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为了鉴定引起海南定安县番茄叶片呈蕨叶型或变窄成线条状,叶片表面凹凸不平、生长不规则等症状的病原物,对该病样的叶片提取总RNA,反转录成第一链c DNA,然后根据CMV RNA1保守序列设计2对特异引物,再利用常规PCR方法鉴定其病原是否为CMV。利用两对特异引物进行PCR方法扩增后,分别获得大小约为500 bp和700 bp的目的 DNA条带,经序列相似性比对,发现海南定安县番茄的病原物为CMV。构建系统进化树和同源性分析,进一步确定了引起番茄叶片蕨叶和生长不规则的CMV为重花叶株系(包括蕨叶症状),属CMV IB亚组。本研究鉴定了CMV是引起番茄叶片蕨叶和生长不规则等症状的重要病原物,通过序列比较和同源分析确定了该病原物为CMV的重花叶株系(包括蕨叶症状)。
In order to identify the pathogen causing tomato leaves in Ding’an County of Hainan Province to appear fern leaf type or to narrow into a line shape, the surface of the leaf surface is irregular and irregular in growth, the total RNA was extracted from the diseased leaves and reversely transcribed into the first strand c DNA, and then design two pairs of specific primers according to the conserved sequence of CMV RNA1, and then identify whether the pathogen is CMV by conventional PCR. Two pairs of specific primers were used to amplify the PCR products. The target DNA bands of about 500 bp and 700 bp in size were obtained respectively. The sequence similarity analysis showed that the pathogen of tomato in Ding’an County of Hainan Province was CMV. The phylogenetic tree and homology analysis were constructed to further confirm that CMV, which causes the growth of tomato leaf fern and irregular growth, is a heavy mosaic (including fern leaf symptom), belonging to the CMV IB subgroup. In this study, CMV was identified as an important causative agent causing the growth of tomato leaf fern and its irregular growth. The CMV heavy mosaic (including fern leaf symptoms) was confirmed by sequence comparison and homology analysis.