砂梨上苹果茎痘病毒的调查分析及RT-PCR与TC-RT-PCR检测研究

Investigation and detection of Apple stem pitting virus in Pyrus pyrifolia by RT-PCR and TC-RT-PCR

试验通过西方烟(Nicotianaoccidentalis‘37B’)和杂种(Pyroniaveitchii)、A20接种鉴定及PAS-ELISA检测,对保存于国家砂梨种质资源圃的38份砂梨样品进行了苹果茎痘病毒(Applestempittingvirus,ASPV)带毒率的调查分折。结果表明,生物学鉴定与血清学检测结果一致,ASPV带毒率均为26.3%。应用以dsRNA为模板RT-PCR检测该病毒,在杂种和A20上显症的6个样品均获得316bp目标片段。应用TC-RT-PCR检测显症的西方烟,也获得了预期的目标片段。

In this study,38 samples of pear cultivars belonging to Pyrus pyrifolia were collected in the National Fruit Germplasm Repository of Pyrus pyrifolia for testing and detecting the Apple stem pitting virus (ASPV) by Nicotiana occidentalis '37B',Pyronia veitchii and A20 inoculated and PAS-ELISA. Results showed the detective consequence by PAS-ELISA was consistent with that by biological index, the detection rate reached up to 26.3%. RT-PCR using dsRNA as template was performed to detect ASPV and the expected fragment about 316 bp was amplified in 6 samples on Pyronia veitchii Aspv-affected. The corresponding fragment was also amplified by TC-RT-PCR on Nicotiana occidentalis '37B' Aspv-affected.