新疆喀什辣椒轻斑驳病毒和黄瓜花叶病毒复合侵染的分子鉴定

doi:10.7668/hbnxb.20191552

摘要/Abstract

摘要： 2018-2019年，在新疆喀什地区调查时发现部分种植的辣椒植株上心叶发黄皱缩、卷曲，个别植株叶片呈现斑驳花叶状。为了明确引起新疆喀什辣椒病毒病的病毒种类，提取典型症状样品的总RNA，反转录得到cDNA，分别用辣椒轻斑驳病毒和黄瓜花叶病毒特异性引物进行RT-PCR检测，CMV和PMMoV特异引物分别扩增得到约735，472 bp的特异条带，阴性对照未扩增目的条带，核苷酸序列比对发现，分别与韩国西葫芦CMV分离物（GU327368.1）和日本辣椒PMMoV分离物（AB276030.1）序列同源性最高为95%，99%，结果表明，喀什地区辣椒疑似病毒病样本被CMV与PMMoV复合侵染。

关键词: 辣椒, 黄瓜花叶病毒, 辣椒轻斑驳病毒, 复合侵染, RT-PCR

Abstract: During 2018 to 2019,pepper plants showed typical symptoms of dwarf,yellowing,curling,chlorosis and rolling up of leaves were found in Kashgar,Xinjiang Uygur Autonomous region. To identify the virus that causing pepper virus disease,total RNA was isolated from the diseased tissues with typical virus symptoms,and reverse transcription(RT)-PCR assays were performed with specific primers for Cucumber mosaic virus (CMV)and Pepper mild mottle virus (PMMoV). The expected 735 bp CMV and 472 bp PMMoV fragments were amplified from diseased pepper tissue samples,respectively. Sequence analysis showed 95%,99% similarity with that of CMV(South Korea,GU327368.1)and PMMoV(Japan,AB276030.1),respectively. The results showed that pepper plants were co-infected by CMV and PMMoV in Kashgar,Xinjiang region.

Key words: Pepper, Cucumber mosaic virus, Pepper mild mottle virus, Co-infection, RT-PCR

中图分类号:

S432.4

柴阿丽, 陈利达, 许帅, 帕提古丽·艾斯木托拉, 王利丽, 石延霞, 谢学文, 李磊, 李宝聚. 新疆喀什辣椒轻斑驳病毒和黄瓜花叶病毒复合侵染的分子鉴定[J]. 华北农学报, 2020, 35(S1): 332-337. doi: 10.7668/hbnxb.20191552.

CHAI Ali, CHEN Lida, XU Shuai, PATIGULI Aisimutuola, WANG Lili, SHI Yanxia, XIE Xuewen, LI Lei, LI Baoju. Molecular Identification of Co-infection of Pepper mild mottle virus and Cucumber mosaic virus on Pepper in Kashgar, Xinjiang[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(S1): 332-337. doi: 10.7668/hbnxb.20191552.

http://www.hbnxb.net/CN/Y2020/V35/IS1/332

参考文献

[1] Koeda S, Homma K, Kamitani M, Mari Kamitani, Nagano A J, Taniguchi M, Pohan N, Kesumawati E. Pepper vein yellows virus 9:a novel polerovirus isolated from chili pepper in Indonesia[J]. Archives of Virology Volume,2020,165(12):3017-3021. doi:10.1007/s00705-020-04838-6.
[2] Liu M Y,Liu X N,Li X,Zhang D Y,Dai L Y,Tang Q J. Complete genome sequence of a Chinese isolate of Pepper vein yellows virus and evolutionary analysis based on the CP,MP and RdRp coding regions[J]. Archives of Virology,2016,161(3):677-683. doi:10.1007/s00705-015-2691-9.
[3] Li Y Y,Wang Y,Hu J,Xiao L,Tan G L,Lan P X,Liu Y,Li F. The complete genome sequence,occurrence and host range of Tomato mottle mosaic virus Chinese isolate[J]. Virology Journal,2017,14:15. doi:10.1186/s12985-016-0676-2.
[4] 冯耿,辛敏,曹孟籍,王莉爽,李莉,王锡锋. 深度测序发现贵阳发生的辣椒病毒病由多种病毒复合侵染所致[J]. 植物病理学报, 2017,47(5):591-597. doi:10.13926/j.cnki.apps.000113.
Feng G,Xin M,Cao M J,Wang L S,Li L,Wang X F. Identification of multiple viruses infecting hot pepper in Guiyang by deep sequencing[J]. Acta Phytopathologica Sinica,2017,47(5):591-597.
[5] 赵尊练,史联联,谭根堂,严小良. 陕西省辣椒主产区辣椒病毒病病原种类鉴定及其分布研究[J]. 中国农业科学,2004,37(11):1738-1742. doi:10.3321/j.issn:0578-1752.2004.11.028.
Zhao Z L, Shi L L, Tan G T, Yan X L. Study on pepper (Capsicum annuum L.) virus disease and its distribution in the Guanzhong area of Shaanxi Province[J].Scientia Agricultura Sinica,2004,37(11):1738-1742.
[6] 陈青,廖富荣,陈红运,谢毅璇,陈加福,蔡金镭,林石明. 辣椒上烟草轻型绿花叶病毒的鉴定[J]. 植物病理学报,2013,43(6):651-654. doi:10.13926/j.cnki.apps.2013.06.012.
Chen Q,Liao F R,Chen H Y,Xie Y X,Chen J F,Cai J L,Lin S M. Identification of Tobacco mild green mosaic virus infecting pepper[J]. Acta Phytopathologica Sinica,2013,43(6):651-654.
[7] 龚明霞,赵虎,王萌,吴星,赵曾菁,黄金梅,何志,王日升. 广西辣椒病毒病调查及病原种类初步鉴定[J]. 中国蔬菜, 2020(4):74-79.
Gong M X,Zhao H,Wang M,Wu X,Zhao Z J,Huang J M,He Z,Wang R S. Investigation and preliminary identification of pathogeny species of pepper virus disease in Guangxi[J]. China Vegetables,2020(4):74-79.
[8] 姚玉荣,陈国华,冯兰香,谢丙炎. 北运蔬菜基地辣椒病毒病病原种类的分子检测[J]. 中国蔬菜,2013(10):84-89. doi:10.3969/j.issn.1000-6346.2013.10.014.
Yao Y R,Chen G H,Feng L X,Xie B Y. Molecular detection of pepper viruses in southern vegetable production bases[J]. China Vegetables, 2013(10):84-89.
[9] 郭思瑶,童艳,黄娅,罗信福,青玲. 重庆辣椒病毒病病原初步鉴定和分析[J]. 园艺学报,2015,42(2):263-270. doi:10.16420/j.issn.0513-353x.2014-0774.
Guo S Y,Tong Y,Huang Y,Luo X F,Qing L. Preliminary identification and analyses of viruses causing pepper virus disease in Chongqing,China[J]. Acta Horticulturae Sinica,2015,42(2):263-270.
[10] 高苇,王勇,张春祥,张安盛,竺晓平. 天津地区辣椒病毒病调查及毒源种类初步鉴定[J]. 山东农业科学,2016,48(3):91-94. doi:10.14083/j.issn.1001-4942.2016.03.023.
Gao W,Wang Y,Zhang C X,Zhang A S,Zhu X P. Investigation and pathogen preliminary identification of pepper virus disease in Tianjin[J]. Shandong Agricultural Sciences,2016,48(3):91-94.
[11] 王少立,谭玮萍,杨园园,代惠洁,孙晓辉,乔宁,竺晓平. 山东省辣椒主要病毒种类的分子检测与鉴定[J]. 中国农业科学,2017,50(14):2728-2738. doi:10.3864/j.issn.0578-1752.2017.14.009.
Wang S L,Tan W P,Yang Y Y,Dai H J,Sui X H,Qiao N,Zhu X P. Molecular detection and identification of main viruses on pepper in Shandong Province[J]. Scientia Agricultural Sciences,2017,50(14):2728-2738.
[12] Ge B B, Liu G J, Wang H Q. First report of Tomato mosaic virus infecting pepino in China[J]. Plant disease,2012,96(11):1704. doi:10.1094/PDIS-05-12-0475-PDN.
[13] Niu Y B, Qing L, Yao M, Wang, D F, Liu J D, Wang J S.First report of Tobacco mosaic virus in Abutilon theophrasti in China[J]. Plant Disease,2009,93(11):1221. doi:10.1094/PDIS-93-11-1221B.
[14] 宋婷婷, 陈荣溢, 杨雷亮.应用多重RT-PCR检测烟草上的TMV和CMV[J]. 辽宁农业科学,2007(1):53-54. doi:10.3969/j.issn.1002-1728.2007.01.019.
Song T T,Chen R Y,Yang L L. Detection of TMV and CMV in tobacco by multiplex RT-PCR[J]. Liaoning Agricultural Sciences,2007(1):53-54.
[15] 陈利达,石延霞,谢学文,曹金强,柴阿丽,李宝聚. 我国不同地区番茄主要病毒病种类的分子检测与分析[J]. 华北农学报,2020,35(1):185-193. doi:10.7668/hbnxb.20190230.
Chen L D,Shi Y X,Xie X W,Cao J Q,Chai A L,Li B J. Molecular detection and analyses of main viruses on tomato in China[J]. Acta Agriculturae Boreali-Sinica,2020,35(1):185-193.
[16] Dovas C I,Katis N I,Avgelis A D. Multiplex detection of criniviruses associated with epidemics of a yellowing disease of tomato in Greece[J]. Plant Disease,2002,86(12):1345-1349. doi:10.1094/PDIS.2002.86.12.1345.
[17] 孙琦,张春庆. 马铃薯X病毒的RT-PCR检测[J]. 园艺学报,2003,30(6):687-689. doi:10.3321/j.issn:0513-353X.2003.06.011.
Sun Q,Zhang C Q. Detection of Potato virus X by reverse transcription polymerase chain reaction[J]. Acta Horticulturae Sinica,2003,30(6):687-689.
[18] 刘勇,李凡,李月月,张松柏,高希武,谢艳,燕飞,张安盛,戴良英,程兆榜,丁铭,牛颜冰,王升吉,车海彦,江彤,史晓斌,何自福,吴云锋,张德咏,青玲,严婉荣,杨学辉,汤亚飞,郑红英,唐前君,章松柏,章东方,蔡丽,陶小荣. 侵染我国主要蔬菜作物的病毒种类、分布与发生趋势[J].中国农业科学,2019,52(2):239-261. doi:10.3864/j.issn.0578-1752.2019.02.005.
Liu Y,Li F,Li Y Y,Zhang S B,Gao X W,Xie Y,Yan F,Zhang A S,Dai L Y,Cheng Z B,Ding M,Niu Y B,Wang S J,Che H Y,Jiang T,Shi X B,He Z F,Wu Y F,Zhang D Y,Qing L,Yan W R,Yang X H,Tang Y F,Zheng H Y,Tang Q J,Zhang S B,Zhang D F,Cai L,Tao X R. Identification,distribution and occurrence of viruses in the main vegetables of China[J]. Scientia Agricultura Sinica,2019,52(2):239-261.
[19] 张建云,许文博,刘升学,黄家风. 侵染加工番茄的BBWV2的分子鉴定及田间检测[J]. 石河子大学学报(自然科学版),2013,31(6):675-679. doi:10.3969/j.issn.1007-7383.2013.06.003.
Zhang J Y,Xu W B,Liu S X,Huang J F. ldentification and detection of Broad bean wilt virus 2 (BBWV2)infecting processing tomato[J]. Journal of Shihezi University (Natural Science),2013,31(6):675-679.
[20] 于海龙,张正海,曹亚从,张宝玺,王立浩. 辣椒抗黄瓜花叶病毒病研究进展[J]. 园艺学报,2019,46(9):1813-1824. doi:10.16420/j.issn.0513-353x.2019-0489.
Yu H L,Zhang Z H,Cao Y C,Zhang B X,Wang L H. Progress of resistance to Cucumber mosaic virus in pepper[J]. Acta Horticulturae Sinica,2019,46(9):1813-1824.
[21] 向本春, 谢浩, 崔星明, 李春, 刘素萍, 席德慧, 尹玉琦. 新疆辣椒轻微斑驳病毒的分离鉴定[J]. 病毒学报, 1994, 10(3):240-245. doi:10.13242/j.cnki.bingduxuebao.000860.
Xiang B C, Xie H, Cui X M, Li C, Liu S P, Xi D H, Yin Y Q.Isolation and identification of Pepper mildmottle tobamo virus in Xinjiang[J]. Chinese Journal of Virology, 1994, 10(3):240-245.
[22] 郭广君,刁卫平,刘金兵,潘宝贵,戈伟,王述彬. 辣椒抗黄瓜花叶病毒病研究进展[J]. 华北农学报,2014,29(S1):77-84. doi:10.7668/hbnxb.2014.S1.017.
Guo G J,Diao W P,Liu J B,Pan B G,Ge W,Wang S B. Research process of resistance to Cucumber mosaic virus in pepper[J]. Acta Agriculturae Boreali-Sinica,2014,29(S1):77-84.
[23] 李晓冬,吴元华. 辣椒轻斑驳病毒研究进展[J]. 生态学杂志,2016,35(10):2831-2837.doi:10.13292/j.1000-4890.201610.018.
Li X D,Wu Y H. Research progress on Pepper mild mottle virus[J]. Chinese Journal of Ecology,2016,35(10):2831-2837.
[24] 于海龙,靳远,刘婧,张宝玺,张正海,曹亚从,王立浩. 我国辣椒病毒病发生情况及发展趋势-基于2018年和2019年辣椒主产区的调查[J]. 中国蔬菜, 2020(9):25-30.
Yu H L, Jin Y, Liu J, Zhang B X, Zhang Z H, Cao Y C, Wang L H. Occurrence and development trend of pepper virus disease in China-based on main pepper producing areas investigation in 2018 and 2019[J]. China Vegetables, 2020(9):25-30.
[25] Chai A L,Chen L D,Li B J,Xie X W,Shi Y X. First report of a mixed infection of Tomato mottle mosaic virus and Tobacco mild green mosaic virus on eggplants in China[J]. Plant Disease,2018,102(12):2668. doi:10.1094/PDIS-04-18-0686-PDN.
[26] 鲁瑞芳, 李为民, 彭学贤. 植物病毒协生作用及其分子机理[J]. 中国病毒学,2001,16(3):195-201.
Lu R F,Li W M,Peng X X. Molecular mechanism of synergism of plant virus[J]. Virologica Sinica, 2001,16(3):195-201.

选择文件类型/文献管理软件名称

选择包含的内容