[1]赵小慧,刘冲,郁凯,等.利用小RNA深度测序技术鉴定江苏盐城辣椒病毒种类[J].江苏农业学报,2023,(01):37-43.[doi:doi:10.3969/j.issn.1000-4440.2023.01.005]
 ZHAO Xiao-hui,LIU Chong,YU Kai,et al.Identification of viruses from peppers in Yancheng, Jiangsu province by small RNA deep sequencing[J].,2023,(01):37-43.[doi:doi:10.3969/j.issn.1000-4440.2023.01.005]
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利用小RNA深度测序技术鉴定江苏盐城辣椒病毒种类()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2023年01期
页码:
37-43
栏目:
植物保护
出版日期:
2023-02-28

文章信息/Info

Title:
Identification of viruses from peppers in Yancheng, Jiangsu province by small RNA deep sequencing
作者:
赵小慧1刘冲1郁凯1钟明娟2郑佳秋1邢锦城1
(1.江苏沿海地区农业科学研究所,江苏盐城224002;2.诸城市农业农村局, 山东诸城262200)
Author(s):
ZHAO Xiao-hui1LIU Chong1YU Kai1ZHONG Ming-juan2ZHENG Jia-qiu1XING Jin-cheng1
(1.Institute of Agricultural Sciences in the Coastal District of Jiangsu Province, Yancheng 224002, China;2.Zhucheng Agricultural and Rural Bureau, Zhucheng 262200, China)
关键词:
辣椒小RNA深度测序病毒病
Keywords:
peppersmall RNA deep sequencingviral disease
分类号:
S436.418.1+2
DOI:
doi:10.3969/j.issn.1000-4440.2023.01.005
文献标志码:
A
摘要:
2019年6月江苏省盐城市发生了较为严重的辣椒病毒病,危害症状表现为植株重度矮化、叶片黄化、蕨叶甚至畸形。将采集的11株疑似感染病毒的辣椒植株叶片研磨成浆液,再用其摩擦接种本氏烟植株,发现用其中3株辣椒的叶片浆液摩擦接种本氏烟植株后,本氏烟植株出现病毒侵染的典型症状,初步判断这些辣椒被病毒感染,但不能确定其种类和归属。为了进一步明确辣椒感染的病毒类型,将3株辣椒样品混成2份样品利用小RNA深度测序技术和生物信息学分析法进行检测,结果发现样品1被蚕豆萎蔫病毒2号(Broad bean wilt virus 2,BBWV2)、苜蓿花叶病毒(Alfalfa mosaic virus, AMV)和辣椒脉斑驳病毒(Chilli veinal mottle virus, ChiVMV)3种病毒复合侵染,从样品2中检测到BBWV2 1种病毒,通过RT-PCR检测验证了这一结果的可靠性。最后利用RT-PCR方法对田间采集的11份辣椒样品和接种的本氏烟植株进行毒源鉴定,其中2株辣椒样品及其接种的本氏烟叶片上鉴定出了BBWV2、ChiVMV和AMV,这3种病毒侵染辣椒在江苏省均为首次发现。研究结果为对江苏省辣椒构成潜在严重威胁的病毒的早期诊断和及时防控提供了参考。
Abstract:
In June 2019, an incidence of relatively serious pepper virus disease was reported in Yancheng City, Jiangsu province. The symptoms included plant dwarf stature, yellowing and deformities of pepper leaves. Eleven strains collected from pepper leaves infected with suspected virus were inoculated into Nicotiana benthamiana leaves by surface rubbing and it was found that three strains of peppers had typical symptoms of virus infection after surface inoculation of N. benthamiana. It was preliminarily suggested that these peppers were infected by a virus with unknown identity. In order to further clarify the virus type, two samples obtained by mixing three pepper standard samples were detected by small RNA deep sequencing technology and bioinformatics analysis. It was found that sample 1 was co-infected by broad bean wilt virus 2 (BBWV2), alfalfa mosaic virus (AMV) and chilli veinal mottle virus (ChiVMV). One virus of BBWV2 was detected in sample 2, and RT-PCR detection verified the reliability of the results. Finally, 11 pepper samples collected from the field and inoculated N. benthamiana were identified by RT-PCR. This is the first report of BBWV2, ChiVMV and AMV viruses infecting peppers in Jiangsu. The current study acts as a base line for early diagnosis and timely prevention and control of novel viral strains that pose a potential threat to peppers in Jiangsu province.

参考文献/References:

[1]王立浩,马艳青,张宝玺. 我国辣椒品种市场需求与育种趋势[J].中国蔬菜,2019(8):1-4.
[2]龚明霞,赵虎,王萌,等. 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.
[3]刘勇,李凡,李月月,等. 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.
[4]于海龙,靳远,刘婧,等. 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.
[5]吴贺,荆诗韵,刘丹,等. 苏南五地(市)主要蔬菜作物的病毒种类、区域分布和发生趋势[J].植物病理学报,2021,51(3):325-333.
[6]谢慧婷,崔丽贤,秦碧霞,等. 侵染广西甜椒的西瓜银斑驳病毒分子鉴定[J].植物病理学报,2021,51(3):474-477.
[7]龚明霞,赵虎,王萌,等. 广西辣椒病毒的sRNA深度测序和RT-PCR鉴定[J].园艺学报,2022,49(5):1060-1072.
[8]汪沛,汤琳菲,雷艳,等. 辣椒病毒病研究进展[J].湖南农业科学,2015(7):151-154.
[9]吴淑华,赵文浩,李廷芳,等. 南京辣椒上一种斑驳类型病毒病的分子鉴定[J].江苏农业学报,2015,31(6):1284-1290.
[10]乔俊卿,罗德旭,孙玉东,等. 江苏省淮安市设施辣椒病虫害的发生及防治现状调研分析[J].江苏农业科学,2019,47(16):127-132.
[11]王少立,谭玮萍,杨园园,等. 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.
[12]陈丽,樊民周,卫军锋,等. 陕西辣椒病毒病的毒源鉴定及化学防治药剂筛选[J].西北农林科技大学学报(自然科学版),2007,35(1):121-126.
[13]吉训聪,王健华,王运勤,等. 海南黄灯笼辣椒种传病毒的初步鉴定[J].海南大学学报(自然科学版),2006,24(3):284-288.
[14]赵尊练,史联联,谭根堂,等. 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.
[15]李兴红,严红,郭京泽,等. 种传辣椒轻斑驳病毒病DAS-ELISA的检测[J].植物保护,2005,31(3):66-68.
[16]高苇,王勇,张春祥,等. 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.
[17]严婉荣,赵志祥,肖彤斌,等. 海南辣椒、樱桃番茄病毒病调查及病原dot-ELISA鉴定[J].广东农业科学,2021,48(1):119-125.
[18]KUMAR S, PRAKASH H S. Detection of tobacco mosaic virus and tomato mosaic virus in pepper seeds by enzyme linked immunosorbent assay (ELISA)[J]. Archives of Phytopathology and Plant Protection, 2016,49(1/2/3/4):56-63.
[19]李月月,王前进,田珊,等. 黄瓜花叶病毒在洛阳地区的发生及亚组鉴定[J].广东农业科学,2020,47(7):114-120.
[20]张永江,辛言言,李桂芬,等. 实时荧光RT-PCR方法检测马铃薯V病毒[J].江苏农业科学,2013,41(6):36-38.
[21]NEMES K, SALNKI K. A multiplex RT-PCR assay for the simultaneous detection of prevalent viruses infecting pepper (Capsicum annuum L.)[J]. Journal of Virological Methods,2020,278(78):113838.
[22]韩帅,张河庆,吴婕,等. 侵染四川辣椒的番茄斑萎病毒和辣椒轻斑驳病毒的小RNA测序检测及鉴定[J].植物病理学报,2020,50(2):147-154.
[23]汤亚飞,裴凡,李正刚,等. 基于小RNA深度测序技术鉴定侵染广东辣椒的病毒种类[J].中国农业科学,2019,52(13):2256-2267.
[24]冯耿,辛敏,曹孟籍,等. 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.
[25]王红艳. 烟草脉带花叶病毒和马铃薯X病毒的协生作用及二者侵染本氏烟的转录组学分析[D].泰安:山东农业大学,2017.
[26]杨华兵,刘勇,李文正,等. 烟草品种辣椒叶脉斑驳病毒病的症状与抗病性鉴定[J].云南农业大学学报(自然科学),2014,29(1):22-26.
[27]岳阳,梁巧兰,魏列新,等. 苜蓿花叶病毒和白三叶草花叶病毒复合侵染对本氏烟中5种激素含量的影响[J].草业科学,2021,38(11):2255-2265.
[28]GONZLEZ-JARA P, TENLLADO F, MARTNEZ-GARCA B, et al. Host-dependent differences during synergistic infection by Potyviruses with potato virus X[J]. Molecular Plant Pathology,2004,5(1):29-35.
[29]GONZLEZ-JARA P, ATENCIO F A, MARTNEZ-GARCA B, et al. A single amino acid mutation in the plum pox virus helper component-proteinase gene abolishes both synergistic and RNA silencing suppression activities[J]. Phytopathology,2005,95(8):894-901.
[30]刘湘宁. 湖南辣椒病毒病毒原鉴定和进化分析[D].长沙:湖南农业大学, 2016.
[31]丁亚东. 中国辣椒ChiVMV抗性遗传及HSP70基因家族分析[D].海口:海南大学, 2021.
[32]何宛芹. 凤果花叶病毒和苜蓿花叶病毒单克隆抗体的创制及其应用[D].杭州:浙江大学,2021.

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备注/Memo

备注/Memo:
收稿日期:2022-06-24基金项目:国家自然科学基金项目(31901856);江苏省农业重大新品种创制项目(PZCZ201715)作者简介:赵小慧(1989-),女,山东莱阳人,博士,助理研究员,主要从事植物病毒的检验和检疫研究。(E-mail)zhaoxiaohui_g@163.com通讯作者:邢锦城,(E-mail)sdauxxx@163.com
更新日期/Last Update: 2023-03-21