplified by primers P1∕P2,which is specific to V.dahliae,three sets of primer pairs were designed and screened in PCR assay.The results showed that the lowest detectable DNA concentration of this nested PCR was 2.325 ×10^-4 ng∕μL using primer pair P1∕P2 as the first-step PCR amplification primer and PN1∕PN2 as the second-step（nested） amplification primer and its sensitivity was 100 times higher than that of ordinary PCR.The established nested PCR approach could achieve specific detection of V.dahliae with higher sensitivity and it could provide an effective method for rapid detection and early diagnosis of the verticillium wilt of Cotinus coggygria.更多还原