Asad Zohaib, Ashfaq Muhammad, Iqbal Naeem, Parvaiz Fahed, Mehmood Mirza Abid, Hameed Akhtar, Malik Amir Humayun, Kayani Samah Bashir, Al-Kahtani Mohamed A, Ahmad Zubair
Department of Plant Pathology, PMAS- Arid Agriculture University Rawalpindi, Pakistan.
Plant Pathology, Institute of Plant Protection, MNS- University of Agriculture Multan, Pakistan.
Saudi J Biol Sci. 2022 May;29(5):3577-3585. doi: 10.1016/j.sjbs.2022.02.027. Epub 2022 Feb 19.
(CGMMV), a well-known , infects cucurbits across the globe. To determine its current status, molecular characterization, genetic recombination, gene flow and selection pressure, 10 districts from Punjab province of Pakistan were surveyed and a total of 2561 cucurbits samples were collected during 2019-2020. These samples were subjected to virus-specific double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA) for the detection of CGMMV. The results revealed that viral disease was prevalent in all surveyed districts of Punjab with an overall 25.69% disease incidence. ELISA positive samples were further confirmed through RT-PCR and sequencing of coat protein (CP) cistron. Sequence analysis showed that the present studied CGMMV isolates have 96-99.5% nucleotide and 94.40-99.50% amino acid identities with those already available in GenBank. Phylogenetic analysis also revealed that understudied isolates were closely related with South Korean (AB369274) and Japanese (V01551) isolates and clustered in a separate clad. Sequence polymorphisms were observed in 663 bp of sequence within 31 CGMMV isolates covering complete CP gene. Total number of sites were 662, of which 610 and 52 sites were monomorphic and polymorphic (segregating), respectively. Of these polymorphic, 24 were singleton variable and 28 were parsimony informative. Overall nucleotide diversity (π) in all the understudied 31 isolates was 0.00010 while a total of 1 InDel event was observed and InDel Diversity (k) was 0.065. Haplotype diversity analysis revealed that there was a total 29 haplotypes with haplotype diversity (Hd) of 0.993458 in all the 31 isolates which provide evidence of less diversity among Pakistani isolates. The statistical analysis revealed the values 2.568, 5.31304 and 4.86698 of Tajima's D, Fu, & Li's F* and D*, respectively, which witnessed the population of CGMMV was under balanced selection pressure.
黄瓜绿斑驳花叶病毒（CGMMV）是一种广为人知的病毒，可感染全球范围内的葫芦科植物。为了确定其当前状况、分子特征、基因重组、基因流动和选择压力，对巴基斯坦旁遮普省的10个地区进行了调查，并在2019年至2020年期间共收集了2561份葫芦科植物样本。这些样本采用病毒特异性双抗体夹心酶联免疫吸附测定（DAS-ELISA）检测CGMMV。结果显示，旁遮普省所有调查地区均普遍存在病毒性疾病，总体发病率为25.69%。ELISA阳性样本通过RT-PCR和外壳蛋白（CP）顺反子测序进一步确认。序列分析表明，本研究的CGMMV分离株与GenBank中已有的分离株具有96 - 99.5%的核苷酸同一性和94.40 - 99.50%的氨基酸同一性。系统发育分析还表明，所研究的分离株与韩国（AB369274）和日本（V01551）的分离株密切相关，并聚集在一个单独的分支中。在覆盖完整CP基因的31个CGMMV分离株的663 bp序列中观察到序列多态性。位点总数为662个，其中610个和52个位点分别为单态和多态（分离）。在这些多态性位点中，24个为单例变异，28个为简约信息位点。所有31个研究分离株的总体核苷酸多样性（π）为0.00010，同时观察到总共1个插入缺失事件，插入缺失多样性（k）为0.065。单倍型多样性分析表明，所有31个分离株共有29个单倍型，单倍型多样性（Hd）为0.993458，这表明巴基斯坦分离株之间的多样性较低。统计分析显示，Tajima's D、Fu & Li's F和D的值分别为2.568、5.31304和4.86698，这表明CGMMV群体处于平衡选择压力之下。
