辣椒ToMMV的分子鉴定

Molecular Identification of Tomato mottle mosaic virus

采用双抗体夹心酶联免疫吸附法(DAS-ELISA)和RT-PCR法,对甘肃省农业科学院蔬菜研究所试验地的辣椒病毒病病原进行检测。结果表明:在72份病样中,烟草花叶病毒(Tobacco mosaic virus,TMV)的总检出率为66.7%,没有检出黄瓜花叶病毒(Cucumber mosaic virus,CMV)。设计3对TMV引物对阳性样品进行扩增,其中引物TMV1扩增出相似片段目的条带,经BLASTN比对与番茄斑驳花叶病毒(Tomato mottle mosaic virus,To MMV)的序列一致性为97%。针对To MMV设计3对引物进行扩增,均扩增出单一且与预期大小相同的目标条带,经测序后与To MMV的序列一致性为98%以上,证明发生在甘肃省农业科学院蔬菜研究所试验地的辣椒病毒病病原为番茄斑驳花叶病毒。

Using DAS-ELISA and RT-PCR methods,this study identified pepper pathogenetic virus in experimental fields of Vegetable Insititute,Gansu Academy of Agricultural Sciences. The results showed that in 72 disease infected samples,the total TMV detection rate was 66.7%,whereas CMV had not been detected as positive. Three pairs of primers were designed to amplify positive samples. A strip with similar segments was amplified only from primer TMV1,which had 97% sequence similarity with ToMMV by BLASTN comparison. Aiming at ToMMV,3 pairs of primers ToMMV1,ToMMVcp and ToMMVspe were designed to amplify ToMMV. A single target band with same size as expected was amplified. Its sequence was 98% accorded with ToMMV by sequencing,proving the virus found in the experimental fields of Vegetable Insititute was truly ToMMV.