滇重楼辣椒轻斑驳病毒分离物的鉴定及全基因组序列分析  

Identification and genome sequence analysis of pepper mild mottle virus isolate from Paris polyphylla var.yunnanensis

作　　者：杨林毅 陈潞 陈泽历 飞进强 王喆 徐绍忠[2] 赵明富[1] 文国松[2] YANG Lin-yi;CHEN Lu;CHEN Ze-li;FEI Jin-qiang;WANG Zhe;XU Shao-zhong;ZHAO Ming-fu;WEN Guo-song(Key Laboratory of Agrobiodiversity and Pest Management,Ministry of Education,Yunnan Agricultural University,Kunming 650201,China;College of Agronomy and Biotechnology,Yunnan Agricultural University,Kunming 650201,China)

机构地区：[1]云南农业大学农业生物多样性与病害控制教育部重点实验室,昆明650201 [2]云南农业大学农业与生物技术学院,昆明650201

出　　处：《湖北农业科学》2020年第13期154-157,162,共5页Hubei Agricultural Sciences

基　　金：国家自然科学基金项目(81860774)。

摘　　要：滇重楼(Paris polyphylla var.yunnanensis)是多年生草本植物,其药用部位为地下根茎,是一种重要的中药材。为明确侵染滇重楼的病毒病原,对采自云南省曲靖市的滇重楼进行透射电镜观察、DASELISA和RT-PCR技术检测。在透射电镜下观察到杆状病毒粒子,大小为(200~300)nm×(20~36)nm。对采集到的7份病样进行DAS-ELISA检测,结果表明,其中5份病样呈阳性。通过RT-PCR技术扩增、克隆获得1个全长为6357 bp的曲靖分离物PMMoV-QJ的全基因组序列。将PMMoV-QJ的全基因组序列与国内外已经报道的10个PMMoV分离物进行同源性分析比对,其同源性为99.42%~99.81%。基于全基因序列的系统发育分析表明,PMMoV-QJ与韩国分离物亲缘关系密切。本研究明确了该分离物的亲缘关系,从分子水平鉴定该分离物,为后续滇重楼病毒病的预防提供理论依据。Paris polyphylla var.yunnanensis is a perennial herb,and its medicinal part is underground rhizomes.It is an important chi⁃nese medicinal material.In order to clarify the viral pathogens infecting the Paris polyphylla,the transmission electron microscopy,DAS-ELISA and RT-PCR techniques were performed on the Paris polyphylla collected from Qujing,Yunnan province.Baculovirus particles were observed under transmission electron microscopy and were(200~300)nm×(20~36)nm in size.The results of DAS-ELI⁃SA on 7 samples collected showed that 5 of them were positive.A whole genome sequence of a 6357 bp Qujing isolate(PMMoV-QJ,accession number MK784568)was cloned by RT-PCR amplification.The homologous sequence of PMMoV-QJ were analyzed by ho⁃mology with 10 PMMoV isolates reported at home and abroad,and the homology was between 99.42%and 99.81%.Phylogenetic analy⁃sis based on the whole gene sequence showed that PMMoV-QJ was closely related to Korean isolates.This study clarifies the genetic re⁃lationship of the isolate,and identifies the isolate from the molecular level,which provides a theoretical basis for the prevention of the subsequent viral disease.

关 键 词：滇重楼(Paris polyphylla var.yunnanensis) 辣椒轻斑驳病毒(PMMoV) 鉴定 序列分析 

分 类 号：S432.41[农业科学—植物病理学]