大豆疫霉的分子检测

Molecular detection of Phytophthora sojae

利用核糖体内转录间隔区(internaltranscribedspacer,ITS)序列通用引物ITS1/ITS4分别获得了大豆疫霉(Phytophthorasojae)和辣椒疫霉(P.capsici)的ITS序列,通过获得序列设计了大豆疫霉的特异性引物PS1和PS2,并建立了分子检测方法。该方法对大豆疫霉(P.sojae)菌丝体和土壤中卵孢子检测的灵敏度分别为10-5ng/μL和每克土壤0.05个卵孢子。

Genetic PCR （polymerase chain reaction） products from the internal transcribed spacer （ITS） regions of the ribosomal DNA of P. sojae and P. capsici were cloned and sequenced. A pair of primers PS1/SP2 specific for P. sojae were designed according to multiple sequences analysis. PCR based on PS1/ SP2 can amplify a 96 bp product from genomic DNA of P. sojae strains ,but not other pathogens. PCR with the primers PS1/SP2 detected pathogen at a level of 10 ^-5 ng/μL or 0.05 oospore theoretically. P. sojae in soils and tissues of diseased sobean can also be targeted by the molecular probe. The PCR protocol provides a rapid and reliable tool to detect P. sojae and identify diseases.