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1.
双链RNA分析技术在鉴定RNA病毒中的应用   总被引:3,自引:0,他引:3  
dsRNA技术是以无RNA病毒或类似病毒因子侵染的植物组织中不含有容易鉴定的同原大分子的dsRNA(>0.1×106)存在为前提。在植物体内dsRNA是RNA病毒和类病毒因子存在的迹象,dsRNA包括dsRNA病毒的基因或ssRNA病的复制中间型。dsRNA在寄主组织相当稳定,可以用物理和化学的方法分离出来。叙述了dsRNA技术用于植物病毒群、成员、同一病毒的不同株系、感染作物的新病毒及其复合侵染、类病毒、卫星病毒等的鉴定。证明该方法使用广泛、快速准确,不需要贵重设备,在一般实验室中即可进行。该方法有重要应用和学术价值,已引起植物病毒和植物病理学工作者广泛兴趣。  相似文献   

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RNA沉默是一种由双链RNA诱导同源RNA降解的现象,是植物的一种天然抗病毒机制.但是由于长期的共进化过程,一些病毒会在siRNA沉默途径以及miRNA沉默途径中编码一些抑制蛋白来对抗这种机制,使其作用受到抑制.文章对RNA沉默的发现、过程、抑制及其由RNA沉默介导的对RNA病毒、DNA病毒的抗性、以及由转病毒基因介导的病毒抗性等几种植物抗病机制.同时,对研究中所存在的一些问题作了分析,并就今后的研究重点和方向作了展望.  相似文献   

4.
Polyadenylic acid in Visna virus RNA   总被引:13,自引:0,他引:13  
Visna virus 70S RNA contains long stretches of polyadenylic acid [poly(A)]. The homogeneity in length of poly(4) regions is observed in 70S RNA from visna virus and all RNA tumor viruses tested, and not with other types of RNA. By this criterion visna virus resembles RNA tumor viruses.  相似文献   

5.
应用PCR技术进行Puumala病毒核酸片段的特异性扩增.将为进一步研究Puumala病毒和相关病毒.以及检测此类病毒感染提供更加特异、敏感、快速的方法。依据已知的Puumala病毒核酸序列,从M节段选出5对寡核苷酸作为PCR反应中的引物,以Puumala病毒RNA作为模板。做PCR。PCR产物分别经凝胶电泳和核酸杂交的方法进行测定,结果表明。目的核酸片段得到了很好的扩增。  相似文献   

6.
In an examination of the molecular basis of oral transmission of bunyaviruses by mosquitoes., La Crosse (LAC), snowshoe hare (SSH), and LAC-SSH reassortant viruses were compared in their ability to be transmitted to laboratory mice by the natural mosquito vector of LAC virus, Aedes triseriatus. Both LAC virus and the reassortment viruses containing the middle-sized (M) segment from the LAC parent were efficiently transmitted. In contrast, SSH virus and reassortment viruses containing the M RNA from the SSH parent were inefficiently transmitted. Thus, the M RNA segment, which codes for the virion glycoproteins, may be a major determinant of oral transmission of bunyaviruses by mosquitoes.  相似文献   

7.
The RNA interference (RNAi) pathway is found in most eukaryotic lineages but curiously is absent in others, including that of Saccharomyces cerevisiae. We show that reconstituting RNAi in S. cerevisiae causes loss of a beneficial double-stranded RNA virus known as killer virus. Incompatibility between RNAi and killer viruses extends to other fungal species in that RNAi is absent in all species known to possess double-stranded RNA killer viruses, whereas killer viruses are absent in closely related species that retained RNAi. Thus, the advantage imparted by acquiring and retaining killer viruses explains the persistence of RNAi-deficient species during fungal evolution.  相似文献   

8.
用小RNA深度测序鉴定广西冬种马铃薯病毒   总被引:3,自引:0,他引:3  
【目的】对广西冬种马铃薯病毒进行鉴定,为无病毒种薯选择和大田防治提供依据。【方法】2012年在马铃薯主要产区采集具有明显病毒病症状的样品,在血清学ELISA检测的基础上,进一步用小RNA深度测序对按症状分类的样品进行混合样本的病毒种类鉴定,再用RT-PCR方法对分组混合样本进行验证。【结果】在109个样本中检测到马铃薯卷叶病毒(Potato leafroll virus,PLRV)、马铃薯A病毒(Potato virus A,PVA)、马铃薯H病毒(Potato virus H,PVH)、马铃薯M病毒(Potato virus M,PVM)、马铃薯S病毒(Potato virus S,PVS)、马铃薯Y病毒(Potato virus Y,PVY)和马铃薯纺锤块茎类病毒(Potato spindle tuber viroid,PSTVd),同时发现PVS有丰富的株系分化。【结论】近年广西冬种马铃薯病毒种类增多,病症多样,亟需加强种薯管理,培育无毒健康种薯。  相似文献   

9.
Human sarcomas contain RNA related to the RNA of a mouse leukemia virus   总被引:20,自引:0,他引:20  
Labeled DNA complementary to the RNA of the Rauscher leukemia virus was hybridized with RNA from the polysome fraction of human sarcomas. Eighteen out of 25 specimens contained RNA possessing homology to the RNA of the mouse leukemia virus but not to that of the unrelated viruses causing mammary tumors in mice or myeloblastosis in chickens. Further, no normal adult or fetal tissues showed significant amounts of RNA specific to mouse leukemia virus. It appears that human sarcomas contain RNA sequences homologous to those found in an agent related to a virus known to cause sarcomas in mice.  相似文献   

10.
【Objective】 The viral disease is one of the major diseases of pepper production in Guangdong Province. The disease incidence is generally 5%-30% in the field, which can up to 100% in serious field. The objective of this study is to identify virus species infecting pepper in Guangdong Province, and to provide a theoretical basis for the prevention and control of pepper virus disease. 【Method】 From 2013 to 2016, a total of 125 susceptible pepper plant samples were collected from 8 major pepper growing areas of Guangzhou, Foshan, Huizhou, Jiangmen, Meizhou, Zhanjiang, Maoming and Shaoguan in Guangdong Province. Total RNA was extracted respectively from the leaves of 125 pepper samples. The 7 mixed samples collected from Maoming, Meizhou and Shaoguan were analyzed by small RNA deep sequencing. According to the results of small RNA deep sequencing analysis, two pairs of specific primers of each virus were designed to RT-PCR. The first pair of specific primers was designed according to the sequences of spliced gene fragments from small RNA deep sequencing. The second pair of specific primers was designed according to the conserved region sequences of viral genome in the GenBank database with the highest homology to sequences of spliced gene fragments from small RNA deep sequencing. Using disease samples RNA involved in small RNA deep sequencing as a template, the RT-PCR amplification was carried out with the two pairs of primers. Based on RT-PCR amplification results, the better pair of specific primers of each virus was selected. Furthermore, all 125 samples collected from Guangdong Province were subjected to detect viruses with the better pair of specific primers by RT-PCR. 【Result】 Fourteen viruses were identified in 125 samples collected from major pepper growing areas in 8 cities of Guangdong Province. According to the order of detection rate from high to low, they were Pepper mild mottle virus (PMMoV) (44.0%), Bell pepper endornavirus (BPEV) (32.8%), Tobacco mild green mosaic virus (TMGMV) (31.2%), Chilli veinal mottle virus (ChiVMV) (29.6%), Pepper vein yellow virus 1 (PeVYV-1) (26.4%), Pepper veinal mottle virus (PVMV) (25.6%), Cucumber mosaic virus (CMV) (18.4%), Chilli ringspot virus (ChiRSV) (16.8%), Pepper vein yellow virus 6 (PeVYV-6) (16.8%), Potato virus Y (PVY) (15.2%), Capsicum chlorosis virus (CaCV) (14.4%), Broad bean wilt virus 2 (BBWV-2) (9.6%), Pepper cryptic virus 1 (PCV1) (8.8%), Tobacco mosaic virus (TMV) (4.0%). The detection rates of PMMoV, BPEV, TMGMV, ChiVMV, PeVYV-1 and PVMV were over 25%. PMMoV, ChiVMV and PVMV were widely distributed in Guangdong Province. PMMoV (except Maoming), ChiVMV (except Shaoguan) were detected in pepper producing areas of other 7 cities. PVMV was detected in pepper producing areas of 8 cities. According to detection rate and distribution range, it was concluded that PMMoV, ChiVMV and PVMV were the dominant viruses infecting pepper in Guangdong Province. The mixed infection phenomenon was common on peppers in Guangdong Province. The detection rate of mixed infection was up to 88.0% in 125 samples. Among them, the mixed detection rates of 2 kinds, 3 kinds, 4 kinds, 5 kinds, 6 kinds, 7 kinds and 8 kinds of viruses were 28.0%, 25.6%, 12.0%, 9.6%, 6.4%, 1.6%, 2.4%, respectively. So, 2 kinds and 3 kinds of viruses mixed infection were the main infection forms on peppers in Guangdong Province. 【Conclusion】 There are 14 kinds of viruses endangering pepper plants in Guangdong Province, among which PMMoV, ChiVMV and PVMV are the dominant viruses. The phenomenon of mixed infection is common. The main infection forms on peppers are 2 kinds and 3 kinds of viruses mixed infection in Guangdong Province.  相似文献   

11.
基于小RNA深度测序技术鉴定侵染广东辣椒的病毒种类   总被引:1,自引:1,他引:0  
【目的】病毒病是广东省辣椒生产上主要病害之一,田间病株率一般为5%—30%,严重时可达100%。本研究旨在探明危害广东辣椒的病毒种类,为辣椒病毒病的防控提供理论依据。【方法】2013—2016年,从广东省广州、佛山、惠州、江门、梅州、湛江、茂名和韶关8市辣椒主要种植区采集疑似病毒病辣椒样品125份,分别提取每份辣椒病样总RNA,对从茂名、梅州、韶关3市采集的病样按地点和症状混合成7份混合病样进行小RNA深度测序分析,根据小RNA深度测序分析结果,对每种病毒分别根据小RNA深度测序拼接的基因片段序列和GenBank数据库中与该拼接序列同源性最高的病毒基因组序列保守区设计2对特异性引物,以小RNA深度测序的病样RNA为模板进行RT-PCR扩增,根据扩增效果对引物进行筛选,进一步应用筛选出的引物,对采集于广东省的125份辣椒病样分别进行RT-PCR检测,根据检测结果明确危害广东辣椒的病毒种类。【结果】从采集于广东省8市的辣椒主要种植区的125份病样中检测到14种病毒,按照检出率从高到低依次为辣椒轻斑驳病毒(Pepper mild mottle virus,PMMoV)(44.0%)、甜椒内源RNA病毒(Bell pepper endornavirus,BPEV)(32.8%)、烟草轻型绿花叶病毒(Tobacco mild green mosaic virus,TMGMV)(31.2%)、辣椒脉斑驳病毒(Chilli veinal mottle virus,ChiVMV)(29.6%)、辣椒黄脉病毒1(Pepper vein yellow virus 1,PeVYV-1)(26.4%)、甜椒斑驳病毒(Pepper veinal mottle virus,PVMV)(25.6%)、黄瓜花叶病毒(Cucumber mosaic virus,CMV)(18.4%)、辣椒环斑病毒(Chilli ringspot virus,ChiRSV)(16.8%)、辣椒黄脉病毒6(Pepper vein yellow virus 6,PeVYV-6)(16.8%)、马铃薯 Y 病毒(Potato virus Y,PVY)(15.2%)、辣椒褪绿病毒(Capsicum chlorosis virus,CaCV)(14.4%)、蚕豆萎蔫病毒2号(Broad bean wilt virus 2,BBWV-2)(9.6%)、辣椒隐症病毒1(Pepper cryptic virus 1,PCV1)(8.8%)、烟草花叶病毒(Tobacco mosaic virus,TMV)(4.0%)。其中,PMMoV、BPEV、TMGMV、ChiVMV、PeVYV-1和PVMV 6种病毒的检出率在25%以上,但PMMoV、ChiVMV和PVMV分布广泛,PMMoV除了茂名外,ChiVMV除了韶关外,其他7市辣椒产区均有分布;PVMV广泛分布于8市辣椒产区,根据检出率和分布范围,得出PMMoV、ChiVMV和PVMV是危害广东辣椒的优势病毒。同时发现,广东辣椒上多种病毒复合侵染现象普遍,在本研究125份病样中病毒复合侵染率达到88.0%,其中2种、3种、4种、5种、6种、7种和8种病毒复合侵染检出率分别为28.0%、25.6%、12.0%、9.6%、6.4%、1.6%和2.4%,由此可知,2种和3种病毒复合侵染是主要侵染形式。【结论】危害广东辣椒的病毒有14种,其中PMMoV、ChiVMV和PVMV为优势病毒,且复合侵染普遍,2种和3种病毒复合侵染是主要侵染形式。  相似文献   

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Aedes triseriatus mosquitoes became dually infected after ingesting two mutants of LaCrosse (LAC) virus simultaneously or after ingesting, by interrupted feeding, the two viruses sequentially within a 2-day period. After 2 weeks of incubation, approximately 25 percent of the vectors contained new virus genotypes as the result of RNA segment reassortment. New viruses were transmitted when the mosquitoes fed on mice. Viruses ingested more than 2 days after the initial infecting virus did not cause superinfection of the mosquito vectors.  相似文献   

13.
甘肃省河西地区菜豆病毒病分子检测   总被引:1,自引:1,他引:0  
以采集自甘肃省河西地区表现病毒病症状的菜豆病株叶组织总RNA为模板,进行RT-PCR扩增,鉴定出苜蓿花叶病毒、小西葫芦黄花叶病毒、菜叶普通花叶病毒、黑眼豇豆花叶病毒4种病毒.对预期大小的扩增产物进行直接测序.结果表明:该地区菜豆病毒病的发生以苜蓿花叶病毒、黑眼豇豆花叶病毒和小西葫芦黄花叶病毒复合侵染为主.  相似文献   

14.
【目的】探究狂犬病病毒HEP-Flury M基因重排对基因转录和蛋白表达的影响,揭示病毒在小鼠神经母细胞瘤(NA)细胞中的表型变化与M基因重排的相关性。【方法】通过荧光定量PCR、Western blot以及病毒在NA细胞中的生长和扩散试验,对亲本毒株rHEP-Flury和M基因重排毒株M2、M4在NA细胞中的基因转录、表达、生长和扩散进行比较。【结果】狂犬病病毒结构基因的转录和表达主要受病毒基因组RNA合成的影响,但是在一个完整转录过程中单个结构基因的转录比例与其所在位置相关,M基因重排病毒的Leader RNA (LeRNA)和L mRNA的转录比例显著高于亲本毒株rHEP-Flury。M基因重排病毒在NA细胞中的生长和扩散都劣于亲本毒株rHEPFlury。【结论】狂犬病病毒亲本毒株rHEP-Flury具有狂犬病病毒原始的基因组顺序,在NA细胞中的生长和扩散都明显优于M基因重排病毒。结构基因在基因组中的位置主要决定其在一次转录过程中的转录比例,进而影响病毒在NA细胞中的生长和扩散。  相似文献   

15.
Virazole is a synthetic nucleoside active in tissue culture against at least 16 DNA and RNA viruses. Applied topically, it inhibits herpetic keratitis in rabbits and tail lesions induced by herpes, vaccinia, and vesicular stomatitis viruses in mice. Injected intraperitoneally into mice, it inhibits splenomegaly and hepatomegaly induced by Friend leukemia virus and respiratory infections caused by influenza A(O), A(2), and B viruses and parainfluenza 1 virus. infections is also effective.  相似文献   

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The common fig(Ficus carica L.)was one of the earliest horticultural crops to be domesticated.A number of different viruses can infect fig trees including Fig mosaic virus(FMV)that has been detected in several commercial fig trees in Xinjiang,China.However,the distribution of FMV and other fig-infecting viruses in China remains unknown.In the present study,a sample from an ancient fig tree growing in Xinjiang was investigated by electron microscopy(EM)followed by PCR/RT-PCR,and FMV,Fig badnavirus 1(FBV-1)and Fig leaf mottle-associated virus 1(FLMaV-1)were detected.Fig leaf samples(252)from commercial orchards across China were subjected to PCR/RT-PCR,and FMV,FBV-1 and Fig fleck-associated virus(FFka V)were relatively abundant(44.4,48.4 and 44%,respectively),while FLMaV-1 and Fig mild mottle-associated virus(FMMa V)were much scarcer(5.6 and 0.4%,respectively),and FLMaV-2,Fig cryptic virus(FCV),and Fig latent virus(FLV)were not detected.The presence of disease-causing viruses in fig trees presents a significant challenge for fig producers in China.This study may help to promote actions aimed at controlling fig viruses,especially FMV.  相似文献   

18.
张峰  张瑜  王树瑜  张勇 《安徽农业科学》2013,(23):9541-9543
很多小RNA科病毒感染宿主细胞后可通过自身的核酸序列或病毒性蛋白产物控制靶细胞的生命活动,而被感染细胞也会通过自身的调控机制对侵染的病毒进行有限的反击,这种现象被认为是宿主细胞对抗小RNA科病毒侵染的防御机制.这种侵染与抵抗的互作机制可由某些病毒蛋白对细胞产生信号干扰或细胞自身翻译合成的细胞因子对病毒的复制路径进行封堵来实现.虽然这些信号通路的上游事件是不同的,但最后的效应却很统一,即使细胞崩解或者细胞将自身按照一定程序与所侵入的病毒同归于尽.此外,一些病毒蛋白具有抑制细胞程序性自杀的功能,它们能够令感染病毒后的细胞不死亡,形成病毒与宿主细胞共存的持续性感染状态.  相似文献   

19.
Associated with some plant viruses are small satellite RNA's that depend on the plant virus to provide protective coat protein and presumably at least some of the proteins necessary for satellite RNA replication. Multimeric forms of the satellite RNA of tobacco ringspot virus are probable in vivo precursors of the monomeric satellite RNA. Evidence is presented for the in vitro autolytic processing of dimeric and trimeric forms of this satellite RNA. The reaction generates biologically active monomeric satellite RNA, apparently is reversible to form dimeric RNA from monomeric RNA, and does not require an enzyme for its catalysis.  相似文献   

20.
Negative-strand RNA viruses condense their genome into a helical nucleoprotein-RNA complex, the nucleocapsid, which is packed into virions and serves as a template for the RNA-dependent RNA polymerase complex. The crystal structure of a recombinant rabies virus nucleoprotein-RNA complex, organized in an undecameric ring, has been determined at 3.5 angstrom resolution. Polymerization of the nucleoprotein is achieved by domain exchange between protomers, with flexible hinges allowing nucleocapsid formation. The two core domains of the nucleoprotein clamp around the RNA at their interface and shield it from the environment. RNA sequestering by nucleoproteins is likely a common mechanism used by negative-strand RNA viruses to protect their genomes from the innate immune response directed against viral RNA in human host cells at certain stages of an infectious cycle.  相似文献   

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