马铃薯Y病毒pipo基因的分子变异及结构特征分析

doi:10.3724/SP.J.1005.2013.01125

遗传 ›› 2013, Vol. 35 ›› Issue (9): 1125-1134.doi: 10.3724/SP.J.1005.2013.01125

马铃薯Y病毒pipo基因的分子变异及结构特征分析

高芳銮¹, 沈建国², 史凤阳¹, 常飞¹, 谢联辉¹, 詹家绥¹

1. 福建农林大学植物病毒研究所, 福建省植物病毒学重点实验室, 福州 350002;
2. 福建出入境检验检疫局检验检疫技术中心, 福州 350001

收稿日期:2013-04-15 修回日期:2013-07-08 出版日期:2013-09-20 发布日期:2013-09-25
通讯作者: 谢联辉 E-mail:fjxlh@126.com
基金资助:
国家现代农业马铃薯产业技术体系(编号：CARS-10), 福建省自然科学基金项目(编号：2013J01088), 福建省教育厅科技项目(编号：JA12119)和福建出入境检验检疫局科技项目(编号：FK2011-07)资助

Sequence variation and protein structure of pipo gene in Potato virus Y

GAO Fang-Luan¹, SHEN Jian-Guo², SHI Feng-Yang¹, CHANG Fei¹, XIE Lian-Hui¹, ZHAN Jia-Sui¹

1. Fujian Key Laboratory of Plant Virology, Institute of Plant Virology, Fujian Agriculture and Forestry University, Fuzhou 350002, China;
2. Inspection & Quarantine Technology Center, Fujian Entry-Exit Inspection and Quarantine Bureau, Fuzhou 350001, China

Received:2013-04-15 Revised:2013-07-08 Online:2013-09-20 Published:2013-09-25

摘要/Abstract

摘要：

为揭示马铃薯Y病毒(Potato virus Y, PVY) pipo基因的分子变异和结构特征, 文章根据文献报道的马铃薯Y病毒属(Potyvirus) pipo 基因保守区序列设计一对简并引物, 从感染PVY的马铃薯病叶中克隆获得pipo基因的cDNA全长序列, 分析其核苷酸序列和氨基酸序列的特征, 并基于氨基酸序列使用贝叶斯法重建了Potyvirus的系统发育树。结果显示：20个PVY分离物成功扩增出预期大小(约235 bp)的特异性片段, 其核苷酸序列与已报道的其它PVY 株系的pipo基因核苷酸序列一致性均在92%以上; 5′端均含有典型的G1-2A6-7 基序(motif), 无碱基插入/缺失, 所有的核苷酸变异都是碱基置换, 共发现13个多态性位点, 其中4个简约信息位点, 9个单一变异位点, 表明该基因高度保守, 但不同分离物也存在一定的分子变异; PIPO蛋白理论等电点11.26~11.62, 无信号肽和跨膜区, 是可溶的亲水性蛋白; 整个蛋白含有3个保守区, 其中位于10~59aa的基序最为保守。该蛋白主要定位于线粒体中, 可能是线粒体导肽。系统发育分析结果显示, 源于PVY不同株系优先相聚成簇, 而向日葵褪绿斑驳病毒(Sunflower chlorotic mottle virus, SuCMoV)与辣椒重花叶病毒(Pepper severe mosaic virus, PepSMV)的亲缘关系较PVY相比更近, 与前人的结果相一致, 表明PIPO蛋白可以作为研究Potyvirus系统发育关系的新的分子标记。

关键词: 马铃薯Y病毒, pipo基因, 病毒进化

Abstract:

The objectives of this study were to understand the sequence variation and the putative protein structure of pipo gene in the Potato virus Y (PVY) collected from Solanum tuberosum. The pipo gene in PVY was cloned using a pair of degenerate primers designed from its conserved region and its sequences were used to re-construct phylogenetic tree in Potyvirus genera by a Bayesian inference method. An expected fragment of 235 bp was amplified in all 20 samples by RT-PCR and the pipo genes in the 20 samples assayed shared more than 92% nucleotide sequence similarity with the published sequences of PVY strains. Among the 20 pipo gene sequences, 13 polymorphic sites were detected, including 4 parsimony informative sites and 9 singleton variable sites. These results indicate that PVY pipo gene is highly conserved but some sequence variations exist. Further analyses suggest that the pipo gene encodes a hydrophilic protein without signal peptide and transmembrane region. The protein has theoretical isoelectric points (pI) ranging from 11.26 to 11.62 and contains three highly conserved regions, especially between aa 10 and 59. The protein is likely located in the mitochondria and has α-helix secondary structure. Bayesian inference of phylogenetic trees reveals that PVY isolates are clustered in the same branch with high posterior probability, while Sunflower chlorotic mottle virus (SoCMoV) and Pepper severe mosaic virus (PepSMV) are closely related, consisting with the classification of Potyvirus genera using other approaches. Our analyses suggest that the pipo gene can be a new marker for phylogenetic analysis of the genera. The results reported in this paper provide useful insights in the genetic variation and the evolution of PVY and can stimulate further research on structure and function of the PIPO protein.

Key words: evolution of virus, Potato virus Y, pipo gene

高芳銮沈建国史凤阳常飞谢联辉詹家绥. 马铃薯Y病毒pipo基因的分子变异及结构特征分析[J]. 遗传, 2013, 35(9): 1125-1134.

GAO Fang-Luan SHEN Jian-Guo SHI Feng-Yang CHANG Fei XIE Lian-Hui ZHAN Jia-Sui. Sequence variation and protein structure of pipo gene in Potato virus Y[J]. HEREDITAS, 2013, 35(9): 1125-1134.

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马铃薯Y病毒pipo基因的分子变异及结构特征分析

Sequence variation and protein structure of pipo gene in Potato virus Y

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