深度测序技术分析番茄斑萎病毒病害寄主及介体中病毒种类
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摘要
正番茄斑萎病毒属病毒(orthotospoviruses)是严重危害云南蔬菜等重要农业经济作物的病毒病原之一。采用血清学检测、小RNA深度测序以及RT-PCR验证相结合的方法,从云南省昆明市晋宁区的主要作物寄主(番茄、辣椒、油麦菜)、重要中间寄主(鬼针草)和传毒介体(蓟马)中鉴定到TSWV、TZSV、PCSV和INSV 4种病毒,其中TSWV为该地区的主要优势病毒,而PCSV则是首次报道侵染鬼针草。通过对云南番茄斑萎病毒病害重病区作物寄主、中间寄主及蓟马三者进行病毒种类分析研究,明确TSWV为引起云南省昆明市晋宁区作物的主要病毒,TZSV、PCSV和INSV零星发生于不同寄主中。
Orthotospoviruses diseases were the severe vegetables diseases in Yunnan province,China.This study aimed at analysis of the orthotospoviruses species on host and vector in serious area,Kunming county,Yunnan Province.We confirmed the dominant species of orthotospoviruses in vegetable crops (tomatoe,pepper and lettuce) ,weed (Bidens pilosa) and thrips vectors by using ELISA,RT-PCR and deep sequence analysis.The study supported that there were four orthotospoviruses isolated from crops,weeds and thrips,Tomato spotted wilt orthotospovirus (TSWV) ,Tomato zonate spot virus (TZSV) ,Impatiens necrotic spot orthotospovirus (INSV) and Pepper chlorotic spot virus (PCSV) ,respectively.TSWV was a dominant species in this area and host.PCSV was first reported in Bidens pliosa.In conclusion,this paper reveals that TSWV is a major orthotospoviruses to cause the disease and TZSV,INSV,PCSV were sporadic outbreaks reported in Kunming county,Yunnan province.
Orthotospoviruses diseases were the severe vegetables diseases in Yunnan province,China.This study aimed at analysis of the orthotospoviruses species on host and vector in serious area,Kunming county,Yunnan Province.We confirmed the dominant species of orthotospoviruses in vegetable crops (tomatoe,pepper and lettuce) ,weed (Bidens pilosa) and thrips vectors by using ELISA,RT-PCR and deep sequence analysis.The study supported that there were four orthotospoviruses isolated from crops,weeds and thrips,Tomato spotted wilt orthotospovirus (TSWV) ,Tomato zonate spot virus (TZSV) ,Impatiens necrotic spot orthotospovirus (INSV) and Pepper chlorotic spot virus (PCSV) ,respectively.TSWV was a dominant species in this area and host.PCSV was first reported in Bidens pliosa.In conclusion,this paper reveals that TSWV is a major orthotospoviruses to cause the disease and TZSV,INSV,PCSV were sporadic outbreaks reported in Kunming county,Yunnan province.
引文
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[17]Li Y Y,Xiao L,Tan G L,et al.First report of Tomato spotted wilt virus on celery in China[J].Plant disease,2015,99(5):734.
[18]Xiao L,Li Y Y,Lan P X,et al.First report of Tomato spotted wilt virus infecting cowpea in China[J].Plant Disease,2016,100(1):2.
[19]Jiang L,Huang Y,Sun L,et al.Occurrence and diversity of Tomato spotted wilt virus isolates breaking the Tswresistance gene of Capsicumchinense in Yunnan,southwest China[J].Plant Pathology,2017,66(6):980-989.
[20]Lu X,Ding M,Fang Q,et al.Molecular variation of natural populations of Tomato zonate spot virus infecting flue-cured tobacco in Yunnan(in Chinese)[J].Acta Phytopathologica Sinica(植物病理学报).2012,42(2):195-201.
[21]Li Q F,Zhi L,Li M,et al.Tomato zonate spot virus as a pathogen found in iris tectorum(in Chinese)[J].Journal of Yunnan Agricultural University(云南农业大学学报),2014,29(2):167-172.
[22]Wang J G,Chen H R,Su Y S,et al.Natural infection of Bidens pilosa L.by Bidens mottle virus in Yunnan(in Chinese)[J].Scientia Agriculture Sinica(中国农业科学),2009,42(5):1849-1853.
[23]Dong J H,Zhang Z K,Yin Y Y,et al.Natural host ranges of Tomato zonate spot virus in Yunnan[J].Journal of Insect Science,2010,10(166):12.
[24]Zheng X,Chen Y D,Wu K,et al.Occurrence characteristics of tospoviruses and thrips vectors on tomato and pepper of Yunnan in 2014(in Chinese)[J].Journal of Southern Agriculture(西南农业学报),2015,46(3):428-432.
[25]Zhu X J,Zhang Z J,Lu Y B.Effects of inoculation of host plants with Tomato spotted wilt virus on populations of the western flower thrips,Frankliniella occidentalis(Pergande)(Thysanoptera:Thripidae)(in Chinese)[J].Acta Entomo Logica Sinica(昆虫学报),2011,54(4):425-431.
[26]Kumar S,Stecher G,Tamura K.MEGA7:Molecular evolutionary genetics analysis version 7.0 for bigger datasets[J].Molecular Biology and Evolution,2016,33(7):1870-1874.
[27]Parrella G,Gognalons P,Gebre-Selassie K,et al.An update of the host range of Tomato spotted wilt virus[J].Journal of Plant Pathology,2003,85(4:Special issue):227-264.
[28]Zheng X,Zhang J,Chen Y D,et al.Effects of tomato zonate spot virus infection on the development and reproduction of its vector Frankliniella occidentalis[J].Florida Entomologist,2014,97(2):549-554.
[2]Adams M J,Lefkowitz E J,King A M Q,et al.Changes to taxonomy and the international code of virus classification and nomenclature ratified by the International Committee on Taxonomy of Viruses[J].Archieve of Virology,2017,162(8):2505-2538.
[3]Whitfi H R,Jones R A C,Jain R K.Global status of Tospovirus epidemics indiverse cropping systems:Successes achieved and challenges ahead[J].Virus Research,2009,141(2):219-236.
[4]Elliott R M.Molecular biology of the Bunyaviridae[J].Journal of General Virology,1990,71(3):501-522.
[5]Adkins S.Tomato spotted wilt virus positive steps towards negative success[J].M olecular Plant Pathology,2000,1(3):151-157.
[6]Pappu H R,Jones R A C,Jain R K.Global status of Tospovirus epidemics indiverse cropping systems:successes achieved and challenges ahead[J].Virus Research,2009,141(2):219-236.
[7]Ding M,Luo Y Q,Fang Q,et al.First report of G roundnut yellowspot virus infecting Capsicumannuumin China[J].Journal of Plant Pathology,2007,89(2):305.
[8]Dong J H,Cheng X F,Yin Y Y,et al.Characterization of Tomato zonate spot virus,a newTospovirus species in China[J].Archives of Virology,2008,153(5):855-864.
[9]Chen X F,Dong J H,Fang Q,et al.Detection of Impatiens necrotic spot virus infecting Phalaenopsis in Yunnan[J].Journal of Plant Pathology,2010,92(2):545.
[10]Dong J H,Yin Y Y,Xu X Y,et al.First report of Tomato spotted wilt virus in tomato and tobacco in China[J].Journal of Plant Pathology,2010,92(S4):121.
[11]Liu Y T,Lu X P,Zhi L,et al.Calla lily chlorotic spot virus fromSpider Lily(Hymenocallis litteralis)and Tobacco(Nicotiana tabacum)in the South-west of China[J].Journal of Phytopathology,2012,160(4):201-205.
[12]Dong J H,Yin Y Y,Fang Q,et al.A newTospovirus causing chlorotic ringspot on Hippeastrumsp.in China[J].Virus Genes,2013,46(3):567-570.
[13]Yin Y Y,Fang Q,Lu X,et al.Detection Watermelon silver mottle virus infecting watermelon in Yunnan,southwest of China[J].Journal of Plant Pathology,2014,93(S4):123.
[14]Yin Y Y,Li T T,Lu X,et al.First report of capsicumchlorosis virus infecting tomato in Yunnan,southwest of China[J].Plant Disease,2016,10(1),230.
[15]Zheng K Y,Chen T C,Yeh S D,et al.Characterization of a newisolate of pepper chlorotic spot virus fromYunnan province,China[J].Archives of Virology,2017,162(9):2809-2814.
[16]Yin Y Y,Dong J H,Xu X Y,et al.Genetic diversity of isolates of Tomato spotted wilt virus in Kunming(in Chinese)[J].Southwest China Journal of Agricultural Sciences(西南农业学报),2013,26(1):159-162.
[17]Li Y Y,Xiao L,Tan G L,et al.First report of Tomato spotted wilt virus on celery in China[J].Plant disease,2015,99(5):734.
[18]Xiao L,Li Y Y,Lan P X,et al.First report of Tomato spotted wilt virus infecting cowpea in China[J].Plant Disease,2016,100(1):2.
[19]Jiang L,Huang Y,Sun L,et al.Occurrence and diversity of Tomato spotted wilt virus isolates breaking the Tswresistance gene of Capsicumchinense in Yunnan,southwest China[J].Plant Pathology,2017,66(6):980-989.
[20]Lu X,Ding M,Fang Q,et al.Molecular variation of natural populations of Tomato zonate spot virus infecting flue-cured tobacco in Yunnan(in Chinese)[J].Acta Phytopathologica Sinica(植物病理学报).2012,42(2):195-201.
[21]Li Q F,Zhi L,Li M,et al.Tomato zonate spot virus as a pathogen found in iris tectorum(in Chinese)[J].Journal of Yunnan Agricultural University(云南农业大学学报),2014,29(2):167-172.
[22]Wang J G,Chen H R,Su Y S,et al.Natural infection of Bidens pilosa L.by Bidens mottle virus in Yunnan(in Chinese)[J].Scientia Agriculture Sinica(中国农业科学),2009,42(5):1849-1853.
[23]Dong J H,Zhang Z K,Yin Y Y,et al.Natural host ranges of Tomato zonate spot virus in Yunnan[J].Journal of Insect Science,2010,10(166):12.
[24]Zheng X,Chen Y D,Wu K,et al.Occurrence characteristics of tospoviruses and thrips vectors on tomato and pepper of Yunnan in 2014(in Chinese)[J].Journal of Southern Agriculture(西南农业学报),2015,46(3):428-432.
[25]Zhu X J,Zhang Z J,Lu Y B.Effects of inoculation of host plants with Tomato spotted wilt virus on populations of the western flower thrips,Frankliniella occidentalis(Pergande)(Thysanoptera:Thripidae)(in Chinese)[J].Acta Entomo Logica Sinica(昆虫学报),2011,54(4):425-431.
[26]Kumar S,Stecher G,Tamura K.MEGA7:Molecular evolutionary genetics analysis version 7.0 for bigger datasets[J].Molecular Biology and Evolution,2016,33(7):1870-1874.
[27]Parrella G,Gognalons P,Gebre-Selassie K,et al.An update of the host range of Tomato spotted wilt virus[J].Journal of Plant Pathology,2003,85(4:Special issue):227-264.
[28]Zheng X,Zhang J,Chen Y D,et al.Effects of tomato zonate spot virus infection on the development and reproduction of its vector Frankliniella occidentalis[J].Florida Entomologist,2014,97(2):549-554.