北方四省区番茄褪绿病毒的分子鉴定  

Molecular identification of Tomato chlorosis virus from four provinces or autonomous region in northern China

作　　者：王翠琳[1] 冯佳[1] 孙晓辉[1] 王少立[1] 赵静[2] 刘金亮[3] 竺晓平[1] 

机构地区：[1]山东农业大学植物保护学院,泰安271018 [2]潍坊科技学院蔬菜研究所,寿光262700 [3]吉林大学农学部植物科学学院,长春130062

出　　处：《植物保护》2017年第2期141-145,共5页Plant Protection

基　　金：山东省科技发展计划项目(2014GNC111008);山东省自然科学基金(ZR2015CQ022)

摘　　要：2015-2016年间,在山西省、内蒙古自治区、辽宁省、吉林省采集到疑似感染番茄褪绿病毒Tomato chlorosis virus(ToCV)的番茄植株。利用扩增ToCV外壳蛋白(coat protein,CP)和类热激蛋白(heat shock protein 70homolog,HSP70h)基因片段的两对特异性引物,通过RT-PCR对疑似样品进行分子检测,得到970bp和1 864bp的特异条带,经测序、比对确定为ToCV。序列分析表明,山西晋中分离物SXJZ(KX853540)的CP核苷酸序列与河南安阳分离物HNAYHX(KP264983)相似性为99.7%,内蒙古呼和浩特分离物NMHHHT(KU204709)和辽宁大连分离物LNDL(KU204707)的CP核苷酸序列与国内已报道的山东寿光分离物SDSG(KC709510)相似性分别为99.5%和99.7%,吉林长春分离物JLCC(KU306111)的CP核苷酸序列与山东聊城分离物SDLC(KC812622)的相似性为99.8%。而HSP70h核苷酸序列的相似性分析表明,山西晋中分离物SXJZ(KX853539)与日本分离物Tochigi(AB513442)相似性为99.4%,内蒙古呼和浩特分离物NMHHHT(KU204710)、辽宁大连分离物LNDL(KU204708)和吉林长春分离物JLCC(KX880384)与山东寿光分离物SDSG(KC709510)相似性分别为99.7%、99.8%和99.7%。试验结果明确了番茄褪绿病毒已蔓延传播到我国山西、内蒙古及东北地区。During 2015 and 2016,tomato plant samples with symptoms similar to Tomato chlorosis virus（ToCV）infection were collected from field in Shanxi,Liaoning,Jilin provinces and Inner Mongolia Autonomous Region.The specific primers for coat protein gene（CP）and heat shock protein70 homolog（HSP70h）gene of ToCV were applied to perform RT-PCR detection.Two specific fragments with length of 970 bp and 1 864 bp respectively,were obtained and sequenced.BLAST results confirmed the presence of ToCV in these samples.The similarity analysis showed that the nucleotide sequence of CP from the isolate SXJZ（KX853540）shared 99.7% sequence identity with reported isolate HNAYHX（KP264983,Anyang,Henan）;the isolate NMHHHT（KU204709,Hohhot,Inner Mongolia）and LNDL（KU204707,Dalian,Liaoning）shared 99.5% and 99.7% sequence identity with reported isolate SDSG（KC709510,Shouguang,Shandong）,respectively.And the nucleotide sequence of CP of the isolate JLCC（KU306111,Changchun,Jilin）shared 99.8% sequence identity with isolate SDLC（KC812622,Liaocheng,Shandong）.As for HSP70 h,the nucleotide sequence of HSP70 hfrom the isolate SXJZ（KX853539,Jinzhong,Shanxi）shared 99.4% sequence identity with isolate Tochigi（AB513442,Japan）.And the isolate NMHHHT（KU204710）,isolate LNDL（KU204708）and isolate JLCC（KX880384）shared 99.7%,99.8% and 99.7% nucleotide sequence identity with isolate SDSG（KC709510）,respectively.These results confirmed that the ToCV has spreaded to the Shanxi,the Inner Mongolia and the northeast of China.

关 键 词：番茄褪绿病毒 CP HSP70h RT-PCR 

分 类 号：S432.412[农业科学—植物病理学]