小RNA深度测序鉴定昭通市烟草脉斑病病毒
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摘要
烟草脉斑病在云南昭通市的烟草上发生危害严重,为明确昭通市烟草脉斑病的病毒种类,本研究采集昭通市的4个烟草种植区症状表现为疑似烟草脉斑病的烟草样品,采用小RNA深度测序技术对不同来源的混合样品进行小RNA测序分析。结果显示混合样品中的病毒分别属于烟草花叶病毒属Tobamovirus、马铃薯Y病毒属Potyvirus和马铃薯卷叶病毒属Polerovirus。根据不同病毒属设计通用引物,分别对不同地区的烟草样品进行RT-PCR验证,结果表明,烟草样品中病毒种类有烟草花叶病毒、烟草脉带花叶病毒、烟草脉扭病毒和马铃薯Y病毒的PVYN、PVYN-Wi和PVYNTN株系。
Flue-cured tobacco vein spot disease causes severe damage to tobacco production in Zhaotong City,Yunnan Province of China.In this study,siRNA deep sequencing was used to analyze the viruses in tobacco samples with necrotic and mosaic symptoms on tobacco leaves.The samples were collected from four different areas in Zhaotong City and mixed into one sample for siRNA sequencing analysis.The results showed that the identified viruses belonged to three genera,including Tobamovirus,Potyvirus and Polerovirus.RT-PCR and sequencing were used to identify the species of the viruses.Four viruses were detected in the samples,including TMV,TVBMV,TVDV and three strains of PVY(PVY~N,PVY~N-Wi,PVY~(NTN)).
Flue-cured tobacco vein spot disease causes severe damage to tobacco production in Zhaotong City,Yunnan Province of China.In this study,siRNA deep sequencing was used to analyze the viruses in tobacco samples with necrotic and mosaic symptoms on tobacco leaves.The samples were collected from four different areas in Zhaotong City and mixed into one sample for siRNA sequencing analysis.The results showed that the identified viruses belonged to three genera,including Tobamovirus,Potyvirus and Polerovirus.RT-PCR and sequencing were used to identify the species of the viruses.Four viruses were detected in the samples,including TMV,TVBMV,TVDV and three strains of PVY(PVY~N,PVY~N-Wi,PVY~(NTN)).
引文
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[14]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.
[15]丁铭,何跃峰,杨莉,等.侵染云南烟草的粉虱传双生病毒卫星分子基因组结构特征[J].西南农业学报,2010,23(5):1515-1521.
[16]卢训,丁铭,方琦,等.马铃薯Y病毒云南昭通烟草分离物的检测及年度间差异比较[J].植物保护,2013,39(3):56-60.
[17]段玉琪,秦西云,莫笑晗,等.烟草丛顶病(TBTV)传毒特性及寄主范围研究[J].中国烟草科学,2003(4):23-26.
[18]莫笑晗,秦西云,杨程,等.烟草脉扭病毒基因组部分序列的克隆和分析[J].中国病毒学,2003,18(1):58-62
[19]TIAN Y P,LIU J L,YU X Q.Molecular diversity of tobacco vein banding mosaic virus[J].Archives of Virology,2007,152(1):1911-1915.
[2]马国胜,郭红,陈娟.浅析皖北烟草脉斑病连年发生并间歇性暴发原因及对策[J].植物保护,2000,26(4):26-28.
[3]程宝玉,吴娟霞,陈卫华,等.豫西烟草脉斑病发生规律及药剂防治研究[J].烟草科技,2002(7):46-48.
[4]陈瑞泰,朱贤朝,王智发,等.全国l6个省(区)烟草侵染性病害调研报告[J].中国烟草科学,1997(4):1-7.
[5]张磊,丁铭,赵声春,等.昭通烟草脉斑病发病规律的研究[J].西南农业学报,2013,26(1):181-186.
[6]张永江,马荣群,辛言言,等.烟草花叶病毒属条形码鉴定方法的建立[J].湖北农业科学,2015,54(11):2624-2627.
[7]ROBERTSON N L,FRENCH R,GRAY S M.Use of groupspecific primers and the polymerase chain reaction for the detection and identification of luteoviruses[J].Journal of General Virology,1991,72(6):1473-1477.
[8]王健华,章绍延,龚殿,等.辣椒环斑病毒分子检测方法的建立及应用[J].热带作物学报,2012,33(2):342-345.
[9]RIGOTTI S,GUGERLI P.Rapid identification of potato virus Y strains by one-step triplex RT-PCR[J].Journal of Virological Methods,2007,140(1):90-94.
[10]TAMURA K,STECHER G,PETERSON D,et al.MEGA6:molecular evolutionary genetics analysis version 6.0[J].Molecular Biology Evolution,2013,30(12):2725-2729.
[11]秦西云,卢训,方琦,等.云南烟草中花叶病毒株系比较分析[J].中国烟草科学,2013,19(6):106-113.
[12]刘勇,莫笑晗,余清,等.云南、福建、湖南烟区烟草花叶病主要病毒种类检测及黄瓜花叶病毒亚组鉴定[J].植物病理学报,2006,36(4):310-313.
[13]秦西云,卢训,方琦,等.烟草脉带花叶病毒云南分离物全基因序列分析[J].中国烟草科学,2015,36(5):44-46.
[14]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.
[15]丁铭,何跃峰,杨莉,等.侵染云南烟草的粉虱传双生病毒卫星分子基因组结构特征[J].西南农业学报,2010,23(5):1515-1521.
[16]卢训,丁铭,方琦,等.马铃薯Y病毒云南昭通烟草分离物的检测及年度间差异比较[J].植物保护,2013,39(3):56-60.
[17]段玉琪,秦西云,莫笑晗,等.烟草丛顶病(TBTV)传毒特性及寄主范围研究[J].中国烟草科学,2003(4):23-26.
[18]莫笑晗,秦西云,杨程,等.烟草脉扭病毒基因组部分序列的克隆和分析[J].中国病毒学,2003,18(1):58-62
[19]TIAN Y P,LIU J L,YU X Q.Molecular diversity of tobacco vein banding mosaic virus[J].Archives of Virology,2007,152(1):1911-1915.