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Garrido-Ramirez, E. R., and R. L. Gilbertson. "First Report of Tomato Mottle Geminivirus Infecting Tomatoes in Yucatan, Mexico." Plant Disease 82, no. 5 (May 1998): 592. http://dx.doi.org/10.1094/pdis.1998.82.5.592b.
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Whitefly-transmitted geminiviruses are a major constraint on tomato production in Mexico (3). In the Yucatan State, these viruses can cause serious losses in late season plantings. As part of an effort to characterize these viruses, leaf samples from four tomato plants showing symptoms of geminivirus infection, such as stunted growth and leaf mottling and deformation, were collected from a single field in the Yucatan State in February, 1996. Geminivirus nucleic acids were detected in leaf samples from all four plants by squash blot hybridization analysis with a general DNA probe for Western Hemisphere whitefly-transmitted geminiviruses (2). Nicotiana benthamiana plants inoculated with sap prepared with leaf tissue from one plant developed stunted growth and leaf mottling and deformation. When graft-transmitted from N. benthamiana to tomato, the geminivirus(es) induced leaf mottling and deformation, which were similar to symptoms in the field-collected tomato plants. The presence of geminivirus DNA in the sap- and graft-inoculated plants was confirmed with the polymerase chain reaction (PCR) and degenerate primers for the DNA-A (PAL1v1978 and PAR1c496) or DNA-B (PBL1v2040 and PCRc1) components of whitefly-transmitted geminiviruses (4). Using PCR and these degenerate primers, approximately 1.1-kb DNA-A and approximately 0.6-kb DNA-B fragments were amplified from DNA extracts prepared from leaves of each of the four Yucatan tomato plants. No DNA fragments were amplified from these extracts with primers for pepper huasteco geminivirus (pAL1c2329 and pAL1v1471, or pBR1c840 and pBL1v1830). To determine the identity of the geminivirus(es) infecting these tomato plants, the PCR-amplified DNA-A and DNA-B fragments from one of the samples were cloned and sequenced. Comparisons made with these sequences revealed two distinct types of DNA-A and DNA-B clones, indicating a mixed infection of at least two bipartite geminiviruses. DNA-A and DNA-B sequences of one set of clones were >97% identical to sequences of tomato mottle geminivirus (ToMoV) from Florida (1). The presence of ToMoV in all four tomato leaf samples was demonstrated by the PCR-mediated amplification of a 0.9-kb DNA-A fragment with ToMoV-specific primers (pAL1v2295 and pAR1c580). The identity of this 0.9-kb DNA fragment was further confirmed based upon its hybridization with a full-length clone of ToMoV DNA-A under high stringency conditions (2). A data base search made with the sequence of the other type of DNA-A clone revealed sequence identities of <70% with various bipartite geminiviruses (e.g., identities of 70% with tomato mottle, 69% with Sida golden mosaic, 67% with bean dwarf mosaic, and 66% with taino tomato mottle and with potato yellow mosaic), which confirmed that a second geminivirus was present in a mixed infection with ToMoV in this tomato leaf sample. To confirm the bipartite nature of this geminivirus, a DNA-B fragment that contained the common region (CR) sequence was amplified from the same sample with PCR and primers PBL1v2040 and PBR1c970 (a degenerate primer that anneals within the BV1 open reading frame; F. M. Zerbini and R. L. Gil-bertson, unpublished data), cloned, and sequenced. The CR sequence of this DNA-B fragment was 96% identical to that of the DNA-A fragment, which establishes the presence of another bipartite geminivirus in this sample. This is the first report of ToMoV in Mexico. These results also suggest that at least two bipartite geminiviruses may infect tomatoes in the Yucatan Peninsula. References: (1) A. M. Abouzid et al. J. Gen. Virol. 73:3225, 1992. (2) R. L. Gilbertson et al. Plant Dis. 75:336, 1991. (3) J. E. Polston and P. K. Anderson. Plant Dis. 81:1358, 1997. (4) M. R. Rojas et al. Plant Dis. 77:340, 1993.
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Potter, J. L., M. M. Roca de Doyle, M. K. Nakhla, and D. P. Maxwell. "First Report and Characterization of Rhynchosia golden mosaic virus in Honduras." Plant Disease 84, no. 9 (September 2000): 1045. http://dx.doi.org/10.1094/pdis.2000.84.9.1045a.
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Rhynchosia minima was suspected to be a weed host of Bean golden yellow mosaic virus (BGYMV, previously designated Bean golden mosaic virus type II). Leaf tissue that exhibited yellow mosaic foliar symptoms characteristic of a geminivirus infection was collected in the Comayagua Valley in Honduras in July 1999. Extraction of viral DNA from the symptomatic leaves was accomplished with the DNeasy Plant Mini Kit (Qiagen Inc., Valencia, CA). Subsequent viral DNA amplification was accomplished with degenerative primers for the cp gene (AV494/AC1048) (4). The 570-bp fragment was cloned into the pGEM T-Easy vector (Promega Corp., Madison, WI) producing the recombinant plasmid pRhyb5. The viral insert was sequenced, and from this sequence, specific primers (RHc549 and RHv29) were designed to amplify the remaining part of DNA-A. The 2.1-kb-amplified polymerase chain reaction (PCR) fragment was cloned into the pGEM T-Easy vector producing the recombinant plasmid (pRhya-sp), and the viral insert was sequenced. Nucleotide sequence comparison (GAP program, Wisconsin Package Version 10.0, Genetics Computer Group, Madison, WI) of the complete 2,624-bp DNA-A (GenBank accession no. AF239671) to geminiviruses representing the major phylogenetic clusters (1) showed nucleotide identities ranging from 63 to 82%. Sequence comparisons for the common region and rep, trap, ren, and cp genes with the most closely related geminivirus, Pepper hausteco virus (PHV, X70418), gave 76, 82, 79, 81, and 82% nucleotide identities, respectively. There is a direct repeat (TATCGGT) of 7 nt 5′ (viral sense polarity) of the conserved TATA box, and this repeat is most analogous to that in PHV (1). Specific primers were designed in the complementary sense (RGBc2414, BGBc2553) from the common region DNA-A sequence and used with a degenerative viral sense primer for the DNA-B (PBC1v2039) (3) to amplify a 647-bp fragment. Sequence comparison for the common region (134 nt from the rep gene start codon toward the 3′ end) from the DNA-B sequence had 88% nt identity to the DNA-A sequence, thus indicating that this geminivirus is bipartite. These sequence analyses indicated that this geminivirus isolated from R. minima is distinct from previously described geminiviruses, and we propose the name Rhynchosia golden mosaic virus (RGMV). From rep gene sequence alignments, RGMV has an apparent genome recombination between Old and New World geminiviruses (Tomato yellow leaf curl virus and Bean dwarf mosaic virus) as previously noted for PHV (2). Our results indicate that RGMV is a distinct geminivirus from BGYMV, and, thus, additional studies are needed to establish the importance of R. minima as a reservoir for vegetable-infecting geminiviruses. This study is the first report of another virus in the PHV phylogenetic cluster and is thus of importance in the understanding of recombinant viruses and their phylogenetic relationship to other characterized geminiviruses. References: (1) J. C. Faria et al. Phytopathology 84:321, 1994. (2) M. Padidam et al. Virology 265:218, 1999. (3) M. R. Rojas et al. Plant Dis. 77:340, 1993. (4) S. Wyatt and J. K. Brown. Phytopathology 86:1288, 1996.
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Ribeiro, S. G., A. C. de Ávila, I. C. Bezerra, J. J. Fernandes, J. C. Faria, M. F. Lima, R. L. Gilbertson, E. Maciel-Zambolim, and F. M. Zerbini. "Widespread Occurrence of Tomato Geminiviruses in Brazil, Associated with the New Biotype of the Whitefly Vector." Plant Disease 82, no. 7 (July 1998): 830. http://dx.doi.org/10.1094/pdis.1998.82.7.830c.
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Although tomato golden mosaic virus (TGMV) was reported in Brazil more than 20 years ago (3), tomato-infecting geminiviruses have not been of economic significance in the country until recently. However, a sharp increase in the incidence of geminivirus-like symptoms in tomatoes has been reported in several areas of Brazil since 1994. This has coincided with the appearance of the B biotype of Bemisia tabaci, which, as opposed to the A biotype, readily colonizes solanaceous plants (2). We have isolated geminiviruses from symptomatic tomato plants in the Federal District, in two different areas of the state of Minas Gerais, and in the state of Pernambuco. Tomato plants in these areas showed a variety of symptoms, including yellow mosaic, severe leaf distortion, down-cupping, and epinasty. Whitefly infestation was high in all fields sampled, and in some fields, particularly in Pernambuco, incidence of virus-like symptoms was close to 100%, and no tomatoes of commercial value were harvested (1). Using primer pairs PAL1v1978/PAR1c496 and PCRc1/PBL1v2040 (4), DNA-A and -B fragments were polymerase chain reaction (PCR)-amplified from total DNA extracted from diseased plants, cloned, and sequenced. Sequence comparisons of the PCR fragments indicated the existence of at least six different geminiviruses. The nucleotide sequence homologies for DNA-A fragments ranged from 67 to 80% for the 5′ end of the cp gene, and from 44 to 80% for the 5′ end of the rep gene. Data base comparisons indicated the viruses are most closely related to TGMV, bean golden mosaic virus from Brazil (BGMV-Br), and tomato yellow vein streak virus (ToYVSV), although homologies were less than 80% for the fragments compared. A similar lack of a close relationship with each other and other geminiviruses was obtained with two DNA-B component PCR products compared, corresponding to the 5′ end of the BC1 open reading frame. Infectious, full-length genomic clones from the tomato viruses are being generated for biological and molecular characterization. References: (1) I. C. Bezerra et al. Fitopatol. Bras. 22:331, 1997. (2) F. H. França et al., Ann. Soc. Entomol. Bras. 25:369, 1996. (3) J. C. Matyis et al. Summa Phytopathol. 1:267, 1975. (4) M. R. Rojas et al. Plant Dis. 77:340, 1993.
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Rojas, Maria R., Monica A. Macedo, Minor R. Maliano, Maria Soto-Aguilar, Juliana O. Souza, Rob W. Briddon, Lawrence Kenyon, et al. "World Management of Geminiviruses." Annual Review of Phytopathology 56, no. 1 (August 25, 2018): 637–77. http://dx.doi.org/10.1146/annurev-phyto-080615-100327.
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Management of geminiviruses is a worldwide challenge because of the widespread distribution of economically important diseases caused by these viruses. Regardless of the type of agriculture, management is most effective with an integrated pest management (IPM) approach that involves measures before, during, and after the growing season. This includes starting with resistant cultivars and virus- and vector-free transplants and propagative plants. For high value vegetables, protected culture (e.g., greenhouses and screenhouses) allows for effective management but is limited owing to high cost. Protection of young plants in open fields is provided by row covers, but other measures are typically required. Measures that are used for crops in open fields include roguing infected plants and insect vector management. Application of insecticide to manage vectors (whiteflies and leafhoppers) is the most widely used measure but can cause undesirable environmental and human health issues. For annual crops, these measures can be more effective when combined with host-free periods of two to three months. Finally, given the great diversity of the viruses, their insect vectors, and the crops affected, IPM approaches need to be based on the biology and ecology of the virus and vector and the crop production system. Here, we present the general measures that can be used in an IPM program for geminivirus diseases, specific case studies, and future challenges.
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Ascencio-Ibáñez, J. T., R. Diaz-Plaza, J. Méndez-Lozano, Z. I. Monsalve-Fonnegra, G. R. Argüello-Astorga, and R. F. Rivera-Bustamante. "First Report of Tomato Yellow Leaf Curl Geminivirus in Yucatán, México." Plant Disease 83, no. 12 (December 1999): 1178. http://dx.doi.org/10.1094/pdis.1999.83.12.1178a.
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Geminiviruses are probably the most important viral pathogen affecting tomatoes and other crops in the Caribbean region. In addition to losses previously caused by native virus populations, the introduction of tomato yellow leaf curl virus (TYLCV) into the area has become a major concern for tomato growers (1). Since the detection of TYLCV in Cuba, and later in Florida (2,3), we have been monitoring the tomato- and pepper-growing areas of the Yucatán Peninsula, México, for TYLCV. We also have reanalyzed samples previously collected. Other geminiviruses (pepper huasteco virus [PHV], Texas pepper virus [TPV], and tomato mottle virus [ToMoV]) in the area can cause symptoms similar to those induced by TYLCV, which led us to refine our analysis of samples, using a polymerase chain reaction (PCR) procedure that can differentiate between monopartite and bipartite begomoviruses based on the size of the amplification product, 750 and 600 bp, respectively. One advantage of using this set of primers is that the PCR product, which includes the amino terminus of the Rep protein, intergenic region, precoat protein, and amino terminus of the coat protein, can be sequenced completely with only one sequencing reaction from each end. Using the primer set, we analyzed samples collected from tomato and pepper fields (as well as from weeds surrounding the fields) from December 1996 until March 1999. In most cases, samples were taken from plants that showed yellowing, curling, and stunting symptoms. Most of the samples that were positive for geminiviruses came from plants infected with PHV or TPV. However, three tomato samples collected during two seasons in Dzidzantun and Yobain counties (northeast of Mérida, Yucatan) produced the larger PCR amplification product (750 bp) expected for monopartite begomoviruses. PCR products were cloned and sequenced to confirm their identity. The sequence was deposited in the GenBank Database (Accession no. AF168709) and compared with all geminivirus sequences deposited in the database. Analysis showed that the amplified fragment from the TYLCV strain present in the Yucatán is 99% identical to the isolate reported in the Dominican Republic and later found in Cuba (2). As previously noted, the isolate is almost identical to TYLCV-Isr (2). In addition to the PCR product, a full-length TYLCV clone was obtained directly from DNA extracts of an infected tomato plant. Further characterization of the full-length clone is underway. The fact that TYLCV was detected in two counties and in samples collected during two seasons confirms the presence of TYLCV in the Yucatán. Interestingly, although the first positive sample for TYLCV was collected during the winter of 1996 and 1997, current incidence is rather low—only two other positive samples have been detected in more recently collected samples. Perhaps the characteristics of the agriculture system in the Yucatán (small, disperse plots) or the presence of other geminiviruses have contributed to a slow spread of the virus. More comprehensive surveys are required to confirm the actual distribution of the pathogen in the area. References: (1) J. E. Polston et al. Plant Dis. 81:1358, 1997. (2) J. E. Polston et al. Plant Dis. 83:984, 1999. (3) P. L. Ramos et al. Plant Dis. 80:1208, 1996.
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Richter, Kathrin S., and Holger Jeske. "KU80, a key factor for non-homologous end-joining, retards geminivirus multiplication." Journal of General Virology 96, no. 9 (September 1, 2015): 2913–18. http://dx.doi.org/10.1099/jgv.0.000224.
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Morilla, Gabriel, Björn Krenz, Holger Jeske, Eduardo R. Bejarano, and Christina Wege. "Tête à Tête of Tomato Yellow Leaf Curl Virus and Tomato Yellow Leaf Curl Sardinia Virus in Single Nuclei." Journal of Virology 78, no. 19 (October 1, 2004): 10715–23. http://dx.doi.org/10.1128/jvi.78.19.10715-10723.2004.
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ABSTRACT Since 1997 two distinct geminivirus species, Tomato yellow leaf curl Sardinia virus (TYLCSV) and Tomato yellow leaf curl virus (TYLCV), have caused a similar yellow leaf curl disease in tomato, coexisted in the fields of southern Spain, and very frequently doubly infected single plants. Tomatoes as well as experimental test plants (e.g., Nicotiana benthamiana) showed enhanced symptoms upon mixed infections under greenhouse conditions. Viral DNA accumulated to a similar extent in singly and doubly infected plants. In situ tissue hybridization showed TYLCSV and TYLCV DNAs to be confined to the phloem in both hosts, irrespective of whether they were inoculated individually or in combination. The number of infected nuclei in singly or doubly infected plants was determined by in situ hybridization of purified nuclei. The percentage of nuclei containing viral DNA (i.e., 1.4% in tomato or 6% in N. benthamiana) was the same in plants infected with either TYLCSV, TYLCV, or both. In situ hybridization of doubly infected plants, with probes that discriminate between both DNAs, revealed that at least one-fifth of infected nuclei harbored DNAs from both virus species. Such a high number of coinfected nuclei may explain why recombination between different geminivirus DNAs occurs frequently. The impact of these findings for epidemiology and for resistance breeding concerning tomato yellow leaf curl diseases is discussed.
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Monci, F., J. Navas-Castillo, and E. Moriones. "Evidence of a Naturally Occurring Recombinant Between Tomato yellow leaf curl virus and Tomato yellow leaf curl Sardinia virus in Spain." Plant Disease 85, no. 12 (December 2001): 1289. http://dx.doi.org/10.1094/pdis.2001.85.12.1289a.
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Tomato yellow leaf curl virus (TYLCV, formerly TYLCV-Is) and Tomato yellow leaf curl Sardinia virus (TYLCSV, formerly TYLCV-Sar) are geminivirus species of the genus Begomovirus that cause the disease known as tomato yellow leaf curl. In Spain, TYLCV and TYLCSV have coexisted in field and greenhouse tomato (Lycopersicon esculentum) crops since 1996 (2). TYLCV is also the causal agent of the leaf crumple disease of common bean (Phaseolus vulgaris) (1), a species that TYLCSV is unable to infect (2). Analysis of field samples from common bean plants affected by leaf crumple disease collected in Almería (southeastern Spain) during 1999 showed that, unexpectedly, several samples hybridized with TYLCV- and TYLCSV-specific probes prepared to the intergenic region (IR) as previously described (1). Polymerase chain reactions (PCR) performed with total nucleic acids extracted from one of these samples (ES421/99) using primer pairs specific to the IR of TYLCV (MA-30/MA-31) or TYLCSV (MA-14/MA-15) (1) gave no amplification product. However, the combination of MA-30 (5′ end of TYLCV IR) and MA-15 (3′ end of TYLCSV IR) produced a PCR DNA product of the expected size (351 bp). Direct DNA sequencing of this product (GenBank Accession No. AF401478) indicated the presence of a chimeric IR in ES421/99. Comparison of the obtained sequence with those available for isolates reported from Spain showed that the 5′ side (149 nt) from the stem-loop structure conserved in the IR of all geminiviruses was 99% identical to the corresponding region of TYLCV (GenBank Accession No. AF071228) and only 62% identical to TYLCSV (GenBank Accession No. Z25751). In contrast, the 3′ side (124 nt) from the stem-loop was 98% identical to the corresponding region of TYLCSV and only 57% identical to TYLCV. The 33-nt region involved in the stem-loop was 100% identical to TYLCV and showed one nucleotide change in the loop with respect to TYLCSV. Therefore, this DNA sequence data showed evidence of the occurrence in ES421/99 of a natural recombination between TYLCV and TYLCSV. The biological and epidemiological consequences of the presence of this new interspecific recombinant have yet to be determined. References: (1) J. Navas-Castillo et al. Plant Dis. 83:29, 1999. (2) S. Sánchez-Campos et al. Phytopathology 89:1038, 1999.
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Caciagli, P., and D. Bosco. "Quantitation Over Time of Tomato Yellow Leaf Curl Geminivirus DNA in Its Whitefly Vector." Phytopathology® 87, no. 6 (June 1997): 610–13. http://dx.doi.org/10.1094/phyto.1997.87.6.610.
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The amount of tomato yellow leaf curl geminivirus (TYLCV) DNA that accumulated in the vector Bemisia tabaci was studied by quantitative chemiluminescent dot-blot assay, using digoxigenin-labeled specific DNA probes. Large groups of female whiteflies were allowed to feed for 4, 12, 24, or 48 h on TYLCV-infected tomato plants and then were transferred to TYLCV-immune cucumber plants. Insects were sampled at different times during and after acquisition access and tested for TYLCV-DNA content. TYLCV-DNA assays were done either on whole insects oron the head plus prothorax (to include salivary glands) and abdomen separately. The maximum amount of TYLCV DNA, averaging from 0.5 to 1.6 ng per insect, was always attained at the end of the acquisition period. The mean amount then decreased by about 1 to 2% per day, remaining clearly detectable up to 20 days after the end of the acquisition period. Only some whiteflies that were TYLCV-positive in the abdomen were positive for head plus prothorax. In both parts of the body, TYLCV DNA remained detectable up to 18 days after the end of the acquisition period, showing that TYLCV DNA remains in insect tissues much longer than infectivity indicates.
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Jackel, Jamie N., R. Cody Buchmann, Udit Singhal, and David M. Bisaro. "Analysis of Geminivirus AL2 and L2 Proteins Reveals a Novel AL2 Silencing Suppressor Activity." Journal of Virology 89, no. 6 (December 31, 2014): 3176–87. http://dx.doi.org/10.1128/jvi.02625-14.
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ABSTRACTBoth posttranscriptional and transcriptional gene silencing (PTGS and TGS, respectively) participate in defense against the DNA-containing geminiviruses. As a countermeasure, members of the genusBegomovirus(e.g.,Cabbage leaf curl virus) encode an AL2 protein that is both a transcriptional activator and a silencing suppressor. The related L2 protein ofBeet curly top virus(genusCurtovirus) lacks transcription activation activity. Previous studies showed that both AL2 and L2 suppress silencing by a mechanism that correlates with adenosine kinase (ADK) inhibition, while AL2 in addition activates transcription of cellular genes that negatively regulate silencing pathways. The goal of this study was to clarify the general means by which these viral proteins inhibit various aspects of silencing. We confirmed that AL2 inhibits systemic silencing spread by a mechanism that requires transcription activation activity. Surprisingly, we also found that reversal of PTGS and TGS by ADK inactivation depended on whether experiments were conducted in vegetative or reproductiveNicotiana benthamianaplants (i.e., before or after the vegetative-to-reproductive transition). While AL2 was able to reverse silencing in both vegetative and reproductive plants, L2 and ADK inhibition were effective only in vegetative plants. This suggests that silencing maintenance mechanisms can change during development or in response to stress. Remarkably, we also observed that AL2 lacking its transcription activation domain could reverse TGS in reproductive plants, revealing a third, previously unsuspected AL2 suppression mechanism that depends on neither ADK inactivation nor transcription activation.IMPORTANCERNA silencing in plants is a multivalent antiviral defense, and viruses respond by elaborating multiple and sometimes multifunctional proteins that inhibit various aspects of silencing. The studies described here add an additional layer of complexity to this interplay. By examining geminivirus AL2 and L2 suppressor activities, we show that L2 is unable to suppress silencing inNicotiana benthamianaplants that have undergone the vegetative-to-reproductive transition. As L2 was previously shown to be effective in matureArabidopsisplants, these results illustrate that silencing mechanisms can change during development or in response to stress in ways that may be species specific. The AL2 and L2 proteins are known to share a suppression mechanism that correlates with the ability of both proteins to inhibit ADK, while AL2 in addition can inhibit silencing by transcriptionally activating cellular genes. Here, we also provide evidence for a third AL2 suppression mechanism that depends on neither transcription activation nor ADK inactivation. In addition to revealing the remarkable versatility of AL2, this work highlights the utility of viral suppressors as probes for the analysis of silencing pathways.
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Yang, Xiaojuan, Surendranath Baliji, R. Cody Buchmann, Hui Wang, John A. Lindbo, Garry Sunter, and David M. Bisaro. "Functional Modulation of the Geminivirus AL2 Transcription Factor and Silencing Suppressor by Self-Interaction." Journal of Virology 81, no. 21 (August 22, 2007): 11972–81. http://dx.doi.org/10.1128/jvi.00617-07.
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ABSTRACT The DNA genomes of geminiviruses have a limited coding capacity that is compensated for by the production of small multifunctional proteins. The AL2 protein encoded by members of the genus Begomovirus (e.g., Tomato golden mosaic virus) is a transcriptional activator, a silencing suppressor, and a suppressor of a basal defense. The related L2 protein of Beet curly top virus (genus Curtovirus) shares the pathogenicity functions of AL2 but lacks transcriptional activation activity. It is known that AL2 and L2 can suppress local silencing by interacting with adenosine kinase (ADK) and can suppress basal defense by interacting with SNF1 kinase. However, how the activities of these viral proteins are regulated remains an unanswered question. Here, we provide some answers by demonstrating that AL2, but not L2, interacts with itself. The zinc finger-like motif (CCHC) is required but is not sufficient for AL2 self-interaction. Alanine substitutions for the invariant cysteine residues that comprise the motif abolish self-interaction or cause aberrant subnuclear localization but do not abolish interaction with ADK and SNF1. Using bimolecular fluorescence complementation, we show that AL2:AL2 complexes accumulate primarily in the nucleus, whereas AL2:ADK and L2:ADK complexes accumulate mainly in the cytoplasm. Further, the cysteine residue mutations impair the ability of AL2 to activate the coat protein promoter but do not affect local silencing suppression. Thus, AL2 self-interaction correlates with nuclear localization and efficient activation of transcription, whereas AL2 and L2 monomers can suppress local silencing by interacting with ADK in the cytoplasm.
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Yang, Y., T. A. Sherwood, C. P. Patte, E. Hiebert, and J. E. Polston. "Use of Tomato yellow leaf curl virus (TYLCV) Rep Gene Sequences to Engineer TYLCV Resistance in Tomato." Phytopathology® 94, no. 5 (May 2004): 490–96. http://dx.doi.org/10.1094/phyto.2004.94.5.490.
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Tomato yellow leaf curl virus (TYLCV), a member of the genus Begomovirus (family Geminiviridae), causes severe losses in tomato production in the tropics and subtropics. In order to generate engineered resistance, eight different constructs of the TYLCV replication-associated protein (Rep) and C4 gene sequences were tested in transformed tomato inbred lines. Transgenic plants were screened for resistance to TYLCV using viruliferous whiteflies. No symptoms were observed and no TYLCV genomic DNA was detected by both hybridization and polymerase chain reaction in progenies of plants transformed with three constructs. This resistance was observed in plants that contained one of the following transgenes: 2/5Rep (81 nucleotides [nt] of the intergenic region [IR] plus 426 nt of the 5′ end of the TYLCV Rep gene), Δ2/5Rep (85 nt of the IR plus 595 nt of the 5′ end of the TYLCV Rep gene in the antisense orientation), and RepΔ2/5Rep (81 nt of the IR, the entire Rep gene, and 41 nt 3′ to the end of the Rep gene fused to Δ2/5Rep). Our study differs from other transgenic Geminivirus resistance reports involving the Rep gene in that viruliferous whiteflies were used for challenge inoculation instead of agroinoculation or biolistic inoculation, and TYLCV resistance was evaluated under field conditions.
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Arif, Muhammad, Saif ul Islam, Saqer S. Alotaibi, Ahmed M. Elshehawi, Mohamed A. A. Ahmed, and Abdullah M. Al-Sadi. "Infectious clone construction and pathogenicity confirmation of Cotton leaf curl Multan virus (CLCuMuV), Ramie mosaic virus (RamV) and Corchorus yellow vein Vietnam virus (CoYVV) by southern blot analysis." PLOS ONE 16, no. 5 (May 14, 2021): e0251232. http://dx.doi.org/10.1371/journal.pone.0251232.
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Geminiviruses are insect-transmissible, economically vital group of plant viruses, which cause significant losses to crop production and ornamental plants across the world. During this study, infectious clones of three devastating begomoviruses, i.e., Cotton leaf curl Multan virus (CLCuMuV), Ramie mosaic virus (RamV) and Corchorus yellow vein Vietnam virus (CoYVV) were constructed by following novel protocol. All infectious clones were confirmed by cloning and sequencing. All of the infectious clones were agro-inoculated in Agrobacterium. After the agro-infiltrations, all clones were injected into Nicotiana benthamiana and jute plants under controlled condition. After 28 days of inoculation, plants exhibited typical symptoms of their corresponding viruses. All the symptomatic and asymptomatic leaves were collected from inoculated plants for further analysis. The southern blot analysis was used to confirm the infection of studied begomoviruses. At the end, all the products were sequenced and analyzed.
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Fondong, V. N., J. S. Pita, C. Rey, R. N. Beachy, and C. M. Fauquet. "First Report of the Presence of East African Cassava Mosaic Virus in Cameroon." Plant Disease 82, no. 10 (October 1998): 1172. http://dx.doi.org/10.1094/pdis.1998.82.10.1172b.
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Cassava mosaic disease (CMD) occurs in all cassava-growing regions of Africa, India, and Sri Lanka. Characterized by mosaic and distortion of cassava leaves and reduced plant growth, causing high yield losses, CMD is caused by geminiviruses (genus Begomovirus, family Geminiviridae) transmitted through infected cuttings or by the whitefly, Bemisia tabaci. Three such geminiviruses have been described: African cassava mosaic virus (ACMV) occurs in most of the cassava-producing zones of Africa; East African cassava mosaic virus (EACMV) in East Africa; and Indian cassava mosaic virus (ICMV) in the Indian subcontinent (1). The two components of ACMV and ICMV genomes, DNA-A and DNA-B, have been sequenced; only DNA-A of EACMV has been identified and sequenced. Variations in symptom expression and severity within the same cassava variety have been observed in Cameroon. To determine the nature of the virus species inducing such variations, 50 samples were collected from CMD-infected plants in the savannah and rainforest zones of Cameroon: 2 from the sahel/savannah plain, 13 from the western highland savannah, and 35 from the main cassava-producing belt of the southwestern rainforest. There is a high incidence of CMD in the rainforest region, with some farms completely infected, while in the savannah regions farms generally have less than 25% incidence. Variation in symptom expression was more common in the rainforest region. Samples were collected from plants with distinct symptoms and/or different extents of symptom severity, then analyzed with the polymerase chain reaction (PCR) with specific primers: JSP1, ATG TCG AAG CGA CCA GGA GAT; JSP2, TGT TTA TTA ATT GCC AAT ACT; and JSP3, CCT TTA TTA ATT TGT CAC TGC. Primer JSP1 anneals to the 5′ end of the coat protein (CP) of ACMV and EACMV; primers JSP2 and JSP3 anneal to the 3′ ends of ACMV and EACMV, respectively. Virus identification was based on presence of an amplified fragment of either virus. ACMV was detected in all 50 samples; EACMV was detected in 8. All samples infected with EACMV were from the southwestern rainforest of Cameroon and were more severely affected by the disease than single infected plants. Previous reports have limited occurrence of EACMV to East Africa (1). This is the first report of the occurrence of EACMV in West Africa. The CP gene of three isolates of EACMV from Cameroon (EACMV/CM) was sequenced from cloned PCR products. There was a high CP nucleotide sequence identity (>99%) with only two amino acid differences among all three EACMV isolates. In contrast, there was a rather low sequence identity (94%) with EACMV/TZ from Tanzania (2), suggesting they may belong to a previously undescribed West African strain of EACMV. This indicates the geminiviruses causing CMD in Africa are more widely distributed than previously reported. None of the Cameroon isolates showed the type of recombination of the EACMV isolate from Uganda (EACMV/ UG) (having the CP core segment the identical to the corresponding ACMV CP sequence) (2). This emphasizes the need for characterization of the viruses causing CMD in different cassava-growing regions of Africa since appropriate control strategies depend on adequate knowledge of disease etiology. References: (1) Y. G. Hong et al. J. Gen. Virol. 74:2437, 1993. (2) X. Zhou et al. J. Gen. Virol. 78:2101, 1997.
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Schuster, David J., and Jane E. Polston. "Management of Whiteflies, Bugs, and Leafminers on Fresh Market Tomatoes, Fall 1996." Arthropod Management Tests 23, no. 1 (January 1, 1998): 156–57. http://dx.doi.org/10.1093/amt/23.1.156.
Full textAbstract:
Abstract Transplants were set 4 Sep, 18 inches apart on raised beds on EauGallie fine sand covered with black polyethylene mulch. Plots were 3-21 -ft-long rows on 5-ft centers and were irrigated by a seepage subirrigation system. Insecticidal treatments were replicated 4 times in a RCB design. Spray treatments were applied with a high-clearance, self-propelled sprayer on 13, 18, 23, 30 Sept, 9, 14, 21, 28 Oct, 4, 18, 25 Nov, 2 and 9 Dec. The sprayer was operated at 200 psi and 3.4 mph and was outfitted with orange Albuz™ ceramic nozzles. The number of nozzles per row was increased from 4 to 8 to increase gallonage as the plants grew. Thus, 60 gpa was applied for the first six sprays (four nozzles), 90 gpa was applied for the seventh spray (six nozzles) and 120 gpa was applied for the remaining six sprays (eight nozzles). Admire treatments were applied in 4 oz water/plant on 4 Sep. All plots were sprayed weekly with Bacillus thuringiensis for control of army worm larvae. The terminal leaflet was collected from the 7-8th leaf from the top of one branch of each of 10 plants in the middle row of each plot on 30 Oct, 22 Nov, and 16 Dec. Numbers of crawlers, sessile nymphs, and pupae of SLWF were counted and the data were averaged over all dates for analysis. All of the plants in the two outer rows of each plot were examined weekly for symptoms of virus. There are two viruses present which have nearly indistinguishable symptoms: tomato mottle virus (ToMoV), a geminivirus vectored by the SLWF, and potato virus Y (PVY), a potyvirus vectored by several species of aphids. All red ripe fruits were harvested from the middle 10 plants of the middle row of each plot on 12 Oct, 20 Nov, 4 and 17 Dec. The fruits were separated by the presence or absence of thrips and damage by the bugs SGS + LF. These fruits were counted and weighed. Each fruit was also rated 1-4 for increasing severity of irregular ripening (IRR), a disorder caused by SLWF feeding. The numbers of leafmines by LM were counted during a 2-min search of the middle row of each plot on 17 Dec. At the end of the experiment, a sample of three terminal leaflets was collected from each plant with ToMoV symptoms in the checks. The samples were frozen for later determination of the presence of ToMoV using nucleic acid spot hybridization and the presence of PVY using ELIS A.
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Arroyo-Mateos, Manuel, Blanca Sabarit, Francesca Maio, Miguel A. Sánchez-Durán, Tabata Rosas-Díaz, Marcel Prins, Javier Ruiz-Albert, Ana P. Luna, Harrold A. van den Burg, and Eduardo R. Bejarano. "Geminivirus Replication Protein Impairs SUMO Conjugation of Proliferating Cellular Nuclear Antigen at Two Acceptor Sites." Journal of Virology 92, no. 18 (June 27, 2018). http://dx.doi.org/10.1128/jvi.00611-18.
Full textAbstract:
ABSTRACTGeminiviruses are DNA viruses that replicate in nuclei of infected plant cells using the plant DNA replication machinery, including PCNA (proliferating cellular nuclear antigen), a cofactor that orchestrates genome duplication and maintenance by recruiting crucial players to replication forks. These viruses encode a multifunctional protein, Rep, which is essential for viral replication, induces the accumulation of the host replication machinery, and interacts with several host proteins, including PCNA and the SUMO E2 conjugation enzyme (SCE1). Posttranslational modification of PCNA by ubiquitin or SUMO plays an essential role in the switching of PCNA between interacting partners during DNA metabolism processes (e.g., replication, recombination, and repair, etc.). In yeast, PCNA sumoylation has been associated with DNA repair involving homologous recombination (HR). Previously, we reported that ectopic Rep expression results in very specific changes in the sumoylation pattern of plant cells. In this work, we show, using a reconstituted sumoylation system inEscherichia coli, that tomato PCNA is sumoylated at two residues, K254 and K164, and that coexpression of the geminivirus protein Rep suppresses sumoylation at these lysines. Finally, we confirm that PCNA is sumoylatedin plantaand that Rep also interferes with PCNA sumoylation in plant cells.IMPORTANCESUMO adducts have a key role in regulating the activity of animal and yeast PCNA on DNA repair and replication. Our work demonstrates for the first time that sumoylation of plant PCNA occurs in plant cells and that a plant virus interferes with this modification. This work marks the importance of sumoylation in allowing viral infection and replication in plants. Moreover, it constitutes a prime example of how viral proteins interfere with posttranslational modifications of selected host factors to create a proper environment for infection.