ZHANG Wei,BAI Yan-ju,SHEN Yu,et al.Cloning and Sequences Analysis of CP Gene ofPotato Virus X Heilongjiang Isolate[J].HEILONGJIANG AGRICULTURAL SCIENCES,2010,(08):1-5.
马铃薯X病毒黑龙江分离物外壳蛋白基因克隆与序列分析
- Title:
- Cloning and Sequences Analysis of CP Gene ofPotato Virus X Heilongjiang Isolate
- 文章编号:
- 1002-2767(2010)08-0001-05
- Keywords:
- potato virus X; coat protein; amino acids; sequence analysis; sibship
- 分类号:
- S435.32
- 文献标志码:
- A
- 摘要:
- 马铃薯X病毒(Potato virus X,PVX)是重要的马铃薯病毒之一,几乎分布于全国所有马铃薯种植区域,严重影响马铃薯的产量和品质。对其建立RT-PCR分子检测体系以及对其外壳蛋白(CP)基因进行序列分析尤为重要。根据已报道的PVX的核苷酸序列设计合成1对引物,以带毒植株总RNA为模板,应用RT-PCR方法,克隆了PVX黑龙江分离物(P2)全长CP基因序列。序列分析表明:P2CP基因全长711 bp,编码237个氨基酸残基,与GenBank中17 个不同分离物的CP核苷酸序列同源性为75.5%~96.3%,氨基酸序列同源性为89.4%~99.2%。依据PVX CP氨基酸序列建立系统进化树,将PVX不同分离物划分为两大类群,P2与PVX中国分离物亲缘关系较近,同属于类型Ⅱ,表现一定的地域相关性。同时,应用该RT-PCR分子检测体系进行马铃薯样品检测。
- Abstract:
- Potato virus X(PVX)is the impotaint virus disease in potato,distributing nearly in all the potato planted area,can affect the yield and quality of potato seriously,so erecting RT-PCR molecule detecting system and CP sequence analysising are important.A pair of primers was designed based on the reported CP gene of PVX the hole CP gene sequence of PVX Heilongjiang isolate was cloned by reverse transcription polymerase chain reaction(RT-PCR).The analysis of sequence showed that:The CP gene of P2 was made up of 711 nucleic acids,coding 237 amino acids.The CP nucleic acid sequence homology of P2 and other 17 isolates of PVX in GenBank was between 75.5%~96.3%,the CP amino acid sequence homology was between 89.4%~99.2%.The phylogenic tree which established based on PVX CP amino acid was clustered into two groups,P2 has nearer sibship with the isolates of China,both in Ⅱgroup,they represented region correlation.Finally,using the RT-PCR detection system to detect potato virus.
参考文献/References:
[1]张威,白艳菊,高艳玲,等.马铃薯主产区病毒病发生情况调查[J].黑龙江农业科学,2010(4) :71 - 73. [2] Slack S A,German T L.Disease caused by viruses and viro-ids[J].Phytopathology Society,2001,8(1):57-62. [3] 李芝芳.中国马铃薯主要病毒图鉴[M].北京:中国农业出版社,2004. [4] 库尔斯塔克.植物病毒比较诊断指南[M].裴美云,译.北京:农业出版社,1991. [5] 耿宏伟,白艳菊,范国权,等.应用RT-PCR技术检测马铃薯A病毒[J].中国马铃薯,2009,23(6):329-332. [6] 侯义龙,张开春,吴禄平,等.果树组织中总RNA提取的新方法[J].沈阳农业大学学报,2002,33(2):122-125. [7] Zhang Yun,Guo Dong-chun,Lin Ming,et al.Characterization of M-class genome segments of Muscovy duck reovirus S14[J].Virus Research,2007,125:42-53. [8] Rouis S,Lafaye P,Jaoua-Aydi L,et al.Cloning and expression of functional single-chain Fv antibodies directed against NIa and coat proteins of potato virus Y[J].Journal of Virological Methods,2006,137:1-6. [9]Nie X,Singh R P.Detection of multiple potato viruses using an oligo(dT)as a common cDNA primer in multiplex RT-PCR[J].Journal of Virological Methodsds,2000,86:179-185. [10]萨姆布鲁克J,拉塞尔D W.分子克隆实验指南(上、下册)[M].3版.黄培堂,王嘉玺,朱厚础,等.译. 北京:科学出版社,2002. [11] Goulden M G,Kohm B A,Cruz S S,et al.A feature of the coat Protein of potato virus X affects both induced virus resistance in potato and viral fitness[J].Virology,1997,231:35-42.[12] 曲静,朱常香,温孚江,等.一个马铃薯X病毒分离物的外壳蛋白基因序列分析与株系鉴定[J].植物保护学报,2003,30(4):358-364. [13]Atreya C D.Application of genome sequence information in potyvirus taxonomy[J].Archives of Virology,1992,5:17-23. [14] Brunt A A.The general properties of potyvirus[J].Archives of Virology,1992,5:3-16. [15]Tsuneyoshi T,Sumi S.Differentiation among garlic viruses in mixed infections based on RT-PCR procedures and direct tissue blotting immunoassay[J].Phytopathology,1996,86:253-259.
相似文献/References:
[1]苗艳梅,赵敏.马铃薯Y病毒属病毒外壳蛋白功能[J].黑龙江农业科学,2019,(03):165.[doi:10.11942/j.issn1002-2767.2019.03.00165]
[J].HEILONGJIANG AGRICULTURAL SCIENCES,2019,(08):165.[doi:10.11942/j.issn1002-2767.2019.03.00165]
备注/Memo
基金项目:948资助项目(2008-Z23);马铃薯产业技术体系资助项目(NYCYTX-15);农业科技成果转化资助项目(2008GB2B200084)