Martin Robert R, Pinkerton Jack N, Kraus Jennifer
USDA-ARS Horticultural Crops Research Laboratory, Corvallis, OR 97330, USA.
J Virol Methods. 2009 Jan;155(1):91-5. doi: 10.1016/j.jviromet.2008.10.002. Epub 2008 Nov 18.
Tomato ringspot virus (ToRSV), Tobacco ringspot virus (TRSV) and Tobacco rattle virus (TRV) are transmitted to healthy plants by viruliferous nematodes in the soil. A method was developed for extraction of genomic viral RNA from virus particles carried within nematodes and a sensitive nested RT-PCR detection assay. The procedure has been adapted to microscale for handling multiple samples. This assay is effective for detection of ToRSV or TRSV in Xiphinema americanum or TRV in Paratrichodorus allius. With this method, viruses can be detected in nematodes fed on infected plants or from field-collected nematodes where the percentage of viruliferous nematodes is unknown. Soil samples from four red raspberry fields infected with ToRSV were collected in 2003 and 2004. Nematodes isolated from these samples were assayed for ToRSV by RT-PCR and compared to cucumber baiting bioassay for virus transmission from the same soil samples. ToRSV was detected in nematodes throughout the season with similar frequencies by the RT-PCR assay and the transmission bioassay.
番茄环斑病毒（ToRSV）、烟草环斑病毒（TRSV）和烟草脆裂病毒（TRV）可通过土壤中携带病毒的线虫传播至健康植株。已开发出一种从线虫体内携带的病毒颗粒中提取基因组病毒RNA的方法以及一种灵敏的巢式逆转录聚合酶链反应（RT-PCR）检测方法。该程序已适用于微量规模以处理多个样本。此检测方法对于检测美洲剑线虫中的ToRSV或TRSV以及葱短体线虫中的TRV有效。通过这种方法，可以在取食受感染植株的线虫中或从田间采集的线虫中检测到病毒，而携带病毒的线虫百分比未知。2003年和2004年从四个感染ToRSV的红树莓田采集了土壤样本。从这些样本中分离出的线虫通过RT-PCR检测ToRSV，并与用于检测相同土壤样本中病毒传播的黄瓜诱饵生物测定法进行比较。通过RT-PCR检测方法和传播生物测定法在整个季节中以相似频率在线虫中检测到ToRSV。
