Takaichi Miyuki, Yamamoto Mika, Nagakubo Takayuki, Oeda Kenji
Biotechnology Laboratory, Sumitomo Chemical Co. Ltd., 4-2-1 Takatsukasa, Takarazuka, Hyogo 6658555, Japan.
Plant Dis. 1998 Jun;82(6):694-698. doi: 10.1094/PDIS.1998.82.6.694.
A specific and highly efficient indexing method for four major garlic viruses, GV1 carlavirus, GV2 potyvirus, onion yellow dwarf virus (OYDV), and mite-borne mosaic virus, was developed using the reverse transcription-polymerase chain reaction method (RT-PCR). When specific primers were synthesized to amplify the coat protein genes of these viruses, the amplified PCR bands had the same expected sizes. This indexing method can be performed using an extremely small amount of leaf tissue (50 mg) to obtain reproducible data. Diseased garlic plants collected from five fields in northern Japan were tested using this method. As GV2 was detected in all the fields, and high frequency of disease symptoms was obtained in the case of heavy infection, GV2 may well be a major garlic virus in this region. By contrast, GV1 was detected in only one of the five fields. OYDV and mite-borne mosaic viruses were also detected in various fields and at different frequencies. Judging from the virus indexing data and the disease symptoms, these two viruses appeared to cause severe symptoms in the case of a mixed infection with GV2. The frequency of virus re-infection to virus-free clones was low in isolated fields, and GV2 was the major virus that re-infected virus-free clones.
利用逆转录-聚合酶链反应法(RT-PCR)开发了一种针对四种主要大蒜病毒的特异性高效检测方法,这四种病毒分别是大蒜卡尔病毒1(GV1)、马铃薯Y病毒大蒜分离物(GV2)、洋葱黄矮病毒(OYDV)和螨传花叶病毒。当合成特异性引物以扩增这些病毒的外壳蛋白基因时,扩增的PCR条带具有预期的相同大小。这种检测方法可以使用极少量的叶片组织(50毫克)来获得可重复的数据。使用该方法对从日本北部五个田地采集的患病大蒜植株进行了检测。由于在所有田地中都检测到了GV2,并且在重度感染的情况下出现了高频率的病害症状,因此GV2很可能是该地区的主要大蒜病毒。相比之下,在五个田地中只有一个检测到了GV1。在不同田地中也检测到了OYDV和螨传花叶病毒,且频率各异。从病毒检测数据和病害症状判断,这两种病毒在与GV2混合感染的情况下似乎会引发严重症状。在隔离田地中,病毒重新感染无病毒克隆的频率较低,GV2是重新感染无病毒克隆的主要病毒。