广西三种甜瓜病毒分离物的分子检测与鉴定

瓜类褪绿黄化病毒Cucurbits chlorotic yellows virus(CCYV)、甜瓜黄化斑点病毒Melon yellow spot virus(MYSV)及甜瓜坏死斑点病毒Melon necrotic spot virus(MNSV)是近年来报道侵染瓜类作物的新病毒,在个别种植区大面积发生,对生产构成严重威胁。为了解3种病毒在广西的发生情况,先后到各西甜瓜种植区进行了调查,并采集疑似CCYV、MYSV及MNSV的甜瓜病叶样品,提取病叶总RNA,通过特异性引物分别进行一步法RTPCR扩增,电泳结果显示RT-PCR产物与预期大小一致的序列条带;将扩增产物分别连接到pMD19-T克隆载体上,挑选阳性克隆子进行序列测定及比对分析。结果表明:CCYV、MYSV和MNSV广西分离物的CP基因序列与其他已报道核苷酸序列一致性分别达95.1%~100%、96.5%~99%和83.7%~92.5%。

Molecular detection and identification of three viruses isolated from melons in Guangxi

Cucurbit chlorotic yellows virus (CCYV), Melon yellow spot virus (MYSV) and Melon necrotic spot virus (MNSV), which were emerging threat to melon plantation, have recently been found infecting melons in some large melongrowing areas. In order to understand the incidences of these three viruses, a survey was conducted on these viruses on melons, and leaves infected by CCYV, MYSV and MNSV were collected from Nanning and Beihai, Guangxi. Total RNAs were extracted from these diseased samples and RTPCR was performed by using specific primers. The PCR products were purified and inserted into pMD19-T cloning vector, and the expected DNA fragment were sequenced and analyzed by BLASTn in GenBank. The results showed that nucleocapsid protein gene sequences of CCYV, MYSV and MNSV isolates from Guangxi had 95.1%-100%, 96.5%-99% and 83.7%-92.5% identities with those of the isolates from other parts of China or some countries reported previously.