云南番茄褪绿病毒和番茄黄化曲叶病毒复合侵染的分子鉴定
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摘要
在云南省元谋县发现大量下部叶片褪绿黄化,顶叶卷曲畸形的番茄植株。将叶片采集带回实验室,采用番茄褪绿病毒(Tomato chlorosis virus,To CV)和番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)的特异性引物分别扩增得到774和400 bp的目标条带,测序比对发现其与To CV的KR184676.1和TYLCV的FJ012359.1序列相似度均为99%,确认这两条带对应的两种病毒分别为To CV和TYLCV,明确该番茄植株被To CV和TYLCV复合侵染。同时发现云南地区有B型和Q型两种烟粉虱存在,且Q型的比例高于B型,二者均可携带To CV和TYLCV。To CV首次在云南省发现,其由沿海向内陆蔓延的过程中与TYLCV的复合侵染已蔓延至西南地区,值得高度重视和警惕。
The tomato leaves that displayed symptoms of the chlorosis and yellowing on the lower leaves and symptoms of the curl and deformity on the upper leaves were found and collected in Yunnan. The target DNA band of Tomato chlorosis virus(To CV)and Tomato yellow leaf curl virus(TYLCV),774 bp and 400 bp,respectively,was amplified using the special primer for virus. The target PCR products were sequenced and compared with the NCBI nucleotide database via the BLAST tools. The result showed that the sample had 99% similarity with the KR184676.1 of To CV and the FJ012359.1 of TYLCV. We can confirm that the tomato was simultaneously infected by the To CV and TYLCV. Meanwhile,we found that in Yunnan Province Bemisia tabaci B and Q coexisted,and the proportion of Q was higher than B. Both of B and Q could carry To CV and TYLCV. This is the first report of To CV in Yunnan Province. In the spread process from coast to interior areas,the co-infection of To CV and TYLCV has transmitted to the Southwest of China,and we should pay high attention to the trend of the virus.
The tomato leaves that displayed symptoms of the chlorosis and yellowing on the lower leaves and symptoms of the curl and deformity on the upper leaves were found and collected in Yunnan. The target DNA band of Tomato chlorosis virus(To CV)and Tomato yellow leaf curl virus(TYLCV),774 bp and 400 bp,respectively,was amplified using the special primer for virus. The target PCR products were sequenced and compared with the NCBI nucleotide database via the BLAST tools. The result showed that the sample had 99% similarity with the KR184676.1 of To CV and the FJ012359.1 of TYLCV. We can confirm that the tomato was simultaneously infected by the To CV and TYLCV. Meanwhile,we found that in Yunnan Province Bemisia tabaci B and Q coexisted,and the proportion of Q was higher than B. Both of B and Q could carry To CV and TYLCV. This is the first report of To CV in Yunnan Province. In the spread process from coast to interior areas,the co-infection of To CV and TYLCV has transmitted to the Southwest of China,and we should pay high attention to the trend of the virus.
引文
Barbosa J C,Teixeira A P M,Moreira A G,Camargo L E A,Bergamin Filho A,Kitajima E W,Rezende J A M.2008.First report of Tomato chlorosis virus infecting tomato crops in Brazil.Plant Disease,92(12):1709-1709.
Dietzgen R G,Mann K S,Johnson K N.2016.Plant virus-insect vector interactions:current and potential future research directions.Viruses,8(11):303.
Fang Y,Jiao X,Xie W,Wang S,Wu Q,Shi X,Chen G,Su Q,Yang X,Pan H P,Zhang Y.2013.Tomato yellow leaf curl virus alters the host preferences of its vector Bemisia tabaci.Scientific Reports,3:2876.
García-Cano E,Resende R O,Fernández-Mu?oz R,Moriones E.2006.Synergistic interaction between Tomato chlorosis virus and Tomato spotted wilt virus results in breakdown of resistance in tomato.Phytopathology,96:1263-1269.
Gilbertson R L,Batuman O,Webster C G,Adkins S.2015.Role of the insect supervectors Bemisia tabaci and Frankliniella occidentalis in the emergence and global spread of plant viruses.Annual Review of Virology,2(1):67-93.
Gil-Salas F M,Petersb J,Boonhamb N,Cuadradoa I M,Janssen D.2012.Co-infection with Cucumber vein yellowing virus and Cucurbit yellow stunting disorder virus leading to synergism in cucumber.Plant Pathology,61:468-478.
Hanssen I M,Lapidot M,Thomma B P H J.2010.Emerging viral diseases of tomato crops.Molecular Plant-Microbe Interactions,23(23):539-548.
Hirota T,Natsuaki T,Murai T,Nishigawa H,Niibori K,Goto K,Hartono S,Suastika G,Okuda S.2010.Yellowing disease of tomato caused by Tomato chlorosis virus newly recognized in Japan.Journal of General Plant Pathology,76(2):168-171.
Kil E J,Lee Y J,Cho S,Auh C K,Kim D,Lee K Y,Kim M K,Choi H S,Kim C S,Lee S.2015.Identification of natural weed hosts of Tomato chlorosis virus in Korea by RT-PCR with root tissues.European Journal of Plant Pathology,142(2):419-426.
Lozano G,Moriones E,Navas-Castillo J.2006.Complete nucleotide sequence of the RNA2 of the Crinivirus Tomato chlorosis virus.Archives of Virology,151(3):581-587.
Lozano G,Moriones E,Navas-Castillo J.2007.Complete sequence of the RNA1 of a European isolate of Tomato chlorosis virus.Archives of Virology,152(4):839-841.
Moreno-Delafuente,Garzo A E,Moreno A,Fereres A.2013.A plant virus manipulates the behavior of its whitefly vector to enhance its transmission efficiency and spread.PLo S ONE,8(4):e61543.
Orfanidou C G,Pappi P G,Efthimiou K E,Katis N I,Maliogka V I.2016.Transmission of Tomato chlorosis virus(To CV)by Bemisia tabaci biotype Q and evaluation of four weed species as viral sources.Plant Disease,100(10):2043-2049.
Pan H P,Preisser E L,Chu D,Wang S L,Wu Q J,Carriere Y,Zhou X G,Zhang Y J.2015.Insecticides promote viral outbreaks by altering herbivore competition.Insecticides promote viral outbreaks by altering herbivore competition.Ecological Applications,25(6):1585-1595.
PicóBelén,Díez María José,Nuez Fernando.1996.Viral diseases causing the greatest economic losses to the tomato crop II.The Tomato yellow leaf curl virus-a review.Scientia Horticulturae,67(3-4):151-196.
Segev L,Wintermantel W M,Polston J E,Lapidot M.2004.First report of Tomato chlorosis virus in Israel.Plant Disease,88(10):1160.
Shi X B,Chen G,Tian L X,Peng Z K,Xie W,Wu Q J,Wang S L,Zhou X G,Zhang Y J.2016.The salicylic acid-mediated release of plant volatiles affects the host choice of Bemisia tabaci.International Journal of Molecular Sciences,17(7):1048.
Shi X B,Pan H P,Xie W,Jiao X G,Fang Y,Chen G,Yang X,Wu Q J,Wang S L,Zhang Y J.2014a.Three-way interactions between the tomato plant,Tomato yellow leaf curl virus,and Bemisia tabaci(Hemiptera:Aleyrodidae)facilitate virus spread.Journal of Economic Entomology,107(3):920-926.
Shi X B,Pan H P,Xie W,Wu Q J,Wang S L,Liu Y,Fang Y,Chen G,Gao X,Zhang Y J.2013.Plant virus differentially alters the plant’s defense response to its closely related vectors.PLo S ONE,8(12):e83520.
Shi X B,Pan H P,Zhang H,Jiao X G,Xie W,Wu Q J,Wang S L,Fang Y,Chen G,Zhou X G.2014b.Bemisia tabaci Q carrying Tomato yellow leaf curl virus strongly suppresses host plant defenses.Scientific Reports,4:5230.
Song Xi,Shi Ying-chun,Zhang Shi-chen,Liang Yan,Wang Nian,Chen Shan-yi,Zhou Tao.2013.Determination of Tomato pseudosciaena crocea virus isolate in Beijing and preliminary identification of its strains.Journal of Plant Pathology,43(2):113-119.(in Chinese)宋晰,师迎春,张世晨,梁彦,王廿,陈善义,周涛.2013.北京地区番茄黄化曲叶病病毒分离物测定及株系的初步鉴定.植物病理学报,43(2):113-119.
Sun X H,Gao L L,Wang C L,Wang S L,Zhao J,Qiao N,Zhu X P.2016.Natural occurrence of Tomato chlorosis virus in pumpkin in China.Plant Disease,100(12):2543.
Tang X,Shi X B,Zhang D Y,Li F,Yan F,Zhang Y J,Liu Y,Zhou X G.2017.Detection and epidemic dynamic of To CV and CCYV with Bemisia tabaci and weed in Hainan of China.Virology Journal,14:169.
Tsai W,Shih S,Green S,Hanson P,Liu H.2004.First report of the occurrence of Tomato chlorosis virus and Tomato infectious chlorosis virus in Taiwan.Plant Disease,88(3):311.
Wintermantel W M,Wisler G C.2006.Vector specificity,host range,and genetic diversity of Tomato chlorosis virus.Plant Disease,90(6):814-819.
Wintermantel W M,Wisler G C,Anchieta A G,Liu H Y,Karasev A V,Tzanetakis I E.2005.The complete nucleotide sequence and genome organization of Tomato chlorosis virus.Archives of Virology,150(11):2287-2298.
Wisler G C,Li R H,Liu H Y,Lowry D S,Duffus J E.1998.Tomato chlorosis virus:a new whitefly-transmitted,phloem-limited,bipartite closterovirus of tomato.Phytopathology,88(5):402-409.
Wu J B,Dai F M,Zhu X P.2006.First report of Tomato yellow leaf curl virus in China.Plant Disease,90(10):1359.
Wu Shu-hua,Li Ting-fang,Zhao Wen-hao,Cheng Zhao-bang,Guo Qing-yun,Zhao Tong-min,Yu Wen-gui,Zhu Ye-qin,Ji Ying-hua.2016.Molecular detection of Tomato chlorosis virus and Tomato yellow leaf curl virus complex infection in Jiangsu Province.Acta Horticulturae Sinica,43(1):89-99.(in Chinese)吴淑华,李廷芳,赵文浩,程兆榜,郭青云,赵统敏,余文贵,朱叶芹,季英华.2016.江苏省番茄黄化曲叶病毒和褪绿病毒复合侵染的分子检测.园艺学报,43(1):89-99.
Zhao R N,Wang R,Wang N,Fan Z,Zhou T,Shi Y,Chai M.2013.First report of Tomato chlorosis virus in China.Plant Disease,100(10):2043-2049.
Zhao L M,Li G,Gao Y,Liu Y J,Sun G Z,Zhu X P.2014.Molecular detection and complete genome sequences of Tomato chlorosis virus isolates from infectious outbreaks in China.Journal of Phytopathology,162:627-634.
Zhao Li-ming,Li Gang,Liu Yong-guang,Guo Jia-jin,Wei Jia-peng,Zhu Xiao-ping.2014.Molecular identification of Tomato chlorosis virus and Tomato yellow leaf curl virus complex infection.China Vegetables,(12):15-20.(in Chinese)赵黎明,李刚,刘永光,国家进,魏家鹏,竺晓平.2014.番茄褪绿病毒与番茄黄化曲叶病毒复合侵染的分子鉴定.中国蔬菜,(12):15-20.
Zheng Hui-xin,Xia Ji-xing,Zhou Xiao-mao,Zhang You-jun.2016.Vigilance of Bemisia tabaci transmission of Tomato chlorosis virus disease in China rapid spread.China Vegetables,(4):22-26.(in Chinese)郑慧新,夏吉星,周小毛,张友军.2016.警惕烟粉虱传播的番茄褪绿病毒病在我国快速扩散.中国蔬菜,(4):22-26.
Zhou Tao,Yang Pu-yun,Zhao Ru-na,Shi Ying-chun,Yuan Kai,Fan Zai-feng.2014.Be alert to the spread and harm of Tomato chlorotic virus in China.Plant Protection,40(5):196-199.(in Chinese)周涛,杨普云,赵汝娜,师迎春,原锴,范在丰.2014.警惕番茄褪绿病毒在我国的传播和危害.植物保护,40(5):196-199.
Dietzgen R G,Mann K S,Johnson K N.2016.Plant virus-insect vector interactions:current and potential future research directions.Viruses,8(11):303.
Fang Y,Jiao X,Xie W,Wang S,Wu Q,Shi X,Chen G,Su Q,Yang X,Pan H P,Zhang Y.2013.Tomato yellow leaf curl virus alters the host preferences of its vector Bemisia tabaci.Scientific Reports,3:2876.
García-Cano E,Resende R O,Fernández-Mu?oz R,Moriones E.2006.Synergistic interaction between Tomato chlorosis virus and Tomato spotted wilt virus results in breakdown of resistance in tomato.Phytopathology,96:1263-1269.
Gilbertson R L,Batuman O,Webster C G,Adkins S.2015.Role of the insect supervectors Bemisia tabaci and Frankliniella occidentalis in the emergence and global spread of plant viruses.Annual Review of Virology,2(1):67-93.
Gil-Salas F M,Petersb J,Boonhamb N,Cuadradoa I M,Janssen D.2012.Co-infection with Cucumber vein yellowing virus and Cucurbit yellow stunting disorder virus leading to synergism in cucumber.Plant Pathology,61:468-478.
Hanssen I M,Lapidot M,Thomma B P H J.2010.Emerging viral diseases of tomato crops.Molecular Plant-Microbe Interactions,23(23):539-548.
Hirota T,Natsuaki T,Murai T,Nishigawa H,Niibori K,Goto K,Hartono S,Suastika G,Okuda S.2010.Yellowing disease of tomato caused by Tomato chlorosis virus newly recognized in Japan.Journal of General Plant Pathology,76(2):168-171.
Kil E J,Lee Y J,Cho S,Auh C K,Kim D,Lee K Y,Kim M K,Choi H S,Kim C S,Lee S.2015.Identification of natural weed hosts of Tomato chlorosis virus in Korea by RT-PCR with root tissues.European Journal of Plant Pathology,142(2):419-426.
Lozano G,Moriones E,Navas-Castillo J.2006.Complete nucleotide sequence of the RNA2 of the Crinivirus Tomato chlorosis virus.Archives of Virology,151(3):581-587.
Lozano G,Moriones E,Navas-Castillo J.2007.Complete sequence of the RNA1 of a European isolate of Tomato chlorosis virus.Archives of Virology,152(4):839-841.
Moreno-Delafuente,Garzo A E,Moreno A,Fereres A.2013.A plant virus manipulates the behavior of its whitefly vector to enhance its transmission efficiency and spread.PLo S ONE,8(4):e61543.
Orfanidou C G,Pappi P G,Efthimiou K E,Katis N I,Maliogka V I.2016.Transmission of Tomato chlorosis virus(To CV)by Bemisia tabaci biotype Q and evaluation of four weed species as viral sources.Plant Disease,100(10):2043-2049.
Pan H P,Preisser E L,Chu D,Wang S L,Wu Q J,Carriere Y,Zhou X G,Zhang Y J.2015.Insecticides promote viral outbreaks by altering herbivore competition.Insecticides promote viral outbreaks by altering herbivore competition.Ecological Applications,25(6):1585-1595.
PicóBelén,Díez María José,Nuez Fernando.1996.Viral diseases causing the greatest economic losses to the tomato crop II.The Tomato yellow leaf curl virus-a review.Scientia Horticulturae,67(3-4):151-196.
Segev L,Wintermantel W M,Polston J E,Lapidot M.2004.First report of Tomato chlorosis virus in Israel.Plant Disease,88(10):1160.
Shi X B,Chen G,Tian L X,Peng Z K,Xie W,Wu Q J,Wang S L,Zhou X G,Zhang Y J.2016.The salicylic acid-mediated release of plant volatiles affects the host choice of Bemisia tabaci.International Journal of Molecular Sciences,17(7):1048.
Shi X B,Pan H P,Xie W,Jiao X G,Fang Y,Chen G,Yang X,Wu Q J,Wang S L,Zhang Y J.2014a.Three-way interactions between the tomato plant,Tomato yellow leaf curl virus,and Bemisia tabaci(Hemiptera:Aleyrodidae)facilitate virus spread.Journal of Economic Entomology,107(3):920-926.
Shi X B,Pan H P,Xie W,Wu Q J,Wang S L,Liu Y,Fang Y,Chen G,Gao X,Zhang Y J.2013.Plant virus differentially alters the plant’s defense response to its closely related vectors.PLo S ONE,8(12):e83520.
Shi X B,Pan H P,Zhang H,Jiao X G,Xie W,Wu Q J,Wang S L,Fang Y,Chen G,Zhou X G.2014b.Bemisia tabaci Q carrying Tomato yellow leaf curl virus strongly suppresses host plant defenses.Scientific Reports,4:5230.
Song Xi,Shi Ying-chun,Zhang Shi-chen,Liang Yan,Wang Nian,Chen Shan-yi,Zhou Tao.2013.Determination of Tomato pseudosciaena crocea virus isolate in Beijing and preliminary identification of its strains.Journal of Plant Pathology,43(2):113-119.(in Chinese)宋晰,师迎春,张世晨,梁彦,王廿,陈善义,周涛.2013.北京地区番茄黄化曲叶病病毒分离物测定及株系的初步鉴定.植物病理学报,43(2):113-119.
Sun X H,Gao L L,Wang C L,Wang S L,Zhao J,Qiao N,Zhu X P.2016.Natural occurrence of Tomato chlorosis virus in pumpkin in China.Plant Disease,100(12):2543.
Tang X,Shi X B,Zhang D Y,Li F,Yan F,Zhang Y J,Liu Y,Zhou X G.2017.Detection and epidemic dynamic of To CV and CCYV with Bemisia tabaci and weed in Hainan of China.Virology Journal,14:169.
Tsai W,Shih S,Green S,Hanson P,Liu H.2004.First report of the occurrence of Tomato chlorosis virus and Tomato infectious chlorosis virus in Taiwan.Plant Disease,88(3):311.
Wintermantel W M,Wisler G C.2006.Vector specificity,host range,and genetic diversity of Tomato chlorosis virus.Plant Disease,90(6):814-819.
Wintermantel W M,Wisler G C,Anchieta A G,Liu H Y,Karasev A V,Tzanetakis I E.2005.The complete nucleotide sequence and genome organization of Tomato chlorosis virus.Archives of Virology,150(11):2287-2298.
Wisler G C,Li R H,Liu H Y,Lowry D S,Duffus J E.1998.Tomato chlorosis virus:a new whitefly-transmitted,phloem-limited,bipartite closterovirus of tomato.Phytopathology,88(5):402-409.
Wu J B,Dai F M,Zhu X P.2006.First report of Tomato yellow leaf curl virus in China.Plant Disease,90(10):1359.
Wu Shu-hua,Li Ting-fang,Zhao Wen-hao,Cheng Zhao-bang,Guo Qing-yun,Zhao Tong-min,Yu Wen-gui,Zhu Ye-qin,Ji Ying-hua.2016.Molecular detection of Tomato chlorosis virus and Tomato yellow leaf curl virus complex infection in Jiangsu Province.Acta Horticulturae Sinica,43(1):89-99.(in Chinese)吴淑华,李廷芳,赵文浩,程兆榜,郭青云,赵统敏,余文贵,朱叶芹,季英华.2016.江苏省番茄黄化曲叶病毒和褪绿病毒复合侵染的分子检测.园艺学报,43(1):89-99.
Zhao R N,Wang R,Wang N,Fan Z,Zhou T,Shi Y,Chai M.2013.First report of Tomato chlorosis virus in China.Plant Disease,100(10):2043-2049.
Zhao L M,Li G,Gao Y,Liu Y J,Sun G Z,Zhu X P.2014.Molecular detection and complete genome sequences of Tomato chlorosis virus isolates from infectious outbreaks in China.Journal of Phytopathology,162:627-634.
Zhao Li-ming,Li Gang,Liu Yong-guang,Guo Jia-jin,Wei Jia-peng,Zhu Xiao-ping.2014.Molecular identification of Tomato chlorosis virus and Tomato yellow leaf curl virus complex infection.China Vegetables,(12):15-20.(in Chinese)赵黎明,李刚,刘永光,国家进,魏家鹏,竺晓平.2014.番茄褪绿病毒与番茄黄化曲叶病毒复合侵染的分子鉴定.中国蔬菜,(12):15-20.
Zheng Hui-xin,Xia Ji-xing,Zhou Xiao-mao,Zhang You-jun.2016.Vigilance of Bemisia tabaci transmission of Tomato chlorosis virus disease in China rapid spread.China Vegetables,(4):22-26.(in Chinese)郑慧新,夏吉星,周小毛,张友军.2016.警惕烟粉虱传播的番茄褪绿病毒病在我国快速扩散.中国蔬菜,(4):22-26.
Zhou Tao,Yang Pu-yun,Zhao Ru-na,Shi Ying-chun,Yuan Kai,Fan Zai-feng.2014.Be alert to the spread and harm of Tomato chlorotic virus in China.Plant Protection,40(5):196-199.(in Chinese)周涛,杨普云,赵汝娜,师迎春,原锴,范在丰.2014.警惕番茄褪绿病毒在我国的传播和危害.植物保护,40(5):196-199.