摘要
应用改进的SDS-酚氯仿法,在沉淀RNA前先加入1/4体积无水乙醇、1/10体积5mol/L乙酸钾沉淀多糖,然后再用异丙醇沉淀RNA,成功地从大豆干种子中提取了大豆花叶病毒(SMV)总RNA;应用RT-PCR技术对大豆种子中携带的SMV进行了检测,同时以DAS-ELISA方法作比较,建立了能直接以大豆干种子为检测对象的SMV快速、灵敏、特异的RT-PCR检测技术。
Total RNA of Soybean mosaic virus (SMV) was extracted successfully from soybean dry seed with improved SDS-phenol-chloroform procedure. The improvement of procedure lay in 1/4 volume of 100% ethanol and 1/10 volume of 5 mol/L potassium acetate were added to pellet polysaccharides selectively before pelleting RNA. A pair of specific primers was designed based on the nucleotide sequence of coat protein (CP) gene of SMV G2 strain, and then it was used to detect SMV in soybean dry seed by reverse transcription-polymerase chain reaction (RT-PCR). Meanwhile, DAS-ELISA was performed for testing same samples. The result showed that a rapid, sensitive and specific RT-PCR assay was established for detection of SMV in soybean dry seed.
出处
《植物病理学报》
CAS
CSCD
北大核心
2005年第3期214-220,共7页
Acta Phytopathologica Sinica
基金
国家科技攻关项目(2004BA525B01)
