天津地区番茄褪绿病毒的分子检测和鉴定

doi:10.7668/hbnxb.2015.03.036

摘要/Abstract

摘要： 为明确2014年夏天在天津番茄种植区采集到疑似感染番茄褪绿病毒的番茄样品的侵染病原,利用ToCV外壳蛋白和热激蛋白的特异引物进行反转录PCR (RT-PCR)检测,分别扩增得到特异核苷酸片段。序列分析表明,HSP70核苷酸序列与已登录的番茄褪绿病毒日本分离物Tochigi(AB513442)相似性为99.5%,CP氨基酸和核苷酸序列与已登录的番茄褪绿病毒日本分离物Tochigi(AB513443)相似性分别为99.2%和99.3%,表明天津地区的番茄受到ToCV侵染,且天津与日本ToCV分离物序列相似性最高。由于调查地区番茄黄化曲叶病毒发生普遍,针对ToCV阳性样品利用TYLCV特异性引物进行了分子检测,扩增产物序列与TYLCV以色列株系分离物的相似性均在99.0%以上,表明2种病毒在田间发生复合侵染,该研究首次明确天津地区的番茄受到ToCV侵染和ToCV与TYLCV的复合侵染。

关键词: 番茄褪绿病毒, 番茄黄化曲叶病毒, 复合侵染, 分子检测

Abstract: In the summer of 2014,tomato plants showed suspected symptoms caused by Tomato chlorosis virus (ToCV) infection collected in greenhouse in Tianjin.In order to clarify the Pathogen of tomato plants,specific primer pairs of ToCV coat protein (CP) and heat shock protein 70 (HSP70) were used for reversing transcription PCR (RT-PCR) detection,specific fragments were obtained and sequenced.HSP70 nucleotide sequences obtained 99.5% similarity with ToCV isolate Tochigi (AB513442),CP amino acid and nucleotide sequences obtained shared 99.2% and 99.3% similarity with ToCV isolate Tochigi (AB513443),indicating the tomato was infected by ToCV in Tianjin,the sequences of ToCV isolate between Tianjin and Japan with a highest similarity.As Tomato yellow leaf curl virus (TYLCV) were widespread in survey areas,the ToCV positive samples were also tested for TYLCV by specific primers.The result was positive and gene sequence analysis showed that the similarity between amplification products and TYLCV-IL isolates was more than 99.0%,indicating mixed infections in the field.It is the first report that ToCV infect tomato,ToCV and TYLCV mixed infections tomato in Tianjin.

Key words: Tomato chlorosis virus, Tomato yellow leaf curl virus, Mixed infections, Molecular detection

中图分类号:

S436.412.1+1

高利利, 孙国珍, 王勇, 高苇, 张春祥, 张安胜, 竺晓平. 天津地区番茄褪绿病毒的分子检测和鉴定[J]. 华北农学报, 2015, 30(3): 211-215. doi: 10.7668/hbnxb.2015.03.036.

GAO Li-li, SUN Guo-zhen, WANG Yong, GAO Wei, ZHANG Chun-xiang, ZHANG An-sheng, ZHU Xiao-ping. Molecular Detection and Identification of Tomato chlorosis virus in Tianjin[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(3): 211-215. doi: 10.7668/hbnxb.2015.03.036.

http://www.hbnxb.net/CN/Y2015/V30/I3/211

参考文献

[1] Wisler G C,Li R H,Liu H Y,et al. Tomato chlorosis virus :A new whitefly-transmitted,phloem-limited,bipartite closterovirus of tomato[J].Phytopathology,1998,88(5):402-409.
[2] Accotto G P,Vaira A M,Vecchiati M,et al.First report of Tomato chlorosis virus in Italy[J].Plant Disease,2001,85(11):1208.
[3] Dalmon A,Bouyer S,Cailly M,et al.First report of Tomato chlorosis virus and Tomato infectious chlorosis virus in tomato crops in France[J].Plant Disease,2005,89(11):1243-1243.
[4] Alvarez-Ruiz P,Jimenez C G,Leyva-López N E,et al.First report of Tomato chlorosis virus infecting tomato crops in Sinaloa Mexico[J].Plant Pathology,2007,56(6):1043.
[5] Martínez-Zubiaur Y,Fiallo-Olivé E,Carrillo-Tripp J,et al.First report of Tomato chlorosis virus infecting tomato in singleand mixed infections with Tomato yellow leaf curl virus in Cuba[J].Plant Disease,2008,92(5):836.
[6] Barbosa J C,Teixeira A P M,Moreira A G,et al.First report of Tomato chlorosis virus infecting tomato crops in Brazil[J].Plant Disease,2008,92(12):1709.
[7] Fiallo-OlivéE,Hamed A A,Moriones E,et al.First report of Tomato chlorosis virus infecting tomato in Sudan[J].Plant Disease,2011,95(12):1592.
[8] Segev L,Wintermantel W M,Polston J E,et al.First report of Tomato chlorosis virus in Israel[J].Plant Disease,2004,88(10):1160.
[9] Cevik B,Erkiβ G.First report of Tomato chlorosis virus in Turkey[J].Plant Pathology,2008,57(4):767.
[10] Hirota T,Natsuaki T,Murai T,et al.Yellowing disease of tomato caused by Tomato chlorosis virus newly recognized in Japan[J].Journal of General Plant Pathology,2010,76(2):168-171.
[11] Tsai W S,Shih S L,Green S K,et al.First report of the occurrence of Tomato chlorosis virus and Tomato infecttions chlorosis virus in Taiwan[J].Plant Disease,2004,88(3):311.
[12] Zhao R N,Wang R,Wang N,et al.First report of Tomato chlorosis virus in China[J].Plant Disease,2013,97(8):1123.
[13] Karwitha M,Feng Z,Yao M,et al.The complete nucleotide sequence of the RNA1 of a Chinese isolate of Tomato chlorosis virus[J].Journal of Phytopathology,2014,162(6):411-415.
[14] Zhao L M,Li G,Gao Y,et al.Molecular detection and complete genome sequences of Tomato chlorosis virus isolates from infectious outbreaks in China[J].Journal of Phytopathology,2014,162(10):627-634.
[15] 刘永光,魏家鹏,乔宁,等.番茄褪绿病毒在山东暴发及其防治措施[J].中国蔬菜,2014(5):67-69.
[16] 周涛,杨普云,赵汝娜,等.警惕番茄褪绿病毒在我国的传播和危害[J].植物保护,2014,40(5):196-199.
[17] Papayiannisa L C,Harkoua I S,Markoua Y M,et al.Rapid discrimination of Tomato chlorosis virus,Tomato infectious chlorosis virus and co-amplification of plant internal control using real-time RT-PCR[J].Journal of Virological Methods,2011,176:53-59.
[18] Zhao L M,Li G,Gao Ying,et al.Reverse transcription loop-mediated isothermal amplification assay for detecting Tomato chlorosis virus [J].Journal of Virological Methods,2015,213:93-97.
[19] Jacquemond M,Verdin E,Dalmon A,et al.Serological and molecular detection of Tomato chlorosis virus and Tomato infectious chlorosis virus in tomato[J].Plant Pathology,2009,58:210-220.
[20] Harrison B D,Liu Y L,Khalid S,et al.Detection and relationships of cotton leaf curl virus and allied whitefly-transmitted geminiviruses occurring in Pakistan[J].Annals of Applied Biology,1997,130(1):61-75.
[21] 赵黎明,李刚,刘永光,等.番茄褪绿病毒与番茄黄化曲叶病毒复合侵染的分子鉴定[J].中国蔬菜,2014,12:15-20.
[22] Wintrmantel W M,Wisler G C.Vector specificity,host range,and genetic diversity of Tomato chlorosis virus[J].Plant Disease,2006,90(6):814-819.
[23] 赵黎明,李刚,刘永杰,等.侵染番茄的番茄褪绿病毒山东泰安分离物的分子鉴定和序列分析[J].植物保护,2014,40(5):34-39.
[24] 赵汝娜,王蓉,师迎春,等.侵染甜椒的番茄褪绿病毒的分子鉴定[J].植物保护,2014,40(1):128-130.
[25] Freitas D M S,Nardin I,Shimoyama N,et al.First report of Tomato chlorosis virus in potato in Brazil[J].European Journal of Plant Pathology,2012,134(1):81-86.
[26] 李洁,李慧,丁天波,等.胶东半岛地区番茄褪绿病毒的快速检测与鉴定[J].山东农业科学,2015,47(2):86-89.
[27] 孙娜,李岩,魏珉,等.补光对日光温室越冬番茄生长及产量品质的影响[J].天津农业科学,2014,20(3):91-93,96.
[28] 郝永娟,王万立,金凤媚,等.天津市番茄黄化曲叶病毒病的发生与防治[J].天津农业科学,2010,16(2):48-50.

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