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[目的]鉴定引起海南水茄叶片呈典型皱缩和轻微花叶症状的病原物。[方法]对该病样提取总RNA,反转录成第一链c DNA,然后设计WTMV(Wild tomato mosaic virus)cp基因的特异引物,再利用常规PCR方法鉴定其病原是否为WTMV。[结果]通过PCR扩增获得一段约900 bp的DNA序列,包含WTMV全部CP(813 bp)蛋白基因编码区。分析表明该片段编码270个氨基酸,与WTMV-Laichua分离物核酸序列同源性为89%,氨基酸序列同源性为94%,且在所有的WTMV分离物中保守,可形成一个小的group。与其它马铃薯Y病毒属病毒的同源关系依次是PVMV、Chi VMV-Ⅰ、其它16个potyviruses(包括8个属于Chi VMV-Ⅱ的Chi VMV)。[结论]确定了引起海南水茄叶片皱缩和轻微花叶症状的病原物为野番茄花叶病毒(WTMV)。海南WTMV该分离物CP蛋白基因的Gen Bank登录号为KR781519。据我们所知,这是国内首次发现WTMV自然侵染水茄。
[Objective] The aim was to identify the pathogen that caused the typical shrinkage and mild mosaic symptoms in the leaves of Hainan watermelon. [Method] Total RNA was extracted from the sample and reversely transcribed into the first strand of c DNA. Then the specific primers of cp gene of WTMV (Wild tomato mosaic virus) were designed and the pathogen was identified as WTMV by conventional PCR. [Result] A DNA sequence of about 900 bp was obtained by PCR amplification and contained the coding region of all CP (813 bp) protein gene of WTMV. The result showed that the fragment encoded 270 amino acids. The fragment shared 89% homology with WTMV-Laichua isolate and 94% amino acid sequence identity. The fragment was conserved in all WTMV isolates and formed a small group. Homology to other viruses of the genus V virus in turn was PVMV, Chi VMV-I and 16 other potyviruses (including 8 Chi VMVs belonging to Chi VMV-II). [Conclusion] The pathogen that caused the shrinkage of cucumber leaves and mild mosaic symptoms was wild tomato mosaic virus (WTMV). The Gen Bank Accession No. of the CP protein gene of Hainan isolate WTMV is KR781519. To the best of our knowledge, this is the first time that WTMV has been found naturally infected with watermelon.