共查询到20条相似文献，搜索用时 31 毫秒
1.
The patterns of dsRNA bands separated by gel electrophoresis were used to analyse cryptic viruses in a wide range of beet plants, but uncertainties about the identities of some bands made the method unreliable. Four cDNA clones, each specific to one of the dsRNA genomic components of the beet cryptic viruses (BCVs) in sugar beet cv. Sharpes Klein E, were used in the analysis of Northern blots of a wide range of beet plants for the presence of BCV1 and BCV2. The presence of both viruses was demonstrated in sugar beetcvs Regina, Amethyst, Salohill and Bravo, red beet cv. Boltardy, leaf beet cv. Perpetual Spinach and fodder beet cv. Monofix, but only BCV1 or a closely related virus was found in Beta maritima. Beet temperate virus (BTV) from Japanese leaf beet was shown to have nucleic acid sequences in common with BCV1 from sugar beet cv. Sharpes Klein E. 相似文献
2.
从市场出售的甜菜(Beta vulgaris)种子中首次得到了直径约30毫微米的等轴病毒粒子的分离物。接种试验证明,它不能由摩擦接种传病,也不能由桃蚜(Myzus persicae)传播,但可由种子传毒,种子带毒率高达87.5%。受侵染的甜菜植株不表现症状,体内含毒量很低,病株汁液须经部分提纯并浓缩后才能在电镜下观察到病毒粒子和用琼脂双扩散法检测到病毒。在病株叶片的超薄切片中,未发现病毒粒子和细胞学的变异。在氯化铯溶液中,病毒粒子的浮力密度为1.37克/毫升左右。该分离物能与甜菜潜隐病毒抗血清产生沉淀反应,但与黄瓜花叶病毒无血清学关系。根据上述基本性状,该分离物归属于甜菜潜隐病毒(Beet Cryptic Virus)。 相似文献
3.
Yellow net virus of sugar beet has, until now, been listed as a tentative member of the luteovirus genus, but this is the first study to show that the yellow net symptom (YN) is always associated with a luteovirus. This virus is related to beet mild yellowing virus (BMYV) and when transmitted alone causes a mild yellowing symptom similar to that caused by BMYV in beet. Plants with YN have been found to contain, in addition to the luteovirus, two prominent, low molecular weight double-stranded (ds)RNA species. This dsRNA is indicative of an extra component in a complex with the luteovirus, which gives rise to the conspicuous symptom. The range of experimental hosts for this virus has been extended to include Capsella bursa-pastoris and Physalis floridana . 相似文献
4.
Sarah Cox M.A. Mayo A. Teifion Jones 《European journal of plant pathology / European Foundation for Plant Pathology》2000,106(4):353-364
Analysis was made of dsRNA in 37 cultivars and species of Ribes, that were healthy, naturally affected with the virus-like diseases, blackcurrant yellows, blackcurrant infectious variegation, gooseberry veinbanding or blackcurrant reversion, or graft-inoculated with scions from such diseased plants. Various dsRNA species, differing in size (from ca. 2 to 11kbp), number and staining intensity in gels, were detected in some or all assays of all plants, including those held as virus-tested stock. In different plant tissues from individual plants, the dsRNA species were usually similar in size and number but, in some sources, the dsRNA profile from flowers and/or bark differed greatly from the profiles of dsRNA obtained from leaves. No dsRNA species were associated consistently with any of these diseases. A 499kbp cDNA probe was obtained that in Northern blot analysis was specific to a ca. 5kbp dsRNA species present in the blackcurrant cv. Baldwin. It also detected a similarly sized dsRNA species in plants of many other blackcurrant cultivars, but it did not react with a similarly sized dsRNA species in redcurrant and gooseberry tissues. The 156 amino acid sequence encoded by the cDNA was very similar to sequences in the RNA-directed RNA polymerases of virus species in the family Totiviridae, especially Saccharomyces cerevisiae viruses L-1 and L-A. The significance of these findings and the possible origin of these dsRNA species are discussed. 相似文献
5.
正向和反向重复RNA介导的抗马铃薯Y病毒基因工程比较研究 总被引：20，自引：2，他引：20
RNA介导的病毒抗性与RNA沉默现象密切相关。反向重复cDNA序列(IR)的转录产物往往形成双链RNA结构,而双链RNA是诱发RNA沉默的有效因子。据此,本研究通过体外合成马铃薯Y病毒坏死株系衣壳蛋白基因(PVYN-CP)5'端反向重复cDNA序列和正向重复cDNA序列(DR),分别构建植物表达载体pROK-IR和pROK-DR,利用农杆菌介导方法转化烟草NC89,比较这2种转基因烟草在RNA介导抗病性方面的差异。抗病性检测表明,转化IR和DR的转基因烟草均可获得抗病程度达到免疫的植株,但转化IR序列可大大提高抗病植株在转基因植株中的比例。分析结果表明所获得的抗病性为RNA介导的抗病性,是RNA沉默的结果。这一研究结果为利用IR策略进行抗病毒遗传育种提供了理论依据,并为讲一步开展RNA介导抗病性的机制研究奠宗了基础。 相似文献
6.
Two viruses, beet yellows virus (BYV) and beet mild yellowing virus (BMW), cause yellowing of sugar beet, their principal vector being Myzus persicae. Although the viruses have different properties which are likely to influence their spread within root crops, for the purpose of control they have been treated in the past as one disease. This paper describes the results of a crop survey carried out in co-operation with agricultural staff of British Sugar plc from 1981 to 1984 in which leaves from plants with symptoms of virus yellows were tested by enzyme-linked immunosorbent assay (ELISA) for BYV and BMW. The two viruses differed in their incidence and distribution within the national sugar beet root crop; BMW was the main cause of yellowing and occurred in all parts of the growing region while BYV, which has the more severe effect on yield, was more limited in distribution. The survey located areas in southern East Anglia which are at greatest risk from BYV infection. The possible need to modify control measures depending on which virus threatens to invade the crops is discusssed. A high proportion of leaves visually identified as infected was found to contain neither BYV nor BMYV, emphasizing the difficulties of identifying virus-infected plants by field symptoms. 相似文献
7.
The cassava common mosaic virus (CsCMV) and the frogskin (FSD) disease agent have been reported to reduce cassava yields significantly in South America. However, little information is available on the distribution and incidence of these and other cassava virus diseases in Colombia. Cassava plants collected in three principal cassava production zones of Colombia were tested for the presence of CsCMV, cassava × virus (CsXV), and the Caribbean mosaic disease (CMD) and FSD agents. Some plants were also tested for the presence of double-stranded RNA (dsRNA). CsCMV was not detected in any of the 870 plants from 86 plantations. CsXV was detected in 51% of the 150 plants collected in the Cauca Department in south-central Colombia. The virus was present on all 15 cassava plantations surveyed. The CMD agent was detected in 17% of the 138 plants sampled in the Department of Magdalena in northern Colombia. FSD root symptoms were observed on 25 and 3% of plants examined in the Departments of Cauca and Magdalena, respectively. None of the 570 plants collected in areas west of the Rio Magdalena in the Departments of Atläntico, Bolivar, Córdoba and Sucre were found to be infected with any of these disease agents. However, some plants sampled in this region were found to contain multiple dsRNA species of unknown origin. 相似文献
8.
表达dsRNA的细菌提取液可抑制黄瓜花叶病毒对烟草的侵染 总被引：7，自引：0，他引：7
利用RT-PCR分别克隆了CMV P3613株系的RNA2片段、MP(movement protein)基因片段及CMV AN株系的CP(coat protein)基因片段。以CP基因为中间间隔序列,分别构建了含有RNA2片段和MP基因反向重复片段的原核表达载体。体外转录试验表明:两个载体转录后都能形成预期大小的dsRNA。经过IPTG诱导,在大肠杆菌HT115(DE3)菌株中可表达产生预期大小的核酸片段,经DNase和RNaseA消化处理,证实为dsRNA。将表达病毒基因dsRNA的细菌超声破碎后处理烟草,进行保护和治疗试验,结果表明:表达CMV MP基因和RNA2片段dsRNA的细菌破碎液能够诱导烟草对CMV产生抗性。接种病毒60d后,保护效果试验病株率分别为45%和60%,治疗效果试验病株率分别为75%和85%,而其他对照发病率均为100%。本研究结果证明了利用RNA沉默的原理,构建具有反向重复序列的原核表达载体,用细菌表达dsRNA的粗提取物可防治CMV对烟草的侵染。 相似文献
9.
ABSTRACT Twenty-one strains of Botrytis cinerea isolated from 13 species of plants grown in China were compared for pathogenicity on Brassica napus, mycelial growth on potato dextrose agar, and presence of double-stranded (ds)RNA. The results showed that the strain CanBc-1 was severely debilitated in pathogenicity and mycelial growth, compared with the 20 virulent strains. A dsRNA of approximately 3.0 kb in length was detected in CanBc-1 and 4 hypovirulent single-conidium (SC) isolates of CanBc-1, but was not detected in the 20 virulent strains of B. cinerea and 4 virulent SC isolates of CanBc-1. Results of the horizontal transmission experiment showed that the hypovirulent trait of CanBc-1 was transmissible and the 3.0-kb dsRNA was involved in the transmission of hypovirulence. Analysis of a 920-bp cDNA sequence generated from the 3.0-kb dsRNA of CanBc-1 indicated that the dsRNA element was a mycovirus, designated as B. cinerea debilitation-related virus (BcDRV). Further analyses showed that BcDRV is closely related to Ophiostoma mitovirus 3b infecting O. novo-ulmi, the causal agent of Dutch elm disease. Mitochondria and cytoplasm in hyphal cells of CanBc-1 became degenerated, compared with the virulent isolate CanBc-1c-66 of B cinerea. This is the first report on the occurrence of Mitovirus-associated hypovirulence in B. cinerea. 相似文献
10.
Albiach-Martí MR Guerri J de Mendoza AH Laigret F Ballester-Olmos JF Moreno P 《Phytopathology》2000,90(2):134-138
ABSTRACT A total of 14 Spanish isolates of Citrus tristeza virus (CTV) and 1 isolate from Japan were transmitted by Aphis gossypii, and the subisolates obtained were compared with the source isolates for symptom expression and double-stranded RNA (dsRNA) pattern. Of the 14 Spanish isolates, 9 showed altered dsRNA patterns after aphid transmission but only minor variations in the intensity of symptoms induced on Mexican lime. Northern blot hybridization with complementary DNA (cDNA) probes corresponding to both the 5' and the 3' termini of the CTV genomic RNA (gRNA) showed that the dsRNA bands that could be used to discriminate between the dsRNA pattern of the source and the aphid-transmitted isolates were the replicative forms of defective RNAs (D-RNAs). Conversely, the Japanese isolate and two subisolates obtained from it by aphid transmission had the same dsRNA pattern, but one of the subisolates induced milder symptoms in several hosts. Dot-blot hybridization with cDNA probes representing several regions of the gRNA showed that most of the aphid-transmitted isolates differed from the corresponding source isolate by their hybridization pattern. Our results indicate that aphid transmission often sorts the populations of gRNA variants and D-RNAs present in CTV isolates. 相似文献
11.
12.
M.E. Rott W. Jelkmann 《European journal of plant pathology / European Foundation for Plant Pathology》2001,107(4):411-420
For the identification and analysis of new RNA plant viruses infecting fruit trees, an initial step often involves the laborious procedure of isolation and cDNA synthesis and cloning from purified viral dsRNA. For subsequent RT-PCR detection of these and other viruses from tissue with high phenolic and polysaccharide concentrations, a simple and efficient extraction protocol for viral nucleic acid is also important. A method for rapid cDNA cloning from small amounts of purified dsRNA using a modification of degenerate oligo primed polymerase chain reaction mbox(DOP-PCR), and a modification of a protocol for effective extraction of viral RNA for use in RT-PCR are presented. Both methods were used to analyze a number of mottling diseases described in cherry. The causal agents for two of these diseases have been previously described, Cherry green ring mottle virus, a tentative member of the foveaviruses, and Cherry mottle leaf virus, a member of the trichoviruses. For the diseases cherry rusty mottle and cherry necrotic rusty mottle, data are presented identifying viruses associated with each disease. Viruses associated with cherry rusty mottle, cherry necrotic rusty mottle and European isolates of cherry mottle leaf diseases, are closely related to Cherry green ring mottle virus and can be tentatively included in the foveavirus genus. An additional virus, related to cherry green ring mottle virus, was discovered by RT-PCR cloning and appears to be a common latent virus of cherry. Finally, isolates of cherry necrotic mottle disease could be assayed positive by RT-PCR for a virus 相似文献
13.
Eduardo Segundo María P. Carmona Elisa Sáez Leonardo Velasco Germán Martín Leticia Ruiz Dirk Janssen Isabel M. Cuadrado 《European journal of plant pathology / European Foundation for Plant Pathology》2008,122(4):579-591
A study was conducted to determine the identity and prevalence of viruses in 455 greenhouses in the main Spanish green bean
growing area. Directed surveys were conducted in 422 crops from 2000–2004 to collect samples from diseased plants displaying
symptoms that could be attributed to viruses. The samples were analysed to detect any virus by means of dsRNA extraction,
mechanical inoculation to test plants, as well as ELISA and/or RT-PCR tests to detect potyviruses, geminiviruses and viruses
previously known to infect beans in Spain. Random surveys were conducted in the years 2002 and 2005 (in 21 and 12 greenhouses,
respectively) to study the actual incidence of known viruses in the area. Symptoms were recorded in 23,108 plants from which
664 plants were collected and analysed by ELISA or RT-PCR. The results of the directed surveys showed that all the analyzed
crops carried the cryptic virus Phaseolus vulgaris endornavirus (PVuV), whereas phytopathogenic viruses appeared in smaller percentages of the crops: Tomato yellow leaf curl virus (TYLCV) 20.4%, Southern bean mosaic virus (SBMV) 9.0%, Tomato spotted wilt virus (TSWV) 4.0%, and the new species Bean yellow disorder virus (BnYDV) that broke out in 2004 with occurrence values higher than 34.3% that year. From 2000–2004 an important decrease in
TYLCV was observed, along with a slight increase in SBMV and a consistently low occurrence of TSWV. The results of the random
surveys confirmed the increased occurrence of virus detected during the directed surveys, and furthermore demonstrated the
percentage of incidence for each virus. 相似文献
14.
Beet yellows virus (BYV), beet mild yellowing virus (BMYV), beet chlorosis virus (BChV), and beet mosaic virus (BtMV) cause virus yellows (VY) disease in sugar beet. The main virus vector is the aphid Myzus persicae. Due to efficient vector control by neonicotinoid seed treatment over the last decades, there is no current knowledge regarding virus species distribution. Therefore, Europe-wide virus monitoring was carried out from 2017 to 2019, where neonicotinoids were banned in 2019. The monitoring showed that closterovirus BYV is currently widely spread in northern Europe. The poleroviruses BMYV and BChV were most frequently detected in the northern and western regions. The potyvirus BtMV was only sporadically detected. To study virus infestation and influence on yield, viruses were transmitted to sugar beet plants using viruliferous M. persicae in quadruplicate field plots with 10% inoculation density simulating natural infection. A plant-to-plant virus spread was observed within 4 weeks. A nearly complete infection of all plants was observed in all treatments at harvest. In accordance with these findings, a significant yield reduction was caused by BMYV and BChV (−23% and −24%) and only a moderate reduction in yield was observed for BYV (−10%). This study showed that inoculation at low densities mimics natural infection, and quick spreading induced representative yield effects. Within the background of a post-neonicotinoid era, this provides the basis to screen sugar beet genotypes for the selection of virus tolerance/resistance and to test the effectiveness of insecticides for the control of M. persicae with a manageable workload. 相似文献
15.
A high incidence (86%) of potyvirus infection was noted in tobacco plants exhibiting a form of leaf curl in South Africa. Despite leaf curl being reported in the literature to be of geminiviral aetiology, no geminiviruses were detected. Furthermore, no other virus particles were detected by virus purification, TEM and serology. Twelve species of dsRNA were consistently isolated from these tobacco plants, but were absent from other forms of leaf curl-affected and healthy tobacco. Aphid and mechanical inoculation demonstrated that the purified potyvirus(es) did not cause leaf curl symptoms, but rather mild mottle and mosaic symptoms in tobacco. Partial characterization of the potyvirus preparation showed a possible relationship to a South African strain of potato virus Y. Because potyvirus-inoculated plants did not manifest leaf curl symptoms, and because leaf curl symptoms were noted in some plants not infected with a potyvirus, it was concluded that the potyvirus is not involved in the leaf curl aetiology, but causes a latent infection, the symptoms of which are masked. The pattern of the dsRNA banding, induction of enations and lack of mechanical and seed transmission are common to plant reoviruses. The possibility of a phytoreovirus involvement in this form of leaf curl is currently being investigated. The results from this study suggest that tobacco leaf curl disease worldwide, with regard to geminiviruses, be re-evaluated. 相似文献
16.
17.
ABSTRACT Comparison of a sampling of complementary DNA (cDNA) sequences from the Florida citrus tristeza virus (CTV) isolates T3 and T30 to the sequence of the genome of the Israeli isolate VT showed a relatively consistent or symmetrical distribution of nucleotide sequence identity in both the 5' and 3' regions of the 19.2-kb genome. In contrast, comparison of these sequences to the sequence of isolate T36 showed a dramatic decrease in sequence identity in the 5' proximal 11 kb of the genome. A cDNA probe derived from this region of the T36 genome hybridized to double-stranded RNA (dsRNA) of only 3 of 10 different Florida CTV isolates. In contrast, analogous probes from T3 and T30 hybridized differentially to the seven isolates not selected by the T36 probe. Primers designed from cDNA sequence for polymerase chain reaction (PCR) selectively amplified these 10 isolates, allowing them to be classified as similar to T3, T30, or T36. In contrast, individual cDNA probes derived from the 3' terminal open reading frames of the T3, T30, and T36 genomes all hybridized to dsRNA from all Florida CTV isolates tested, and PCR primers designed from the T36 capsid protein gene sequence amplified successfully from all isolates. Based on these data, we propose the creation of two groups of CTV, exemplified by the VT and T36 isolates, respectively. Isolates in the VT group, which include isolates VT, T3, and T30, have genomic sequence divergence that is relatively constant in proportion and distribution throughout the genome, and candidate isolates for that group could be considered strains of the same virus. The T36 group is differentiated from the VT group by the highly divergent 5' genomic sequence. This 5' region of the CTV genome, thus, can serve as a measure of the extent of sequence divergence and can be used to define new groups and group members in the CTV complex. 相似文献
18.
N. L. Robertson 《Plant pathology》2004,53(5):569-576
A new virus named Nootka lupine vein-clearing virus (NLVCV) was isolated from Lupinus nootkatensis plants that were confined to a relatively small area in the Talkeetna mountains of south-central Alaska. Annual surveys (2000–03) consistently found leaf symptoms of pronounced vein clearing and mosaic on 3- to 4-week-old plants in late June. Spherical particles ≈30 nm in diameter were isolated from these leaves. Virions contained a single-stranded RNA of ≈4·0–4·2 kb and one species of capsid protein estimated to be ≈40 kDa. The double-stranded RNA profile from naturally infected leaves consisted of three major bands ≈4·2, 1·9 and 1·5 kbp. Protein extractions from either sap or virions of diseased plants reacted to polyclonal antiserum made against the virions in Western blot assays. A predicted PCR product ≈500 bp was synthesized from virion RNA using primers specific to the carmovirus RNA-dependent RNA polymerase (RDRP) gene. The nucleotide sequence of the amplified DNA did not match any known virus, but contained short regions of identity to several carmoviruses. Only species belonging to the Fabaceae were susceptible to NLVCV by mechanical inoculation. Based on dsRNA profile, size of virion RNA genome and capsid protein, and similarity of the RDRP gene to that of other carmoviruses, it is suggested that NLVCV is a member of the family Tombusviridae , and tentatively of the genus Carmovirus . As the host range, RDRP gene and dsRNA profile of NLVCV are different from those of known viruses, this is a newly described plant virus. 相似文献
19.
甜菜花叶病毒病的研究Ⅰ.病毒的分离鉴定 总被引：1，自引：0，他引：1
1986-1987年对黑龙江、内蒙古部分甜菜原料产区与采种区进行了初步调查,采种区甜菜花叶病毒病发病率达100%,与其邻近的原料产区有零星发病,远离采种区则没有发病的迹象。田间采集的病叶,用磨擦、蚜传接种方法均能在甜菜上再现症状。经昆诺藜(Chenopodium quinoa)分离,普通电镜及乳胶电镜负染均能看到典型的马铃薯Y组病毒的粒体。通过鉴别寄主、甜菜上的症状表现,分离物的物理特性及血清学试验,证明此分离物为甜菜花叶病毒。在提纯方面,摸索了不同方法,认为匀浆前液氮或冷冻处理,二次PEG沉淀后过Sepharose4B柱,分离病毒效果较好,还可省去超速离心步骤。以内蒙五号甜菜品种作为毒源的繁殖寄主,能使提纯病毒产量达0.6mg/100g叶片,耳静脉与多点皮下注射相结合免疫兔子,微量沉淀测定效价为1:1024。 相似文献
20.
百合黄瓜花叶病毒及其检测 总被引：3，自引：0，他引：3
应用酶联免疫吸附法(ELISA)检测了浙江丽水和杭州的东方百合、亚洲百合等栽培品种上黄瓜花叶病毒(CMV),64个田间样品中有26个带毒,检出率为40.6%;dsRNA检测结果与已知CMV-FQS株系的电泳条带相同,但百合组织中CMVdsRNA含量较低;电镜观察,受CMV侵染的样品中大多含有线状病毒,复合侵染率为35.9%;寄主反应测定显示从百合植株上获得的CMV株系均不能通过汁液摩擦接种侵染昆诺藜、苋色藜、普通烟、心叶烟等6科10种指示植物。 相似文献