共查询到20条相似文献，搜索用时 612 毫秒
1.
2.
3.
Irena Mavrič Pleško Mojca Viršček Marn Saša Širca Gregor Urek 《European journal of plant pathology / European Foundation for Plant Pathology》2009,123(3):261-268
In 2003, Raspberry bushy dwarf virus was found for the first time in grapevine. Since this was the first report of a non-Rubus natural host, information about it is sparse. During this study the grapevine isolates were characterised biologically, serologically
and genetically and compared with known information about Rubus isolates. Infected plant material was used for mechanical inoculation of test plants, and for serological characterisation
with monoclonal antibodies. Most of RNA 2 was sequenced and compared with other sequences from the database. Grapevine isolates
infected Chenopodium murale systemically, which is not known for raspberry isolates. They were differentiated from Slovenian raspberry isolates with
three monoclonal antibodies using TAS-ELISA. Phylogenetic analyses clustered grapevine isolates separately from raspberry
isolates. Additionally, the virus was detected using nested RT-PCR in Longidorus juvenilis nematodes collected in an infected vineyard. Grapevine isolates of RBDV are distinct from raspberry isolates. 相似文献
4.
Pasquale Saldarelli Adib Rowhani Geoffrey Routh Angelantonio Minafra Michele Digiaro 《European journal of plant pathology / European Foundation for Plant Pathology》1998,104(9):945-950
RT-PCR with degenerate primers was used for the screening of the genome of some members of the Closterovirus, Vitivirus and Trichovirus genera. Two sets of primers, targeted to conserved sequences of the heat shock protein 70 homologue of closteroviruses or to the RNA dependent RNA polymerase genes of tricho- and vitiviruses, amplified the expected fragments from total RNA extracts or double-stranded RNAs of infected plants. Amplified cDNAs were cloned, sequenced and phylogenetically analyzed. Results support the allocation of grapevine viruses A, B, D and heracleum latent virus (HLV) in the genus Vitivirus, whereas, the detection of a HSP70 homologue in grapevine leafroll-associated viruses agrees with their assignment in the genus Closterovirus. The use of degenerate primers for the identification of grapevine viruses belonging to Vitivirus and Closterovirus genera is envisaged. 相似文献
5.
R. L. Lamprecht G. Pietersen G. G. F. Kasdorf L. H. Nel 《European journal of plant pathology / European Foundation for Plant Pathology》2009,123(1):105-110
A previously uncharacterised plant rhabdovirus, infecting Bermuda grass (Cynodon dactylon) in the North West Province, South Africa, has been found. To determine the morphology and virion size of this virus, embedded
ultra-thin sections of infected plant samples were observed under a transmission electron microscope. The virion distribution
within the cell, its bullet-shaped morphology and its size (240 × 63 nm) indicated that this might be a rhabdovirus of the
genus Nucleorhabdovirus. Degenerate polymerase chain reaction (PCR) primers were designed by alignment of the polymerase gene sequences of several
plant rhabdoviruses in order to identify conserved regions. Standard PCR and sequencing protocols were used to determine a
partial polymerase gene sequence of this virus sample which was then compared to the most closely related sequences available
on Genbank. The analysis indicated that the virus was indeed most closely related to known nucleorhabdoviruses, with the highest
nucleotide sequence similarities being to Maize mosaic virus and Taro vein chlorosis virus (70% and 69.7% respectively). Serological testing indicated that the South African Cynodon rhabdovirus had a close serological relationship with the nucleorhabdovirus Cynodon chlorotic streak virus. 相似文献
6.
7.
为明确水茄Solanum torvum植株叶片邹缩、褪绿是否由菜豆金色花叶病毒属病毒侵染引起,从云南省西双版纳傣族自治州田间采集具有疑似感染症状的水茄植株叶片样品,应用菜豆金色花叶病毒属病毒简并引物和特异性引物进行PCR扩增、克隆和测序,通过生物信息软件分析比较其核苷酸序列特征,并对其进行系统发育分析。结果显示,从采集的疑似病叶中共克隆获得了5条菜豆金色花叶病毒属病毒DNA-A全序列和3条DNA-B全序列,经全序列分析发现,侵染水茄的2种菜豆金色花叶病毒属病毒分离物分别属于中国南瓜曲叶病毒(squash leaf curl China virus,SLCCNV)和野茼蒿黄脉病毒(Crassocephalum yellow vein virus,CraYVV)。SLCCNV水茄分离物的基因组具有典型的菜豆金色花叶病毒属病毒双组分结构特征,与来自泰国的SLCCNV分离物(AB330078)亲缘关系最近,相似性最高达到99.0%;CraYVV水茄分离物的基因组具有典型的菜豆金色花叶病毒属病毒单组分结构特征,与来自云南省景洪市的CraYVV分离物(EF165536)亲缘关系最近,相似性最高达到97.6%。表明水茄是这2种菜豆金色花叶病毒属病毒的新寄主,并首次发现双组分和单组分菜豆金色花叶病毒属病毒可复合侵染水茄。 相似文献
8.
9.
10.
11.
12.
应用TaqMan MGB探针技术检测菜豆荚斑驳病毒 总被引：3，自引：1，他引：2
根据菜豆荚宽驳病毒(Beanpod mottle virus,BPMV)不同分离株外壳蛋白基因(coat protein,CP)的保守序列,设计了特异性引物与TaqMan MGB荧光探针,建立了BPMV的实时荧光RT-PCR检测方法.该方法与酶联免疫检测方法相比,灵敏度提高了100倍,具有快速、灵敏和高特异性的优点,适于对BPMV的快速检测.应用该方法,对来自美国不同地区的携带有BPMV的大豆样品进行检测,均能够得到BPMV的典型扩增曲线,表明引物与探针对BPMV不同分离株具有良好的简并性. 相似文献
13.
14.
侵染扶桑的烟草花叶病毒分离物鉴定 总被引：5，自引：0，他引：5
从表现叶斑驳症状的扶桑病株上获得一病毒分离物,电镜下可见约300 nm×18 nm的杆状粒子,其与烟草花叶病毒抗血清呈明显的阳性反应,dsRNA约为6.4 kbp。根据烟草花叶病毒(tobacco.mosaic virus,TMV)的RNA序列设计引物,进行RT-PCR检测,扩增出约800 bp的预期特异片段。将PCR产物连接pMD18-T载体,转化大肠杆菌DH5α,得到了含有目的片段的重组子。序列分析表明,与周雪平等报道的序列(GenBank AJ011933.1)同源性达99％。通过生物学、病毒粒子观察、血清学以及分子生物学实验结果,确定该病毒分离物为TMV。 相似文献
15.
Grapevine leafroll‐associated virus 3 (GLRaV‐3) is associated with grapevine leafroll disease, one of the most economically important viral diseases of grapevines. This disease impacts on both vine health and grape quality; reduction in yield, brix and wine colour are among its detrimental effects. Many methods, including serological and molecular procedures, have been developed for the detection of GLRaV‐3; however, there is no PCR‐based assay available to quantify virus populations within plant tissues. A real‐time RT‐PCR assay with TaqMan probe was developed for specific and reliable quantitative detection of GLRaV‐3 in infected tissues. The designed primers and probes target the conserved sequence in the RNA‐dependent RNA polymerase (RdRp) domain of the viral genome to prevent amplification of most subgenomic and defective RNAs. This protocol was used to examine the seasonal dynamics and translocation of GLRaV‐3 in field‐grown grapevines. The results showed that the virus spread quickly from trunks to new growing shoots and leaves early in the growing season, and most samples still harboured detectable virus during late summer and autumn. The seasonal progress of one GLRaV‐3 isolate was compared in four grapevine cultivars (Chardonnay, Cabernet Sauvignon, Italia and Thompson Seedless). Within cultivars there was little variability in the distribution and translocation of GLRaV‐3, except for in Thompson Seedless. This quantitative detection assay will be a valuable tool for GLRaV‐3 diagnosis, disease monitoring and population ecology studies. 相似文献
16.
为获得兼抗马铃薯X病毒(Potato virus X,PVX)、马铃薯Y病毒(Potato virus Y,PVY)、马铃薯卷叶病毒(Potato leaf roll virus,PLRV)和马铃薯潜隐花叶病毒(Potato virus S,PVS)4种病毒的转基因马铃薯新材料,分别以这4种病毒全长CP基因为模板,通过设计PCR引物和亚克隆获得4种病毒CP基因相对保守区段的基因片段,并将其拼接成融合基因,以载体pHANNIBAL和pBI121为基础,构建RNA干扰(RNA interference,RNAi)载体,利用农杆菌介导的转基因体系进行马铃薯遗传转化,并对获得的转基因马铃薯进行病毒抗性检测。结果表明,所获得的融合基因片段RH1和RH2,酶切鉴定分别得到长度为1 200 bp的条带,与预期片段相符;构建了含pdk内含子和RH1、RH2融合基因的RNAi植物表达载体,经Bam H I/Sac I双酶切,获得长度约3 200 bp的片段,表明RNAi植物表达载体pBI121-pRH构建成功;转化易感病毒马铃薯品种陇薯11号,PCR检测和PCRSouthern杂交分析表明融合基因已整合到陇薯11号马铃薯基因组中;抗病性检测显示4株转基因马铃薯植株对4种病毒均免疫。表明利用RNAi可筛选出抗多种病毒的转基因马铃薯新种质。 相似文献
17.
18.
Ho-Hsiung Chang Hsin-Mei Ku Wen-Shi Tsai Rui-Che Chien Fuh-Jyh Jan 《European journal of plant pathology / European Foundation for Plant Pathology》2010,127(2):219-228
Oriental melon plants, Cucumis melo var. makuwa cv. Silver Light, showing virus-induced symptoms of mosaic, leaf curl and puckering were observed in the fields of eastern
Taiwan in 2007. A virus culture, designated as SL-1, isolated from the diseased melon was established in systemic host plants,
Nicotiana benthamiana and oriental melon, by mechanical inoculation. SL-1 did not react to the antisera against common cucurbit-infecting RNA viruses.
Viral DNAs extracted from the diseased plant were amplified with the degenerate primers for begomoviruses. The full-length
genomic DNA-A and DNA-B of SL-1 were sequenced and found to be closest, with 97.7% and 90.6% nucleotide identity, respectively,
to Tomato leaf curl New Delhi begomovirus (ToLCNDV) cucumber isolate from a group of cucurbit-infecting begomoviruses. The virus SL-1 was designated as ToLCNDV oriental
melon isolate (ToLCNDV-OM). The pathogenicity of ToLCNDV-OM was confirmed by agroinfection. Progeny virus from the agroinfected
N. benthamiana plants was able to infect oriental melon by mechanical inoculation and caused symptoms similar to the original diseased melon
in the field. The ToLCNDV-OM also infected five other species of cucurbitaceous plants by mechanical inoculation. This is
the first report of a new ToLCNDV isolate causing severe disease on oriental melon in Taiwan. 相似文献
19.