基于细胞色素氧化酶基因和激发素基因的大豆疫霉菌特异性PCR引物

Specific PCR primers of Phytophthora sojae based on its cytochrome oxidase and elicitin genes

引起大豆疫霉根腐病的大豆疫霉菌(Phytophthora sojae)是危害大豆的破坏性病原菌之一,也是我国重要的检疫性植物病原菌。简单、快速、准确的鉴定和检测技术是阻止大豆疫霉菌传入和病害早期诊断的有效工具。本研究从大豆疫霉菌细胞色素氧化酶基因Ⅱ(coxⅡ)序列和两个激发素(elicitin)家族基因EST序列中开发了3对大豆疫霉菌特异引物:Cox3-F/Cox3-R、PSEL1-F/PSEL1-R和PSEL2-F/PSEL2-R。这3对引物在大豆疫霉菌中分别扩增出450、289bp和370bp的特异性片段,其检测大豆疫霉菌基因组DNA的灵敏度分别为20、2pg/μL和2pg/μL。3对引物能够有效检测大豆疫霉菌侵染的大豆病株,可以用于病害诊断和鉴别。

Phytophthora sojae causing soybean Phytophthora root rot is one of the most devastating pathogens of soybean, It is also an important quarantine plant pathogen in China. For quarantine detection and early diagnosis of the disease, it is necessary to develop simple, fast and effective methods for identification and detection of P. sojae. In this study, based on cox Ⅱ and elicitin gene sequences of P. sojae, 3 specific primer pairs, namely Cox3-F/Cox3-R, PSEL1-F/PSEL1-R and PSEL2-F/PSEL2-R, were developed. They could produce 450 bp, 289 bp and 370 bp fragments from P. sojae, respectively. The sensitivities of the 3 specific primer pairs for detection of P. sojae genomic DNA were determined to be 20 pg/uL, 2 pg/uL and 2 pg/uL, respectively. They could be used to detect P. sojae in infected soybean plants.