论文部分内容阅读
黄瓜细菌性角斑病是黄瓜上的一种重要细菌病害,其病原为丁香假单胞菌黄瓜致病变种(Pseudomonas syringae pv.lachrymans),目前未见到该病害特异性PCR检测方法的报道。通过分析丁香假单胞菌(P.syringae)不同致病变种glyceraldehyde-3-phosphate dehydrogenase 1(gap1)基因序列设计得到一对Psl特异性PCR引物。利用该引物对丁香假单胞菌不同致病变种、假单胞菌属其他种及其他属的共46株菌株进行了PCR扩增,结果表明,所有不同来源的12株黄瓜细菌性角斑病菌均得到179bp的目标片段,而所有其他参试菌株均无扩增条带,PCR检测的灵敏度为7.5×103cfu/mL。利用该方法可从接种后发病的黄瓜叶片总DNA中检测到特异条带,而健康叶片无条带。该引物的PCR检测方法可直接用于植株总DNA的检测,无需进行病原菌的分离培养,快速简便,适用于进出境检验检疫及种苗健康检测等。
Cucumber bacterial angular leaf spot is an important bacterial disease on cucumber, the pathogen is Pseudomonas syringae pv. Lachrymans, and no specific PCR detection method has been reported yet. A pair of Psl-specific PCR primers was designed by analyzing the sequence of the gene of glyceraldehyde-3-phosphate dehydrogenase 1 (gap1) in different pathogenic strains of P. syringae. A total of 46 strains of Pseudomonas syringae different pathogenic species, other Pseudomonas species and other genera were amplified by PCR. The results showed that all of the 12 Culex pusillus isolates The target fragment of 179bp was obtained, while all the other tested strains had no amplification bands. The sensitivity of PCR detection was 7.5 × 103cfu / mL. Using this method, specific bands can be detected from the total DNA of cucumber leaves after inoculation, while healthy leaves have no bands. The primer PCR detection method can be directly used for detecting the total DNA of the plant without the isolation and culture of the pathogen, which is quick and easy, and is suitable for the inbound and outbound inspection and quarantine and seedling health detection.