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云南农业大学学报(自然科学) 2017, Vol. 32 Issue (2) :218-225    DOI: 10.16211/j.issn.1004-390X(n).2017.02.005
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云南不同地区蜘蛛兰上朱顶红褪绿环斑病毒RNA S序列分析
高君直1, 吴保为1, 高雪1, 白继龙2, 李文贵1, 飞进强1, 刘雅婷1, 李永忠3
1. 云南农业大学 农学与生物技术学院, 云南 昆明 650201;
2. 云南红河绿春县农业技术推广中心 云南 红河 662599;
3. 云南高原特色农业产业研究院, 云南 昆明 650201
S RNA Analysis of Hippeastrum Chlorotic Ringspot Virus on Spider Lily from Different Areas in Yunnan Province
GAO Junzhi1, WU Baowei1, GAO Xue1, BAI Jilong2, LI Wengui1, FEI Jinqiang1, LIU Yating1, LI Yongzhong3
1. College of Agronomy and Biotechnology, Yunnan Agricultural University, Kunming 650201, China;
2. Agricultural Technology Promotion Center of Lyuchun County, Honghe 662599, China;
3. Yunnan Plateau Characteristic Agricultural Industry Research Institute, Kunming 650201, China

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摘要 朱顶红褪绿环斑病毒(Hippeastrum chlorotic ringspot virus,HCRV)是2013年首次发现并报道的番茄斑萎病毒属(Tospovirus)的新种,该病毒可引起蜘蛛兰产生同心环纹、叶片畸形、坏死斑等症状。本研究从云南省不同地区选出6份HCRV单独侵染的蜘蛛兰样本,通过RT-PCR、克隆、转化、测序和拼接,获得6份样本的HCRV分离物RNA S序列。6个分离物S序列长度在2 714~2 747 nt之间,其上编码的非结构蛋白NSs(Nonstructural protein by the S RNA)的NSs基因是1 332 nt,编码N(Nucleocapsid)蛋白的N基因是825 nt。通过生物信息学分析,确定了不同HCRV株系间的关系。同时对S RNA序列及其所编码的非结构蛋白NSs及核壳体蛋白N进行突变和重组分析,确定这6个HCRV分离物的RNA S序列均存在突变和重组事件。结合6个分离物采集地的环境信息,可以看出气候类型、温度等环境差异很大,因此推测云南省多样的气候类型跟HCRV在云南省的广泛分布有关,在这6个HCRV分离物中检测到的快速突变和重组可能是导致HCRV快速适应不同环境,分布范围广泛的原因之一。最后,通过系统进化分析,推测采集自云南省保山市的HLS-TCRH是6个HCRV分离物中最先开始进化的,由它分为两个进化支,一支包括玉溪的YN-YX-HT-1和德宏瑞丽的HLS-YNRL-1,另一支包括昆明的HLS-YL,红河的HLS-HHXY以及临沧的HLS-LC-YLHW。
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高君直
吴保为
高雪
白继龙
李文贵
飞进强
刘雅婷
李永忠
关键词蜘蛛兰   番茄斑萎病毒属   朱顶红褪绿环斑病毒   突变   重组     
Abstract: Hippeastrum chlorotic ringspot virus(HCRV)is a new Tospovirus species which was first discovered and reported in 2013 in Yunnan Province. HCRV induced concentric ring lines, leaf malformation, and necrotic spots symptoms on spider lily. Six samples infecting HCRV were selected by RT-PCR from different regions of Yunnan Province in this study. Six samples of the RNA S sequences were obtained through cloning, transformation and sequencing. The length of S sequence of the six isolates ranged from 2 714 to 2 747 nt, and the coding gene NSs coding the nonstructural protein by the S RNA(NSs)is 1 332 nt, N gene coding the nucleocapsid protein(N)is 825 nt. The relationships among different HCRV strains were determined by bioinformatics analysis. And we did mutation analysis and recombination detection of RNA S sequence and the protein coding. The results showed that mutation and recombination events were detected in all S RNA sequences of the six HCRV isolates. Combined with the environmental information collected from six separate objects, it was seen that there were great difference between the climate type, temperature and other environmental information in six collection cites, which suggested that the various climate types of Yunnan might relate to HCRV's wide distribution in Yunnan. Rapid mutation and recombination in these six HCRV isolates may be one of the reasons for the rapid adaptation of HCRV to different environments and a wide range of distribution. Finally, through the systematic evolution analysis, it is inferred that the HLS-TCRH collected from Baoshan, Yunnan was the first to evolve in the six isolates, it is divided into two clades, one branch includes YN-YX-HT-1 in Yuxi and HLS-YNRL-1 in Dehong Ruili, the other branch includes HLS-YL in Kunming, HLS-HHXY in Honghe and HLS-LC-YLHW in Lincang.
Keywordsspider lily   Tospovirus   Hippeastrum chlorotic ringspot virus   mutation   recombination     
Received 2016-05-24;
Fund:国家自然科学基金项目(31260451,31471828);云南省重点新产品开发计划项目(2014BB005);国家级大学生创新项目(201510676002);云南省大学生创新项目(2016106760015);云南农业大学研究生创新项目(2015ykc16)。
Corresponding Authors: 刘雅婷(1971-),女,湖南祁东人,博士,教授,主要从事植物病原微生物与寄主互作研究。E-mail:liuyating66@163.com;李永忠(1969-),男,云南龙陵人,教授,主要从事烟草科学研究。E-mail:liyongzhong168@163.com     Email: liuyating66@163.com;liyongzhong168@163.com
About author: 高君直(1985-),女,北京人,硕士,主要从事植物与病原物的互作研究。E-mail:gaojunzhi127@sina.com;吴保为(1988-),男,湖北黄冈人,硕士,主要从事植物与病原物的互作研究。E-mail:wubaowei250@163.com
引用本文:   
高君直, 吴保为, 高雪, 白继龙, 李文贵, 飞进强, 刘雅婷, 李永忠.云南不同地区蜘蛛兰上朱顶红褪绿环斑病毒RNA S序列分析[J]  云南农业大学学报(自然科学), 2017,V32(2): 218-225
GAO Junzhi, WU Baowei, GAO Xue, BAI Jilong, LI Wengui, FEI Jinqiang, LIU Yating, LI Yongzhong.S RNA Analysis of Hippeastrum Chlorotic Ringspot Virus on Spider Lily from Different Areas in Yunnan Province[J]  Journal of Yunnan Agricultural University, 2017,V32(2): 218-225
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