土壤中大豆疫霉检测技术的改进  

Improved detecting technology of Phytophthora sojae in soil

作　　者：吴翠萍[1] 郑斯竹[1] 李彬[1] 粟寒[1] 李敏[1] 安榆林[1] 

机构地区：[1]江苏出入境检疫局动植物与食品检测中心,江苏南京210001

出　　处：《植物检疫》2010年第5期34-38,共5页Plant Quarantine

摘　　要：目前在中国口岸检疫部门使用的大豆疫霉(Phytophthorasojae)常规PCR检测方法存在稳定性不好、检出率不高的局限性。通过在常规PCR的引物两端各增加一条引物设计成套式PCR,并通过对涉及卵孢子DNA提取的各个步骤如取样、细胞破壁、离心等的改进,充分保证了DNA的最高扩增效率和最大提取量。这些步骤的改进使得口岸送检大豆疫霉检出率比常规方法提高了3～4倍。Currently conventional PCR detecting methods for Phytophthora sojae,which is generally used in quarantine department in China,has disadvantages of poor stability and low detecting rate.Designing nested-PCR by adding a primer to the both ends of the conventional PCR primer respectively,and improving various steps of extracting DNA of oospores such as sampling,cell wall breaking and centrifugating,which guarantee the highest amplifying efficiency and maximum recovering of DNA.These improvement steps make P.sojae in soil detecting rate of port increase 3~4 times than conventional methods.

关 键 词：土样制备 大豆疫霉 套式PCR DNA提取 

分 类 号：S41[农业科学—植物保护] S435.651