Bean Dwarf Mosaic Virus Research Articles

Interactions between Common Bean Viruses and Their Whitefly Vector.

Common bean (Phaseolus vulgaris L.) is a widely cultivated crop, representing an important protein source in the human diet in developing countries. The production of this crop faces serious challenges, such as virus diseases transmitted by the whitefly Bemisia tabaci. Although there is a lot of information about some of these viruses, most of what we know has been developed using model systems, such as tomato plants and tomato yellow leaf curl virus (TYLCV). There is still very little information on the most relevant common bean viruses, such as bean golden mosaic virus (BGMV), bean golden yellow mosaic virus (BGYMV), bean dwarf mosaic virus (BDMV), cowpea mild mottle virus (CPMMV), and bean yellow disorder virus (BnYDV). In this review, we discuss the available data in the most up-to-date literature and suggest future research avenues to contribute to the development of management tools for preventing or reducing the damage caused by viruses in this important crop.

Bean dwarf mosaic virus

This datasheet on Bean dwarf mosaic virus covers Identity, Distribution, Vectors & Intermediate Hosts, Further Information.

Fine-tuning expression of begomoviral movement and nuclear shuttle proteins confers cell-to-cell movement to mastreviral replicons in Nicotiana benthamiana leaves.

Geminiviruses are a group of small plant viruses responsible for devastating crop damage worldwide. The emergence of agricultural diseases caused by geminiviruses is attributed in part to their high rates of recombination, leading to complementary function between viral components across species and genera. We have developed a mastreviral reporter system based on bean yellow dwarf virus (BeYDV) that replicates to high levels in the plant nucleus, expressing very high levels of GFP. To investigate the potential for complementation of movement function by other geminivirus genera, the movement protein (MP) and nuclear shuttle protein (NSP) from the bipartite begomovirus Bean dwarf mosaic virus (BDMV) were produced and characterized in Nicotiana benthamiana leaves. While overexpression of MP and NSP strongly inhibited GFP expression from the mastreviral reporter and caused adverse plant symptoms, optimizing the expression levels of MP and NSP allowed functional cell-to-cell movement. Hybrid virus vectors were created that express BDMV MP and NSP from mastreviral replicons, allowing efficient cell-to-cell movement comparable to native BDMV replicons. We find that the expression levels of MP and NSP must be fine-tuned to provide sufficient MP/NSP for movement without eliciting the plant hypersensitive response or adversely impacting gene expression from viral replicons. The ability to confer cell-to-cell movement to mastrevirus replicons depended strongly on replicon size: 2.1-2.7 kb replicons were efficiently moved, while 3 kb replicons were inhibited, and 3.9 kb replicons were very strongly inhibited. Optimized expression of MP/NSP from the normally phloem-limited Abutilon mosaic virus (AbMV) allows efficient movement in non-phloem cells.

First Report of Tobacco curly shoot virus Infecting Phaseolus vulgaris in China

Common bean (Phaseolus vulgaris) is one of the most economically important vegetable crops in China. It was reported that begomoviruses could infect P. vulgaris, including tomato yellow leaf curl virus (TYLCV), tomato yellow leaf curl China virus, bean dwarf mosaic virus, and cotton leaf crumple virus. In 2016, P. vulgaris with leaf crumple, thickening, and curling symptoms was found in Sichuan province, China. To identify potential begomoviruses infecting P. vulgaris, five samples of P. vulgaris were collected, and total DNA was extracted from symptomatic leaves. An approximately 500-bp DNA fragment was amplified from all the samples using the degenerate primer pair PA/PB detecting members of the genus Begomovirus (Deng et al. 1994). The fragment of isolate SC740 was cloned, and four clones were randomly chosen to be sequenced. Sequence alignments showed that three clones shared over 99% identity to tobacco curly shoot virus (TbCSV-[China:Yunnan Y98:03]), and one clone was most closely related to TYLCV-[China:Yunnan 3560:15] (98.5%). Based on the obtained sequences, two pairs of primers TbCSV-full-F (5′-AGACTTATTCGCCTGACAC-3′)/TbCSV-full-R (5′-TCTCTACTAACTGCAGATAT-3′) and TYLCV-full-F (5′-GATTTCGTTGTATGTTAGCT-3′)/TYLCV-full-R (5′-CGTGAACAGATTCAGGAAAT-3′) were designed for amplification of full-length DNA of TbCSV and TYLCV, respectively. The full length of TbCSV (accession no. MH165181) was 2,743 nt and shared 99.6% identity with TbCSV-[China:Yunnan 3560:15], and the complete DNA sequence of TYLCV (accession no. MH205950) was determined to be 2,781 nt and had 99.7% identity with TYLCV-[Australia: AU1923:06]. A fragment of approximately 600 bp was obtained from all samples using TYLCV-specific primers TYT-F/TYT-R (Ruan et al. 2013) by polymerase chain reaction, and an approximately 1.0-kb fragment was obtained from one (SC740) out of the five samples using TbCSV-specific primer pair Y35F1/Y35+10R (Qing et al. 2009). With the universal abutting primers (Briddon et al. 2002) for betasatellite, an amplicon of approximately 1,300 bp was obtained from sample SC740, but no amplification was obtained from the other four samples. Sequence alignment showed the betasatellite was 1,354 nt (accession no. MH165182) and shared the highest identity (99.8%) with tobacco curly shoot betasatellite (TbCSB-[China:Sichuan 776:16]). The infectious clones of TYLCV, TbCSV, and TbCSB were constructed and coinoculated to P. vulgaris. Typical leaf crumple, thickening, and curling symptoms were observed at 25 days postinoculation, which were indistinguishable from those observed on P. vulgaris in the field. In Sichuan, TbCSV has been reported infecting Malvastrum coromandelianum and caused severe leaf curl disease on pepper but has not been found on P. vulgaris (Qing et al. 2010). To our knowledge, this is the first report of TbCSV associated with its betasatellite infecting P. vulgaris in China, and coinfection of TYLCV and TbCSV/TbCSB.

Molecular identification of a new begomovirus infecting yellow passion fruit (Passiflora edulis) in Colombia.

The complete genome sequence of a bipartite begomovirus (genus Begomovirus, family Geminiviridae) infecting yellow passion fruit (Passiflora edulis) in the state of Valle del Cauca (Colombia) has been determined. The complete DNA-A and DNA-B components were determined to be 2600 and 2572 nt in length, respectively. The DNA-A showed the highest nucleotide sequence identity (87.2%) to bean dwarf mosaic virus (M88179), a begomovirus found in common bean crops in Colombia, and only 77.4% identity to passion fruit severe leaf distortion virus (FJ972767), a begomovirus identified infecting passion fruit in Brazil. Based on its sequence identity to all other begomoviruses known to date and in accordance with the ICTV species demarcation criterion for the genus Begomovirus (≥91% sequence identity for the complete DNA-A), the name passion fruit leaf distortion virus is proposed for this new begomovirus. To our knowledge, this is the first report of a bipartite begomovirus affecting passion fruit in Colombia and the second report of a geminivirus affecting this crop worldwide.

Blocking single-stranded transferred DNA conversion to double-stranded intermediates by overexpression of yeast DNA REPLICATION FACTOR A.

Agrobacterium tumefaciens delivers its single-stranded transferred DNA (T-strand) into the host cell nucleus, where it can be converted into double-stranded molecules. Various studies have revealed that double-stranded transfer DNA (T-DNA) intermediates can serve as substrates by as yet uncharacterized integration machinery. Nevertheless, the possibility that T-strands are themselves substrates for integration cannot be ruled out. We attempted to block the conversion of T-strands into double-stranded intermediates prior to integration in order to further investigate the route taken by T-DNA molecules on their way to integration. Transgenic tobacco (Nicotiana benthamiana) plants that overexpress three yeast (Saccharomyces cerevisiae) protein subunits of DNA REPLICATION FACTOR A (RFA) were produced. In yeast, these subunits (RFA1-RFA3) function as a complex that can bind single-stranded DNA molecules, promoting the repair of genomic double strand breaks. Overexpression of the RFA complex in tobacco resulted in decreased T-DNA expression, as determined by infection with A. tumefaciens cells carrying the β-glucuronidase intron reporter gene. Gene expression was not blocked when the reporter gene was delivered by microbombardment. Enhanced green fluorescent protein-assisted localization studies indicated that the three-protein complex was predominantly nuclear, thus indicating its function within the plant cell nucleus, possibly by binding naked T-strands and blocking their conversion into double-stranded intermediates. This notion was further supported by the inhibitory effect of RFA expression on the cell-to-cell movement of Bean dwarf mosaic virus, a single-stranded DNA virus. The observation that RFA complex plants dramatically inhibited the transient expression level of T-DNA and only reduced T-DNA integration by 50% suggests that double-stranded T-DNA intermediates, as well as single-stranded T-DNA, play significant roles in the integration process.

Tomato chlorotic leaf distortion virus, a new bipartite begomovirus infecting Solanum lycopersicum and Capsicum chinense in Venezuela

Virus isolate T217L was obtained from a diseased tomato (Solanum lycopersicum) plant showing leaf deformation and chlorotic mottle symptoms near Maracaibo in the state of Zulia, Venezuela. Full-length DNA-A and DNA-B molecules of T217L were cloned and sequenced. The genome organization of T217L was identical to the bipartite genomes of other begomoviruses described from the Americas. Characteristic disease symptoms were reproduced in S. lycopersicum and Capsicum annum plants inoculated using the cloned viral DNA-A and DNA-B components, confirming disease aetiology. A sequence analysis of DNA-A showed that the T217L isolate has the highest sequence identity (84%) with sida yellow mosaic Yucatan virus (SiYMYuV), sida golden mosaic Honduras virus (SiGMHV) and bean dwarf mosaic virus (BDMV) isolates. This is less than the 89% identity in the DNA-A component that has been defined as the threshold value for the demarcation of species in the genus Begomovirus. The molecular data show that isolate T217L belongs to a novel tentative begomovirus species, for which the name tomato chlorotic leaf distortion virus is proposed. TCLDV was also detected in symptomatic C. chinense plants growing near the T217L-infected plant.

Bean dwarf mosaic virus: a model system for the study of viral movement

Bean dwarf mosaic virus-[Colombia:1987] (BDMV-[CO:87]) is a single-stranded plant DNA virus, a member of the genus Begomovirus of the family Geminiviridae. BDMV virions are twinned incomplete isosahedra measuring 18 x 30 nm. The viral particle is composed of 110 subunits of coat protein, organized as 22 pentameric capsomers. Each subunit has a molecular mass of approximately 29 kDa. BDMV possesses two DNA components (designated DNA-A and DNA-B), each approximately 2.6 kb in size. The natural and most important host of BDMV is the common bean (Phaseolus vulgaris). Nicotiana benthamiana is often used as an experimental host. Common bean germplasm can be divided into two major gene pools: Andean materials, which are mostly susceptible to BDMV, and Middle American materials, which are mostly resistant to BDMV. The symptom intensity in common bean plants depends on the stage of infection. Early infection of susceptible bean seedlings will result in severe stunting and dwarfing, leaf distortion and mottling or mosaic, as well as chlorotic or yellow spots or blotches. BDMV-infected plants usually abort their flowers or produce severely distorted pods. Late infection of susceptible plants or early infection of moderately resistant genotypes may show a mild mosaic, mottle and crumpling or an irregular distribution of variegated patches. BIOLOGICAL PROPERTIES: As a member of the Begomovirus group, BDMV is transmitted from plant to plant by the whitefly Bemisia tabaci. BDMV is a nonphloem-limited virus and can replicate and move in the epidermal, cortical and phloem cells. As a nonphloem-limited virus, it is sap-transmissible.

Tomato yellow leaf curl virus infection of a resistant tomato line with a silenced sucrose transporter gene LeHT1 results in inhibition of growth, enhanced virus spread, and necrosis

To identify genes involved in resistance of tomato to Tomato yellow leaf curl virus (TYLCV), cDNA libraries from lines resistant (R) and susceptible (S) to the virus were compared. The hexose transporter LeHT1 was found to be expressed preferentially in R tomato plants. The role of LeHT1 in the establishment of TYLCV resistance was studied in R plants where LeHT1 has been silenced using Tobacco rattle virus-induced gene silencing (TRV VIGS). Following TYLCV inoculation, LeHT1-silenced R plants showed inhibition of growth and enhanced virus accumulation and spread. In addition, a necrotic response was observed along the stem and petioles of infected LeHT1-silenced R plants, but not on infected not-silenced R plants. This response was specific of R plants since it was absent in infected LeHT1-silenced S plants. Necrosis had several characteristics of programmed cell death (PCD): DNA from necrotic tissues presented a PCD-characteristic ladder pattern, the amount of a JNK analogue increased, and production of reactive oxygen was identified by DAB staining. A similar necrotic reaction along stem and petioles was observed in LeHT1-silenced R plants infected with the DNA virus Bean dwarf mosaic virus and the RNA viruses Cucumber mosaic virus and Tobacco mosaic virus. These results constitute the first evidence for a necrotic response backing natural resistance to TYLCV in tomato, confirming that plant defense is organized in multiple layers. They demonstrate that the hexose transporter LeHT1 is essential for the expression of natural resistance against TYLCV and its expression correlates with inhibition of virus replication and movement.

Quantitative Resistance to Bean dwarf mosaic virus in Common Bean Is Associated with the Bct Gene for Resistance to Beet curly top virus.

The dominant resistance gene, Bct, in common bean (Phaseolus vulgaris) confers qualitative resistance to Beet curly top virus, a leafhopper-transmitted geminivirus in the genus Curtovirus. To determine whether this gene confers resistance to other geminiviruses, bean plants of a recombinant inbred population were sap-inoculated with Bean dwarf mosaic virus (BDMV), a whitefly-transmitted bipartite begomovirus in the genus Begomovirus. Results indicated that Bct (or tightly linked gene) is associated with quantitative resistance to BDMV; thus, the Bct locus is associated with resistance to a bean-infecting begomovirus and curtovirus. The difference in the nature of the resistance to these geminiviruses may indicate a role for minor genes in begomovirus resistance or differences in the virus-host interaction. The Bct locus, whether it acts alone or represents a cluster of tightly linked genes, will be useful in breeding for broad-spectrum begomovirus resistance in common bean.

Geminivirus resistance breeding in common bean.

Abstract Common bean ( Phaseolus vulgaris L.) is among the most important food staples worldwide, mainly because of its relatively high protein content and other valuable nutritional qualities. Unfortunately, common beans are highly susceptible to plant viruses in general, including one of the largest families of plant viruses known as the Geminiviridae . These viruses, unlike most RNA plant viruses, possess a DNA genome that allows these pathogens to replicate in the nucleus of their plant host cells, thus, causing major damage to susceptible legume crops. Although some geminiviruses, belonging to two ( Mastrevirus and Curtovirus ) of the four genera that compose the Geminiviridae , can be transmitted by leafhoppers (Cicadellidae) to common bean, most geminiviruses that attack legumes belong to the genus Begomovirus , and are transmitted by the whitefly Bemisia tabaci (Genn.). In fact, the type species of this genus is bean golden yellow mosaic virus (BGYMV), which attacks over a million hectares of common bean grown in the lowlands and mid-altitude valleys of Central America, southern Mexico, the Caribbean region and northern South America. Another begomovirus, bean golden mosaic virus (BGMV), affects over two million hectares of common bean in Brazil, northern Argentina and the lowlands of Bolivia, in South America. In this region, bean dwarf mosaic virus (BDMV) has also caused the loss of thousands of hectares of highly valuable common bean genotypes in northwestern Argentina. In northwestern Mexico, thousands of hectares of common bean have been under the constant attack of a begomovirus related to a virus originally described in cucurbits, squash leaf curl virus (SLCV), in southwestern USA, currently known in Mexico as bean calico mosaic virus (BCaMV). Fortunately, none of these viruses is present in Eastern Africa, where the common bean is also a major food staple. Another Old World begomovirus reported to attack common bean both in the Old and New World, is tomato yellow leaf curl virus (TYLCV). This review describes the different breeding approaches followed since the 1970s to introduce resistance to begomoviruses infecting important legume crops, particularly common bean. This has been a challenging process, considering that no immune genotypes have been identified in more than 30000 common bean genotypes exposed to begomoviruses, such as BGMV or BGYMV.

The N-terminus of the Begomovirus Nuclear Shuttle Protein (BV1) Determines Virulence or Avirulence in Phaseolus vulgaris

The BV1 gene of the bipartite Begomovirus genome encodes a nuclear shuttle protein (NSP) that is also an avirulence determinant in common bean. The function of the NSP of two common bean-infecting bipartite begomoviruses, Bean dwarf mosaic virus (BDMV) and Bean golden yellow mosaic virus (BGYMV), was investigated using a series of hybrid DNA-B components expressing chimeric BDMV and BGYMV NSP, and genotypes of the two major common bean gene pools: Andean (cv. Topcrop) and Middle American (cvs. Alpine and UI 114). BDMV DNA-A coinoculated with HBDBG4 (BDMV DNA-B expressing the BGYMV NSP) and HBDBG9 (BDMV DNA-B expressing a chimeric NSP with the N-terminal 1 to 42 amino acids from BGYMV) overcame the BDMV resistance of UI 114. This established that the BDMV NSP is an avirulence determinant in UI 114, and mapped the domain involved in this response to the N-terminus, which is a variable surface-exposed region. BDMV DNA-A coinoculated with HBDBG10, expressing a chimeric NSP with amino acids 43 to 92 from BGYMV, was not infectious, revealing an essential virus-specific domain. In the BGYMV background, the BDMV NSP was a virulence factor in the Andean cv. Topcrop, whereas it was an avirulence factor in the Middle American cultivars, particularly in the absence of the BGYMV NSP. The capsid protein (CP) also played a gene pool-specific role in viral infectivity; it was dispensable for infectivity in the Andean cv. Topcrop, but was required for infectivity of BDMV, BGYMV, and certain hybrid viruses in the Middle American cultivars. Redundancy of the CP and NSP, which are nuclear proteins involved directly or indirectly in viral movement, provides a masking effect that may allow the virus to avoid host defense responses.

Characterization of a novel Toll/interleukin‐1 receptor (TIR)‐TIR gene differentially expressed in common bean (Phaseolus vulgaris cv. Othello) undergoing a defence response to the geminivirus Bean dwarf mosaic virus

SUMMARY Common bean (Phaseolus vulgaris L.) cultivar (cv.) Othello develops a hypersensitive response-associated vascular resistance to infection by Bean dwarf mosaic virus (BDMV), a single-stranded DNA virus (genus Begomovirus, family Geminiviridae). A PCR-based cDNA subtraction approach was used to identify genes involved in this resistance response. Eighteen clones, potentially involved with BDMV resistance, were identified based upon being up-regulated in BDMV-infected tissues and/or having sequence similarity with known resistance-associated genes. Analysis of these clones revealed potential genes involved in pathogen defence, including pathogenesis-related protein genes and resistance gene analogues (RGAs). Further characterization of one RGA, F1-10, revealed that it encodes a predicted protein with a double Toll/interleukin-1 receptor (TIR) motif. Full-length (F1-10) and spliced (F1-10sp) forms of the RGA were strongly up-regulated in BDMV-infected cv. Othello hypocotyl tissues by 4 days post-inoculation, but not in equivalent mock-inoculated tissues. In agroinfiltration experiments, F1-10, but not F1-10sp, mediated resistance to BDMV in the susceptible common bean cv. Topcrop. By contrast, transgenic Nicotiana benthamiana lines expressing F1-10 or F1-10sp were not resistant to BDMV. Interestingly, when these transgenic lines were inoculated with the potyvirus Bean yellow mosaic virus, some F1-10 lines showed a more severe symptom phenotype compared with non-transgenic control plants. Based on these findings, F1-10 was named: Phaseolus vulgaris VIRUS response TIR-TIR GENE 1 (PvVTT1).

Permeabilized mammalian cells as an experimental system for nuclear import of geminiviral karyophilic proteins and of synthetic peptides derived from their nuclear localization signal regions

The plant-infecting geminiviruses deliver their genome and viral proteins into the host cell nucleus. Members of the family Geminiviridae possess either a bipartite genome composed of two approximately 2.6 kb DNAs or a monopartite genome of approximately 3.0 kb DNA. The bipartite genome of Bean dwarf mosaic virus (BDMV) encodes several karyophilic proteins, among them the capsid protein (CP) and BV1 (nuclear shuttle protein). A CP is also encoded by the monopartite genome of Tomato yellow leaf curl virus (TYLCV). Here, an in vitro assay system was used for direct demonstration of nuclear import of BDMV BV1 and TYLCV CP, as well as synthetic peptides containing their putative nuclear localization signals (NLSs). Full-length recombinant BDMV BV1 and TYLCV CP mediated import of conjugated fluorescently labelled BSA molecules into nuclei of permeabilized mammalian cells. Fluorescently labelled and biotinylated BSA conjugates bearing the synthetic peptides containing aa 3-20 of TYLCV CP (CP-NLS) or aa 84-106 of BDMV BV1 (BV1-NLS) were also imported into the nuclei of permeabilized cells. This import was blocked by the addition of unlabelled BSA-NLS peptide conjugates or excess unlabelled free NLS peptides. The CP- and BV1-NLS peptides also mediated nuclear import of fluorescently labelled BSA molecules into the nuclei of microinjected mesophyll cells of Nicotiana benthamiana leaves, demonstrating their biological function in intact plant tissue. BV1-NLS and CP-NLS were shown to mediate specific binding to importin alpha, both in vitro and in vivo. These results are consistent with a common nuclear-import pathway for CP and BV1, probably via importin alpha.

A viral resistance gene from common bean functions across plant families and is up-regulated in a non-virus-specific manner.

Genes involved in a viral resistance response in common bean (Phaseolus vulgaris cv. Othello) were identified by inoculating a geminivirus reporter (Bean dwarf mosaic virus expressing the green fluorescent protein), extracting RNA from tissue undergoing the defense response, and amplifying sequences with degenerate R gene primers. One such gene (a TIR-NBS-LRR gene, RT4-4) was selected for functional analysis in which transgenic Nicotiana benthamiana were generated and screened for resistance to a range of viruses. This analysis revealed that RT4-4 did not confer resistance to the reporter geminivirus; however, it did activate a resistance-related response (systemic necrosis) to seven strains of Cucumber mosaic virus (CMV) from pepper or tomato, but not to a CMV strain from common bean. Of these eight CMV strains, only the strain from common bean systemically infected common bean cv. Othello. Additional evidence that RT4-4 is a CMV R gene came from the detection of resistance response markers in CMV-challenged leaves of RT4-4 transgenic plants, and the identification of the CMV 2a gene product as the elicitor of the necrosis response. These findings indicate that RT4-4 functions across two plant families and is up-regulated in a non-virus-specific manner. This experimental approach holds promise for providing insights into the mechanisms by which plants activate resistance responses against pathogens.

REPLACING THE AC2/AC3 GENES OF ABUTILON MOSAIC VIRUS (ABMV) WITH THOSE OF BEAN DWARF MOSAIC VIRUS ENHANCES ABMV ACCUMULATION, MOVEMENT, AND SYMPTOM SEVERITY IN BEAN

SUMMARY Abutilon mosaic virus (AbMV) and Bean dwarf mosaic virus (BDMV) are phylogenetically related, bipartitegenome begomoviruses. AbMV is limited to the plant host phloem but BDMV is not. We have previously provided evidence that the genomic DNA-A component of BDMV contains determinants involved in movement (Levy and Czosnek, 2003). We report here that the DNA-A-encoded genes AC2 and AC3 are involved in virus accumulation and spread. To follow AbMV and BDMV movement in inoculated bean plants, we replaced their coat protein genes (CP) with DNA encoding the green or the red fluorescent proteins (GFP, RFP) to create BDMV-CP:GFP, BDMV-CP:RFP, and AbMV-CP:GFP. Frame-shift mutations in BDMV AC2 and AC3 to produce BDMV-CP:GFP-mC23 resulted in inhibition of BDMV movement when co-inoculated with BDMV DNA-B. The mutation reverted to wildtype and movement was restored when BDMVCP:GFP-mC23 was co-inoculated with BDMV-CP:RFP and BDMV DNA-B, which strongly suggests that the AC2/AC3 region is important for BDMV movement. Consequently we replaced the AC2/AC3 region of AbMV-CP:GFP with that of BDMV to create AbMVCP:GFP-C23: BDMV. AbMV-CP: GFP-C23: BDMV inoculated together with AbMV DNA-B moved from cell to cell in the epidermis towards the phloem, and long-distance in the entire plant, even though inocula lacked part of all of BDMV DNA-B. AbMV-CP:GFP with its cognate DNA-B was unable to move. Inoculation of bean with AbMV-CP:GFP-C23:BDMV and BDMV DNA-B resulted in the accumulation of very large amounts of viral DNA, in remarkably fast systemic movement, and in the early appearance of severe symptoms, which in most cases, resulted in an inhibition of germination. These results suggest that interactions between AC2/AC3 of BDMV and other proteins encoded by AbMV DNA-A and DNA-B influence the ability of

Interaction of DNA with the movement proteins of geminiviruses revisited.

Geminiviruses manage the transport of their DNA within plants with the help of three proteins, the coat protein (CP), the nuclear shuttle protein (NSP), and the movement protein (MP). The DNA-binding capabilities of CP, NSP, and MP of Abutilon mosaic virus (AbMV; family Geminiviridae; genus Begomovirus) were scrutinized using gel mobility shift assays and electron microscopy. CP and NSP revealed a sequence-independent affinity for both double-stranded and single-stranded DNA, as has been previously reported for other begomoviruses. MP interacted selectively with dimeric supercoiled plasmid DNA in the electrophoretic assay. Further apparent size- and form-selective binding capacities of MP have been previously reported for another geminivirus (Bean dwarf mosaic virus), but in the case of AbMV, they have been identified as the result of electrophoretic interference rather than of complex formation. Without these complications, electron microscopy confirmed the assembly of double-stranded supercoiled DNA with NSP and MP into conspicuous structures and provided the first direct evidence for cooperative interaction of MP, NSP, and DNA. Based on these results and previous ones, a transport model of geminiviruses is discussed in which NSP packages DNA and MP anchors this complex to the protoplasmic leaflets of plasma membranes and microsomes for cell-to-cell movement.

The DNA-B of the non-phloem-limited bean dwarf mosaic virus (BDMV) is able to move the phloem-limited Abutilon mosaic virus (AbMV) out of the phloem, but DNA-B of AbMV is unable to confine BDMV to the phloem.

Abutilon mosaic virus (AbMV) and bean dwarf mosaic virus (BDMV) are two phylogenetically related bipartite begomoviruses. While AbMV is restricted to phloem, BDMV spreads to non-phloem tissues. Cell-to-cell and long-distance movement of AbMV and BDMV were investigated after replacing the coat protein (CP) gene with the reporter gene encoding the green fluorescence protein (GFP). The DNA-A and DNA-B genomic components of AbMV and BDMV, and their pseudorecombinants (PR), were delivered to bean (Phaseolus vulgaris) seedlings and detached leaves with DNA-coated microprojectiles. Virus-associated fluorescence was observed with the confocal microscope. Delivery of AbMV and BDMV GFP reporters showed that the epidermal tissue was the main recipient of the viral DNA; the DNA-A of the two viruses was unable to move out of the recipient cells. AbMV DNA-A co-inoculated with AbMV DNA-B did not move from cell to cell in the epidermis and did not reach the phloem. However, co-inoculation of AbMV DNA-A with BDMV DNA-B resulted in PR cell-to-cell movement out of the epidermis and long-distance movement in the phloem. In contrast, BDMV DNA-A moved from cell to cell and over a long distance when co-inoculated with either its own DNA-B or with the DNA-B of AbMV. Thus, the DNA-B of the non-phloem-limited BDMV overcame the phloem limitation of AbMV. In the reciprocal case, the DNA-B of the phloem-limited AbMV did not confine the non-phloem limited BDMV to the phloem. Hence, we assume that the DNA-A component of BDMV includes determinants involved in the movement pattern of the virus in addition to the DNA-B-encoded BC1 and BV1 which have previously been shown to be involved in virus movement. The results also confirm that the CP is not necessary for virus movement; however, replacing the CP of AbMV and BDMV with GFP resulted in a decrease in symptom severity. DNA-B was involved in symptom severity; the B component of BDMV produced symptoms more severe than those induced by that of AbMV, whether in wild-type PRs or in PRs with CP-GFP replacement. It is interesting to note that when the GFP gene under the control of the CaMV 35S promoter (35S-GFP) was delivered to the bean tissue, with or without the DNA-B component of BDMV, GFP was expressed but did not move from cell to cell. However, when the 35S-GFP was delivered together with BDMV DNA-A and DNA-B, GFP showed cell-to-cell movement in the epidermis but was restricted to these cells. Hence, infection of cells with a functional bipartite begomovirus may facilitate cell-to-cell movement of macromolecules.

Genetics of resistance to the geminivirus, Bean dwarf mosaic virus, and the role of the hypersensitive response in common bean.

Bean dwarf mosaic virus (BDMV) is a single-stranded DNA virus (genus: Begomovirus, family: Geminiviridae) that infects common bean ( Phaseolus vulgaris L.) and causes stunted plant growth, and mosaic and mottle symptoms in leaves. BDMV shows differential pathogenicity in common bean, infecting germplasm of the Andean gene pool (e.g., the snap bean cultivar Topcrop), but not that of the Middle American gene pool (e.g., the pinto bean cultivar Othello). Resistance to BDMV in Othello is associated with development of a hypersensitive response (HR) in vascular (phloem) tissues. In this study, Middle American germplasm representing the four recognized races (i.e., Durango, Guatemala, Jalisco, and Mesoamerica) and the parents of Othello were inoculated with BDMV and a BDMV-green fluorescent protein (GFP) reporter. All genotypes showed partial or complete resistance to BDMV and BDMV-GFP, indicating the widespread distribution of resistance in the Middle American gene pool. A number of BDMV-resistant germplasm did not show the HR, indicating it is not correlated with resistance. In the F(1), F(2), and F(3) of reciprocal crosses between Othello and Topcrop, a single dominant allele, Bdm, conferred BDMV resistance.

Tomato mottle Taino virus pseudorecombines with PYMV but not with ToMoV: implications for the delimitation of cis- and trans-acting replication specificity determinants.

Over the last decade, the tomato production in Cuba has been affected by new whitefly-associated diseases. In addition to the well-documented presence of Tomato yellow leaf curl virus (TYLCV) along the island, the occurrence of bipartite begomoviruses has also been reported. One of them, tentatively named Tomato mottle Taino virus (ToMoTV), has now been cloned and characterized at the molecular level. Its genomic organization is similar to other bipartite geminiviruses. Phylogenetic analyses placed ToMoTV in a subcluster with other geminiviruses isolated in the Caribbean Basin: Tomato mottle virus (ToMoV), Bean dwarf mosaic virus, Abutilon mosaic virus, Sida golden mosaic virus and Potato yellow mosaic virus (PYMV). Biolistic inoculation of tobacco and tomato plants with cloned viral DNA showed that ToMoTV pseudorecombines with PYMV-GP as predicted by the identity of their iterative elements, whereas it does not show the same ability with ToMoV, even when their replication-associated proteins (Rep and REn) show the highest percentage of similarity. A comparative analysis of Rep proteins from begomoviruses that are able to produce viable reassortants suggests that some key elements for virus replication specificity are located in the first ten amino acids of this protein.