Rasoulpour Rasoul, Afsharifar Alireza, Izadpanah Keramat
Plant Virology Research Center, College of Agriculture, Shiraz University, Shiraz, Iran.
Virusdisease. 2016 Jun;27(2):193-7. doi: 10.1007/s13337-016-0310-3. Epub 2016 Mar 29.
Melon seedlings showing systemic chlorotic spots and mosaic symptoms were collected in central part of Iran, and a virus was isolated from diseased plants by mechanical inoculation. The virus systemically infected the most inoculated test plants by inducing mosaic symptoms, while, in the members of Fabaceae family and Chenopodium quinoa induced local lesions. Agar gel diffusion test using a polyclonal antiserum against a squash Cucumber mosaic virus (CMV) isolate showed the presence of CMV in the mechanically inoculated plants (designated CMV-Me). The virus was purified by polyethylene glycol precipitation and differential centrifugation. A polyclonal antiserum was produced against the virus that reacted specifically with virus antigen in ELISA and agar gel diffusion tests. The virus was molecularly characterized by PCR amplification of the full length of the coat protein gene using cucumovirus genus specific primer pair CPTALL-3/CPTALL-5 and sequence analysis of the resulting product. No RNA satellite was detected using the primer pair CMVsat3H/sat5T7P. Phylogenetic analysis based on the coat protein amino acid sequences showed that CMV-Me belongs to Subgroup IB. These results may be helpful in melon breeding programs, focusing on plant resistance to plant viruses including CMV.
在伊朗中部采集了表现出系统性褪绿斑点和花叶症状的甜瓜幼苗,并通过机械接种从患病植株中分离出一种病毒。该病毒通过诱导花叶症状系统性地感染了大多数接种的试验植株,而在豆科植物和藜麦中则诱导产生局部病斑。使用针对南瓜黄瓜花叶病毒(CMV)分离株的多克隆抗血清进行的琼脂凝胶扩散试验表明,在机械接种的植株(命名为CMV-Me)中存在CMV。该病毒通过聚乙二醇沉淀和差速离心进行纯化。制备了针对该病毒的多克隆抗血清,其在ELISA和琼脂凝胶扩散试验中与病毒抗原发生特异性反应。使用黄瓜花叶病毒属特异性引物对CPTALL-3/CPTALL-5对衣壳蛋白基因全长进行PCR扩增,并对所得产物进行序列分析,从而对该病毒进行分子特征鉴定。使用引物对CMVsat3H/sat5T7P未检测到RNA卫星。基于衣壳蛋白氨基酸序列的系统发育分析表明,CMV-Me属于亚组IB。这些结果可能有助于甜瓜育种计划,重点关注植物对包括CMV在内的植物病毒的抗性。