Tomato Yellow Leaf Curl Virus Research Articles

AmiRNA Technology Enhances Tomato Disease Resistance by Suppressing Plant-Pathogen Interaction Pathways through Inhibiting TYLCV Replication.

Tomato yellow leaf curl virus disease has seriously threatened the quality and yield of tomatoes. In this study, we investigated the role of amiRNA technology in disease resistance in tomatoes (cherry tomato and large-fruited tomato) and analyzed the physiological and molecular mechanisms of disease resistance in transgenic plants. TYLCV contains six functional genes, of which the C1, C2, and V1 genes have more phosphorylation sites and glycosylation sites, and the protein structure is more complex. The virus replication was inhibited, the peroxidation of membrane lipids was reduced, and disease resistance was enhanced in all transgenic cherry tomato (J6) plants in which the C1, C2, and V1 genes were silenced, respectively. Similarly, silencing of the C1 gene enhanced disease resistance in large-fruited tomatoes. In conclusion, amiRNA technology hinders viral replication, leading to reduced activity of the tomato plant-pathogen interaction pathway and weakening tomato-virus interactions, thereby improving disease resistance.

First report of paprika as a natural host plant for tomato chlorosis virus in Korea.

Paprika (Capsicum annuum var. grossum (L.) Sendtn.), also called bell pepper or sweet pepper, is a valuable greenhouse crop that is mostly consumed as fresh fruit in Korea. In 2022, it was cultivated on 726 ha in Korea and 82,042 tons were produced (KOSIS, 2023). In April 2023, interveinal chlorosis and yellowing were observed on the lower and middle leaves of greenhouse paprika in Jinju and Haman, South Korea, and many whiteflies (Bemisia tabaci) were present. The disease incidence was 30-40% on five farms (approximately 4.5 ha) in the two areas. The typical symptoms and presence of whiteflies in greenhouses indicated potential infection by the whitefly-transmitted crinivirus, tomato chlorosis virus (ToCV). Six samples collected from symptomatic plants were examined by leaf dip using transmission electron microscopy (TEM) and revealed filamentous virus particles about 800 nm long. To confirm the TEM results, six symptomatic paprika leaf samples were subject to RT-PCR using the specific primers ToCV-M-4F and ToCV-M-4R to detect ToCV (Choi, 2023). All tested samples were positive for ToCV. To confirm the presence of other viruses and obtain the complete genome sequences, one of the six ToCV-positive samples (ToCV-PAP-JJ6) was subject to high-throughput sequencing (HTS). Total RNA was extracted from symptomatic leaves using the RNeasy Plant Mini Kit (QIAGEN, Germany) and a transcriptome library was generated using the TruSeq Stranded Total RNA LT Sample Prep Kit (Illumina, San Diego, CA, USA) according to standard protocols. HTS was performed on an Illumina NovaSeq 6000 system (Macrogen, Korea). De novo transcriptome assembly of the 85,668,854 reads with Trinity software (r20140717) yielded 817,303 contigs of 201 to 38,987 nucleotides (nt). BLASTn and BLASTx analysis of the contigs against the NCBI viral reference database showed that three large contigs were virus sequences, two of which were homologous to ToCV-RNA1 and -RNA2 and one to bell pepper alphaendornavirus (BPEV). The ToCV-RNA1 (genome 8,594 nt, 11,212 mapped reads, mean read coverage 196.2 times) and ToCV-RNA2 (genome 8,242 nt, 40,122 mapped reads, mean read coverage 734.9 times) contigs showed 100% coverage and 99% base pair matching (8,583/8,594 and 8,233/8,242, respectively) with known genome sequences of ToCV-RNA1 (MG813908) and ToCV-RNA2 (KP114534) isolated from tomato in Korea (Lee et al. 2018). The sequences were deposited in GenBank as isolates ToCV-PAP-JJ6 RNA1 (OR865222) and ToCV-PAP-JJ6 RNA2 (OR865223). The BPEV contig (genome 14,728 nt, 2,928,519 mapped reads, mean read coverage 15,012.4 times) was detected at very high read depth and RT-PCR for BPEV confirmed that all six samples were coinfected with ToCV. BPEV was recently reported in paprika and red pepper in Korea (Jo et al., 2022). The BPEV-infected pepper cultivars have been reported not to produce any symptoms (Escalante & Valverde 2019). ToCV was first reported in tomato greenhouses in Korea in 2013 and is a problematic virus in tomato crops, along with tomato yellow leaf curl virus (Jo et al. 2023; Lee et al. 2018). Due to the continuous presence of ToCV, whiteflies, and natural weed hosts (Kil et al. 2015) in tomato greenhouses, ToCV has spread to other crops, including paprika. To the best of our knowledge, this is the first report of ToCV infecting paprika in Korea. A nationwide study is needed to prevent the spread of ToCV, a new threat to paprika farms.

Commodity risk assessment of Petunia spp. and Calibrachoa spp. unrooted cuttings from Costa Rica.

The European Commission requested the EFSA Panel on Plant Health to evaluate the probability of entry of pests (likelihood of pest freedom at entry), including both regulated and non-regulated pests, associated with unrooted cuttings of the genera Petunia and Calibrachoa produced under physical isolation in Costa Rica. The relevance of any pest for this opinion was based on evidence collected according to specific criteria, following the methodology used for high-risk plants adapted for the specificity of this assessment. Twenty-two EU regulated pests (beet curly top virus, Bemisia tabaci, Chloridea virescens, Eotetranychus lewisi, Epitrix cucumeris, Epitrix tuberis, euphorbia mosaic virus, Helicoverpa zea, Liriomyza huidobrensis, Liriomyza sativae, Liriomyza trifolii, pepper golden mosaic virus, potato spindle tuber viroid, Ralstonia pseudosolanacearum, Ralstonia solanacearum, Spodoptera ornithogalli, squash leaf curl virus, Thrips palmi, tomato golden mosaic virus, tomato leaf curl Sinaloa virus, tomato spotted wilt virus, tomato yellow leaf curl virus) and one pest that is not regulated in the EU (Aleurodicus dispersus) fulfilled all relevant criteria and were selected for further evaluation. For these pests, the risk mitigation measures proposed in the technical dossier from Costa Rica were evaluated taking into account possible factors limiting their efficacies. Additionally, an expert judgement is given on the likelihood of pest freedom taking into consideration the risk mitigation measures acting on the pest, including uncertainties associated with the assessment. The estimated degree of pest freedom varies among the pests evaluated, with tomato spotted wilt virus being the pest most frequently expected on the imported cuttings. The expert knowledge elicitation indicated, with 95% certainty that between 9927 and 10,000 bags containing unrooted cuttings of Petunia spp. and Calibrachoa spp. per 10,000 would be free of tomato spotted wilt virus.

Estimating TYLCV resistance level using RGBD sensors in production greenhouse conditions

Automated phenotyping is the task of automatically measuring plant attributes to help farmers and breeders in developing and growing strong robust plants. An automated tool for early illness detection can accelerate the process of identifying plant resistance and quickly pinpoint problematic breeding. Many such phenotyping tasks can be achieved by analyzing images from simple, low cost, RGB-D sensors. In this paper we focused on a particular case study — identifying the resistance level of tomato hybrids to the tomato yellow leaf curl virus (TYLCV) in production greenhouses. This is a difficult task, as separating between resistance levels based on images is difficult even for expert breeders. We collected a large dataset of images from an experiment containing many tomato hybrids with varying resistance levels. We used the depth information to identify the topmost part of the tomato plant. We then used deep learning models to classify the various resistance levels. For identifying plants with visual symptoms, our methods achieved an accuracy of 0.928, a precision of 0.934, and a recall of 0.95. In the multi-class case we achieved an accuracy of 0.76 in identifying the correct level, and an error of 0.278. Our methods are not particularly tailored for the specific task, and can be extended to other tasks that identify various plant diseases with visual symptoms such as ToBRFV,mildew, ToMV and others.

Resistance gene Ty-1 restricts TYLCV infection in tomato by increasing RNA silencing

A major antiviral mechanism in plants is mediated by RNA silencing through the action of DICER-like (DCL) proteins, which cleave dsRNA into discrete small RNA fragments, and ARGONAUTE (AGO) proteins, which use the small RNAs to target single-stranded RNA. RNA silencing can also be amplified through the action of RNA-dependent RNA polymerases (RDRs), which use single stranded RNA to generate dsRNA that in turn is targeted by DCL proteins. As a counter-defense, plant viruses encode viral suppressors of RNA silencing (VSRs) that target different components in the RNA silencing pathway. The tomato Ty-1 gene confers resistance to the DNA virus tomato yellow leaf curl virus (TYLCV) and has been reported to encode an RDRγ protein. However, the molecular mechanisms by which Ty-1 controls TYLCV infection, including whether Ty-1 is involved in RNA silencing, are unknown. Here, by using a transient expression assay, we have confirmed that Ty-1 shows antiviral activity against TYLCV in Nicotiana benthamiana. Also, in transient expression-based silencing assays, Ty-1 augmented systemic transgene silencing in GFP transgenic N. benthamiana plants. Furthermore, co-expression of Ty-1 or other RDRγ proteins from N. benthamiana or Arabidopsis with various proteins resulted in lower protein expression. These results are consistent with a model wherein Ty-1-mediated resistance to TYLCV is due, at least in part, to an increase in RNA silencing activity.

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.

Non-Feeding Transmission Modes of the Tomato Yellow Leaf Curl Virus by the Whitefly Bemisia tabaci Do Not Contribute to Reoccurring Leaf Curl Outbreaks in Tomato.

Tomato yellow leaf curl virus (TYLCV) causes significant yield loss in tomato production in the southeastern United States and elsewhere. TYLCV is transmitted by the whitefly Bemisia tabaci cryptic species in a persistent, circulative, and non-propagative manner. Unexpectedly, transovarial and sexual transmission of TYLCV has been reported for one strain from Israel. In this study, the potential contribution of the B. tabaci B cryptic species transovarial and sexual transmission of TYLCV (Israel strain, Georgia variant, Georgia, USA) to reoccurring outbreaks was investigated by conducting whitefly-TYLCV transmission assays and virus DNA detection using end point PCR, DNA quantitation via real-time PCR, and virion detection by immunocapture PCR. TYLCV DNA was detectable in four, two, and two percent of first-generation fourth-instar nymphs, first-generation adults, and second-generation adults, respectively, following transovarial acquisition. Post-mating between viruliferous counterparts, the virus's DNA was detected in four percent of males and undetectable in females. The accumulation of TYLCV DNA in whiteflies from the transovarial and/or sexual experiments was substantially lower (100 to 1000-fold) compared with whitefly adults allowed a 48-hr acquisition-access period on plants infected with TYLCV. Despite the detection of TYLCV DNA in whiteflies from the transovarial and/or mating experiments, the virions were undetectable by immunocapture PCR-a technique specifically designed to detect virions. Furthermore, tomato test plants exposed to whitefly adults that presumably acquired TYLCV transovarially or through mating remained free of detectable TYLCV DNA. Collectively, the extremely low levels of TYLCV DNA and complete absence of virions detected in whiteflies and the inability of the B. tabaci cryptic species B to transmit TYLCV to test tomato plants following transovarial and mating acquisition indicate that neither transovarial nor sexual transmission of TYLCV are probable or epidemiologically relevant for TYLCV persistence in this pathosystem.

Resistance to the insect vector Bemisia tabaci enhances the robustness and durability of tomato yellow leaf curl virus resistance conferred by Ty-1

Tomato yellow leaf curl virus (TYLCV) is a begomovirus (genus Begomovirus, family Geminiviridae) transmitted persistently by the whitefly Bemisia tabaci. It causes tomato yellow leaf curl disease (TYLCD), resulting in significant yield losses worldwide. TYLCD is controlled mainly by using F1 hybrid tomato cultivars harboring the TYLCV resistance gene Ty-1. However, infected Ty-1-bearing tomato plants accumulate viral DNA, which may eventually lead to the emergence of a resistance-breaking TYLCV variant. Recently, a B. tabaci-resistant tomato line derived from the introgression of type IV leaf glandular trichomes and acylsucrose secretion from wild tomato (Solanum pimpinellifolium) was shown to effectively control the spread of TYLCV. In this study, we combined B. tabaci resistance and Ty-1-based TYLCV resistance to increase the robustness and durability of the TYLCD resistance mediated by Ty-1 in tomato plants. Specifically, we characterized and used a Group 2-like isolate of the Israel strain of TYLCV (TYLCV-IL-G2) that contributes to TYLCD epidemics in southeastern Spain. A comparison with isolates of the previously identified TYLCV variant revealed TYLCV-IL-G2 has a similar host range, but it induces a slightly more severe TYLCD in Ty-1-bearing tomato plants. Moreover, we demonstrated that acylsucrose-producing B. tabaci-resistant tomato plants can limit the spread of TYLCV-IL-G2 better than a near-isogenic line lacking type IV trichomes and unable to secrete acylsucrose. Pyramiding Ty-1-based TYLCV resistance and B. tabaci resistance provided by type IV glandular trichomes helped to decrease the effects of TYLCV on Ty-1-bearing tomato plants as well as the likelihood of TYLCV evolution in infected plants.

Gene Editing for Disease Resistance in Crops: Success Stories and Challenges

Gene editing has emerged as a transformative tool in modern agriculture, offering new avenues for enhancing disease resistance in crops. By precisely modifying the DNA of plants, scientists can develop varieties that are better equipped to withstand the onslaught of pathogens, which pose significant threats to global food security. This article delves into the success stories and challenges associated with gene editing for disease resistance in crops, with a focus on technologies like CRISPR-Cas9, TALENs, and ZFNs. One of the major success stories in this field is the development of disease-resistant varieties of wheat, rice, and tomatoes. For instance, researchers have used CRISPR-Cas9 to create wheat varieties resistant to powdery mildew, a devastating fungal disease. Similarly, gene editing has enabled the development of rice strains with enhanced resistance to bacterial blight, a disease that can lead to significant yield losses. In tomatoes, gene editing has been employed to confer resistance against the Tomato Yellow Leaf Curl Virus, which severely affects tomato production worldwide. These achievements underscore the potential of gene editing to create crops that are not only more resilient but also capable of maintaining high yields under disease pressure. However, the application of gene editing in crop disease resistance is not without challenges. One of the primary concerns is the regulatory landscape, which varies significantly across different countries. While some nations have embraced gene-edited crops, others have imposed strict regulations, treating them similarly to genetically modified organisms (GMOs). This inconsistency can hinder the global deployment of disease-resistant crops and create trade barriers. Additionally, there are concerns related to off-target effects, where unintended changes in the genome may occur, potentially leading to unintended consequences in the plant's growth or ecological interactions.

Characterization of F2 generation tomato plants and marker assisted selection against tomato spotted wilt virus (tswv) and tomato yellow leaf curl virus (tylcv)

Characterization of F2 generation tomato plants and marker assisted selection against tomato spotted wilt virus (tswv) and tomato yellow leaf curl virus (tylcv)

Demonstration of Insect Vector-Mediated Transfer of a Betasatellite between Two Helper Viruses.

Begomoviruses, transmitted by the whitefly Bemisia tabaci, pose significant threats to global agriculture due to their severe impact on various crops. Among the satellite molecules associated with begomoviruses, betasatellites play a crucial role in enhancing disease severity and yield losses. The spread and association of these molecules with helper viruses in host plants are thus matters of concern. Here, we focus on the propagation of betasatellites and, more specifically, on their transfer between different helper viruses and hosts through vector transmission. Our results show that the cotton leaf curl Gezira betasatellite (CLCuGeB), initially acquired with its helper virus cotton leaf curl Gezira virus (CLCuGeV) from an okra plant, can be transmitted and assisted by a different helper virus, tomato yellow leaf curl virus (TYLCV), in a different host plant (tomato plant). The new association can be formed whether TYLCV and CLCuGeB encounter each other in a host plant previously infected with TYLCV or in whiteflies having acquired the different components separately. Our findings reveal two pathways by which betasatellites can be transferred between helper viruses and host plants and highlight the ability of betasatellites to spread in begomovirus-infected environments.

Field Screening of tomato breeding lines for improved resistance against Tomato Yellow Leaf Curl Virus (TYLCV)

The tomato represents an extensively cultivated crop within tropical and subtropical regions of the world for their fresh market and processing attribute. However, its production frequently encounters substantial setbacks due to notable losses associated with diseases such as Tomato Yellow Leaf Curl Virus (TyLCV). To address this challenge, the current study aimed to assess the resistance or susceptibility of selected 25 tomato breeding lines for TyLCV under natural field screenings to mimic real-world scenarios in accordance with the disease reaction score of Banerjee and Kalloo (1987). The field screening results showed that, the check Arka Vishes line demonstrated high resistance (HR) to TLCV, with reduced PDS, PDI values and a low coefficient of infection (CI). Several lines, including CBE SL 101, CBE SL 105, CBE SL 108, CBE SL 110, and CBE SL 114, exhibited a resistant (R) reaction, while others, such as CBE SL 102, CBE SL 107, CBE SL 112, CBE SL 120, and Arka Rakshak, displayed moderate resistance (MR). Conversely, lines CBE SL 103, CBE SL 104, CBE SL 106, CBE SL 109, CBE SL 111, CBE SL 115, CBE SL 117, and CBE SL 123 showed moderate susceptibility (MS), and the remaining lines, namely CBE SL 113, CBE SL 116, CBE SL 118, CBE SL 119, CBE SL 121, and CBE SL 122, were deemed susceptible to TLCV. The varying disease responses observed across these lines provide valuable insights into the complex dynamics of host-pathogen interactions in tomato plants, informing strategies for disease management and breeding efforts aimed at enhancing resistance to TLCV.

Geminivirus C4/AC4 proteins hijack cellular COAT PROTEIN COMPLEX I for chloroplast targeting and viral infections.

Geminiviruses infect numerous crops and cause extensive agricultural losses worldwide. During viral infection, geminiviral C4/AC4 proteins relocate from the plasma membrane to chloroplasts, where they inhibit the production of host defense signaling molecules. However, mechanisms whereby C4/AC4 proteins are transported to chloroplasts are unknown. We report here that tomato (Solanum lycopersicum) COAT PROTEIN COMPLEX I (COPI) components play a critical role in redistributing Tomato yellow leaf curl virus C4 protein to chloroplasts via an interaction between the C4 and β subunits of COPI. Coexpression of both proteins promotes the enrichment of C4 in chloroplasts that is blocked by a COPI inhibitor. Overexpressing or downregulating gene expression of COPI components promotes or inhibits the viral infection, respectively, suggesting a proviral role of COPI components. COPI components play similar roles in C4/AC4 transport and infections of two other geminiviruses: Beet curly top virus and East African cassava mosaic virus. Our results reveal an unconventional role of COPI components in protein trafficking to chloroplasts during geminivirus infection and suggest a broad-spectrum antiviral strategy in controlling geminivirus infections in plants.

Herbivore-induced volatiles reduce the susceptibility of neighboring tomato plants to transmission of a whitefly-borne begomovirus.

Plant viruses exist in a broader ecological community, with key components include non-vector herbivores that can impact vector abundance, behavior, and virus transmission within shared host plants. However, little is known about the effects of non-vector herbivores infestation on the virus transmission by vector insects on the neighboring plants through inter-plant airborne chemicals. In this study, we investigated how volatiles emitted from tomato plants infested with the two-spotted spider mite (Tetranychus urticae) affect the infection of Tomato yellow leaf curl virus (TYLCV) transmitted by the whitefly (Bemisia tabaci) in the neighboring plants. Exposure of neighboring tomato plants to volatiles released from T. urticae-infested tomato plants reduced subsequent herbivory as well as TYLCV transmission and infection, and JA signaling pathway was essential for generation of the inter-plant defense signals. We also demonstrated that (E)-β-Ocimene and MeSA were two volatiles induced by T. urticae that synergistically attenuated TYLCV transmission and infection in tomato. Thus, our findings suggest that plant-plant communication via volatiles likely represents a widespread defensive mechanism that substantially contributes to plant fitness. Understanding such phenomena may help us to predict the occurrence and epidemic of multiple herbivores and viruses in the agroecosystem, ultimately to manage pest and virus outbreaks.

Tomato yellow leaf curl virus manipulates Bemisia tabaci, MEAM1 both directly and indirectly through changes in visual and volatile cues.

The sweetpotato whitefly, Bemisia tabaci MEAM1, is one of the most devastating pests of row-crop vegetables worldwide, damaging crops directly through feeding and indirectly through the transmission of many different viruses, including the geminivirus Tomato yellow leaf curl virus (TYLCV). Y-tube olfactometer tests were conducted at different stages of TYLCV infection in tomatoes to understand how TYLCV affects B. tabaci behavior. We also recorded changes in tomato hosts' color and volatile profiles using color spectrophotometry and gas chromatography-mass spectrometry (GC-MS). We found that the infection status of B. tabaci and the infection stage of TYLCV influenced host selection, with uninfected whiteflies showing a preference for TYLCV-infected hosts, especially during the late stages of infection. Viruliferous B. tabaci attraction to visual targets significantly differed from non-viruliferous B. tabaci. Late-stage infected hosts had larger surface areas reflecting yellow-green wavelengths and higher emissions of methyl salicylate in their volatile profiles. These findings shed new light on several critical mechanisms involved in the viral manipulation of an insect vector and its economically important host.

High production of recombinant protein using geminivirus-based deconstructed vectors in Nicotiana benthamiana.

We focused on the geminiviral vector systems to develop an efficient vector system for plant biotechnology. Begomoviruses and curtoviruses, which belong to the Geminiviridae family, contain an intergenic region (IR) and four genes involved in replication, including replication-associated protein (Rep, C1), transcriptional activator (TrAP, C2), and replication enhancer (REn, C3). Geminiviruses can amplify thousands of copies of viral DNA using plant DNA polymerase and viral replication-related enzymes and accumulate viral proteins at high concentrations. In this study, we optimized geminiviral DNA replicon vectors based on tomato yellow leaf curl virus (TYLCV), honeysuckle yellow vein virus (HYVV), and mild curly top virus (BMCTV) for the rapid, high-yield plant-based production of recombinant proteins. Confirmation of the optimal combination by co-delivery of each replication-related gene and each IR harboring the Pontellina plumata-derived turbo green fluorescence protein (tGFP) gene via agroinfiltration in Nicotiana benthamiana leaves resulted in efficient replicon amplification and robust protein production within 3 days. Co-expression with the p19 protein of the tomato bush stunt virus, a gene-silencing suppressor, further enhanced tGFP accumulation by stabilizing mRNA. With this system, tGFP protein was produced at 0.7-1.2 mg/g leaf fresh weight, corresponding to 6.9-12.1% in total soluble protein. These results demonstrate the advantages of rapid and high-level production of recombinant proteins using the geminiviral DNA replicon system for transient expression in plants.

Effect of Introgression of Ty-1 and ty-5 Genes on Productivity, Quality, and Antioxidant Compounds in De la Pera Tomato Breeding Lines

Tomato (Solanum lycopersicum L.) is a crop that is affected by more than a hundred viral species. De la pera is a local varietal type of tomato that is very popular in southeastern Spain. However, it is highly susceptible to several viruses, such as Tomato yellow leaf curl virus (TYLCV), which is considered one of the most important diseases of tomato crops and is a limiting factor for production in both outdoor and protected crops, making it difficult to eradicate. This study shows the effect of gene introgression on the performance of traditional lines of De la pera by combining two genes that offer tolerance to TYLCV, Ty-1 and ty-5, on some yield and quality traits and on the antioxidant capacity of tomato fruits. Two pear tomato breeding families, UMH175 and UMH220, were evaluated. Four lines from each of the families with all homozygous combinations of the Ty-1 and ty-5 genes were studied. The results showed that the introgression of the ty-5 allele produced a slight negative effect on yield, mean fruit weight, total soluble solids, and titratable acidity, in contrast to Ty-1, which produced a large negative effect. None of the introgressions showed a negative effect on the antioxidant compounds. ty-5 is a promising gene for use in breeding programs.

Molecular Diagnosis and Genetic Diversity of Tomato Yellow Leaf Curl Virus in Basra Governorate

Molecular Diagnosis and Genetic Diversity of Tomato Yellow Leaf Curl Virus in Basra Governorate

A simplified single-leaflet graft inoculation method and a Duplex PCR assay for the identification of the Tomato Yellow Leaf Curl Virus infection in tomatoes

Tomato yellow leaf curl virus (TYLCV), a member of the Geminivirus genus, has caused an epidemic disease in tomato worldwide, including in Azerbaijan. TYLCV is a phloem-limited virus that is transmitted in a semi-persistent manner by whiteflies (Bemisia tabaci). Experimental propagation of TYCV has been performed primarily by using the single-leaflet grafting inoculation method. To develop a simple and effective method for transmission of TYLCV, we investigated grafting single-leaflets from tomato plants infected with TYLCV to recipient tomato seedlings. Fifty-one (51) tomato seedlings were tested and forty-eight (48) were grafted successfully with single-leaflets infected with TYLCV. Among them, forty-six (46) seedlings (92%) were systemically infected with TYLCV and developed typical symptoms. In our study, we found that single-leaflet grafting could provide a sufficient amount of inoculum for the transmission of TYLCV to the grafted tomato seedlings. The reported results regarding the development of the TYLCV inoculation and monitoring method and its application to study host plant-virus interaction showed that it could be used as an efficient alternative inoculation method.

Infection of tomato in Iraq with tomato leaf curl Palampur virus and multiple variants of tomato yellow leaf curl virus

Tomato yellow leaf curl disease (TYLCD) and tomato leaf curl disease (TLCD) cause serious losses in tomato production, especially in tropical and sub-tropical regions. In 2014–2015, tomato samples with TYLCD/TLCD-like symptoms were collected from different provinces of Iraq and infection with tomato yellow leaf curl virus (TYLCV) identified. To study the diversity of TYLCV, DNA of eight positive samples from this survey was used for rolling-circle amplification, cloning and sequencing. Pairwise nucleotide sequence comparisons with complete genomes showed that the Iraqi TYLCV isolates belonged to the strains TYLCV-IL and TYLCV-Mld. In a phylogenetic analysis, the Iraqi TYLCV-IL isolates grouped into three distinct clades, consisting of TYLCV-IL (A) and the two new variants TYLCV-IL (D) and TYLCV-IL (E). The Iraqi isolate of TYLCV-Mld grouped into the newly proposed TYLCV-Mld (D) variant. For one sample, sequencing also revealed co-infection with tomato leaf curl Palampur virus (ToLCPalV). The phylogenetic tree of ToLCPalV DNA-A showed a close relationship between the isolates of different hosts from Iraq and Iran. No evidence of recombination was detected in ToLCPalV DNA-A, but recombination was observed for the TYLCV isolates. The results indicate that there is a high diversity of TYLCV in Iraq, including new variants, that is partly shared with Kuwait and countries in the Eastern Mediterranean Region. Occurrence of multiple TYLCV variants and ToLCPalV can act as a potential threat to tomato production in Iraq.