甘蔗白叶病植原体PCR检测方法的建立与应用

The Establishment and Application of a PCR Method for Sugarcane White Leaf Phytoplasma Detection

为建立简便、快速、特异性的甘蔗白叶病植原体检测方法,本研究根据甘蔗白叶病植原体secA基因设计特异引物secA-F/secA-R,建立了甘蔗白叶病植原体PCR检测方法。使用该方法可以从含有甘蔗白叶病植原体的甘蔗样品中稳定扩增出目标基因片段,片段长度为350 bp。本方法的最低检测浓度为50 fg/µL。应用该方法对云南耿马蔗区采集的7份甘蔗白叶病样品进行了检测鉴定,结果表明,所有样品均为阳性。以上结果表明,本研究建立的甘蔗白叶病植原体PCR检测方法特异性和敏感性好,操作简便,适用于甘蔗白叶病植原体的快速检测。

In order to establish a simple,rapid and specific PCR detection method for sugarcane white leaf phytoplasma,the experiment designed specific primers secA-F/secA-R based on the secA gene of sugarcane white leaf phytoplasma and established a detection PCR method for sugarcane white leaf phytoplasma.Using this method,the target gene fragment with a fragment length of 350 bp can be stably amplified from sugarcane samples containing the sugarcane white leaf phytoplasma.The minimum detection concentration for this method is 50 fg/µL.The method was applied to test seven sugarcane white leaf samples collected from the Gengma sugarcane planting area in Yunnan,and the results showed that all samples were positive.The above results showed that the PCR method established in this study had good specificity and sensitivity,easy operation and was suitable for the rapid detection of sugarcane white leaf phytoplasma.