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马铃薯Y病毒福建分离物P1基因的分子变异和结构特征 预览 被引量:5

Sequence variation of P1 gene in Potato virus Y isolated from Fujian Province
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摘要 为揭示马铃薯Y病毒(Potato virus y,PVY)P1基因的分子变异及结构特征,并查明福建分离物P1基因的变异来源.文章设计一对简并引物从感染PVY的马铃薯病叶扩增、克隆获得福建分离物P1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,并采用贝叶斯法重建P1基因的系统发育树.结果显示,12个福建分离物均成功扩增出预期大小(约915 bp)的特异性片段,P1基因的核苷酸序列与已报道的PVY不同株系的核苷酸序列一致性为73%~99%; QK44,XT02,XT08,LH05分离物的309 nt位置均检测到一个强烈的重组信号.12个PVY分离物中,P1蛋白有85个变异的氨基酸位点,表明其高度变异;P1蛋白的第41~275位是一个高度保守的结构功能域,含有3个保守的活性位点(H192、D201、V235);系统发育分析显示,福建分离物形聚为3种不同的簇,其对应的卷曲结构(Coiled-coil domain)和空间结构也不相同,表明不同株系型的P1基因在系统发育关系上分化较大.该研究结果表明PVY P1基因在核苷酸序列、氨基酸序列以及空间结构具有高度变异性,但行使P1蛋白功能的活性位点所在的氨基酸(H192、D201和V235)均高度保守;福建分离物P1基因的变异源主要来自碱基突变和基因重组. To understand the sequence variation and the putative protein structure of P1 gene in Potato virus Y (PVY) and to identify the sources of the variation, P1 gene in PVY isolated from Fujian Province was amplified by re- verse-transcription polymerase chain reaction (RT-PCR) using a pair of degenerate primers designed from the conserved regions of published sequences. Sequence variation and putative protein structure were analyzed, and phylogenetic tree was reconstructed using Bayesian inference method. Expected fragments of 915 bp in size were amplified t~om 12 samples col- lected from Fujian Province by RT-PCR. The 12 sequences shared 73%--99% nucleotide identity with the reference se- quences from GenBank. A strong recombination signal was identified at position 309 in sequences of isolates QK44, XT02, XT08 and LH05. Among the 12 sequences, 85 amino acid variants were detected, indicating high sequence variation in the P1 protein. However, positions 41-275 in the protein were highly conserved, especially in three active sites (Ht92, Dz01 and V235). Phylogenetic analysis grouped the sequences into three clades, each with different Coiled-coil domains and 3D-structures, suggesting divergent phylogenetic relationship among the groups. The above results show P1 gene in PVY is highly variable but contains 3 conserved active sites (H192, D201, V235) and the high genetic variation in the gene is primarily due to mutation and recombination.
作者 史凤阳 高芳銮 沈建国 常飞 詹家绥 Fengyang Shi, Fangluan Gao, Jianguo Shen, Fei Chang, Jiasui Zhan ( 1. Key Laboratory of Plant Virology of Fujian Province, Institute of Plant Virology, Fujian Agriculture and Forestry University, Fuzhou 350002, China; 2. Inspection & Quarantine Technology Center, Fujian Exit-Entry Inspection and Quarantine Bureau, Fuzhou 350001, China)
出处 《遗传》 CAS CSCD 北大核心 2014年第7期713-722,共10页 Hereditas
基金 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),国家质检总局科技计划项目(编号:2013IK299,2013IK094)和福建出入境检验检疫局科技项目(编号:FK2011-07,FK2007-07)资助
关键词 马铃薯Y病毒 P1基因 变异 重组 Potato virus Y P1 gene variation recombination
作者简介 史凤阳,在读硕士研究生,专业方向:马铃薯病毒.E-mail:shifyang@126.com. 高芳銮,在读博士研究生,专业方向:马铃薯病毒.E-mail:raindy@fafu.edu.cn. 通汛作者:詹家绥,教授,研究方向:病原群体遗传与进化。E-mail:jiasui.zhan@fafu.edu.cn
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