Mali Virus Research Articles

Screening of Solanaceous Plants for Resistance and Tolerance to Widespread Begomoviruses in Burkina Faso

Solanaceous fruits are economically important and contribute to food security worldwide. Despite this importance, solanaceous crops are confronted with constraints, including begomoviruses. In Burkina Faso, the pepper yellow vein Mali virus (PepYVMLV) and the tomato leaf curl Mali virus (ToLCMLV) have been reported to be the most widespread begomoviruses. They are responsible for leaf deformation and yellowing in Solanaceae crops. The use of resistant/tolerant accessions would be of help in finding suitable solutions. This study aims to contribute to the appropriate and effective management of solanaceous diseases due to begomoviruses. To achieve this objective, an inventory of solanaceous accessions sold and produced in the western region of Burkina Faso and a screening of the preferred accessions were conducted in semi controlled conditions. Our results showed that twenty-seven accessions, including two newly introduced ones, were identified, with Mongal F1, Jaune du Burkina, and Simbad F1 being the most preferred. Preferred solanaceous plants were monitored for 39 days after whitefly-mediated transmission. The hot pepper Jaune du Burkina exhibited mild symptoms with an infection rate of 66%, while the tomato Mongal F1 and the sweet pepper Simbad F1 showed disease with severe symptoms with infection rates ranging from 33% to 100%. When the behaviors of newly introduced accessions were compared to most preferred accessions, the results indicated that the newly introduced accessions (Mona F1 and Ganga) showed mild symptoms, suggesting potential tolerance despite high infection rates. Howerever, it is suitable to access the impact of viruses on the productivity of these accessions.

A new monopartite begomovirus infecting Melochia tomentosa in Burkina Faso.

This is the first description of the complete genome sequence of a newly characterized monopartite begomovirus isolated from an asymptomatic uncultivated plant, Melochia tomentosa, collected in Burkina Faso. The sequence was obtained through rolling-circle amplification, cloning, and Sanger sequencing. The provisional species name "Begomovirus melochiae" and common virus name "melochia associated virus" (MeAV) are proposed. The MeAV genome was found to share the most nucleotide sequence similarity with three African monopartite begomoviruses: tomato curly stunt virus (74%), pepper yellow vein Mali virus (73%), and tomato leaf curl Cameroon virus (73%). Phylogenetic analysis confirmed its relationship to Old World monopartite begomoviruses. The discovery of MeAV in an uncultivated and asymptomatic plant provides a further example of the high diversity of begomoviruses in sub-Saharan African ecosystems.

Screening of widely used accessions of tomato for resistance/tolerance to the <i>Pepper yellow vein Mali virus</i> in the central region of Burkina Faso

Tomatoes are a widely consumed fruit, important economically and in terms of food security. Despite this importance, tomato crops are confronted with constraints, including begomoviruses, such as the Pepper yellow vein Mali virus (PepYVMLV). Responsible for leaf deformation and yellowing in cultivated Solanaceae, it is the most virulent begomovirus infecting tomatoes in Burkina Faso. The use of resistant/tolerant accessions would be of great help in finding suitable solutions. This study aims to contribute to the appropriate and effective management of tomato diseases due to PepYVMLV. To achieve this objective, an inventory of tomato accessions sold and produced in the central region of Burkina Faso and a screening of the preferred accessions of tomatoes in semi-control conditions were conducted. Our results showed that sixteen varieties are sold in shops, with Mongal F1, Emerald F1, and Cobra 26 F1 reported to be the most preferred by farmers. Monitoring of the tomato plants inoculated using whitefly-mediated transmission during 32 days showed that Diva F1, a newly introduced accession, was the least susceptible to PepYVMLV, followed by Emerald F1, Cobra 26 F1, and Mongal F1, with yield losses ranging from 25.38 to 359.9 g/plant. Our results suggest that even if the productivity of Diva F1 seems to be slightly impacted by the virus, it is unsuitable for farmers because of its lower yield.

First report of a naturally occurring isolate of Pepper yellow vein Mali virus causing tobacco yellow leaf curl disease in Burkina Faso

First report of a naturally occurring isolate of <i>Pepper yellow vein Mali virus</i> causing tobacco yellow leaf curl disease in Burkina Faso

Synergy between an emerging monopartite begomovirus and a DNA-B component

In recent decades, a legion of monopartite begomoviruses transmitted by the whitefly Bemisia tabaci has emerged as serious threats to vegetable crops in Africa. Recent studies in Burkina Faso (West Africa) reported the predominance of pepper yellow vein Mali virus (PepYVMLV) and its frequent association with a previously unknown DNA-B component. To understand the role of this DNA-B component in the emergence of PepYVMLV, we assessed biological traits related to virulence, virus accumulation, location in the tissue and transmission. We demonstrate that the DNA-B component is not required for systemic movement and symptom development of PepYVMLV (non-strict association), but that its association produces more severe symptoms including growth arrest and plant death. The increased virulence is associated with a higher viral DNA accumulation in plant tissues, an increase in the number of contaminated nuclei of the phloem parenchyma and in the transmission rate by B. tabaci. Our results suggest that the association of a DNA-B component with the otherwise monopartite PepYVMLV is a key factor of its emergence.

Diversity, distribution and prevalence of vegetable‐infecting geminiviruses in Burkina Faso

In West and Central Africa, as in many regions of the world, vegetables are severely affected by geminivirus diseases. In Burkina Faso, observation of various virus‐like symptoms, especially on tomato, suggests the involvement of several geminiviruses and underlines the pressing need for additional information on their diversity, distribution, prevalence and host plant reservoirs. Large‐scale surveys conducted in Burkina Faso confirmed the presence of tomato (yellow) leaf curl diseases (ToLCD‐TYLCD) and geminiviruses in all localities with mean prevalences of 25% and 45%, respectively. Five geminiviruses including four begomoviruses (pepper yellow vein Mali virus (PepYVMLV), tomato leaf curl Burkina Faso virus, tomato leaf curl Mali virus and tomato leaf curl Ghana virus) and a dicot‐infecting mastrevirus (chickpea chlorotic dwarf virus) were characterized on tomato. In addition, PepYVMLV and cotton leaf curl Gezira virus (CLCuGeV) were characterized on pepper and okra, respectively, in combination or not with alphasatellites and betasatellites for CLCuGeV. The most severe, prevalent and widely distributed virus on vegetables was PepYVMLV, which was characterized for the first time in combination with a genetically divergent DNA‐B component that may constitute a key factor of PepYVMLV pathogenicity. Of the eight weeds identified as potential reservoir hosts of begomoviruses, four host PepYVMLV. The results confirm the importance of geminivirus diseases on vegetable crops in Burkina Faso and highlight the complex association of geminiviruses and satellites. The detection of begomoviruses in weeds growing close to crops points to the increasing necessity to consider reservoir plants and virus communities in the control of virus diseases.

Detection of Pepper Yellow Vein Mali Virus associated with pepper in Yamoussoukro, Côte d’Ivoire

Detection of Pepper Yellow Vein Mali Virus associated with pepper in Yamoussoukro, Côte d’Ivoire

English

Geminiviruses, in particular the members of the genus begomovirus, are considered to be a major phytosanitary problem for tomato crops production in the world. They are responsible for yield losses of up to 20 to 100% and Togo is not spared from this situation. The present study aimed to investigate the genetic diversity of the begomoviruses affecting tomato crops production in Togo and their relationship with other begomoviruses. To achieve these objectives, 307 samples of tomato leaves and wild plant species with typical virus symptoms were collected in the Maritime, Plateau, Central, Kara and Savannah regions and subjected to polymerase chain reaction (PCR) analysis. The results revealed the presence of begomovirus in 25.40% of the analyzed samples. The PCR products obtained were subjected to direct sequencing. Phylogenetic analysis of the sequences of the different regions of the DNA-A of the begomovirus identified in this study with that of other begomoviruses showed a nucleotide identity of 96% respectively to Tomato leaf curl Togo virus-Fontem, Tomato Leaf Curl Togo Virus, and Ageratum leaf curl Cameroon Alphasatellite; 98% respectively to Tomato leaf curl Nigeria virus, Ageratum leaf curl Cameroon virus, Tomato leaf curl Cameroon virus-Fontem, Ageratum leaf curl Cameroon virus and 99% respectively to Tomato leaf curl Kumasi virus, Pepper yellow vein Mali virus Bazegahot, and Pepper yellow vein Mali virus-Ouaga. These results suggest a high degree of genetic diversity of tomato begomoviruses identified in Togo. Key words: Begomoviruses, genetic diversity, phylogenetic relationships, tomato, wild plants.

Tomato leaf curl Burkina Faso virus: a novel tomato-infecting monopartite begomovirus from Burkina Faso.

In this report, we present the first description of the complete genome sequence of a new monopartite begomovirus isolated from tomatoes collected in Burkina Faso and presenting with symptoms of tomato leaf curl disease. We propose the tentative name "tomato leaf curl Burkina Faso virus'' (ToLCBFV). DNA-A-like nucleotide sequence of ToLCBFV shares the highest nucleotide sequence identity (85%) with the pepper yellow vein Mali virus (PepYVMLV). Phylogenetic analysis confirmed the affiliation of ToLCBFV to Old World monopartite begomoviruses. This discovery of a new species confirms the existence of high genetic diversity in monopartite begomoviruses in sub-Saharan Africa and particularly in West Africa.

Genetic Diversity of Tomato (&amp;lt;i&amp;gt;Solanum lycopersicum&amp;lt;/i&amp;gt; L.) Begomovirus in Togo

Geminiviruses, in particular the members of the genus Begomovirus , are considered to be a major phytosanitary problem for tomato crops production in the world. They are responsible for yield losses of up to 20% to 100%. Regrettably, Togo is not spared from this situation. This work aims to show the genetic diversity of the begomoviruses affecting tomato crops production in Togo and their relationship with other begomoviruses. To achieve these objectives, 307 samples of tomato leaves and wild plant species with typical virus symptoms were collected in the Maritime, Plateaus, Central, Kara and Savannah regions and submitted to PCR analysis. The results revealed the presence of begomovirus in 25.40% of the analyzed samples. The PCR products obtained were submitted to direct sequencing. Phylogenetic analysis of sequences of DNA-A different regions of begomovirus identified in this work with that of other begomoviruses showed a nucleotide identity of 96% respectively for Tomato leaf curl Togo virus-Fontem, Tomato Leaf Curl Togo Virus , Ageratum leaf curl Cameroon Alphasatellite; 98% respectively to Tomato leaf curl Nigeria virus , Ageratum leaf curl Cameroon virus , Tomato leaf curl Cameroon virus-Fontem, Ageratum leaf curl Cameroon virus and 99% respectively to Tomato leaf curl Kumasi virus , Pepper yellow vein Mali virus Bazegahot and Pepper yellow vein Mali virus-Ouaga. These results suggest a high degree of genetic diversity of tomato begomoviruses identified in Togo.

First report of Pepper yellow vein Mali virus associated with pepper yellow vein disease in Cote d'Ivoire

In Mali (Zhou et al ., 2008) and Burkina Faso (Tiendrebeogo et al ., 2008), pepper yellow vein disease (PYVD) is associated with the African monopartite begomovirus Pepper yellow vein Mali virus (PepYVMLV). In January 2012 and August…

Serological Detection of Tobacco Mosaic Virus and Cucumber Mosaic Virus Infecting Tomato (Solanum Lycopersicum) Using a Lateral Flow Immunoassay Technique

A study was conducted to determine the prevalence of Cucumber mosaic virus (CMV) and Tobacco mosaic virus (TMV) in some major tomato growing areas in Ghana as part of a comprehensive strategy for the management of viral diseases on tomato. The lateral flow immunoassay technique (the immunostrip test of Agdia Inc.) was used to assay for TMV and CMV in samples with symptoms of virus infection collected from some major tomato growing areas in Ghana; Vea, Tono and Pwalugu (Upper East region-UE), Agogo and Akumadan (Ashanti region-AR) and Tanoso and Tuobodom (Brong-Ahafo region-BA) in 2011 and 2012. In the UE, TMV and CMV were both detected at Vea, Tono and Pwalugu. In the AR, CMV and TMV were both detected at Akumadan while only CMV was detected at Agogo. In BA, TMV and CMV were both detected at Tanoso and Tuobodom. Field incidence of TMV was higher in the UE which had higher average daily temperatures. The serological identification of TMV in the three regions is the first of this virus in Ghana. This is also the first report of the use of this lateral flow immunoassay technique to detect these viruses in the country. The identification of TMV and CMV in Ghana gives an indication of the need to focus efforts for virus disease management on these virus in addition to the already known begomoviruses like Tomato yellow leaf curl Mali virus (TYLCMV)(Delete), Tomato leaf curl Kumasi virus and Tomato leaf curl Ghana virus.

Molecular and biological characterization of Pepper yellow vein Mali virus (PepYVMV) isolates associated with pepper yellow vein disease in Burkina Faso

Yellow vein disease (YVD) is a major problem in pepper in West Africa. Despite the recent implication of a begomovirus in YVD in Mali and in Burkina Faso, the aetiology of the disease remains unclear. Using symptomatic samples from the main vegetable cultivation regions in Burkina Faso, 10 full-length DNA-A-like begomovirus sequences were obtained, each showing 98% nucleotide identity to pepper yellow vein Mali virus (PepYVMV). The host range was determined after construction of a viral clone for agroinfection. Severe symptoms developed in tomato and Nicotiana benthamiana. By contrast, no symptoms developed in either commercial or local pepper cultivars, demonstrating that the aetiology of YVD is not only associated with the presence of PepYVMV.

A severe symptom phenotype in tomato in Mali is caused by a reassortant between a novel recombinant begomovirus (Tomato yellow leaf curl Mali virus) and a betasatellite

Tomato production in West Africa has been severely affected by begomovirus diseases, including yellow leaf curl and a severe symptom phenotype, characterized by extremely stunted and distorted growth and small deformed leaves. Here, a novel recombinant begomovirus from Mali, Tomato yellow leaf curl Mali virus (TYLCMLV), is described that, alone, causes tomato yellow leaf curl disease or, in combination with a betasatellite, causes the severe symptom phenotype. TYLCMLV is an Old World monopartite begomovirus with a hybrid genome composed of sequences from Tomato yellow leaf curl virus-Mild (TYLCV-Mld) and Hollyhock leaf crumple virus (HoLCrV). A TYLCMLV infectious clone induced leaf curl and yellowing in tomato, leaf curl, crumpling and yellowing in Nicotiana benthamiana and common bean, mild symptoms in N. glutinosa, and a symptomless infection in Datura stramonium. In a field-collected sample from a tomato plant showing the severe symptom phenotype in Mali, TYLCMLV was detected together with a betasatellite, identified as Cotton leaf curl Gezira betasatellite (CLCuGB). Tomato plants co-agroinoculated with TYLCMLV and CLCuGB developed severely stunted and distorted growth and small crumpled leaves. These symptoms were more severe than those induced by TYLCMLV alone, and were similar to the severe symptom phenotype observed in the field in Mali and in other West African countries. TYLCMLV and CLCuGB also induced more severe symptoms than TYLCMLV in the other solanaceous hosts, but not in common bean. The increased symptom severity was associated with hyperplasia of phloem-associated cells, but relatively little increase in TYLCMLV DNA levels. In surveys of tomato virus diseases in West Africa, TYLCMLV was commonly detected in plants with leaf curl and yellow leaf curl symptoms, whereas CLCuGB was infrequently detected and always in association with the severe symptom phenotype. Together, these results indicate that TYLCMLV causes tomato yellow leaf curl disease throughout West Africa, whereas TYLCMLV and CLCuGB represent a reassortant that causes the severe symptom phenotype in tomato.

Complete genomic sequences of Tomato yellow leaf curl Mali virus isolates infecting tomato and pepper from the North Province of Cameroon

Complete genomic sequences of Tomato yellow leaf curl Mali virus isolates infecting tomato and pepper from the North Province of Cameroon.

First Report and Molecular Characterization of DNA A of Three Distinct Begomoviruses Associated with Tomato Leaf Curl Disease in Ghana.

Tomato leaf curl disease is reported to be widespread in Ghana and to cause severe yield losses (4). So far, the causal agent has not been identified. Thirty-three tomato (Solanum lycopersicum L.) samples with symptoms such as curling, yellowing, small leaves, and stunting were collected from the Ashanti Region, the main tomato-production area in Ghana, including three samples from Akumandan in the autumn of 2007 and 30 samples from Kumasi in the spring of 2008. The observed leaf curl disease incidence in the farmer's field in Kumasi was approximately 75%. Viral DNAs were extracted from the 33 samples and tested for the presence of begomoviral DNA-A, DNA-B, and associated satellite DNA by PCR with previously described primers (1,3). The expected 1.4-kb DNA-A begomovirus fragment was obtained from one of the samples from Akumadan and from 25 samples from Kumasi. DNA-B and DNA-beta were not detected by PCR. The 1.4-kb PCR products from all positive samples were cloned and sequenced. Sequence comparison by MegAlign software (DNASTAR, Inc., Madison, WI) showed three distinct virus groups. One isolate from each group was selected and specific primers were designed to complete the DNA-A sequence. The DNA-As of GH5-3 (group 1), GOTB2-2 (group 2), and GHK2 (group 3) isolates consisted of 2,803 (GenBank Accession No. EU350585), 2,794 (GenBank Accession No. EU847739), and 2,792 nt (GenBank Accession No. EU847740) respectively. All contain the geminiviral conserved nonanucleotide sequence TAATATTAC in the intergenic region and the six predicted open reading frames (ORFs V1, V2, C1, C2, C3, and C4). BLASTn analysis was conducted with geminivirus sequences available in the GenBank database at National Center for Biotechnology Information (Bethesda, MD). Further sequence comparisons were performed by Clustal V algorithm of MegAlign software with the representative isolates of begomovirus species reported by Fauquet et al (2) and the sequences that showed high scores in BLASTn search. The DNA-A sequence of isolate GHK2 from Kumasi showed highest sequence identity (96.5%) with Tomato yellow leaf curl Mali virus (TYLCMLV; GenBank Accesssion No. AY502934). The DNA-A sequence of GH5-3 and GOTB2-2 isolates had 87.5% sequence identity with each other. Both had highest sequence identities of 76.7 and 77.6%, respectively, with Tomato leaf curl Antsiranana virus, Madagascar (GenBank Accession No. AM701764). They constitute two distinct begomovirus species based on DNA-A sequence comparisons and the International Committee on Taxonomy of Viruses proposed species demarcation of 89% sequence identity. The names Tomato leaf curl Ghana virus for isolate GH5-3 and Tomato leaf curl Kumasi virus for isolate BOTB2-2 are proposed, respectively. To our knowledge, this is the first report of molecular characterization of begomoviruses associated with tomato leaf curl disease in Ghana and of the presence of three distinct tomato begomoviruses. This presence should be considered for recommending or developing stable begomovirus resistant tomato cultivars for Ghana.

Characterization of Pepper yellow vein mali virus in Capsicum sp. in Burkina Faso

Characterization of Pepper yellow vein mali virus in Capsicum sp. in Burkina Faso

Evidence of local evolution of tomato-infecting begomovirus species in West Africa: characterization of tomato leaf curl Mali virus and tomato yellow leaf crumple virus from Mali

Tomato yellow leaf curl (TYLC) and tomato leaf curl (ToLC) diseases are serious constraints to tomato production in Mali and other countries in West Africa. In 2003 and 2004, samples of tomato showing virus-like symptoms were collected during a survey of tomato virus diseases in Mali. Three predominant symptom phenotypes were observed: (1) TYLC/ToLC (stunted upright growth and upcurled leaves with interveinal yellowing and vein purpling), (2) yellow leaf crumple and (3) broccoli or bonsai (severe stunting and distorted growth). Squash blot (SB) hybridization with a general begomovirus probe and/or SB/PCR analyses revealed begomovirus infection in plants with each of these symptom phenotypes and no evidence of phytoplasma infection. Sequence analysis of PCR-amplified begomovirus fragments revealed two putative new begomovirus species associated with the TYLC/ToLC and yellow leaf crumple symptom phenotypes, respectively. Full-length clones of these begomoviruses were obtained using PCR and overlapping primers. When introduced into N. benthamiana and tomato plants, these clones induced upward leaf curling and crumpling (the TYLC/ToLC-associated begomovirus) or downward leaf curl/yellow mottle (yellow leaf crumple-associated begomovirus) symptoms. Thus, these begomoviruses were named tomato leaf curl Mali virus (ToLCMLV) and tomato yellow leaf crumple virus (ToYLCrV). The genome organization of both viruses was similar to those of other monopartite begomoviruses. ToLCMLV and ToYLCrV were most closely related to each other and to tobacco leaf curl Zimbabwe virus (TbLCZV-[ZW]) and tomato curly stunt virus from South Africa (ToCSV-ZA). Thus, these likely represent tomato-infecting begomoviruses that evolved from indigenous begomoviruses on the African continent. Mixed infections of ToLCMLV and ToYLCrV in N. benthamiana and tomato plants resulted in more severe symptoms than in plants infected with either virus alone, suggesting a synergistic interaction. Agroinoculation experiments indicated that both viruses induced symptomatic infections in tomato and tobacco, whereas neither virus induced disease symptoms in pepper, common bean, small sugar pumpkin, African eggplant, or Arabidopsis. Virus-specific PCR primers were developed for detection of ToLCMLV and ToYLCrV and will be used to further investigate the distribution and host range of these viruses.

First report of a distinct begomovirus associated with okra yellow crinkle disease in Mali

In November of 2004 and July of 2005, typical symptoms of begomovirus infection such as yellow veining, leaf yellowing, crinkling and cupping were observed in okra (Abelmoschus esculentus) variety trials near Bamako, Mali. Thirty-one samples from seven varieties with symptoms were collected and tested for the presence of begomovirus by polymerase chain reaction (PCR), using the begomovirus-specific degenerate primer pair PAL1v1978/PAR1c715 (Rojas et al., 1993). The expected 1·4-kb PCR product for begomoviruses was detected from seventeen samples from four varieties with yellow veining, leaf yellowing, crinkling or cupping symptoms. One PCR product from each of the 2004 and 2005 collection was cloned and sequenced. Based on the sequence of the 1·4-kb DNA product, specific primers were designed to complete the DNA-A sequence. DNA-B was not detectable in the two samples with DNA-B specific primer pairs (Green et al., 2001). However, a DNA-beta of 1·3-kb was detected in both samples with the DNA-beta primer pair Beta01/Beta02 (Briddon et al., 2002). The DNA-A of the two virus isolates shared 99·2% sequence identity and are considered the same virus. Each consisted of 2792 nucleotides (Acc. Nos. DQ902715 and DQ875879) and contained the six predicted open reading frames (ORFs V1, V2, C1, C2, C3 and C4). A BLAST analysis was conducted with geminivirus sequences available in the GenBank database and MegAlign software (DNASTAR) was used for further sequence comparisons. Highest sequence identity (76·7%) was found with Tobacco leaf curl Zimbabwe virus (AF350330). Only low sequence identities (70·9 to 72·5%) were obtained with full length DNA-A sequences of other okra-infecting begomoviruses available in the GenBank from African and Asian countries, including Sudan (AY036006 and AY036008), Egypt (AY036010), India (AF241479) and Pakistan (AJ002451, AJ002453 and AJ002459). Even lower sequence identities (63·1 to 63·9%) were found with okra-infecting begomoviruses from New World countries, such as Mexico (AY751753 and DQ022611). Sequence identities with begomoviruses reported from other vegetable crops in Mali, including two from tomato (AY502934 and AY503936) and one from pepper (AY502935) were also low, ranging from 73·4 to 76·4%. This is the first report of a begomovirus associated with okra yellow crinkle disease in Mali. Based on the DNA-A sequence comparison and the ICTV demarcation of species at 89% sequence identity, the okra begomovirus from Mali constitutes a distinct begomovirus and the name Okra yellow crinkle Mali virus is proposed.

First Report of a Begomovirus Associated with Tomato Yellow Leaf Curl Disease in Ethiopia.

During December 2003, severe leaf yellowing, leaf curling, and stunting symptoms were observed in tomato (Lycopersicon esculentum) plantings in Melkassa (1,550 m above sea level), Ethiopia. Eleven symptomatic samples were collected and tested for the presence of a begomovirus using polymerase chain reaction (PCR) with the begomovirus-specific degenerate primer pair PAL1v1978/PAR1c715 (3). Samples were also tested for Cucumber mosaic virus (CMV), Potato virus Y (PVY), Tobacco etch virus (TEV), Pepper veinal mottle virus (PVMV), and Tomato mosaic virus (ToMV) using enzyme-linked immunosorbent assay (ELISA). All samples were negative for CMV, PVY, TEV, PVMV, and ToMV. However, the expected 1.4-kb PCR product for begomoviruses was obtained from all samples. DNA-B and DNA-beta were not detectable using PCR with the DNA-B specific primer pairs DNABLC1/DNABLV2 and DNABLC2/ DNABLV2 (2) and the DNA-beta primer pair Beta01/Beta02 (1), respectively. The 1.4-kb PCR product of one sample was cloned and sequenced. On the basis of the sequence of the 1.4-kb DNA product, specific primers were designed to complete the DNA-A sequence. The DNA-A consisted of 2,785 nucleotides (GenBank Accession No. DQ358913) and was found to contain the six predicted open reading frames (ORFs V1, V2, C1, C2, C3, and C4). A BLAST analysis was conducted with geminivirus sequences available in the GenBank database at the National Center for Biotechnology Information (Bethesda, MD), and DNAMAN software (Lynnon Corporation, Quebec, Canada) was used for further comparisons. The DNA-A sequence of the virus associated with yellow leaf curl disease of tomato from Ethiopia showed highest sequence identity (92%) with Tomato yellow leaf curl Mali virus (TYLCMLV; GenBank Accession No. AY502934). On the basis of the DNA-A sequence comparison and the ICTV demarcation of species at 89% sequence identity, the Ethiopian virus is a provisional strain of TYLCMLV described from Mali. To our knowledge, this is the first report of a begomovirus associated with tomato yellow leaf curl disease in Ethiopia.