Complete Genome Sequence of a Novel Wild tomato mosaic virus Isolate Infecting Nicotiana tabacum in China

Virus particles of approximately 740–760 nm in length and 13 nm in diameter were observed from a diseased Nicotiana tabacum (tobacco) plant in Sichuan Province, China. The complete genomic sequence of the virus isolate XC1 was determined to contain 9659 nucleotides without 3′ terminal poly(A) tail. XC1 has a genome typical of members of the genus Potyvirus, encoding a large polyprotein of 3075 amino acids. Putative proteolytic cleavage sites and a number of well characterized functional motifs were identified by sequence comparisons with those of known potyviruses. Sequence comparison revealed that XC1 shared the highest level of nucleotide sequence identity (76.5%) with Wild tomato mosaic virus (WTMV). Phylogenetic analysis showed that XC1 was closely related to the WTMV Guangdong isolate with an identity of 94.3% between CP gene sequence of the two viruses. We thus named XC1 WTMV‐XC‐1 as a novel isolate of WTMV. The full sequence of WTMV‐XC‐1 may serve as a basis for future investigations on the gene diversity of WTMV.     

Current status of Potyvirus in India

Potyvirus particles are flexuous rods of 700–900?nm in length and contain one positive sense single-stranded genomic RNA molecule of approximately 10?kb, which is encapsidated by a single type of coat protein. According to available NCBI database, Potyvirus infection is prominently present in Solanaceae, Leguminosae and Cucurbitaceae families in India. Potyviruses can induce a wide range of different symptoms in infected host plants including mosaic, stripe, mottling, vein clearing, vein banding, ringspots, necrotic or chlorotic lesions, flower breaking, stunting, wilting, and most commonly lead tostunting and yield losses. PCR-based methods for the detection and identification of potyviruses rely on degenerate primers designed for conserved regions. Potyvirus infection requires the interaction of host factors with viral proteins and RNA for its replication and systemic spread, i.e interaction between VPg and eIF4E is required for Potyvirus genome translation. Mutations in host translational initiation factor eIF4E cause the conformational shift in encoded proteins which are unable to bind with viral protein (VPg), resulting in broad-spectrum Potyvirus resistance.     

The incidence and distribution of viruses infecting lettuce, cultivated Brassica and associated natural vegetation in Spain

A virus survey was conducted during the spring and autumn of 2001 and 2002 to determine the presence, prevalence and distribution in Spain of the viruses that are most commonly found infecting lettuce and Brassica worldwide. Crop plants showing virus symptoms from the principal lettuce and Brassica-growing regions of Spain, and some samples of the annual and perennial flora nearby, were tested by enzyme-linked immunosorbent assays using specific commercial antibodies against the following viruses: Alfalfa mosaic virus (AMV), Broad bean wilt virus 1 (BBWV-1), Beet western yellows virus (BWYV), Cauliflower mosaic virus (CaMV), Cucumber mosaic virus (CMV), Lettuce mosaic virus (LMV), Pea seed-borne mosaic virus (PSbMV), Turnip mosaic virus (TuMV) and Tomato spotted wilt virus (TSWV). Samples were also tested with a Potyvirus genus antibody. Virus incidence was much lower in spring than in autumn, especially in 2001. In spring 2002, CMV and LMV were the most prevalent viruses in lettuce, while CaMV was the most important virus present in Brassica crops grown in Navarra, followed by CMV and BWYV. In the autumn, the spectrum of viruses was different; potyviruses were widespread in lettuce grown in Madrid, but TSWV and BWYV were predominant in the Murcia region. The prevalent Potyvirus detected in lettuce fields was LMV, but none of the samples collected were positive for PSbMV or TuMV. In Brassica crops, TSWV was the most abundant in autumn-sown crops, especially in the Navarra region. All of the viruses present in lettuce and Brassica were also frequently detected in their associated natural vegetation at the same time, suggesting that they probably play an important role as virus reservoirs. Sonchus spp. were particularly common and were frequently infected with CMV, LMV and BWYV. Another common species, Chenopodium album, was often infected with TSWV and BWYV. Multiple infections were common, especially in non-crop plants, and the most common combination was BWYV and TSWV. The role of weeds in the epidemiology of viruses that infect lettuce and Brassica crops in Spain is discussed.     

Incidence and diversity of viruses in cowpeas and weeds in the unmanaged farming systems of savanna zones in Nigeria

Fields surveys were conducted to assess the incidence of commonly known legume viruses on cowpeas and weed hosts within and around the cowpeas farms in nine locations across the three agro-ecological zones of Nigeria. Of 315 cowpea leaf samples collected and tested for eight viruses, 69.5% were found to be infected. Bean common mosaic virus-blackeye mosaic (BCMV-BlCM), genus Potyvirus had the highest incidence (70%) and was also the most prevalent (78%). Cowpea aphid-borne mosaic virus (CABMV, genus Potyvirus) had 64% as incidence, incidence of Southern bean mosaic virus (SBMV, genus Sobemovirus) was 21%. Bean pod mosaic virus (BPMV, genus Comovirus) was detected in 1% of the samples tested. Cowpea mosaic virus (CPMV, genus Comovirus) was undetected. Other viruses tested included Cowpea mottle virus (CPMoV, genus Carmovirus), Cucumber mosaic virus (CMV, genus Cucumovirus), and Cowpea mild mottle virus (CPMMV, genus Carlavirus). Multiple virus infections were detected in 68.0% of the infected cowpea leaf samples. The combination of BCMV-BlCM and CABMV was the most common, occurring in 76.4% of all samples. Virus incidence in weeds around the cowpea plots was 2.5% (9 out of 356) whereas 1.5% (5 out of 332) of the weeds collected within the cowpea plots were infected. Some of the weeds infected were Chromoleana odorata, Centrosema sp., Thithonia diversifolia and Talinum triangulare.     

Occurrence of latent virus infection in visually-rated cowpea (Vigna unguiculata L. Walp) seedlings

Abstract

The presence of latent infections was studied in five cowpeas varieties. Seeds of the varieties were planted and the seedlings inoculated with antigens from Cucumber mosaic virus (CMV) genus Cucumovirus, Bean common mosaic virus (BCMV) genus Potyvirus (Blackeye cowpea mosaic virus strain), Southern bean mosaic virus (SBMV) genus Sobemovirus and Cowpea mottle virus (CPMoV) genus Carmovirus seven days after planting. Seedlings expressing symptoms were rouged at two weeks after inoculation, while asymptomatic ones were subjected to serological indexing to detect the presence/absence of latent infection. Protein A-sandwich enzyme-linked immunosorbent assay (PAS ELISA) was employed for the serological detection of CMV, SBMV and CPMoV, while antigen-coated plate (ACP) ELISA was used to detect BCMV in the asymptomatic plants. Cowpea seedlings without virus symptoms but with positive serological reactions were considered as being latently infected. All of the inoculated TVu 1272 and SuVita-2 plants showed symptoms consistent with CMV and CPMoV infections, respectively. The rate of CMV latent infection was high in TVu 1179 (14.5%), low in SuVita-2 (1.3%) but not recorded in TVu 1272.     

Antibody-mediated improved resistance to ClYVV and PVY infections in transgenic tobacco plants expressing a single-chain variable region antibody

Transgenic Nicotiana tabacum plants expressing a single-chain variable region antibody fragment derived from a broad-spectrum monoclonal antibody 3-17 showed suppression of virus infection following challenge by two distinct potyviruses: potato virus Y strain D, and clover yellow vein virus strain 300. Monoclonal antibody 3-17, which was raised against the potyvirus Johnsongrass mosaic virus, was shown to react strongly with 14 potyvirus species. Two different single-chain antibody constructs were used to produce chimeric genes encoding recombinant proteins designed to be targeted either to the apoplasm or to the cytoplasm. Transgenic plant lines showed reduced numbers of local lesions and systemic symptoms when challenged with potato virus Y, strain D and reduced local lesions following challenge with clover yellow vein virus, strain 300. The level of suppression conferred by the transgene when plants were challenged under laboratory conditions with high concentrations of virus, together with the ability of the transgene to partially protect plants against distinct viruses suggest that one single-chain gene construct might be used to protect plants from distinct potyviruses.     

Taxonomy of the family potyviridae*

The paper contains the literature review of the family Potyviridae demonstrating the current state of the problem on classification of the most numerous taxonomic family among the phytopathogenic viruses. The data accumulated by the nineties allowed to transform the group of potyviruses into the family Potyviridae consisting of 4 genera bymo‐, rymo‐, ipomo‐ and potyviruses. The main phenotypic features and molecular parameters of separation of this family are given. The paper presents data of the authors on biological, physico‐chemical and antigenic characteristics of 10 viruses identified in the south of the Russian Far East and referred to the genus of potyviruses. Among them are three new, previously not described viruses in the literature: Tradescantia albiflora virus, Hippeastrum mosaic virus and Trifolium montanwn virus.     

The incidence of viruses in wild Brassica nigra in Dorset (UK)

Using enzyme‐linked immunosorbent assays, the frequency of occurrence of six viruses was determined in Brassica nigra collected from five coastal sites in Dorset, spanning approximately 24 km. During 1998–2000, the viruses detected were: Turnip mosaic virus (genus Potyvirus) (TuMV), Turnip yellow mosaic virus (genus Tymovirus) (TYMV), Turnip crinkle virus (genus Carmovirus) (TCV), Turnip rosette virus (genus Sobemovirus) (TRoV), Beet western yellows virus (genus Polerovirus) (BWYV) and Cauliflower mosaic virus (genus Caulimovirus) (CaMV). Multiple infections were detected in some individuals (48/447). TuMV was detected infrequently over the three‐year period (5/597). A representative isolate of each virus was tested for its effects on glasshouse‐grown individuals from different half‐sib families of B. nigra from four of the sites. Whether inoculated manually or via aphids (Myzus persicae), TuMV caused a rapid (within 10 days) lethal systemic necrosis in the B. nigra seedlings except when they were near flowering at the time of inoculation. Each of the other viruses invaded systemically but were not lethal. Indeed, BWYV systemically invaded 13/19 glasshouse‐grown B. nigra seedlings but did not produce any visible symptoms. Otherwise, the isolates tested differed in their pathogenicity and in the symptoms they produced in infected B. nigra. With TYMV or TCV viral antigen concentration was closely linked to pathogenicity; for TRoV or CaMV, there was little or no difference in virus concentration between plants with and without symptoms. Substantial and reproducible differences were observed in sensitivity/susceptibility among B. nigra genotypes from different sites in Dorset challenged with the same virus isolate.     

Soybean RNA interference lines silenced for eIF4E show broad potyvirus resistance

Soybean mosaic virus (SMV), a potyvirus, is the most prevalent and destructive viral pathogen in soybean-planting regions of China. Moreover, other potyviruses, including bean common mosaic virus (BCMV) and watermelon mosaic virus (WMV), also threaten soybean farming. The eukaryotic translation initiation factor 4E (eIF4E) plays a critical role in controlling resistance/susceptibility to potyviruses in plants. In the present study, much higher SMV-induced eIF4E1 expression levels were detected in a susceptible soybean cultivar when compared with a resistant cultivar, suggesting the involvement of eIF4E1 in the response to SMV by the susceptible cultivar. Yeast two-hybrid and bimolecular fluorescence complementation assays showed that soybean eIF4E1 interacted with SMV VPg in the nucleus and with SMV NIa-Pro/NIb in the cytoplasm, revealing the involvement of VPg, NIa-Pro, and NIb in SMV infection and multiplication. Furthermore, transgenic soybeans silenced for eIF4E were produced using an RNA interference approach. Through monitoring for viral symptoms and viral titers, robust and broad-spectrum resistance was confirmed against five SMV strains (SC3/7/15/18 and SMV-R), BCMV, and WMV in the transgenic plants. Our findings represent fresh insights for investigating the mechanism underlying eIF4E-mediated resistance in soybean and also suggest an effective alternative for breeding soybean with broad-spectrum viral resistance.     

Response of maize (Zea mays L.) lines carrying Wsm1, Wsm2, and Wsm3 to the potyviruses Johnsongrass mosaic virus and Sorghum mosaic virus

Maize dwarf mosaic disease is one of the most important viral diseases of maize (Zea mays L.) throughout the world. It is caused by several virus species in the family Potyviridae, genus Potyvirus, including Maize dwarf mosaic virus (MDMV), Sugarcane mosaic virus (SCMV), Johnsongrass mosaic virus (JGMV) and Sorghum mosaic virus (SrMV). Resistance to another member of the family Potyviridae, Wheat streak mosaic virus (WSMV), is conferred by three alleles (Wsm1, Wsm2, Wsm3) in the maize inbred line Pa405, and these or closely linked genes were previously shown to confer resistance to the potyviruses MDMV and SCMV. In this study, we assessed whether Wsm alleles are linked to resistance to JGMV and SrMV. Near isogenic lines (NILs) carrying one or two of the Wsm alleles introgressed into the susceptible line Oh28 and F1 progeny from NIL × Oh28 were tested for their response to JGMV and SrMV. Our results indicate that Wsm1 provides resistance to both JGMV and SrMV in a dose-dependent manner. Wsm2 and Wsm3 each provide limited resistance, and combining Wsm alleles enhances that resistance.     

Nucleotide sequence of Chinese rape mosaic virus (oilseed rape mosaic virus), a crucifer tobamovirus infectious on Arabidopsis thaliana

The complete nucleotide sequence of Chinese rape mosaic virus has been determined. The virus is a member of the tobamovirus genus of plant virus and is able to infect Arabidopsis thaliana (L.) Heynh systemically. The analysis of the sequence shows a gene array that seems to be characteristic of crucifer tobamoviruses and which is slightly different from the one most frequently found in tobamoviruses. Based on gene organization and on comparisons of sequence homologies between members of the tobamoviruses, a clustering of crucifer tobamoviruses is proposed that groups the presently known crucifer tobamovirus into two viruses with two strains each. A name change of Chinese rape mosaic virus to oilseed rape mosaic virus is proposed.Abbreviations 2-ME 2-mercaptoethanol - EDTA ethylenediaminetetraacetic acid - SDS sodium dodecyl sulfate - UTR untranslated region - MP movement protein - CP capsid protein - CRMV Chinese rape mosaic virus - TVCV turnip vein clearing virus - PaMMV paprika mild mottle virus - PMMV-I pepper mild mottle virus (Italian isolate) - PMMV-S pepper mild mottle virus (Spanish isolate) - ToMV tomato mosaic virus - TMV tobacco mosaic virus - TMGMV tobacco mild green mosaic virus - ORSV odontoglossum ringspot virus - SHMV sunn hemp mosaic virus - CGMMV cucumber green mottle mosaic virus - ORMV oilseed rape mosaic virus     

Translationally controlled tumour protein: A protein necessary for potyvirus intracellular multiplication that supports plant infection by unrelated viruses

The multifunctional protein translationally controlled tumour protein (TCTP) was previously identified as necessary for infection by the potyvirus pepper yellow mosaic virus. Using turnip mosaic virus (TuMV) as a model to study potyvirus biology, we confirmed that TCTP has a positive effect on virus infection. Living cell confocal microscopy demonstrated that TCTP colocalises with 6K2-tagged replication vesicles and with a perinuclear globular structure typically observed during potyvirus infection. Also, TCTP silenced protoplasts showed reduced virus accumulation, quantified by qRT-PCR, which suggests an effect on virus replication, translation or other intracellular process. Finally, TCTP silencing in plants reduced the accumulation of two species belonging to Orthotospovirus and a Begomovirus genus, which are not closely related to potyviruses. The results suggest that TCTP is a general susceptibility factor to several unrelated viruses.     

Host Ranges, Serology and Cytopathology of Eggplant and Tomato Strains of Eggplant Severe Mottle Virus, a New Potyvirus from Nigeria

Two potyvirus isolates from the eggplant (Solanum melongena L.) cultivars ‘Ex Benin’ and ‘Ex Jos’, respectively, in Nigeria proved to be almost identical in host range, symptomatology and reactivity with antisera to various potyviruses. In eggplant they caused a severe systemic mottle, blistering and malformation of leaves and an abnormal serration of the leaf margins. A potyvirus isolate from tomato showing mosaic symptoms was similar, but not identical to the eggplant isolates. In the slide, precipitin test the serological differentiation indices were between 1 and 3 for the eggplant and tomato isolates. In the immunoelectron microscopical decoration test all three virus isolates showed some reactivity with antisera to the following potyvir, uses: dioscorea green banding mosaic, groundnut eyespot, a mungbean isolate of peanut stripe, pepper veinal mottle, telfairia mosaic and a tomato isolate from Taiwan. No reactions were observed with antisera to other potyviruses. Cytopathogenic effects w,ere similar for all three isolates in the arrangement of virus particles, the structure of the cylindrical inclusions and the occurrence of clusters of small vesicles. However, other cytological alterations like accumulations of rod-shaped aggregates of,granular material, formation of giant mitochondria, degeneration of mitochondria and occurrence of a nucleolar inclusion differentiated the isolates.     

Cell-to-cell movement of three genera (+) ss RNA plant viruses

The current investigations of three genera plant virus cell-to-cell movement were presented. Viruses reveal different local transport strategies, but all of them are the results of virus factors–host components interactions. The Tobacco mosaic virus (TMV) does not require capsid protein for translocation through plasmodesmata but 30 K movement protein participates in this process. It was found direct or indirect TMV movement proteins host partners in Tobamovirus movement like: pectin methylesterase, movement protein binding 2C, chaperones or cytoskeleton components and endoplasmatic reticulum membranes. The Potex- and Potyvirus cell-to-cell movement is closely related to replication network. The PVX capsid protein and triple gene block protein system are responsible for efficient local transport. Potyviruses move through the plasmodesmata by involving viral encoded proteins but not specific movement proteins. While the Potyvirus is the biggest known plant virus genus, host components participating in or regulating directly its plasmodesmata-movement are still not clear.     

Inheritance of resistance to potato y viruses in Phaseolus vulgaris L

Summary Resistance to watermelon mosaic virus-2 in Phaseolus vulgaris L. is conferred by two distinct dominant alleles at independent loci. Based on segregation data one locus is designated Wmv, the other, Hsw. The dominant allele Wmv from cv. Great Northern 1140 prevents systemic spread of the virus but viral replication occurs in inoculated tissue. In contrast, Hsw confers both local and systemic resistance to WMV-2 below 30C. At higher temperatures, plants that carry this allele in the absence of modifying or epistatic factors develop systemic veinal necrosis upon inoculation with the virus that results in rapid death. Patho-type specificity has not been demonstrated for either allele; both factors confer resistance to every isolate tested. A temperature-sensitive shift in epistasis is apparent between dominant alleles at these loci. Because Hsw is very tightly linked if not identical to the following genes for hypersensitivity to potyviruses I, (bean common mosaic virus), Bcm, (blackeye cowpea mosaic virus), Cam, (cowpea aphid-borne mosaic virus) and Hss (soybean mosaic virus), parental, reciprocal dihybrid F₁ populations, and selected F₃ families were inoculated with each of these viruses and held at 35 C. F₁ populations developed vascular necrosis completely or primarily limited to inoculated tissue, while F₃ families from WMV-2-susceptible segregates were uniformly susceptible to these viruses. The relationship between Hsw, Wmv and other genes for potyvirus resistance suggest patterns in the evolution of resistance and viral pathogenicity. Characterization of the resistance spectrum associated with each factor provides an additional criterion to distinguish genes for plant virus resistance.     

The incidence of viruses in wild Brassica rapa ssp. sylvestris in southern England

Using enzyme‐linked immunosorbent assays, the frequency of occurrence of six viruses was determined in Brassica rapa ssp. sylvestris collected from two Thameside sites (Abingdon and Culham) in Oxfordshire and one near the Avon (Claverton) in Bath & North East Somerset. During 2000–2001, the viruses detected were: Beet western yellows virus (genus Polerovirus) (BWYV), Cauliflower mosaic virus (genus Caulimovirus) (CaMV), Turnip crinkle virus (genus Carmovirus) (TCV), Turnip rosette virus (genus Sobemovirus) (TRoV), and Turnip yellow mosaic virus (genus Tymovirus) (TYMV). BWYV and TYMV were the most frequently detected viruses at the Oxford shire sites, both as single infections (20/1743 and 66/1743 respectively) and as dual infections (7/1743). Turnip mosaic virus (genus Potyvirus) (TuMV) was not detected in the field‐grown plants assayed from any of the sites. There was a highly significant (x²_[1]=30.07, P<0.001) difference in the proportion of plants at each Oxfordshire site in which one or more viruses were detected, and essentially the same pattern of virus infection was observed in tests on B. rapa from the site near Claverton. At least one representative isolate of each detected virus was tested for its morphological and serological effects on glasshouse‐grown individuals from different half‐sib families of B. rapa from both Oxfordshire sites. Except for TRoV, where there was a large difference in the frequency of successful infection in B. rapa from the two locations (1/15 vs 11/15), no clear evidence of resistance or immunity to challenge was observed, although tolerance (virus invasion without symptoms) was frequent. Fewer of the plants from Abingdon were infected than those from Culham, when mechanically challenged with TRoV, but the two B. rapa populations were not otherwise consistently different, either in their infectibility by this virus or in their responses to challenge. However, with TCV, viral antigen concentration was closely linked to the severity of disease and the B. rapa from both Oxfordshire sites segregated into two classes: those with symptoms and most viral antigen, and those without symptoms and least viral antigen. These results suggest that generic risk assessments cannot be made due to differences in the way distinct B. rapa populations react to virus challenge.