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1.
ABSTRACT A polymerase chain reaction (PCR)-based method for the detection of the curtovirus Beet mild curly top virus (BMCTV, previously named the Worland strain of Beet curly top virus) was developed and used to investigate the BMCTV-beet leafhopper interaction. Using PCR and a BMCTV-specific primer pair, an approximately 1.1-kb BMCTV DNA fragment was amplified from adult leafhoppers and from the organs involved in circulative transmission: the digestive tract, hemolymph, and salivary glands. The temporal distribution of BMCTV in the leafhopper was determined using insects given acquisition access periods (AAPs) ranging from 1 to 48 h on BMCTV-infected shepherd's purse plants. BMCTV was detected in the digestive tract after all AAPs, in the hemolymph after AAPs of 3 h or greater, and in the salivary glands after AAPs of 4 h or greater. The amount of virus detected in the hemolymph and salivary glands increased with AAP length. The virus persisted for up to 30 days in leafhoppers (given a 3-day AAP on BMCTV-infected plants) maintained on corn plants, a nonhost for BMCTV, but transovarial transmission was not detected. These results are consistent with a persistent but nonpropagative mode of circulative transmission. 相似文献
2.
很多植物病毒经介体昆虫以持久循回型的方式水平传播至寄主韧皮部致病，而唾液腺是介体昆虫持久传毒的重要器官，也是植物病毒在介体昆虫内循回需要克服的最后一道防线。持久性植物病毒要完成水平传播，必须突破昆虫唾液腺屏障的阻碍，因此病毒和介体昆虫间形成了“攻”与“守”的较量与对决。揭示持久性植物病毒克服昆虫唾液腺屏障，实现水平传播的机制，对病害控制具有重要意义。该文着眼于介体昆虫唾液腺在持久传毒过程中的重要功能，回顾了虫传植物病毒突破介体昆虫唾液腺侵入屏障和释放屏障的分子机制，探讨了昆虫唾液蛋白通过调节植物或昆虫的适应性和行为促进或抑制病毒水平传播的功能，为制定阻断介体昆虫传播植物病毒途径的防控策略提供理论依据。 相似文献
3.
Jun Ohnishi Toshio Kitamura Fumihiro Terami Ken-ichiro Honda 《Journal of General Plant Pathology》2009,75(2):131-139
We studied the presence of a potential transmission barrier that blocks Tomato yellow leaf curl virus in the nonvector greenhouse whitefly, Trialeurodes vaporariorum. Because T. vaporariorum can ingest and retain the virus after acquisition feeding on an infected plant, comparable to the vector whitefly Bemisia tabaci, circumstance evidence suggested that a transmission barrier presents at location(s) where the virus moves from the digestive
tract lumen to the hemolymph. To provide direct evidence for the site of a transmission barrier in the nonvector insect, we
compared the accumulation levels and localization of the virus between the two species of whiteflies. Quantitative real-time
and conventional PCR analysis showed that accumulation of the virus during acquisition feeding and retention after a short
acquisition period were indistinguishable between the two species, but the circulation of the virus within the whiteflies
differed significantly between the species. In an immunofluorescence analysis using an antibody specific to the coat protein
of the virus, the virus was restricted to the luminal surface of the midgut epithelial cells and did not enter their cytoplasm
or that of the salivary glands in T. vaporariorum. In contrast, the virus was localized within the cytoplasm of the midgut epithelial cells and in the paired salivary glands
of B. tabaci adults. This direct evidence shows that a selective transmission barrier at the luminal membrane surface of midgut epithelial
cells in the nonvector whitefly blocks entrance of the virus into the midgut epithelial cells, resulting in incompetence as
a vector of the virus. 相似文献
4.
Paulo Gouveia Susana Dandlen ?ngela Costa Natália Marques Gustavo Nolasco 《European journal of plant pathology / European Foundation for Plant Pathology》2012,133(1):237-245
GLRaV-3, a member of the Closteroviridae family and type member of the genus Ampelovirus, is involved in the grapevine leafroll disease. Until now no RNA silencing suppressor has been found among viruses of this
genus, contrary to what happens with a large number of other viral genera. In the sister genus Closterovirus, RNA silencing suppressors are present in the 3’ end of the genome and have molecular weights close to 20 KDa. To test for
RNA suppressing activity screening of p21, p19.6 and p19.7 proteins, coded for in an analogous genomic location of the GLRaV-3
was undertaken. Only p19.7 revealed suppressor activity demonstrated in diverse silencing inducing systems. This suppressor
is able to overcome strong silencing inducers and shares several properties with the BYV p21-like family of suppressors of
the closteroviruses. This is the first report of an RNA silencing suppressor in the genus Ampelovirus. 相似文献
5.
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
6.
René Sforza Elisabeth Boudon-Padieu Charles Greif 《European journal of plant pathology / European Foundation for Plant Pathology》2003,109(9):975-981
Many grape viruses, such as filamentous Grapevine leafroll-associated viruses in the Closteroviridae family, are spread primarily through infected propagating material. However, there is increasing evidence that leafroll disease are spread in the field by insect vectors, namely mealybugs and other scale insects. This study was carried out in the northern wine-growing regions of France where Grapevine leafroll-associated virus-1 and -3 (GLRaV-1 and -3) are the most widespread grape Ampelovirus species. The vineyards were inspected for presence of mealybug and scale insects and grapes infected by GLRaV-1 and -3. Mealybugs, Heliococcus bohemicus, Phenacoccus aceris (Pseudococcidae) and the soft scale insect Parthenolecanium corni (Coccidae), were capable of a transmission efficiency of 14%, 23% and 29% respectively. GLRaV-1 and -3 infections that resulted from virus transmission were confirmed with DAS-ELISA using polyclonal antibodies. This is the first report of GLRaV-1 and -3 transmission by mealybug and coccid species in France, and the first report of the ability of H. bohemicus and Phenacoccus aceris to transmit these viruses to grapevines. The relevance of these findings with regards to maintenance of virus-free grapevine stocks and to control leafroll spread in commercial vineyards is discussed. 相似文献
7.
X. Y. Wang C. W. Zhang W. T. Huang J. Yue J. J. Dou L. Y. Wang Q. Wang Y. Q. Cheng 《Plant pathology》2020,69(1):149-158
Efforts to control viral diseases of grapevine include the production of certified material and development of virus-resistant transgenic grapevines. However, effective antiviral agents, once the viruses have infected the plants, are still lacking. This study shows that a crude garlic extract has significant antiviral activity against grapevine viruses. Replication of grapevine leafroll-associated virus 2 (GLRaV-2) was obviously inhibited in grapevine cv. Cabernet Sauvignon calli treated with diluted (1:100) garlic extract. The relative RNA levels of GLRaV-2 and grapevine fleck virus (GFkV) in cv. Summer Black grapevine in in vitro-grown plantlets 10 days after treatment with diluted (1:100) garlic extract were about 22% and 20%, respectively, of that in controls. The viral RNA accumulation of GLRaV-2, GFkV, grapevine virus A (GVA), grapevine fanleaf virus (GFLV) and grapevine rupestris stem pitting-associated virus (GRSPaV) in field-grown grapevine cv. Centennial Seedless plants sprayed with diluted (1:100) garlic extract were about 31–40%, 26–38%, 18–31%, 17–42% and 15–18%, respectively, of that in controls. Moreover, the garlic extract treatment led to a significant decrease in viral RNA accumulation of GLRaV-3, GLRaV-2, GVA, GFkV, GFLV, GRSPaV and grapevine Pinot Gris virus in pot-grown grapevine cv. Shine Muscat plants, and viral disease symptoms in these plants were obviously attenuated. In addition, this extract significantly induced expression of pathogenesis-related protein genes and stimulated activity of antioxidant enzymes in grapevines. Taken together, these results indicate that the crude garlic extract acts as a significant inhibitor against a broad range of grapevine viruses. 相似文献
8.
9.
Gérard Hommay Véronique Komar Olivier Lemaire Etienne Herrbach 《European journal of plant pathology / European Foundation for Plant Pathology》2008,121(2):185-188
Grapevine virus A (GVA, Vitivirus) was transmitted experimentally by first and second instars of the scale insect Parthenolecanium corni from grapevine to grapevine and to the herbaceous host Nicotiana benthamiana. This is the first report of GVA transmission by P. corni. Grapevine leafroll-associated virus-1 (Ampelovirus) was always present in the donor grapevines and, in every case, GVA was transmitted simultaneously with this
ampelovirus from grapevine to grapevine, suggesting possible interactions between the two viruses for transmission. 相似文献
10.
11.
The transmission of two closteroviruses associated with grapevine leafroll, GLRaV-1 and GLRaV-3, from grapevine to grapevine by the mealybugs, Pseudococcus longispinus and P. calceolariae (Homoptera: Pseudococcidae) was studied. Controlled transmission experiments using the first and third instars of each insect were conducted twice during the 1993–94 growing season to investigate the consequence of virus accumulation within the donor vine leaf tissue on the incidence of virus transmission to healthy recipient vines. Transmission of GLRaV-1 and GLRaV-3 was determined by ELISA testing recipient vines in July 1994 and March 1995. GLRaV-3 was transmitted to recipient vines by P. longispinus and P. calceolariae first instars only. An increase in virus titre within the season did not significantly alter the transmission rate of GLRaV-3 by either P. longispinus or P. calceolariae first instars. P. longispinus and P. calceolariae failed to transmit GLRaV-1 to recipient vines. 相似文献
12.
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
13.
水稻黑条矮缩病毒在灰飞虱消化系统的侵染和扩散过程 总被引：2，自引：0，他引：2
水稻黑条矮缩病毒 (Rice black streaked dwarf virus, RBSDV) 由介体灰飞虱(Laodelphax striatellus Fallén)以持久增殖型方式传播, 其编码的P9 1蛋白是形成病毒复制和子代病毒粒体装配的场所—病毒原质(viroplasm)的组分之一。为了明确RBSDV在介体昆虫体内的侵染循回过程, 本研究通过原核表达的P9 1蛋白免疫注射兔子制备P9 1抗体, 应用免疫荧光标记技术研究P9 1在饲毒后不同时期的介体灰飞虱体内的定位。共聚焦显微镜观察到饲毒后3 d, P9 1出现在介体中肠的少数上皮细胞内;饲毒后6 d, 在中肠外表的肌肉细胞分布有P9 1;饲毒后10 d, P9 1分布于中肠和后肠表面的肌肉, 同时在唾液腺也能观察到P9 1的存在。结果表明RBSDV在介体灰飞虱体内首先侵染中肠上皮细胞并复制, 随后扩散到中肠表面的肌肉细胞, 并通过环肌和纵肌扩散到中肠和后肠, 最后扩散到唾液腺。本研究首次直观地阐述了RBSDV在灰飞虱消化系统的侵染和扩散过程, 为有效阻断灰飞虱携带并传播病毒奠定基础。 相似文献
14.
Anna E. C. Jooste Gerhard Pietersen Johan T. Burger 《European journal of plant pathology / European Foundation for Plant Pathology》2011,131(3):371-381
Three genetic variants of Grapevine leafroll-associated virus 3 (GLRaV-3) were identified from vineyards of the Western Cape, South Africa. In a previous study, three full genome sequences
of isolates representing each of the variant groups were determined. The three variant groups were represented by accessions
621, 623 and PL-20, of variant groups I, II and III respectively. A specific single strand conformation polymorphism (SSCP)
profile was assigned to each variant which was used as a quick, reliable detection and differentiation method. In this study
we analysed the occurrence of these three GLRaV-3 variants in mother blocks in different cultivars and from different vine
growing regions using SSCP. The majority of the plants studied, were infected with the group II variant, similar to isolates
623 and GP18. The distribution of three GLRaV-3 variants within a spatio-temporally recorded cluster of diseased plants was
studied by means of SSCP profile analysis of ORF5 amplified PCR products. We showed that different GLRaV-3 variants are transmitted
to adjacent plants in an infection cluster. Results showed that, in some leafroll disease clusters, the variant that was present
in the original GLRaV-3 infected plant of a cluster was transmitted to adjacent plants in a row and across rows. 相似文献
15.
Ramesh R. Chavan Michael N. Pearson Dan Cohen 《European journal of plant pathology / European Foundation for Plant Pathology》2009,124(2):247-259
Actinidia chinensis and A. deliciosa plants from China, showing a range of symptoms, including vein clearing, interveinal mottling, mosaics and chlorotic ring
spots, were found to contain ~300 nm rod-shaped virus particles. The virus was mechanically transmitted to several herbaceous
indicators causing systemic infections in Nicotiana benthamiana, N. clevelandii, and N. occidentalis, and local lesions in Chenopodium quinoa. Systemically- infected leaves reacted with a Tobacco mosaic virus polyclonal antibody in indirect ELISA. PCR using generic and specific Tobamovirus primers produced a 1,526 bp sequence spanning the coat protein (CP), movement protein (MP), and partial RNA replicase genes
which showed a maximum nucleotide identity (88%) with Turnip vein clearing virus and Penstemon ringspot virus. However, when the CP sequence alone was considered the highest CP sequence identity (96% nt and 98% aa) was to Ribgrass mosaic virus strain Kons 1105. The morphological, transmission, serological and molecular properties indicate that the virus is a member
of subgroup 3 of the genus Tobamovirus. 相似文献
16.
Cristina Cabaleiro Antonio Segura 《European journal of plant pathology / European Foundation for Plant Pathology》2006,114(4):441-446
With the aim of investigating the dynamics of transmission of GLRaV-3, we report and analyse time-courses of infection incidence
in various plots in one vineyard in Rías Baixas (Galicia, Spain). GLRaV-3 infection was close to 100% after 15 years monitoring
the epidemic in several plots where P. citri was widely spread although with low density populations. In two plots, virus-free plants were planted close to infected ones
and after 8–9 years more than 80% of the plants tested GLRaV-3 positive (average infection rates of 7.8–12.4% per year). The
Gompertz model best fitted the epidemiological data. 相似文献
17.
Kai-Shu Ling Hai-Ying Zhu Zhao-Yuan Jiang Dennis Gonsalves 《European journal of plant pathology / European Foundation for Plant Pathology》2000,106(4):301-309
A polyclonal antiserum (As163) specific to grapevine leafroll associated closterovirus-3 (GLRaV-3) was developed using a recombinant coat protein expressed in E. coli from a cDNA clone identified after immunoscreening of a cDNA library. Specificity of the antiserum to GLRaV-3 was shown by Western blot and immunosorbent electron microscopy. With this antiserum, an effective double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) was developed for GLRaV-3 detection. To evaluate the sensitivity of the antiserum in DAS-ELISA for virus detection, different combinations of antibodies were compared. Although best results were obtained when As163 was used for coating and a monoclonal antibody (MabNY1.1) was used as an enzyme conjugate, good results were also obtained when As163 was used both for coating and as an enzyme conjugate. Using this As163–Mab system in DAS-ELISA, we confirmed the presence of GLRaV-3 in a diverse collection of leafroll infected vines. 相似文献
18.
Lett JM Granier M Hippolyte I Grondin M Royer M Blanc S Reynaud B Peterschmitt M 《Phytopathology》2002,92(1):65-74
ABSTRACT Spatial and temporal distribution of Maize streak virus (MSV, family Geminiviridae, genus Mastrevirus) was monitored in the vector species Cicadulina mbila and the nonvector species C. china? using conventional and real-time quantitative polymerase chain reaction. Sustained feeding on MSV-infected plants showed that virus accumulation reaches a maximum in C. china?, but not in C. mbila. After a 3-day acquisition access feeding period (AAP), MSV was detected in the gut, the hemolymph, and the head of C. mbila, but only in the gut of C. china?. Similarly, Digitaria streak virus (genus Mastrevirus), which is not transmitted by either of the two species, was only detected in the gut. MSV was detected in the hemolymph of C. mbila 3 h after the beginning of the AAP. Although viral DNA progressively decreases in the vector and nonvector species after a 3-day AAP, MSV DNA remained stable in the salivary glands of C. mbila. 相似文献
19.
Grapevine leafroll disease is caused by grapevine leafroll-associated viruses (GLRaVs). These viruses are common in vineyards worldwide and often associated with vitiviruses that are involved in the rugose wood complex of grapevine. Ten mealybug species are known as vectors of one or several of these grapevine viruses, including the apple mealybug Phenacoccus aceris which is widespread in Holarctic regions and able to transmit Grapevine leafroll-associated virus-1 and -3 (GLRaV-1 and -3). Our aim was to characterize the transmission features of leafroll viruses by Phenacoccus aceris in order to better understand the contribution of this mealybug to leafroll epidemics. Results showed that Phenacoccus aceris is able to transmit GLRaV-1, -3, -4, -5, -6, and -9 to grapevine but not GLRaV-7. This is the first report of GLRaV-6 transmission by a mealybug. Also, for the first time it was shown that Phenacoccus aceris could vector vitiviruses Grapevine virus A (GVA) and Grapevine virus B (GVB). First instar nymphs were the most efficient stage in transmitting GLRaV-1, -3, and GVA. This research sheds light on the transmission biology of grapevine viruses by Phenacoccus aceris and represents a step forward to leafroll disease management. 相似文献
20.
A. Rowhani Steve Daubert K. Arnold M. Al Rwahnih V. Klaassen D. Golino J. K. Uyemoto 《European journal of plant pathology / European Foundation for Plant Pathology》2018,151(4):919-925
An interactive relationship between vitiviruses and grapevine leafroll viruses was characterized in grapevine. Grapevine viruses A and B (GVA and GVB) were found more frequently in the presence of co-infecting Grapevine leafroll associated viruses (GLRaV-1, ?2 or ?3) than in their absence. The titers of the vitiviruses in co-infection with leafroll viruses were found to be higher than were their titers in the absence of leafroll virus infection. The occurrence of vitivirus-associated stem-pitting symptoms was correlated with leafroll virus co-infection. Specific pairing associations on the species level were found between different viti- and leafroll virus species: GVB was associated preferentially with GLRaV-2; GVA was associated preferentially with GLRaV-1 and GLRaV-3. In contrast to the increase in vitivirus titer seen with leafroll virus co-infection, the incidence and titer of grapevine leafroll virus appeared to be unaltered by vitivirus co-infection. The potential for a synergistic enhancement of grapevine disease in co-infected vines is discussed. 相似文献