Tomato Mottle Virus Research Articles

Viruses and viroids in tomato (Solanum lycopersicum) and plant growth promoting rhizobacteria as a management alternative

Viruses and viroids cause several diseases in tomato (Solanum lycopersicum) worldwide, generating important economic losses. About 312 viruses and seven viroids have been associated, of which more than 28 are present in Mexico. Therefore, the use of Plant Growth-Promoting Rhizobacteria (PGPR) can be an effective alternative for the management of viruses and viroids. The genera Pseudomonas, Bacillus, Azospirillum, Anabena and Stenotrophomonas have been implemented against main viruses reported in tomato: Cucumber mosaic virus (CMV), Tobacco mosaic virus (TMV), Tomato chlorotic spot virus (TCSV), Tomato mottle virus (ToMoV), Tomato spotted wilt virus (TSWV), Tomato yellow leaf curl virus (TYLCV), Potato virus Y (PVY), Groundnut bud necrosis virus (GBNV), with benefits in decreased incidence and severity up to 80 % and yield increase over 40 %. In Mexico, only Bacillus has been used. The use of PGPR is a strategy that could mitigate the impact of viral and viroid diseases and can be integrated into integrated management.

Vector acquisition and co-inoculation of two plant viruses influences transmission, infection, and replication in new hosts

This study investigated the role of vector acquisition and transmission on the propagation of single and co-infections of tomato yellow leaf curl virus (TYLCV,) and tomato mottle virus (ToMoV) (Family: Geminiviridae, Genus: Begomovirus) by the whitefly vector Bemisia tabaci MEAM1 (Gennadius) in tomato. The aim of this research was to determine if the manner in which viruses are co-acquired and co-transmitted changes the probability of acquisition, transmission and new host infections. Whiteflies acquired virus by feeding on singly infected plants, co-infected plants, or by sequential feeding on singly infected plants. Viral titers were also quantified by qPCR in vector cohorts, in artificial diet, and plants after exposure to viruliferous vectors. Differences in transmission, infection status of plants, and titers of TYLCV and ToMoV were observed among treatments. All vector cohorts acquired both viruses, but co-acquisition/co-inoculation generally reduced transmission of both viruses as single and mixed infections. Co-inoculation of viruses by the vector also altered virus accumulation in plants regardless of whether one or both viruses were propagated in new hosts. These findings highlight the complex nature of vector-virus-plant interactions that influence the spread and replication of viruses as single and co-infections.

The invasion biology of tomato begomoviruses in Costa Rica reveals neutral synergism that may lead to increased disease pressure and economic loss

Since the late 1980s, tomato production in Costa Rica has been affected by diseases caused by whitefly-transmitted begomoviruses. The first was tomato yellow mottle virus (ToYMoV), a locally evolved New World (NW) bipartite begomovirus associated with the tomato yellow mottle disease (ToYMoD). In the late 1990s, the invasive NW bipartite tomato leaf curl Sinaloa virus (ToLCSiV) was detected in Costa Rica and has become established and associated with ToYMoD. Finally, the invasive Old World (OW) monopartite tomato yellow leaf curl virus (TYLCV) was detected in Costa Rica in 2012 and has also become established and is causing tomato yellow leaf curl disease (TYLCD). In the present study, we investigated the invasion biology of these tomato-infecting begomoviruses in Costa Rica in terms of (i) their biological and genetic properties and (ii) disease symptoms and viral DNA accumulation in tomato plants having single and mixed infections. We first generated infectious DNA-A and DNA-B clones and agroinoculation systems for ToYMoV and ToLCSiV isolates recovered from archival ToYMoD samples collected in Costa Rica in 1990 and 2002, respectively. Tomato plants agroinoculated with the infectious clones of both viruses developed ToYMoD symptoms, completing Koch's postulates for ToYMoV, and showing that ToLCSiV also causes this disease. However, pseudorecombinants formed between the DNA components of these viruses were not infectious, which is consistent with independent evolution in different lineages and limits genetic interactions. Furthermore, ToYMoV is well-adapted to tomato, has a narrow host range and is mechanically transmissible. The DNA-A component has a recombination event in the hot spot area and induced a symptomless infection in agroinoculated Nicotiana benthamiana and tomato plants. Tomato plants co-infected with two or all three viruses developed more severe symptoms compared with plants infected with each virus alone. Symptoms induced by the NW bipartite ToYMoV and ToLCSiV appeared earlier (∼7 d post-inoculation [dpi]) than those induced by TYLCV (∼10 dpi), but TYLCD symptoms became predominant in single and mixed infections by 14 dpi. Viral DNA accumulation was quantified by qPCR and generally revealed a neutral synergistic interaction in which the viruses co-existed in mixed infections. A transient reduction in accumulation of ToYMoV and ToLCSiV was detected in mixed infections at 7 dpi, whereas TYLCV accumulation was not affected in mixed infections and was uniform among treatments and time points. Together our results suggest that this neutral synergistic interaction will lead to increased begomovirus disease severity in Costa Rica. We discuss this in terms of begomovirus invasion biology and disease management.

Artificial microRNA guide strand selection from duplexes with no mismatches shows a purine-rich preference for virus- and non-virus-based expression vectors in plants.

SummaryArtificial microRNA (amiRNA) technology has allowed researchers to direct efficient silencing of specific transcripts using as few as 21 nucleotides (nt). However, not all the artificially designed amiRNA constructs result in selection of the intended ~21‐nt guide strand amiRNA. Selection of the miRNA guide strand from the mature miRNA duplex has been studied in detail in human and insect systems, but not so much for plants. Here, we compared a nuclear‐replicating DNA viral vector (tomato mottle virus, ToMoV, based), a cytoplasmic‐replicating RNA viral vector (tobacco mosaic virus, TMV, based), and a non‐viral binary vector to express amiRNAs in plants. We then used deep sequencing and mutational analysis and show that when the structural factors caused by base mismatches in the mature amiRNA duplex were excluded, the nucleotide composition of the mature amiRNA region determined the guide strand selection. We found that the strand with excess purines was preferentially selected as the guide strand and the artificial miRNAs that had no mismatches in the amiRNA duplex were predominantly loaded into AGO2 instead of loading into AGO1 like the majority of the plant endogenous miRNAs. By performing assays for target effects, we also showed that only when the intended strand was selected as the guide strand and showed AGO loading, the amiRNA could provide the expected RNAi effects. Thus, by removing mismatches in the mature amiRNA duplex and designing the intended guide strand to contain excess purines provide better control of the guide strand selection of amiRNAs for functional RNAi effects.

Tomato mottle virus

This datasheet on Tomato mottle virus covers Identity, Overview, Distribution, Dispersal, Hosts/Species Affected, Vectors & Intermediate Hosts, Diagnosis, Biology & Ecology, Impacts, Prevention/Control, Further Information.

Tomato mottle virus

Tomato mottle virus

First report of Tomato brown rugose fruit virus infecting tomato in Florida, USA

Florida is a leading producer of fresh-market tomatoes (Solanum lycopersicum) in the US, a $262 million industry for the state. In October 2019, eight tomato plants showing chlorotic, cupped leaves and stunted foliage (Fig. 1) were collected from a c. 1.6 ha community garden in Alachua County, Florida. The symptoms observed were characteristic of Tomato yellow leaf curl virus (TYLCV). Samples were tested by PCR using TYLCV-specific primers by Navot et al. (1992). Three samples tested positive for TYLCV. Lateral flow immunoassay tests (“immunostrip assays”; Agdia, USA) for Pepino mosaic virus and Cucumber mosaic virus were negative. However, an immunostrip for Tobacco mosaic virus (TMV) gave a faint positive result for one of three TYLCV-positive sample, suggesting a mixed infection. Tobamovirus-like rigid rod-shaped particles of ∼300 nm in length, were also observed in leaf dips from this plant using an LVEM5 transmission electron microscope (DeLong America, USA). Tests for common tobamoviruses, i.e. TMV, Tomato mosaic virus, Tomato mottle mosaic virus, (Sui et al., 2017), Tobacco mild green mosaic virus (Cohen et al., 2006), and Tomato brown rugose fruit virus (ToBRFV) (Ling et al., 2019) were conducted using specific primers in RT-PCR with total RNA extracted using RNeasy® Plant Mini kits (Qiagen, Germany). Additionally, conventional PCR primers, DPI-782(5859nt-5879nt) = 5′-TAGCGAAGTGTGGAAACCTG-3′ and DPI-783(6154nt-6183nt) = 5′-GGTGCAGAGGACCATTGTAA-3′ (amplicon size of 342 bp), and primers and probes for a real-time RT-PCR Taqman assay DPI-784(6029nt-6049nt) = 5′-CCGAAACGTTAGACGCTACTC-3′, DPI-785(6106nt -6127nt) = ACCTGTTCCTTTGACCAA TTCT-3′, and DPI-787(6060nt-6083nt) TaqMan probe = 5′-ATG CAACGGTGGCTATAAGGA-3’) from the coat protein ∼5800-6130 nt region were developed for ToBRFV. Six of eight samples, including three TYLCV-positive samples, were positive for ToBRFV. The sequenced PCR products (GenBank Accession No. MW322766) were 98 to 99 % identical to ToBRFV isolates from Jordan and Mexico (KT383474, MK319944). The identification of ToBRFV was further confirmed by USDA-APHIS-PPQ-S&T laboratory (Laurel, Maryland, USA) by RT-PCR and sequencing. The Ct values of the real-time assay were in the range of 27 to 31. The weak positive result from the immunostrip, relatively high Ct values and co-infection with TYLCV suggest that ToBRFV was present in a low titre in these samples. ToBRFV was also detected in tomatoes imported from Mexico and for sale in grocery stores in Naples and Gainesville by University of Florida and Florida Department of Agriculture and Consumer Services (FDACS) around the same time (Batuman et al., 2020), Immediately following the survey, all solanaceous plants in the organic garden were destroyed. FDACS is taking all precautionary measures to test tomatoes produced in Florida. Further studies are needed to better understand the risk factors from this virus in Florida. The authors would like to thank Department of Plant Industry (DPI), Florida Department of Agriculture and Consumer Services (FDACS) for all assistance with the project.

Characterization of Local and Systemic Impact of Whitefly (Bemisia tabaci) Feeding and Whitefly-Transmitted Tomato Mottle Virus Infection on Tomato Leaves by Comprehensive Proteomics

Tomato mottle virus (ToMoV) is a single-stranded DNA (ssDNA) begomovirus transmitted to solanaceous crops by the whitefly species complex (Bemisia tabaci), causing stunted growth, leaf mottling, and reduced yield. Using a genetic repertoire of seven genes, ToMoV pathogenesis includes the manipulation of multiple plant biological processes to circumvent antiviral defenses. To further understand the effects of whitefly feeding and whitefly-transmitted ToMoV infection on tomato plants (Solanum lycopersicum ‘Florida Lanai’), we generated comprehensive protein profiles of leaves subjected to feeding by either viruliferous whiteflies harboring ToMoV, or non-viruliferous whiteflies, or a no-feeding control. The effects of whitefly feeding and ToMoV infection were measured both locally and systemically by sampling either a mature leaf directly from the site of clip-cage confined whitefly feeding, or from a newly formed leaf 10 days post feeding (dpf). At 3 dpf, tomato’s response to ToMoV included proteins associated with translation initiation and elongation as well as plasmodesmata dynamics. In contrast, systemic impacts of ToMoV on younger leaves 10 dpf were more pronounced and included a virus-specific change in plant proteins associated with mRNA maturation and export, RNA-dependent DNA methylation, and other antiviral plant processes. Our analysis supports previous findings and provides novel insight into tomato’s local and systemic response to whitefly feeding and ToMoV infection.

Tomato mottle mosaic virus intercepted by Australian biosecurity in Capsicum annuum seed

The Tobamovirus, Tomato mottle mosaic virus (ToMMV), was first reported in Mexico in 2013. The virus is thought to pose a serious risk to capsicum (Capsicum annuum) and tomato (Solanum lycopersicum) crops as it is may break resistance. In May 2019 a shipment of imported capsicum seeds was submitted for testing on-shore and using a one-step RT-PCR which detects Solanaceous tobamoviruses, an amplicon of 811 base pairs (bp) was detected and direct sequencing of this amplicon indicated that it had 98% - 99% nucleotide (nt) identity to the same region in ToMMV isolates. A cDNA library was generated using the Illumina TruSeq Stranded Total RNA and sequenced using Illumina HiSeq 3000 technology, bioinformatic analysis confirmed the arrangement of a 6398 nt genome which was 98% - 99% nt identity with the type strain of ToMMV. This is the first report of ToMMV in Capsicum annuum seed.

Ty-6, a major begomovirus resistance gene on chromosome 10, is effective against Tomato yellow leaf curl virus and Tomato mottle virus

Key messageTy-6 is a major resistance gene on chromosome 10 of tomato that provides resistance against monopartite and bipartite begomoviruses and complements resistance conferred by the known Ty-3 and ty-5 genes.Resistance to monopartite and bipartite begomoviruses is an important breeding objective for cultivated tomato. Several begomovirus resistance genes have been introgressed from related Solanum species and are available for breeding purposes. In the present study, we mapped an additional locus, Ty-6, to chromosome 10 of tomato. Ty-6 is effective against both monopartite Tomato yellow leaf curl virus (TYLCV) and bipartite Tomato mottle virus (ToMoV). Gene action is incomplete dominance, with an intermediate resistance response when Ty-6 is heterozygous. Analysis of populations segregating for Ty-6 along with Ty-3 or ty-5 indicates that the highest level of resistance against TYLCV is attained when Ty-6 is combined with an additional resistance allele. Our results also demonstrate that ty-5 is ineffective against ToMoV. Although multiple SNPs linked to Ty-6 were identified and can be used for breeding purposes, none of these were consistently polymorphic between Ty-6 and ty-6 breeding lines. Further research is underway to generate resequencing data for several Ty-6 inbred lines for the discovery of additional sequence polymorphisms that can be used for fine mapping and characterizing the Ty-6 locus.

Sucrose Nonfermenting 1-Related Protein Kinase 1 Phosphorylates a Geminivirus Rep Protein to Impair Viral Replication and Infection.

Geminiviruses are single-stranded DNA viruses that infect a wide variety of plants and cause severe crop losses worldwide. The geminivirus replication initiator protein (Rep) binds to the viral replication origin and catalyzes DNA cleavage and ligation to initiate rolling circle replication. In this study, we found that the Tomato golden mosaic virus (TGMV) Rep is phosphorylated at serine-97 by sucrose nonfermenting 1-related protein kinase 1 (SnRK1), a master regulator of plant energy homeostasis and metabolism. Phosphorylation of Rep or the phosphomimic S97D mutation impaired Rep binding to viral DNA. A TGMV DNA-A replicon containing the Rep S97D mutation replicated less efficiently in tobacco (Nicotiana tabacum) protoplasts than in wild-type or Rep phosphorylation-deficient replicons. The TGMV Rep-S97D mutant also was less infectious than the wild-type virus in Nicotiana benthamiana and was unable to infect tomato (Solanum lycopersicum). Nearly all geminivirus Rep proteins have a serine residue at the position equivalent to TGMV Rep serine-97. SnRK1 phosphorylated the equivalent serines in the Rep proteins of Tomato mottle virus and Tomato yellow leaf curl virus and reduced DNA binding, suggesting that SnRK1 plays a key role in combating geminivirus infection. These results established that SnRK1 phosphorylates Rep and interferes with geminivirus replication and infection, underscoring the emerging role for SnRK1 in the host defense response against plant pathogens.

Detection of tobamoviruses by RT-PCR using a novel pair of degenerate primers

A generic RT-PCR assay was developed for the universal detection of viruses of the genus Tobamovirus using a novel pair of degenerate primers designed based on conserved regions on replicase genes of 32 tobamoviruses. The assay detected nine tobamoviruses, including six Solanaceae-infecting subgroup tobamoviruses of Tobacco mosaic virus (TMV), Tomato mosaic virus (ToMV), Tomato mottle mosaic virus (ToMMV), Tobacco mottle green mosaic virus (TMGMV), Pepper mild mottle virus (PMMoV), Paprika mild mottle virus (PaMMV), one Orchidaceae-infecting tobamovirus of Odontoglossum ringspot virus (ORSV) and two Cucurbitaceae-infecting subgroup tobamoviruses of Cucumber green mottle mosaic virus (CGMMV) and Zucchini green mottle mosaic virus (ZGMMV), with high amplification efficiency, specificity and sensitivity. The assay was applied to detect tobamoviruses in pepper and tomato fields. Five tobamoviruses, PMMoV, TMV, ToMV, ToMMV and TMGMV, were detected from the pepper fields in single and mixed infections. Single infections of PMMoV, ToMV and ToMMV and mix-infection of ToMV + PMMoV were detected from the tomato fields. Among these viruses, PMMoV was first detected from tomato worldwide, while ToMMV was first detected from tomato plants in China. This generic assay is simple, cost-effective and has great potential to detect more tobamoviruses in the field.

Lanai: A small, fast growing tomato variety is an excellent model system for studying geminiviruses

Geminiviruses are devastating single-stranded DNA viruses that infect a wide variety of crops in tropical and subtropical areas of the world. Tomato, which is a host for more than 100 geminiviruses, is one of the most affected crops. Developing plant models to study geminivirus-host interaction is important for the design of virus management strategies. In this study, “Florida Lanai” tomato was broadly characterized using three begomoviruses (Tomato yellow leaf curl virus, TYLCV; Tomato mottle virus, ToMoV; Tomato golden mosaic virus, TGMV) and a curtovirus (Beet curly top virus, BCTV). Infection rates of 100% were achieved by agroinoculation of TYLCV, ToMoV or BCTV. Mechanical inoculation of ToMoV or TGMV using a microsprayer as well as whitefly transmission of TYLCV or ToMoV also resulted in 100% infection frequencies. Symptoms appeared as early as four days post inoculation when agroinoculation or bombardment was used. Symptoms were distinct for each virus and a range of features, including plant height, flower number, fruit number, fruit weight and ploidy, was characterized. Due to its small size, rapid growth, ease of characterization and maintenance, and distinct responses to different geminiviruses, “Florida Lanai” is an excellent choice for comparing geminivirus infection in a common host.

Recombinase polymerase amplification applied to plant virus detection and potential implications

Several isothermal techniques for the detection of plant pathogens have been developed with the advent of molecular techniques. Among them, Recombinase Polymerase Amplification (RPA) is becoming an important technique for the rapid, sensitive and cost-effective detection of plant viruses. The RPA technology has the advantage to be implemented in field-based scenarios because the method requires a minimal sample preparation, and is performed at constant low temperature (37–42 °C). The RPA technique is rapidly becoming a promising tool for use in rapid detection and further diagnostics in plant clinics and monitoring quarantine services. This paper presents a review of studies conducted using RPA for detection/diagnosis of plant viruses with either DNA genomes (Banana bunchy top virus, Bean golden yellow mosaic virus, Tomato mottle virus, Tomato yellow leaf curl virus) or RNA genomes (Little Cherry virus 2, Plum pox virus and Rose rosette virus).

Molecular and biological characterization of an isolate of Tomato mottle mosaic virus (ToMMV) infecting tomato and other experimental hosts in eastern Spain

Tomato is known to be a natural and experimental reservoir host for many plant viruses. In the last few years a new tobamovirus species, Tomato mottle mosaic virus (ToMMV), has been described infecting tomato and pepper plants in several countries worldwide. Upon observation of symptoms in tomato plants growing in a greenhouse in Valencia, Spain, we aimed to ascertain the etiology of the disease. Using standard molecular techniques, we first detected a positive sense single-stranded RNA virus as the probable causal agent. Next, we amplified and sequenced its full-length genomic RNA which identified the virus as a new ToMMV isolate. Through extensive assays on distinct plant species, we investigated the host range of the Spanish ToMMV isolate. Several plant species were locally and/or systemically infected by the virus, some of which had not been previously reported as ToMMV hosts despite they are commonly used in research greenhouses. Finally, two reliable molecular diagnostic techniques were developed and used to assess the presence of ToMMV. This is the first observation of ToMMV in tomato plants in Europe. We discuss the possibility that, given the high sequence homology between ToMMV and Tomato mosaic virus, the former may have been mistakenly diagnosed as the latter by serological methods.

Subcellular Localization and Detection of Tobacco mosaic virus ORF6 Protein by Immunoelectron Microscopy.

Members of the genus Tobamovirus represent one of the best-characterized groups of plant positive, single stranded RNA viruses. Previous studies have shown that genomes of some tobamoviruses contain not only genes coding for coat protein, movement protein, and the cistron coding for different domains of RNA-polymerase, but also a gene, named ORF6, coding for a poorly conserved small protein. The amino acid sequences of ORF6 proteins encoded by different tobamoviruses are highly divergent. The potential role of ORF6 proteins in replication of tobamoviruses still needs to be elucidated. In this study, using biochemical and immunological methods, we have shown that ORF6 peptide is accumulated after infection in case of two isolates of Tobacco mosaic virus strain U1 (TMV-U1 common and TMV-U1 isolate A15). Unlike virus particles accumulating in the cytoplasm, the product of the ORF6 gene is found mainly in nuclei, which correlates with previously published data about transient expression of ORF6 isolated from TMV-U1. Moreover, we present new data showing the presence of ORF6 genes in genomes of several tobamoviruses. For example, in the genomes of other members of the tobamovirus subgroup 1, including Rehmannia mosaic virus, Paprika mild mottle virus, Tobacco mild green mosaic virus, Tomato mosaic virus, Tomato mottle mosaic virus, and Nigerian tobacco latent virus, sequence comparisons revealed the existence of a similar open reading frame like ORF6 of TMV.

Genetic structure of populations of the begomoviruses Tomato mottle leaf curl virus and Sida mottle Alagoas virus infecting tomato (Solanum lycopersicum) and Sida spp., respectively

Begomoviruses are among the most damaging pathogens infecting cultivated hosts worldwide, and are considered an important limiting factor in tomato fields. In the present study, foliar samples of tomato and Sida spp. were collected in Boqueirao, state of Paraiba, Brazil, in May 2014. Viral genomes were amplified, cloned and sequenced. A total of 30 DNA-A components were obtained. Pairwise comparisons indicated the presence of only two begomovirus species: Tomato mottle leaf curl virus (ToMoLCV) from tomato samples and Sida mottle Alagoas virus (SiMoAV) in Sida spp. ToMoLCV and SiMoAV subpopulations showed high genetic variability, with the ToMoLCV Rep gene being the most variable. Recombination events were detected only among ToMoLCV isolates, with recombination breakpoints occurring in the common region and in the Rep gene. Negative or purifying selection seems to be among the most important evolutionary mechanisms acting on the CP and Rep genes in both ToMoLCV and SiMoAV. Our results indicate that ToMoLCV remains as the main tomato-infecting begomovirus in northeastern Brazil, and that Sida spp. do not contribute to ToMoLCV epidemics in tomato.

Evaluation of recombinase polymerase amplification for detection of begomoviruses by plant diagnostic clinics.

BackgroundPlant viruses in the genus Begomovirus, family Geminiviridae often cause substantial crop losses. These viruses have been emerging in many locations throughout the tropics and subtropics. Like many plant viruses, they are often not recognized by plant diagnostic clinics due in large part to the lack of rapid and cost effective assays. An isothermal amplification assay, Recombinase polymerase amplification (RPA), was evaluated for its ability to detect three begomoviruses and for its suitability for use in plant diagnostic clinics. Methods for DNA extraction and separation of amplicons from proteins used in the assay were modified and compared to RPA manufacturer’s protocols. The modified RPA assays were compared to PCR assays for sensitivity, use in downstream applications, cost, and speed.ResultsRecombinase polymerase amplification (RPA) assays for the detection of Bean golden yellow mosaic virus, Tomato mottle virus and Tomato yellow leaf curl virus (TYLCV) were specific, only amplifying the target viruses in three different host species. RPA was able to detect the target virus when the template was in a crude extract generated using a simple inexpensive extraction method, while PCR was not. Separation of RPA-generated amplicons from DNA-binding proteins could be accomplished by several methods, all of which were faster and less expensive than that recommended by the manufacturer. Use of these modifications resulted in an RPA assay that was faster than PCR but with a similar reagent cost. This modified RPA was the more cost effective assay when labor is added to the cost since RPA can be performed much faster than PCR. RPA had a sensitivity approximate to that of ELISA when crude extract was used as template. RPA-generated amplicons could be used in downstream applications (TA cloning, digestion with a restriction endonuclease, direct sequencing) similar to PCR but unlike some other isothermal reactions.ConclusionsRPA could prove useful for the cost effective detection of plant viruses by plant diagnostic clinics. It can be performed in one hour or less with a reagent cost similar to that of PCR but with a lower labor cost, and with an acceptable level of sensitivity and specificity.Electronic supplementary materialThe online version of this article (doi:10.1186/s12985-016-0504-8) contains supplementary material, which is available to authorized users.

Evaluation of a SUMO E2 Conjugating Enzyme Involved in Resistance to Clavibacter michiganensis Subsp. michiganensis in Solanum peruvianum, Through a Tomato Mottle Virus VIGS Assay.

Clavibacter michiganensis subsp. michiganensis (Cmm) causes bacterial wilt and canker of tomato. Currently, no Solanum lycopersicum resistant varieties are commercially available, but some degree of Cmm resistance has been identified in Solanum peruvianum. Previous research showed up-regulation of a SUMO E2 conjugating enzyme (SCEI) transcript in S. peruvianum compared to S. lycopersicum following infection with Cmm. In order to test the role of SCEI in resistance to Cmm, a fragment of SCEI from S. peruvianum was cloned into a novel virus-induced gene-silencing (VIGS) vector based on the geminivirus, Tomato Mottle Virus (ToMoV). Using biolistic inoculation, the ToMoV-based VIGS vector was shown to be effective in S. peruvianum by silencing the magnesium chelatase gene, resulting in leaf bleaching. VIGS with the ToMoV_SCEI construct resulted in ~61% silencing of SCEI in leaves of S. peruvianum as determined by quantitative RT-PCR. The SCEI-silenced plants showed unilateral wilting (15 dpi) and subsequent death (20 dpi) of the entire plant after Cmm inoculation, whereas the empty vector-treated plants only showed wilting in the Cmm-inoculated leaf. The SCEI-silenced plants showed higher Cmm colonization and an average of 4.5 times more damaged tissue compared to the empty vector control plants. SCEI appears to play an important role in the innate immunity of S. peruvianum against Cmm, perhaps through the regulation of transcription factors, leading to expression of proteins involved in salicylic acid-dependent defense responses.

COMBINATIONS OF TY RESISTANCE GENES GENERALLY PROVIDE MORE EFFECTIVE CONTROL AGAINST BEGOMOVIRUSES THAN DO SINGLE GENES

Begomoviruses are a major threat to tropical and subtropical tomato production, worldwide. Host resistance provides an effective strategy for reducing yield losses associated with these viruses. Resistance has been introgressed into cultivated tomato (Solanum lycopersicum L.) primarily from a number of wild tomato relatives, and the major resistance genes available to breeders today include Ty-1 (and its allele, Ty-3), Ty-2, Ty-4, ty-5, and a recently-identified gene tentatively named “Ty-6”. Important to the effective and efficient deployment of resistant cultivars is an understanding of each resistance allele’s effect when used alone and when combined with other resistance genes. Ty-1 and Ty2 are both dominant and, where effective, provide a high level of resistance; whereas the other genes provide partial resistance and are additive to partially dominant or recessive as with ty-5. Two- and three-gene combinations of Ty-3, Ty-4, and ty-5 in semi-isogenic backgrounds provided high levels of resistance to Tomato yellow leaf curl virus (TYLCV). Tomato mottle virus (ToMoV) trials in Florida using the same materials were severe, but indicated that Ty-3 and Ty-4 had small but significant effects in lowering ToMoV disease severity but, ty-5 did not. In two additional populations segregating for both ty-5 and “Ty-6” that were inoculated with ToMoV, ty-5 had no significant effect on resistance. Combining “Ty-6” and ty-5 homozygously provided a high level of resistance to TYLCV while the combination of homozygous ty-5 with heterozygous “Ty-6” was also good, but not as resistant as the former.