马铃薯Y病毒两个黑龙江烟草分离物的全基因组序列分析
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摘要
为明确黑龙江烟区马铃薯Y病毒(Potato virus Y,PVY)分子株系和进化特征,从哈尔滨和牡丹江感病烟草采集到Harbin和MDJ两个样品,通过RT-PCR扩增了其全基因组序列,并进行了一致率、重组、系统发育和选择压力等分析。结果表明,2个分离物(GenBank登录号分别为MH933741和MH933742)基因组全长均为9699核苷酸。Harbin与MDJ和比利时分离物GBVC 26(JQ969039)一致率最高,为98.5%;MDJ与Harbin和GBVC 26一致率最高,为98.5%。Harbin是美国分离物Oz和瑞士分离物N-605的重组体,MDJ是美国分离物CW和N-605的重组体。利用全基因组序列构建的系统进化树显示,PVY分离物分为9个组,Harbin和MDJ均属PVY~(N-Wi)组。PVY的11个基因均处于负选择,其中NIa蛋白基因的选择压力最大。
To elucidate the molecular strains and evolution characteristics of Potato virus Y(PVY) from tobacco plants in Heilongjiang,two samples were collected from Harbin and Mudanjiang tobacco fields and were, designated as Harbin and MDJ. Their complete genomic sequences were amplified via RT-PCR. Sequence identities, recombination, phylogenetic relationship and selection pressure were analyzed.Results showed that the full-length genome of isolates Harbin(Accession no. MH933741) and MDJ(Accession no. MH933742) were both9699 nucleotides(nt). Harbin had the highest identities of 98.5% with isolate GBVC 26(JQ969039) from USA and MDJ, while MDJ had the highest identity of 98.5% with isolate Harbin and GBVC 26. Harbin was a recombinant of isolate Oz from USA and isolate N-605 from Switzerland, while MDJ was a recombinant of N-605 and isolate CW from USA. From the phylogenetic tree constructed with the complete genomic sequences, PVY was divided into nine groups, among which Harbin and MDJ belonged to group PVYN-Wi. All the eleven proteins of PVY were under negative selection, with NIa gene under the highest selection pressure.
To elucidate the molecular strains and evolution characteristics of Potato virus Y(PVY) from tobacco plants in Heilongjiang,two samples were collected from Harbin and Mudanjiang tobacco fields and were, designated as Harbin and MDJ. Their complete genomic sequences were amplified via RT-PCR. Sequence identities, recombination, phylogenetic relationship and selection pressure were analyzed.Results showed that the full-length genome of isolates Harbin(Accession no. MH933741) and MDJ(Accession no. MH933742) were both9699 nucleotides(nt). Harbin had the highest identities of 98.5% with isolate GBVC 26(JQ969039) from USA and MDJ, while MDJ had the highest identity of 98.5% with isolate Harbin and GBVC 26. Harbin was a recombinant of isolate Oz from USA and isolate N-605 from Switzerland, while MDJ was a recombinant of N-605 and isolate CW from USA. From the phylogenetic tree constructed with the complete genomic sequences, PVY was divided into nine groups, among which Harbin and MDJ belonged to group PVYN-Wi. All the eleven proteins of PVY were under negative selection, with NIa gene under the highest selection pressure.
引文
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[2]Valkonen J P T.Chapter 28-Viruses:Economical Losses and Biotechnological Potential[J].Potato Biology&Biotechnology,2007:619-641.
[3]万秀清,乔婵,赵淑娟,等.黑龙江烟区烟草马铃薯Y病毒株系的分子鉴定[J].烟草科技,2015,48(10):13-18,25.WAN Xiuqing,QIAO Chan,ZHAO Shujuan,et al.Molecular Identification of Tobacco Potato Virus Y Strains in Heilongjiang Tobacco Planting Areas[J].Tobacco Science&Technology,2015,48(10):13-18,25.
[4]Scottolavino E,Du G,Frohman M A.5'end cDNA amplification using classic RACE[J].Nature Protocols,2006,1(6):2555-2562
[5]王凤龙,钱玉梅,王劲波,等.烟草马铃薯Y病毒病国内外研究现状及今后我国的防治研究对策[J].中国烟草学报,1998,4(2):49-55.WANG Fenglong,QIAN Yvmei,WANG Jinbo,et al.Present Situation of Research on Potato Virus Y and the Strategy of Research and Control Method[J].Acta Tabacaria Sinica,1998,4(2):49-55.
[6]陈瑞泰,韩晓东,史万华,等.五省烟区烟草病毒类型的初步研究[J].中国烟草科学,1986,(01):1-4.CHEN Ruitai,HAN Xiaodong,SHI Wanhua,et al.Preliminary study on tobacco virus types in five provinces[J].Acta Tabacaria Sinica,1986,(01):1-4.
[7]韩晓东,陈瑞泰,李林森,等.我国烟草的一个重要病毒--马铃薯Y病毒[J].上海农业科技,1985,(02):20-21.HAN Xiaodong,CHEN Ruitai,LI Linsen,et al.An important virus of tobacco in China-potato y virus[J].Shanghai Agricultural Science and Technology,1985,(02):20-21.
[8]王劲波,王凤龙,钱玉梅,等.山东烟区主要病毒的株系鉴定[J].中国烟草学报,1998,4(1):24-32.WANG Jinbo,WANG Fenglong,QIAN Yvmei,et al.Identification of main viruses in Shandong tobacco region[J].Acta Tabacaria Sinica,1998,4(1):24-32.
[9]蔡伟,杨宏凯,沈建国,等.全基因组序列的系统发育与重组分析鉴定我国马铃薯Y病毒株系[J].激光生物学报,2017,26(4):350-359.CAI Wei,YANG Hongkai,SHEN Jianguo,et al.Determination of Potato virus Y strain in China based on phylogenetic and recombination analyses of the complete genomic sequences[J].Acta Laser Biology Sinica,2017,26(4):350-359.
[10]Tian YP,Liu JL,Zhang CL,et al.Genetic diversity of Potato virus Y infecting tobacco crops in China[J].Phytopathology,2011,101(3):377-387.
[11]Lorenzen J H,Meacham T,Berger P H,et al.Whole genome characterization of Potato virus Y,isolates collected in the western USA and their comparison to isolates from Europe and Canada[J].Archives of Virology,2006,151(6):1055-1074.
[12]Hu X,Karasev A V,Brown C J,et al.Sequence characteristics of potato virus Y recombinants.[J].Journal of General Virology,2009,90(12):3033-3041.
[13]Kamangar S B,Smagghe G,Maes M,et al.Potato virus Y,(PVY)strains in Belgian seed potatoes and first molecular detection of the N-Wi strain[J].Journal of Plant Diseases&Protection,2014,121(1):10-19.
[14]孙剑萍.黑龙江省烟草马铃薯Y病毒病发生规律及防治技术研究[D].东北农业大学,2003.SUN Jianping.Studies on epidemiology and control of potato virus Y on tobacco in Heilongjiang province[D].Northeast Agricultural University,2003.