利用3对番茄褐色皱纹果病毒(Tomato brown rugose fruit virus,ToBRFV)特异性引物对云南红河、宁夏银川、北京顺义疑似病毒病样本进行RT-PCR检测,分别扩增出大小为812、842、1053 bp的特异性条带。对扩增产物进行测序及系统进化分析,发...利用3对番茄褐色皱纹果病毒(Tomato brown rugose fruit virus,ToBRFV)特异性引物对云南红河、宁夏银川、北京顺义疑似病毒病样本进行RT-PCR检测,分别扩增出大小为812、842、1053 bp的特异性条带。对扩增产物进行测序及系统进化分析,发现云南、宁夏、北京的ToBRFV分离物与约旦、意大利、荷兰的ToBRFV分离物聚在同一分支上,亲缘关系较近。致病性结果显示,接种后的番茄植株表现出与田间相同的症状,且在发病组织中检测到病毒ToBRFV。表明来自云南红河、宁夏银川、北京顺义的番茄植株均被ToBRFV侵染。展开更多
Tomato brown rugose fruit virus(ToBRFV)causes severe fruit loss in tomato(Solanum lycopersicum)and pepper(Capsicum annuum)plants.It is an emerging Tobamovirus that is spreading globally.The major challenge is to devel...Tomato brown rugose fruit virus(ToBRFV)causes severe fruit loss in tomato(Solanum lycopersicum)and pepper(Capsicum annuum)plants.It is an emerging Tobamovirus that is spreading globally.The major challenge is to develop a reliable method for the detection of the virus,and to better characterize the symptoms it causes.The aims of this study,therefore,were to characterize the symptom development on tomato and pepper plants,and to establish a reliable detection method for the virus.Following infection of the tomato and pepper plants with ToBRFV,the leaves turned chlorotic,mosaic or mottled,while the fruit became rugose,necrotic and marbled,and showed discoloration with yellow or brown spots.Transmission electron microscopy(TEM)revealed single rod-like virus particles characteristic of the Tobamoviruses.Classical reverse transcription PCR(RT-PCR)and quantitative PCR(qPCR)with specific primers and probes confirmed that the virus is ToBRFV.We found that the resistance genes from tomato,Tm-2^(2),and pepper,L1,L2,L3 and L4,did not confer resistance to ToBRFV.Here,we present a PCR-based method as a diagnostic test for detecting ToBRFV in infected seeds.This method will help to prevent further spread of the virus in commercial seeds.展开更多
番茄褐色皱纹果病毒(Tomato brown rugose fruit virus,ToBRFV)是烟草花叶病毒属(Tobamovirus)的新成员,主要危害番茄、辣椒等茄科作物。2014年在以色列番茄上首次发现该病毒,随后快速扩展至欧洲、美洲的茄科作物主产区。2019年我国山...番茄褐色皱纹果病毒(Tomato brown rugose fruit virus,ToBRFV)是烟草花叶病毒属(Tobamovirus)的新成员,主要危害番茄、辣椒等茄科作物。2014年在以色列番茄上首次发现该病毒,随后快速扩展至欧洲、美洲的茄科作物主产区。2019年我国山东发现ToBRFV侵染番茄,要警惕该病毒在我国暴发危害。本文综述了国内外ToBRFV的分布、为害症状、传播途径、鉴定方法以及致病性等方面的研究进展,并结合烟草花叶病毒属病毒的侵染循环特征,提出防控对策;此外,对未来的研究重点、防控技术研发等进行了展望。展开更多
本研究旨在建立番茄和辣椒种子携带番茄褐色皱果病毒(Tomato brown rugose fruit virus,ToBRFV)快速、精准的核酸检测技术。选用ToBRFV的MP蛋白基因和CP蛋白基因保守序列作为靶标区域设计了1对特异性引物ToBRFV⁃F⁃5506/ToBRFV⁃R⁃6186,...本研究旨在建立番茄和辣椒种子携带番茄褐色皱果病毒(Tomato brown rugose fruit virus,ToBRFV)快速、精准的核酸检测技术。选用ToBRFV的MP蛋白基因和CP蛋白基因保守序列作为靶标区域设计了1对特异性引物ToBRFV⁃F⁃5506/ToBRFV⁃R⁃6186,建立了ToBRFV的RT⁃PCR检测方法。用ToBRFV阳性样品对该方法的灵敏度和特异性进行了系统分析,最后以疑似ToBRFV感染的实际种子样品进行验证,并与已报道的9对ToBRFV特异性引物对实际种子样品的检测效率进行比较分析。结果表明,采用本研究建立的RT⁃PCR检测方法检测限值至2.5ng/μL,且与番茄和辣椒的其他主要病毒无交叉反应,特异性良好,对50份疑似ToBRFV感染的番茄和辣椒种子样品的阳性检出率高达40%,要优于已报道的9对ToBRFV特异性引物。本研究建立的番茄褐色皱果病毒RT⁃PCR检测方法具有较高的灵敏度和良好的特异性,适用于ToBRFV感染的番茄和辣椒种子样品检测。展开更多
文摘利用3对番茄褐色皱纹果病毒(Tomato brown rugose fruit virus,ToBRFV)特异性引物对云南红河、宁夏银川、北京顺义疑似病毒病样本进行RT-PCR检测,分别扩增出大小为812、842、1053 bp的特异性条带。对扩增产物进行测序及系统进化分析,发现云南、宁夏、北京的ToBRFV分离物与约旦、意大利、荷兰的ToBRFV分离物聚在同一分支上,亲缘关系较近。致病性结果显示,接种后的番茄植株表现出与田间相同的症状,且在发病组织中检测到病毒ToBRFV。表明来自云南红河、宁夏银川、北京顺义的番茄植株均被ToBRFV侵染。
文摘Tomato brown rugose fruit virus(ToBRFV)causes severe fruit loss in tomato(Solanum lycopersicum)and pepper(Capsicum annuum)plants.It is an emerging Tobamovirus that is spreading globally.The major challenge is to develop a reliable method for the detection of the virus,and to better characterize the symptoms it causes.The aims of this study,therefore,were to characterize the symptom development on tomato and pepper plants,and to establish a reliable detection method for the virus.Following infection of the tomato and pepper plants with ToBRFV,the leaves turned chlorotic,mosaic or mottled,while the fruit became rugose,necrotic and marbled,and showed discoloration with yellow or brown spots.Transmission electron microscopy(TEM)revealed single rod-like virus particles characteristic of the Tobamoviruses.Classical reverse transcription PCR(RT-PCR)and quantitative PCR(qPCR)with specific primers and probes confirmed that the virus is ToBRFV.We found that the resistance genes from tomato,Tm-2^(2),and pepper,L1,L2,L3 and L4,did not confer resistance to ToBRFV.Here,we present a PCR-based method as a diagnostic test for detecting ToBRFV in infected seeds.This method will help to prevent further spread of the virus in commercial seeds.
文摘番茄褐色皱纹果病毒(Tomato brown rugose fruit virus,ToBRFV)是烟草花叶病毒属(Tobamovirus)的新成员,主要危害番茄、辣椒等茄科作物。2014年在以色列番茄上首次发现该病毒,随后快速扩展至欧洲、美洲的茄科作物主产区。2019年我国山东发现ToBRFV侵染番茄,要警惕该病毒在我国暴发危害。本文综述了国内外ToBRFV的分布、为害症状、传播途径、鉴定方法以及致病性等方面的研究进展,并结合烟草花叶病毒属病毒的侵染循环特征,提出防控对策;此外,对未来的研究重点、防控技术研发等进行了展望。
文摘本研究旨在建立番茄和辣椒种子携带番茄褐色皱果病毒(Tomato brown rugose fruit virus,ToBRFV)快速、精准的核酸检测技术。选用ToBRFV的MP蛋白基因和CP蛋白基因保守序列作为靶标区域设计了1对特异性引物ToBRFV⁃F⁃5506/ToBRFV⁃R⁃6186,建立了ToBRFV的RT⁃PCR检测方法。用ToBRFV阳性样品对该方法的灵敏度和特异性进行了系统分析,最后以疑似ToBRFV感染的实际种子样品进行验证,并与已报道的9对ToBRFV特异性引物对实际种子样品的检测效率进行比较分析。结果表明,采用本研究建立的RT⁃PCR检测方法检测限值至2.5ng/μL,且与番茄和辣椒的其他主要病毒无交叉反应,特异性良好,对50份疑似ToBRFV感染的番茄和辣椒种子样品的阳性检出率高达40%,要优于已报道的9对ToBRFV特异性引物。本研究建立的番茄褐色皱果病毒RT⁃PCR检测方法具有较高的灵敏度和良好的特异性,适用于ToBRFV感染的番茄和辣椒种子样品检测。
