红掌根腐病病原鉴定及其PCR 检测方法

 利用真菌通用引物ITS1 和ITS4 扩增红掌根腐病菌转录间隔区并进行序列测定，通过序列比较，设计了1 对红掌根腐病菌的特异引物SF1/SR2，对30 个红掌根腐病病原菌、8 种其它真菌和2 种细菌基因组DNA 进行PCR 扩增。结果表明，只有红掌根腐病菌获得572 bp 的特异带。使用引物SF1/SR2对华丽腐霉进行PCR 扩增，其检测灵敏度在DNA 水平上可达1 pg。运用设计的引物从红掌根腐病菌基因组DNA 以及人工接种和自然发病的红掌植株中扩增到572 bp 的特异片段，实现了对红掌根腐病菌的快速可靠的检测。

Identification and PCR Detection of the Pathogen Causing Root Rot of Anthurium andraeanum

Based on the difference in internal transcribed spacer （ITS）sequences of Pythium splendensand other Pythium spp.，a specific pair of primers SF1/SR2 was designed. Among 30 P. splendens isolatescausing root rot of Anthurium andraeanum，and other eight fungi and two bacteria species，the primer pairamplified a single 572 bp product from all isolates of P. splendens，but not from any other isolates tested.The sensitivity of detection of the pathogen P. splendens with primers SF1/SR2 was 1 pg genomic DNA. Itcould amplified a specific single product from natural infected A. andraeanum plants，that was notamplified from healthy tissue. The results showed that the PCR protocol provides a rapid，sensitive andreliable tool routine detection and identification of P. splendens. In addition，this study is beneficial tocontrol root rot disease of A. andraeanum.