百合三种病毒的多重RT-PCR检测  

Detection of Three Lily Viruses by Multiplex RT-PCR

作　　者：徐榕雪[1] 明军[1] 穆鼎[1] 刘春[1] 汤庚国[2] 王晓武[1] 

机构地区：[1]中国农业科学院蔬菜花卉研究所,北京100081 [2]南京林业大学森林资源与环境学院,南京210037

出　　处：《园艺学报》2007年第2期443-448,共6页Acta Horticulturae Sinica

基　　金：国家社会公益研究项目(2005DIB022);国家科技支撑项目(2006BAD01A18);北京市花卉重点项目(YL-HH2006001;YLHH2006002)

摘　　要：根据基因库中黄瓜花叶病毒(Cucumber mosaic virus,CMV)、百合斑驳病毒(Lily mottle virus,LMoV)、百合无症病毒(Lily symptomless virus,LSV)的外壳蛋白基因序列,设计了3对特异引物,通过对扩增条件进行优化,建立了同时检测CMV、LMoV和LSV的多重RT-PCR检测方法。该方法可以从带有CMV、LMoV和LSV的样品中同时扩增出3条大小与试验设计相符的657bp(CMV)、428bp(LMoV)、171bp(LSV)的特异性多重RT-PCR扩增带。扩增产物测序表明,CMV、LMoV和LSV三种病毒与GenBank中登录的亚洲和荷兰多数分离物核苷酸同源性在90%以上,地域差异不明显,外壳蛋白序列高度保守。A multiplex RT - PCR method was developed to simultaneously detect three viruses of Lily by using 3 sets of specific primers designed according to CMV （ Cucumber mosaic virus）, LMoV （ Lily mottle virus） and LSV （ Lily symptomless virus）. All samples infected by CMV, LMoV and LSV could be amplified by multiplex PCR, and yielding three special bands of CMV （657 bp）, LMoV （428 bp） and LSV （ 171 bp）. Sequence identities of the three amplified virus fragments correlated well with other isolates from Asia and the Netherlands registered in the GenBank. Remarkably high homology in the coat protein sequences of the three viruses has been observed respectively despite their different origins.

关 键 词：百合 多重RT—PCR 病毒检测 黄瓜花叶病毒 百合斑驳病毒 百合无症病毒 

分 类 号：S682.2[农业科学—观赏园艺]