建兰花叶病毒广东分离物的全基因组序列分析

    Complete Genome Sequence Analysis of Guangdong Isolates of Cymbidium Mosaic Virus

    • 摘要:
      目的 建兰花叶病毒(Cymbidium mosaic virus,CymMV)是最主要的兰花病毒病原之一,其广泛传播对兰花产业发展造成严重威胁。探究CymMV遗传信息和进化,为广东省兰花病毒病的监测预警和兰花抗病毒病基因工程提供重要科学依据。
      方法 对采自广州地区的墨兰疑似病毒病叶片进行CymMV的RT-PCR及DA-SELISA检测鉴定,利用相关分子生物学软件对CymMV分离物进行基因组序列组装、注释、系统进化及选择压力分析。
      结果 首次在广东省发现2个CymMV分离物GZV013和ZC-29,基因组全长均为6 227 nt,编码5个功能蛋白;GZV013与台湾分离物M2的核苷酸序列一致性为97.03%;ZC29与南京分离物NJ-1的核苷酸序列一致性为97.11%;CymMV各分离物的全基因组序列一致性为86.85%~98.31%,且多样性种群的分化与寄主种类和地理隔离关系密切;CymMV基因组每个区域均受到负选择的影响,并符合中性进化模型;编码RdRpTGB1TGB2的基因在基因组中变异率最高。
      结论 GZV013与台湾分离物M2的亲缘关系最近,ZC29与南京分离物NJ -1的亲缘关系最近,同属于一个分支。

       

      Abstract:
      Objective Cymbidium mosaic virus(CymMV)is one of the most important orchid viruses.The wide spread of CymMV poses a serious threat to the development of orchid industry.The study is conducted to explore the genetic information and evolution of CymMV, which will provide an important scientific basis for monitoring and early warning of orchid virus disease and genetic engineering of orchid virus disease in Guangdong Province.
      Method RT-PCR and DASELISA were used to detect and identify CymMV from leaves with suspected virus disease of Cymbidium sinense collected from Guangzhou area.The genome sequence assembly, annotation, phylogeny and selection p ressure analysis of CymMV isolates were performed with related molecular biology software.
      Result Two CymMV isolates(GZV013 and ZC29)were found in Guangdong Province for the first time in this study.Both of the genome lengths of GZV013 and ZC29 were 6 227 nt, encoding five functional proteins.The similarity analysis of the full sequence showed that the nucleotide sequence identity of GZV013 and Taiwan isolate M2 was 97.03% and that of ZC29 and Nanjing isolate NJ-1 was 97.11%.The complete genome sequence identity among CymMV isolates ranged from 86.85% to 98.31%, and the differentiation of diverse populations was closely related to host species and geographical isolation.Each region of CymMV genome was affected by negative selection and conformed to the neutral evolution model.The RdRp, TGB1 and TGB2 genes exhibited the highest genome-wide diversity.
      Conclusion The GZV013 was most closely related to Taiwan isolate M2, and ZC29 was most closely related to Nanjing isolate NJ-1, belonging to the same branch of a family.

       

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