2种水稻矮缩病毒一步检测方法的建立

Construction of one-step detection method on two rice dwarf viruses

水稻黑条矮缩病毒(RBSDV)和南方水稻黑条矮缩病毒(SRBSDV)是引起水稻矮缩病的2种主要病毒,这2种病毒病在田间危害症状较相似,难以准确鉴定。为建立这2种病毒的一步快速检测方法,从混合引物配比、退火温度2个方面优化了反应体系与反应程序,并用这2类病毒的总RNA进行了RT-PCR验证,最终建立了准确、灵敏、快速的一步检测方法。利用建立的方法对从江西省大余县和南昌县采集的水稻矮缩病毒叶片样品进行检测。结果表明:大余县10份样品均为南方水稻黑条矮缩病毒(SRBSDV),检出率为100%;南昌县10份样品中有7份样品检出南方水稻黑条矮缩病毒(SRBSDV),检出率为70%,另外3份样品检出水稻黑条矮缩病毒(RBSDV),检出率为30%。

Rice black-streaked dwarf virus（RBSDV） and southern rice black-streaked dwarf virus（SRBSDV） are the main viruses which bring rice dwarf.However,the symptoms of these two viruses are highly similar and it is difficult to distinguish them only by field observation.To construct a fast detection method of these two viruses,the PCR reaction system and program were optimized from two aspects,ie.the mixing ratios of mixed primers and the annealing temperature.Finally,an exact,sensitive and fast one-step detection method was found.At the same time,the rice dwarf virus samples from Dayu County and Nanchang County in Jiangxi Province were detected with this method,and the results revealed that all of the 10 samples from Dayu County were identified as southern rice black-streaked dwarf virus with a detection ratio of 100%,7 out of 10 samples from Nanchang County were identified as southern rice black-streaked dwarf virus with a detection ratio of 70%,and 3 out of 10 samples from Nanchang County were identified as rice black-streaked dwarf virus with a detection ratio of 30%.