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孟加拉国马铃薯X病毒的全基因组测序与分子检测

Whole genome sequencing and molecular detection of potato virus X in Bangladesh.

作者信息

Islam Md Shafiqul, Ahammed Md Akash, Akhter Fahmida, Rahman Mosaddiqur, Molla Md Mosharraf Hossain

机构信息

Tuber Crops Biotechnology and Seed Production Section, Tuber Crops Research Centre, Bangladesh Agricultural Research Institute, Gazipur, Bangladesh.

Breeder Seed Production Centre, Bangladesh Agricultural Research Institute, Debiganj, Panchagarh, Bangladesh.

出版信息

PLoS One. 2025 May 8;20(5):e0322935. doi: 10.1371/journal.pone.0322935. eCollection 2025.

Abstract

Potato Virus X (PVX) is a significant viral pathogen affecting potato (Solanum tuberosum) crops globally, yet its molecular characterization in Bangladesh remains limited. This study presents the first whole genome sequence (WGS) and molecular analysis of PVX isolated from potato plants in Gazipur, Bangladesh. Initial virus detection was performed using DAS-ELISA on symptomatic potato leaves, followed by RT-PCR targeting the coat protein (CP) gene, which confirmed PVX presence in 'Patnai' and 'Challisha' potato varieties through a 562 bp amplicon. The WGS of the Patnai-PVX isolate was determined to be 6,435 nucleotides long and deposited in GenBank (accession: PQ527059). Genome analysis identified five major open reading frames encoding the RNA-dependent RNA polymerase (RdRp), triple gene block proteins (TGBp1, TGBp2, TGBp3), and CP. Basic Local Alignment Search Tool X (BLASTX) analysis revealed high sequence similarity with PVX isolates from neighboring regions, suggesting evolutionary conservation. Mutation analysis identified 265 SNPs, predominantly synonymous mutations, indicating maintained protein-coding integrity despite genetic variation. Fewer non-synonymous mutations were detected, potentially affecting viral protein functions and pathogenicity. Phylogenetic analysis based on the entire genome sequence placed the Bangladeshi isolate (PQ527059.1) in a well-supported clade (bootstrap value 99%) with isolates from Peru (MT752634.1, MT752612.1, MT752621.1), highlighting potential international transmission routes while also exhibiting unique genetic markers indicative of regional specificity. This comprehensive molecular characterization enhances understanding of PVX genetic diversity and evolution in Bangladesh, providing valuable insights for developing effective virus management strategies in potato cultivation.

摘要

马铃薯X病毒(PVX)是一种影响全球马铃薯(Solanum tuberosum)作物的重要病毒病原体,但在孟加拉国其分子特征研究仍然有限。本研究首次报道了从孟加拉国加济布尔的马铃薯植株中分离出的PVX的全基因组序列(WGS)及分子分析。最初使用双抗夹心酶联免疫吸附测定(DAS-ELISA)对有症状的马铃薯叶片进行病毒检测,随后针对外壳蛋白(CP)基因进行逆转录聚合酶链反应(RT-PCR),通过562 bp的扩增子证实了“帕特奈”和“查利沙”马铃薯品种中存在PVX。帕特奈-PVX分离株的WGS长度为6435个核苷酸,并已存入GenBank(登录号:PQ527059)。基因组分析确定了五个主要的开放阅读框,分别编码依赖RNA的RNA聚合酶(RdRp)、三基因块蛋白(TGBp1、TGBp2、TGBp3)和CP。基本局部比对搜索工具X(BLASTX)分析显示,与来自邻近地区的PVX分离株具有高度的序列相似性,表明其进化保守性。突变分析确定了265个单核苷酸多态性(SNP),主要是同义突变,这表明尽管存在遗传变异,但蛋白质编码完整性得以维持。检测到的非同义突变较少,可能会影响病毒蛋白的功能和致病性。基于全基因组序列的系统发育分析将孟加拉国分离株(PQ527059.1)置于一个得到充分支持的分支中(自展值99%),与来自秘鲁的分离株(MT752634.1、MT752612.1、MT752621.1)在一起,突出了潜在的国际传播途径,同时也展示了表明区域特异性的独特遗传标记。这种全面的分子特征分析增进了对孟加拉国PVX遗传多样性和进化的理解,为制定马铃薯种植中有效的病毒管理策略提供了有价值的见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0510/12061425/20380acb9321/pone.0322935.g001.jpg

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