【摘要】 根据烟草根黑腐病菌(Thielaviopsis basicola)与其它烟草病原真菌核糖体基因转录间隔区(internal transcribed spa-cer,ITS)序列间的差异,设计了一对特异性引物TB-5/TB-3,用于T.basicola的分子检测。利用该对引物对包括T.basi-cola在内的13个烟草病原菌菌株的基因组DNA进行PCR扩增,结果表明:只有T.basicola能扩增到一条400bp左右的特异性条带,其它菌株及阴性对照均无扩增产物。对烟草组织和土壤的检测结果也表明,该对引物能特异性的检测到T.basicola基因组DNA的存在。该引物对T.basicola基因组DNA检测的灵敏度为100fg/μL。更多还原

【Abstract】 Based on the differences in internal transcribed spacer(ITS) sequences of T.basicola and other tobacco fungal pathogens,a pair of species-specific primers TB-5/TB-3 was designed for T.basicola detection.After screening 10 isolates of T.basicola and other tobacco fungal pathogens,TB-5/TB-3 primers amplified only a single product of about 400 bp from T.basicola.And T.basicola could be specifically dectec-ted by PCR assay with TB-5/TB-3 from tobacco tissues infected naturally and soil samples.The sensitivity of detection was 100 fg genomic DNA per 25 μL PCR reaction volume.The PCR-based methods developed here could simplify both plant disease diagnosis and pathogen monitoring,as well as guiding plant disease management.更多还原