为了探明郑州地区感病菊花上病毒的种类及来源，采集有明显病症的菊花样品，根据已知的番茄不孕病毒（TAV）和菊花B病毒（CVB）的基因序列设计特异性引物，采用RT-PCR扩增其外壳蛋白（CP）基因的方法对这2种病毒进行了检测，然后进行序列同源性及系统进化分析。结果显示，所得TAV的CP基因片段长度为481 bp，NCBI登录号为KU873003；所得3条CVB的CP基因片段长度均为547 bp，NCBI登录号分别为KU873004、KU873005、KU873006。将郑州分离物的CP基因与其他NCBI上已登录分离物进行比对，TAV核苷酸序列同源性为91%~99%，氨基酸序列同源性为86%~100%；CVB核苷酸序列同源性为76%~99%，氨基酸序列同源性为77%~99%。进化分析表明，菊花TAV郑州分离物与北京分离物KP019201、黑龙江分离物KP019202、河南分离物KP019200和山西分离物KP019204的亲缘关系最近；CVB郑州分离物与印度分离物AJ812569、AJ871367的亲缘关系最近。
[1]刘辉辉,沈学根,毛碧增.菊花病毒病及其防治对策[J].药物生物技术,2015,22(1):91–94.
[2] 赵喜亭,朱玉婷,田莹,等.番茄不孕病毒(TAV)脱除对怀白菊叶绿素含量及氮代谢相关指标的影响[J].南方农业学报,2016,47(7):1095-1099.
[3] Raj S K,Kumar S,Choudhari S.Identification of Tomato aspermy virus as the cause of yellow mosaic and flower deformation of chrysanthemums in India [J].Australasian Plant Disease Notes,2007,2(1):1–2.
[4] Ohkawa A,Yamada M,Sayama H,et al.Complete nucleotide sequence of a Japanese isolate of Chrysanthemum virus B (genus Carlavirus) [J]. Archives of Virology,2007,152(12) : 2253–2258.
[5] Chung B N,Choi G S,Choi Y M.Identification of tomato aspermy virus(TAV) and chrysanthemum virus B (CVB) from Dendranthema indicum in Korea [J].The Plant Pathology Journal,1999,15(2):119–123.
[6] Noordam D.Virus diseases of Chrysanthemum indicum in the Netherlands [J].Tijdschrift Over Plantenziekten, 1952,58:121–190.
[7]蒋细旺,饱满珠,薛东,等.我国菊花病害种类及危害特征[J].甘肃农业大学学报,2002,37(2):185–189.
[8] Singh L,Hallan V,Martin D P,et al.Genomic sequence analysis of four new chrysanthemum virus B isolates: Evidence of RNA recombination [J].Archives of Virology,2012,157:531–537.
[9] Singh L,Hallan V,Jabeen N,et al.Coat protein gene diversity among Chrysanthemum virus B isolates from India [J].Archives of Virology,2007,152(2):405–413.
[10]吴红芝,孔宝华,陈海如,等.昆明地区菊花病毒病的调查与鉴定[J].云南农业大学学报,2002,17(1):24–27.
[11] Mitiouchkina T Y,Firsov A P,Dolgov S V.Transgenic chrysanthemum plants transformed with the gene encoding for the virus B coat protein [J].Russian Agricultural Sciences,2013, 39(5/6): 431–434.
[12] Blencowe J W,Caldwell J.Aspermy－A new virus disease of the tomato[J].Annals of Applied Biololy,1949, 36(3): 320–326.
[13] Raj S K,Kumar S,Choudhari S,et al.Biological and molecular characterization of three isolates of Tomato aspermy virus infecting chrysanthemums in India [J].Journal of Phytopathology,2009,157: 117–125.
[14] Procter C H.Studies on tomato aspermy virus from Chrysanthemum indicum L. in New Zealand [J].New Zealand Journal of Agricultural Research,1975,18(4):387–390.
[15]孔宝华,李德葆.侵染菊花的番茄不孕病毒[J].浙江农业大学学报,1990,16(1):55–60.
[16] Verma N,Kumar K,Kulshrestha S,et al.Molecular studies on Tomato aspermy virus isolates infecting chrysanthemum [J]. Archives of Phytopathology and Plant Protection,2009, 42(2): 99–111.
[17]史步军,李彦勇,张成林,等.侵染药菊的一株病毒分离物的鉴定[J].植物病理学报,1993,23(3):231–236.
[18]赵霜.菊花病毒脱除与检测技术研究[D].北京:北京林业大学,2013:1–2.
[19]雷强,李旻,丁元明,等.菊花上2种病毒的检测及其部分外壳蛋白基因序列分析[J].华中农业大学学报,2012,31(6):704–709.
