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1.
Studies on nepoviruses and tobraviruses, and their relationships with their associated vector nematodes, are scarce in Spain. However, virus disease symptoms have often been detected and their nematode vectors are widespread in Peninsular Spain. Nepovirus vector nematodes (Longidorus attenuatus, L. coespiticola, L. elongatus, L. macrosoma, Xiphinema coxi, X. diversicaudatum, X. index, X. italiae, X. pseudocoxi and X. vuittenezi) have been found associated with fruit and cereal crops. All are also widespread in other crops and uncultivated areas, together with X. rivesi, which has not yet been found associated with fruit and cereal crops in Spain. Tobravirus vectors have been less studied in Spain. Of the five recorded species, Paratrichodorus minor and Trichodorus primitivus are present on maize and wheat respectively. The geographical and host distribution of these nematodes are given and their ecological characteristics are discussed. 相似文献
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Judith Hübschen Lilo Kling Ulrike Ipach Volker Zinkernagel Nathalie Bosselut Daniel Esmenjaud Derek J.F. Brown Roy Neilson 《European journal of plant pathology / European Foundation for Plant Pathology》2004,110(8):779-788
Xiphinema diversicaudatum and X. index are vector nematode species of economic importance in viticulture regions as they can transmit Arabis Mosaic, Grapevine Fanleaf and Strawberry Latent Ringspot viruses to grapevine. Wang et al. (2003) designed species-specific diagnostic primers from ribosomal genes for both these vector species as well as a vector and a non-vector species X. italiae and X. vuittenezi, respectively. Our study aimed to confirm the specificity and determine the sensitivity and reliability of the primers for the two vector species, X. diversicaudatumand X. indexwhen challenged with closely related longidorid species and general nematode communities typical of vineyard soil. With one exception, no PCR product was observed when the primers were tested against six Longidorus, one Paralongidorus and one Xiphinema non-target species. Occasionally (three out of eight replicate PCR reactions) a weak PCR product was noted when primers for X. index were tested with L. elongatus. Furthermore, when challenged with a range of non-target nematode species comprising the nematode community typical of viticulture soil, no PCR product was amplified. An experimental dilution series of extracted DNA rigorously demonstrated that DNA from an equivalent single specimen of the target virus-vector species, X. diversicaudatum and/or X. index, could be detected amongst 1000 equivalent non-targetX. vuittenezi. Also, extracted DNA from an equivalent single target specimen was detected when added to DNA extracted from the overall soil nematode community. The primers were assessed further by using serial mixtures of actual nematodes rather than extracted DNA to simulate field soil. Using this method, a single target nematode could be detected amongst 200 non-target specimens. Given their specificity, sensitivity and reliability, it appears that these diagnostic primers will be of great benefit to phytosanitary/quarantine services related to the viticulture industry. 相似文献
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The dagger nematode Xiphinema index has a high economic impact in vineyards by direct pathogenicity and above all by transmitting the Grapevine fanleaf virus (GFLV). Agrochemicals have been largely employed to restrict the spread of GFLV by reducing X. index populations but are now banned. As an alternative to nematicides, the use of fallow plants between two successive vine crops was assessed. We selected plant species adapted to vineyard soils and exhibiting negative impact on nematodes and we evaluated their antagonistic effect on X. index in greenhouse using artificially infested soil, and in naturally infested vineyard conditions. The screening was conducted with plants belonging to the families Asteraceae (sunflower, marigold, zinnia, and nyjer), Poaceae (sorghum and rye), Fabaceae (white lupin, white melilot, hairy vetch, and alfalfa), Brassicaceae (rapeseed and camelina), and Boraginaceae (phacelia). In the greenhouse controlled assay, white lupin, nyjer, and marigold significantly reduced X. index populations compared with that of bare soil. The vineyard assay, designed to take into account the aggregative pattern of X. index distribution, revealed that marigold and hairy vetch are good candidates as cover crops to reduce X. index populations in vineyard. Moreover, this original experimental design could be applied to manage other soilborne pathogens. 相似文献
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Surveys for virus and virus-like diseases were carried out in commercial vineyards and nurseries in seven different Syrian provinces (Aleppo, Dara'a, As Suwayda, Al Qunaytirah, Homs, Hamah, Tartous). Samples were collected at random from 835 individual vines (735 Vitis vinifera and 100 rootstock accessions) for laboratory testing. Grapevine fanleaf virus (GFLV) , Arabis mosaic virus (ArMV), and Grapevine virus A (GVA) were the only viruses recovered by mechanical transmission to herbaceous hosts. Vein necrosis developed in c. 53% of graft-inoculated 110R indicators and vein mosaic in V. riparia inoculated with material from cv. Corna Alegra. A total of 71% of the ELISA-tested V. vinifera plants (522 out of 735) were infected by one (14.8%) or more (55.8%) viruses. GVA was the most widespread (54.7%), followed by Grapevine leafroll-associated virus 1 (GLRaV-1, 47.3%), Grapevine fleck virus (GFkV, 29.7%), and Grapevine leafroll-associated virus 3 (GLRaV-3, 23.9%). Other economically relevant viruses were scarcer, i.e. Grapevine leafroll-associated virus 2 (GLRaV-2, 9%), GFLV (0.8%) and ArMV (0.1%). The most important Syrian grapevine varieties, i.e. Hellwany, Salty, Balady, and Zeiny, had average infection rates that ranged between 44% and 91%. The highest incidence of infections was observed at Damascus (90%), whereas it ranged between 68% and 79% in the other provinces, except for Hama (36%). Rootstocks were in much better sanitary condition (25% infection). GFkV (22%) was the most common virus, whilst the presence of GLRaV-3 (3%), GLRaV-1, and GFLV (1%) was negligible. Grapevine rupestris stem pitting associated virus (GRSPaV) was detected in 72.3% of the samples by RT-PCR. A high percentage of the GRSPaV-positive vines (80%) induced vein necrosis reactions in 110R, thus confirming the recently established correlation between this virus and vein necrosis. 相似文献
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Edson Bertolini Julio García Alberto Yuste Antonio Olmos 《European journal of plant pathology / European Foundation for Plant Pathology》2010,128(3):283-287
Table grapes from one of the most important growing area in Spain (Vinalopó, Alicante) protected by the Designation of Origin
“Vinalopó bagged table grape”, were surveyed and analysed to determine the prevalence of the five viruses included in the
Spanish certification program: Arabis mosaic virus (ArMV), Grapevine fanleaf virus (GFLV), Grapevine fleck virus (GFkV), Grapevine leafroll associated virus-1 (GLRaV-1) and Grapevine leafroll associated virus-3 (GLRaV-3). Ninety five sampling points were selected and the position of grapevine plants georeferenced. Samples were collected
in two different vegetative periods and analyses were performed by ELISA and real-time RT-PCR. Purified RNA and immobilized
viral targets from plant extracts on nylon membranes were used in parallel assays as templates for PCR assays. In order to
analyse these five viral species by real-time RT-PCR, new specific primers and TaqMan probes were designed for detection of
ArMV and GFkV. Real time RT-PCR from purified RNA was more sensitive than spot version and ELISA tests. The most prevalent
virus was GFLV (95.8%) followed by GLRaV-3 (94.7%), GLRaV-1 (66.3%) and GFkV (65.3%). ArMV was not detected in any sample.
The high level of viral infections and the presence of mixed infections suggest that initial infected plant material and uncontrolled
traffic of propagation material have played an important role in the spread of viruses. 相似文献
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Development of a real‐time PCR method for the detection of the dagger nematodes Xiphinema index,X. diversicaudatum,X. vuittenezi and X. italiae,and for the quantification of X. index numbers 
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下载免费PDF全文 The ectoparasitic dagger nematodes Xiphinema index and Xiphinema diversicaudatum, often at low numbers in the soil, are vectors of grapevine nepoviruses, which cause huge agronomical problems for the vineyard industry. This study reports a method, based on real‐time PCR, for the specific detection of these species and of the closely related non‐vector species Xiphinema vuittenezi and Xiphinema italiae. Specific primers and TaqMan probes were designed from the ribosomal DNA internal transcribed spacer 1 (ITS1), enabling the specific detection of single individuals of each of the X. index, X. diversicaudatum, X. italiae and X. vuittenezi species whatever the nematode population. The specificity of detection and absence of false positive reaction were confirmed in samples of each species mixed with the three other Xiphinema species or mixed with nematodes representative from other genera (non‐plant‐parasitic Dorylaimida, Longidorus sp., Meloidogyne spp., Globodera spp. and Pratylenchus sp.). The method was shown to be valid for the relative quantification of X. index numbers through its use, from crude nematode extracts of soil samples, in a greenhouse assay of grapevine accessions ranging from highly susceptible to resistant. As an alternative to time‐consuming microscopic identification and counting, this real‐time PCR method will provide a fast, sensitive and reliable diagnostic and relative quantification technique for X. index nematodes extracted from fields or controlled conditions. 相似文献
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Villate L Fievet V Hanse B Delemarre F Plantard O Esmenjaud D van Helden M 《Phytopathology》2008,98(8):942-948
The nematode Xiphinema index is, economically, the major virus vector in viticulture, transmitting specifically the Grapevine fanleaf virus (GFLV), the most severe grapevine virus disease worldwide. Increased knowledge of the spatial distribution of this nematode, both horizontally and vertically, and of correlative GFLV plant infections, is essential to efficiently control the disease. In two infested blocks of the Bordeaux vineyard, vertical distribution data showed that the highest numbers of individuals occurred at 40 to 110 cm depth, corresponding to the two layers where the highest densities of fine roots were observed. Horizontal distribution based on a 10 x 15 m grid sampling procedure revealed a significant aggregative pattern but no significant neighborhood structure of nematode densities. At a finer scale ( approximately 2 x 2 m), nematode sampling performed in a third block confirmed a significant aggregative pattern, with patches of 6 to 8 m diameter, together with a significant neighborhood structure of nematode densities, thus identifying the relevant sampling scale to describe the nematode distribution. Nematode patches correlate significantly with those of GFLV-infected grapevine plants. Finally, nematode and virus spread were shown to extend preferentially parallel to vine rows, probably due to tillage during mechanical weeding. 相似文献
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During 2006 and 2007, a survey on the incidence and distribution of fourteen grapevine viruses was carried out in the Skadar Lake basin, one of the two main grapevine‐growing areas of Montenegro. In total 165 samples were collected from four red (‘Vranac’, ‘Krato?ija’, ‘Merlot’ and ‘Cardinal’), two white (‘Chardonnay’ and ‘Rkaciteli’) and a few unknown grapevine varieties in the vicinity of Podgorica and Bar. The phytosanitary status of the collected samples was analysed by DAS‐ELISA and the presence of Grapevine fanleaf virus (GFLV), Grapevine leafroll‐associated virus 1 (GLRaV‐1), Grapevine leafroll‐associated virus 2 (GLRaV‐2) and Grapevine leafroll‐associated virus 3 (GLRaV‐3) was confirmed in some of them. The most frequently found virus in assayed samples was GLRaV‐3 (54.5%), followed by GFLV (23%), GLRaV‐1 (20%) and GLRaV‐2 (0.6%). These serological analyses showed the absence of Grapevine leafroll‐associated virus 6 (GLRaV‐6), Grapevine leafroll‐associated virus 7 (GLRaV‐7), Raspberry bushy dwarf virus (RBDV), Strawberry latent ringspot virus (SLRSV), Tomato ringspot virus (ToRSV), Raspberry ringspot virus (RpRSV), Arabis mosaic virus (ArMV), Tobacco ringspot virus (TRSV), Tomato black ring virus (TBRV) and Cherry leaf roll virus (CLRV) from all tested samples. 相似文献
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Giorgio Gambino Rosalina Vallania Ivana Gribaudo 《European journal of plant pathology / European Foundation for Plant Pathology》2010,127(4):557-570
In grapevine, somatic embryogenesis is particularly effective in eliminating several important virus diseases. However, the
mechanism whereby regenerated somatic embryos are freed of the viruses is not clear. The distribution of Grapevine fanleaf virus (GFLV), Grapevine leafroll-associated virus-3 (GLRaV-3) and Grapevine virus A (GVA) in embryogenic callus of grapevine was investigated by in situ hybridization using digoxygenin-labelled oligonucleotide probes. Four months after culture initiation, in callus originated
by GFLV-infected explants we observed a mosaic of infected and uninfected cells, with high concentrations of viruses in some
cell groups in peripheral zones of the callus. In addition some abnormal somatic embryos showed a high hybridization signal.
In callus originated by GVA- and GLRaV-3-infected explants the viruses were concentrated in few cells surrounded by areas
of virus-free cells. The two viruses were generally localized in different clusters of cells inside the callus and the levels
of infection were lower than those observed in GFLV-infected callus. No virus was detected in callus nor in somatic embryos
after 6 months of culture. The results highlight the difficulties of some viruses at stably invading callus tissues and the
differential ability of GFLV to spread in the callus cells compared to the phloem-limited viruses. 相似文献
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The nematode-borne Grapevine fanleaf nepovirus (GFLV) causes severe degeneration of grapevines in vineyards worldwide. In a recent survey of the sanitary status of grapevine plants in north Tunisian vineyards, we were interested to study the polymorphism of GFLV. Purified virus, from mechanically inoculated Chenopodium quinoa , was used to produce anti-GFLV antiserum, which specifically recognized GFLV in different Tunisian grapevine samples using the DAS-ELISA technique. Positive samples were subjected to oligoprobe-RT-PCR to amplify a 606 bp region of the viral coat protein sequence. PCR products used for RFLP analysis after digestion with endonuclease Alu I produced 3 restriction profiles. RFLP data allowed clear distinction of two GFLV strains in Tunisia. The nucleotide sequence of the PCR-generated amplicons from each strain was determined showing 93.4% identity at the nucleic acid level and 97.5% similarity at the aminoacid sequence level compared to the previously characterized GFLV-F13 French isolate. This paper is the first report on molecular variability of GFLV in Tunisia. 相似文献
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五种烟草病毒TMV、CMV、TEV、PVY及TVBMV的多重RT-PCR同步检测 总被引：4，自引：0，他引：4
我国烟草病毒主要有烟草花叶病毒(TMV)、黄瓜花叶病毒(CMV)、烟草蚀纹病毒(TEV)、马铃薯Y病毒(PVY)和烟草脉带花叶病毒(TVBMV),通常发生复合侵染。本研究对我国5种烟草病毒的外壳蛋白基因部分序列设计引物,通过优化引物和模板浓度,摸索扩增参数,在一个体系中成功对5种病毒复合侵染的烟草材料进行多重RT-PCR扩增,得到237、273、347、456和547 bp共5条特异性条带,建立了能同时检测TMV、CMV、TEV、PVY和TVBMV的多重RT-PCR检测体系。对田间样品检测结果证明,多重RT-PCR体系能够同时检测5种病毒,并且灵敏度高。 相似文献
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Genetic diversity and mixed infections of begomoviruses infecting tomato, pepper and cucurbit crops in Nicaragua 总被引：1，自引：0，他引：1
M. Ala-Poikela E. Svensson ‡ A. Rojas T. Horko L. Paulin J. P. T. Valkonen A. Kvarnheden † 《Plant pathology》2005,54(4):448-459
Begomoviruses were detected in Nicaraguan fields of tomato ( Lycopersicon esculentum ) and adjacently growing plants of pepper ( Capsicum annuum ), chilli pepper ( C . baccatum ), cushaw ( Cucurbita argyrosperma ) and Mexican fireplant ( Euphorbia heterophylla ) using polymerase chain reaction (PCR) and universal begomovirus primers. All tomato and Mexican fireplant plants showing symptoms were infected with begomoviruses, while only 30–46% of the pepper, chilli pepper and cushaw plants showing symptoms tested virus-positive. No begomoviruses were found in potato. The virus species were provisionally identified by sequencing 533 bp of the viral coat protein gene ( AV1 ). Tomato severe leaf curl virus (ToSLCV), Tomato leaf curl Sinaloa virus (ToLCSinV) and Pepper golden mosaic virus (PepGMV) were found to infect both tomato and pepper. A new provisional species designated Tomato leaf curl Las Playitas virus (ToLCLPV) was detected in a tomato plant. Squash yellow mottle virus (SYMoV) and PepGMV were found in cucurbits, the latter for the first time in this host. Euphorbia mosaic virus (EuMV) was detected in Mexican fireplant. Sequencing of a larger number of PCR-amplified clones from selected plants revealed intraspecific viral sequence variability, and also multiple begomovirus infections which could represent up to three species in a single tomato or cushaw plant. Phylogenetic grouping of virus sequences did not correlate with the host of origin. 相似文献
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双重PCR检测马铃薯晚疫病菌和青枯病菌方法的建立及应用 总被引：3，自引：0，他引：3
利用真菌通用引物ITS1和ITS4扩增马铃薯晚疫病菌转录间隔区并进行序列测定,通过序列比较,设计了1对马铃薯晚疫病菌的特异引物INF1/INF2,并对15种不同真菌、细菌和7种疫霉属和腐霉属卵菌基因组DNA进行PCR扩增,结果只有不同来源的马铃薯晚疫病菌株可获得324 bp的特异带。将引物INF1/INF2与卵菌通用引物进行巢式PCR扩增后,其检测灵敏度在DNA水平上可达30 fg。运用设计的引物与马铃薯青枯病菌特异引物结合建立了双重PCR体系,能从马铃薯晚疫病菌和马铃薯青枯病菌总基因组DNA以及人工接种和自然发病的马铃薯植株中分别或同时扩增到324 bp和281 bp的特异片段。实现了同时对马铃薯晚疫病菌和马铃薯青枯病菌的快速可靠检测。 相似文献
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Mature canes were collected from vines in the main grapevine-growing areas in Tunisia (Cape Bon, Bizerte, Ben Arous), from commercial vineyards and mother-plant plots, to assess the presence of virus and virus-like diseases. Biological (mechanical transmission onto herbaceous hosts and grafting onto indicator woody plants) and serological detection (ELISA) methods were applied. ELISA showed that 96.4% of 669 vines tested were infected, most of them (88.1%) by at least two viruses. Grapevine leafroll-associated 3 closterovirus (GLRaV-3) was the most widespread virus (87.9%), followed by grapevine A vitiviras (GVA, 69.4%), grapevine fleck virus (GFkV, 51.9%), grapevine leafroll-associated 1 closterovirus (GLRaV-1, 36.8%), grapevine leafroll-associated 2 closterovirus (GLRaV-2, 19.1%), grapevine fan leaf nepovirus (GFLV, 18.2%) and grapevine B vitiviras (GVB, 14.8%). ELISA tests yielded negative results for grapevine leafroll-associated 7 closterovirus (GLRaV-7) and potato X potexvirus (PVX). The highest infections were found in Bizerte and Cape Bon regions (100 and 99.2%), and in vineyards aged over 20 years (98.5%) as compared with the younger ones (81.1%). Rootstocks in mother-plant plots were practically free from all the viruses tested (1 plant infected out of 81), whereas severe infections were found in Vitis vinifera mother plants (67.4% of 341 samples), in particular table grapes (92.6%) compared with wine grapes (47.9%). In these mother-plant plots, the prevailing viruses were GLRaV-3 (41.3%), followed by GFkV (36.7%), GVA (27.9%), GLRaV-1 (17%) and GLRaV-2 (15.2%). GFLV and GVB were far more limited (1.5 and 0.6%, respectively). The presence of vein necrosis and vein mosaic was ascertained by transmission onto 110R and Vitis riparia indicators, whereas only GFLV was mechanically transmitted onto herbaceous hosts (from about 20% of the samples). 相似文献
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Shesh Kumari Wilfrida Decraemer Francesca De Luca Wolfgang Tiefenbrunner 《European journal of plant pathology / European Foundation for Plant Pathology》2010,127(4):493-499
Genetic analyses using sequences of partial cytochrome c oxidase subunit 1 (cox1) gene of mitochondrial DNA were conducted to determine the extent of genetic variation within and among Xiphinema diversicaudatum, X. pachtaicum, X. simile and X. vuittenezi populations. Pairwise distance among the four species was 22.5 to 31.2%. Four different sequence variants of cox1 were determined among six populations of X. diversicaudatum and three variants among three populations of X. simile. Nucleotide variation was detected at 18 of 414 bp (1.9 to 2.7%) in X. diversicaudatum and 4 of 435 bp (0.2 to 0.9%) in X. simile. All changes were at silent sites. No nucleotide variation was detected within three populations of X. pachtaicum and within three populations of X. vuittenezi. 相似文献
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侵染昆明玫瑰的李坏死环斑病毒的鉴定及其分子检测 总被引：1，自引：0，他引：1
采集昆明地区种植的花叶症状明显的玫瑰样品,运用现代分子生物学技术与常规技术相结合的方法,初步鉴定引起玫瑰花叶病的主要病毒病原为李坏死环斑病毒.该病毒引起玫瑰植株的系统花叶、畸形和皱缩等症状;电镜下病毒粒体为球形,直径为22～23nm;ELISA检测发现该病毒在植株芽、花粉和顶部叶片的浓度最高;同时,根据外壳蛋白的保守区利用Primer 5.0设计该病毒的特异引物,对该病毒进行分子检测,得到450bp的预期DNA片断,并在此基础上,进行了巢式RT-PCR的分子检测,表明巢式RT-PCR的检测能力最强.并通过序列的同源性分析得知该病毒的外壳蛋白与已知PNRSV的同源性为98.0%,进一步证明了该病毒为李坏死环斑病毒. 相似文献