菠菜霜霉病菌快速分子检测方法研究  

Rapid Detection Spinach Downy Mildew Pathogen by Molecular Method

作　　者：李京[1,2] 张祥林[1] 王翀[1] 李亚伟[1] 张小菊[1] 

机构地区：[1]新疆出入境检验检疫局,乌鲁木齐830063 [2]新疆农业大学农学院,乌鲁木齐830052

出　　处：《新疆农业科学》2016年第2期346-351,共6页Xinjiang Agricultural Sciences

基　　金：国家质检总局科研项目(201310091)~~

摘　　要：【目的】菠菜霜霉病,由专性寄生病原菌Peronospora farinosa f.sp.Spinaciae引起,是世界上菠菜Spinacia oleracea种植中最为重要的经济型病害。建立菠菜霜霉病菌快速分子检测方法。【方法】将菠菜霜霉病菌进行显微镜的形态鉴定;提取DNA、PCR扩增、克隆、酶切、测序、序列比对与分析;采用巢式PCR技术检测菠菜霜霉病:第一轮采用通用引物ITS1/ITS4,第二轮采用特异性引物SMPF/SMPR。【结果】该菌与霜霉属的形态特征描述一致,确定菠菜患有霜霉病;测序结果经Blast比对分析与Peronospora effusa同源性达100%;巢式PCR技术也特异性的检测到了菠菜霜霉病菌。【结论】建立针对菠菜霜霉病的快速分子检测技术。【Objective 】Spinach downy mildew disease,caused by the obligate pathogen Peronospora farinosa f. sp. spinaciae,is the most economically important spinach（ Spinacia oleracea） disease. 【Objective】The purpose of this study is to establish the molecular method of rapid detection of spinach downy mildew pathogen.【Method】First of all,the morphology of spinach downy mildew pathogen should be identified by microscope; then the second process was DNA extraction,PCR amplification,cloning,enzyme digestion,sequencing,the sequence alignment and analysis; and finally,the spinach downy mildew was detected by using nested PCR technique. The first amplification was to use the universal primer ITS1 / ITS4 and the second amplification was to use the specific primer SMPF / SMPR. 【Result】Microscopy results showed that the fungus were consistent with the description of Peronospora morphology,so we made sure that spinach were infected with downy mildew disease; The homology of sequencing results of the blast analysis with Peronospora effuse was 100%; nested PCR technology also specially detected spinach downy mildew pathogen. 【Conclusion】This study established the rapid detection spinach downy mildew pathogen by molecular method but the technique of Real- time quantitative PCR needs more exploration.

关 键 词：菠菜霜霉病 酶切 巢式PCR 

分 类 号：S41[农业科学—植物保护]