凤果花叶病毒的RT-PCR和核酸斑点杂交检测研究  

Studies on the detection of pepino mosaic virus using RT-PCR and dot blot methods

作　　者：李倩 张建坤 曾宪东 王琴 王振华 徐文兴[1] LI Qian;ZHANG Jiankun;ZENG Xiandong;WANG Qin;WANG Zhenhua;XU Wenxing(College of Plant Science and Technology,Huazhong Agricultural University,Wuhan 430070,China;Wuhan University of Technology,Wuhan 430070,China;Wuhan Customs Technology Center,Wuhan 430054,China)

机构地区：[1]华中农业大学植物科技学院,武汉430070 [2]武汉理工大学化学化工与生命科学学院,武汉430070 [3]武汉海关技术中心,武汉430054

出　　处：《植物病理学报》2023年第6期1262-1265,共4页Acta Phytopathologica Sinica

基　　金：2021年度海关总署“揭榜挂帅”科研项目-进出境茄科植物种子携带病毒、类病毒快速检测方法研究(项目编号:2021HK008);海关总署科研项目-基于深度测序技术的仁果类病毒检测鉴定研究(项目编号:2020HK175)。

摘　　要：0引言凤果花叶病毒(pepino mosaic virus,PepMV)属于线形病毒科(Flexiviridae)马铃薯X病毒属(Potexvirus),于1974年首次在秘鲁凤果植物内(Solanum muricatum)发现,后来于1999在荷兰发现该病毒还可侵染番茄[1-2]。被PepMV感染后,番茄生长迟缓,顶部叶片会表现皱缩、畸形和暗绿色不规则斑,下部叶片出现褐色坏死斑,有些叶片上出现亮黄色斑块及疱状突起;在一些大红果色番茄品种上,果实常会出现“大理石状”花纹病斑[3]。Pepino mosaic virus(PepMV),belonging to genus Potexvirus of the Flexiviridae family,infects tomato(Solanum lycopersicum)and other solanaceae plants,resulting in severe yield losses in some of these infected plants,and has emerged as a severe problem of greenhouse tomato crops worldwide.It calls for effective detection methods to control the epidemics,but the detection methods remain limited.Here,the sap samples of PepMV-infected tobacco leaves were mechanically inoculated into tomato seedlings which were kept in green house.At 14 days post inoculation,the mosaic symptoms were developed on the seedlings inoculated with the PepMV-infected leaf saps,which were characterized by yellow blocks in irregular sizes and shapes on most of the leaves,similar to the symptoms that were previously described;no symptoms were observed on the control leaves.The inoculated seedlings together with the controls were subjected to detection of PepMV by RT-PCR,and the symptomatic seedlings instead of the controls were confirmed to be infected by PepMV.Moreover,the infection of PepMV in all these symptomatic leaves while not in the controls can detected by using a digoxin-labeled RNA riboprobe with a titer of 10-3.This study provides biological and molecular approaches for the detection of PepMV,and offers a technical support for controlling the important viral disease.

关 键 词：褐色坏死斑 花叶病毒 马铃薯X病毒 顶部叶片 大理石状 番茄生长 核酸斑点杂交 黄色斑块 

分 类 号：S432.44[农业科学—植物病理学]