北京月季病原病毒的高通量测序鉴定和RT-PCR检测  

Virus identification and detection by high-throughput sequencing and RT-PCR from rose plants in Beijing

作　　者：张秀琪 刘松誉 杨一舟 李梦林 胡安安 刘唱 张宗英 韩成贵 王颖 ZHANG Xiu-qi;LIU Song-yu;YANG Yi-zhou;LI Meng-lin;HU An-an;LIU Chang;ZHANG Zong-ying;HAN Cheng-gui;WANG Ying(State Key Laboratory for Agro-biotechnology and Ministry of Agriculture Key Laboratory of Pest Monitoring and Green Management,College of Plant Protection,China Agricultural University,Beijing 100193,China;College of Biological Sciences,China Agricultural University,Beijing 100193,China)

机构地区：[1]中国农业大学植物保护学院,农业生物技术国家重点实验室、农业部病虫害监测与绿色管理重点实验室,北京100193 [2]中国农业大学生物学院,北京100193

出　　处：《植物病理学报》2021年第4期525-535,共11页Acta Phytopathologica Sinica

基　　金：国家级大学生创新训练项目(201910019028)。

摘　　要：本研究利用高通量测序技术对北京地区的月季染病样品进行了病毒鉴定,通过序列比对和拼接获得了李属坏死环斑病毒(prunus necrotic ringspot virus,PNRSV)、苹果茎沟病毒(apple stem grooving virus,ASGV)、柑橘碎叶病毒(citrus tatter leaf virus,CTLV)、月季黄叶病毒(rose yellow leaf virus,RYLV)、月季黄花叶病毒(rose yellow mosaic virus,RoYMV)、月季潜隐病毒1(rose cryptic virus1,RCV1)和玫瑰黄脉病毒(rose yellow vein virus,RYVV)等7种已知病毒和2种未知病毒的序列信息。分别对PNRSV,ASGV和RYLV园博园分离物的外壳蛋白基因进行系统进化分析,结果表明PNRSV-YBY归属于PV32组,ASGV-YBY归属于Ⅰ组,RYLV-YBY为玫瑰叶畸形病毒(Rosa rugosa leaf distortion virus,RrLDV)的一个新的分离物。对中国农业大学校园、北京植物园、北京动物园和北京园博园的37份样品进行了PNRSV、ASGV和RYLV的RT-PCR检测,检出率分别为41.7%、44.4%和10.8%。本研究初步明确了侵染北京月季的病毒种类和侵染情况,为月季病毒病的检测和防控提供参考。In this study,virus-infected rose samples collected from Beijing were analyzed by using high-throughput sequencing technique,seven known viruses,including prunus necrotic ringspot virus(PNRSV),apple stem grooving virus(ASGV),citrus tatter leaf virus(CTLV),rose yellow leaf virus(RYLV),rose yellow mosaic virus(RoYMV),rose cryptic virus1(RCV1)and rose yellow vein virus(RYVV),and two new viruses were identified through sequence alignment and assembly.Phylogenetic analysis of the coat protein genes of PNRSV,ASGV and RYLV Garden Expo Park isolates show that PNRSV-YBY and ASGV-YBY belong to groups PV32 and I,respectively,and we also identified a new isolate of Rosa rugosa leaf distortion virus(RrLDV)designated RYLV-YBY.A total of 37 samples from China Agricultural University campus,Beijing Botanical Garden,Beijing Zoo,and Beijing Garden Expo Park were detected using real-time polymerase chain reaction(RT-PCR)experiments and the positive detection rates of PNRSV,ASGV and RYLV were 41.7%,44.4%and 10.8%,respectively.In general,the virus species and infection situation of rose in Beijing were preliminarily identified,which provided reference for detection,prevention and control of rose virus disease in the future.

关 键 词：月季病毒 高通量测序 李属坏死环斑病毒 苹果茎沟病毒 月季黄叶病毒 RT-PCR检测 

分 类 号：S432.44[农业科学—植物病理学]