红掌根腐病病原鉴定及其PCR检测方法

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

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572bp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1pg。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572bp的特异片段,实现了对红掌根腐病菌的快速可靠的检测。

Based on the difference in intemal transcribed spacer（ITS）sequences of Pythium splendens and other Pythium spp., a specific pair of primers SF1/SR2 was designed. Among 30 P splendens isolates causing root rot ofAnthurium andraeanum, and other eight fungi and two bacteria species, the primer pair amplified 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. It could amplified a specific single product from natural infected A. andraeanum plants, that was not amplified from healthy tissue. The results showed that the PCR protocol provides a rapid, sensitive and reliable tool routine detection and identification of P splendens. In addition, this study is beneficial to control root rot disease ofA. andraeanum.