Population studies of the wild tomato species Solanum chilense reveal geographically structured major gene-mediated pathogen resistance

Natural plant populations encounter strong pathogen pressure and defence-associated genes are known to be under selection dependent on the pressure by the pathogens. Here, we use populations of the wild tomato Solanum chilense to investigate natural resistance against Cladosporium fulvum, a well-known ascomycete pathogen of domesticated tomatoes. Host populations used are from distinct geographical origins and share a defined evolutionary history. We show that distinct populations of S. chilense differ in resistance against the pathogen. Screening for major resistance gene-mediated pathogen recognition throughout the whole species showed clear geographical differences between populations and complete loss of pathogen recognition in the south of the species range. In addition, we observed high complexity in a homologues of Cladosporium resistance (Hcr) locus, underlying the recognition of C. fulvum, in central and northern populations. Our findings show that major gene-mediated recognition specificity is diverse in a natural plant-pathosystem. We place major gene resistance in a geographical context that also defined the evolutionary history of that species. Data suggest that the underlying loci are more complex than previously anticipated, with small-scale gene recombination being possibly responsible for maintaining balanced polymorphisms in the populations that experience pathogen pressure.     

Inoculation of Transgenic Resistant Potato by Phytophthora infestans Affects Host Plant Choice of a Generalist Moth

Pathogen attack and the plant’s response to this attack affect herbivore oviposition preference and larval performance. Introduction of major resistance genes against Phytophthora infestans (Rpi-genes), the cause of the devastating late blight disease, from wild Solanum species into potato changes the plant-pathogen interaction dynamics completely, but little is known about the effects on non-target organisms. Thus, we examined the effect of P. infestans itself and introduction of an Rpi-gene into the crop on host plant preference of the generalist insect herbivore, Spodoptera littoralis (Lepidoptera: Noctuidae). In two choice bioassays, S. littoralis preferred to oviposit on P. infestans-inoculated plants of both the susceptible potato (cv. Desiree) and an isogenic resistant clone (A01-22: cv. Desiree transformed with Rpi-blb1), when compared to uninoculated plants of the same genotype. Both cv. Desiree and clone A01-22 were equally preferred for oviposition by S. littoralis when uninoculated plants were used, while cv. Desiree received more eggs compared to the resistant clone when both were inoculated with the pathogen. No significant difference in larval and pupal weight was found between S. littoralis larvae reared on leaves of the susceptible potato plants inoculated or uninoculated with P. infestans. Thus, the herbivore’s host plant preference in this system was not directly associated with larval performance. The results indicate that the Rpi-blb1 based resistance in itself does not influence insect behavior, but that herbivore oviposition preference is affected by a change in the plant-microbe interaction.     

Reassessment of QTLs for Late Blight Resistance in the Tomato Accession L3708 Using a Restriction Site Associated DNA (RAD) Linkage Map and Highly Aggressive Isolates of Phytophthora infestans

Tomato late blight caused by the oomycete pathogen Phytophthora infestans (Mont.) de Bary is a major threat to tomato production in cool and wet environments. Intensified outbreaks of late blight have been observed globally from the 1980s, and are associated with migration of new and more aggressive populations of P. infestans in the field. The objective of this study was to reassess late blight resistance in the wild tomato accession L3708 (Solanum pimpinellifolium L.) against pathogens of different aggressiveness. An F_2:3 genetic mapping population was developed using L3708 as the paternal parent. Two isolates of P. infestans, Pi39A and Pi733, were used for inoculation. Pi733 is a highly aggressive genotype that defeats three known late blight resistance genes, Ph-1, Ph-2, and Ph-5t in tomato. In contrast, Pi39A is a less aggressive genotype that defeats only Ph-1. Restriction site Associated DNA Sequencing (RAD-Seq) technology was used to massively sequence 90 bp nucleotides adjacent to both sides of PstI restriction enzyme cutting sites in the genome for all individuals in the genetic mapping population. The RAD-seq data were used to construct a genetic linkage map containing 440 single nucleotide polymorphism markers. Quantitative trait locus (QTL) analysis identified a new disease-resistant QTL specific to Pi733 on chromosome 2. The Ph-3 gene located on chromosome 9 could be detected whichever isolates were used. This study demonstrated the feasibility and efficiency of RAD-Seq technology for conducting a QTL mapping experiment using an F_2:3 mapping population, which allowed the identification of a new late blight resistant QTL in tomato.     

Distribution of Pyrethroid Resistant Populations of Triatoma infestans in the Southern Cone of South America

BackgroundA number of studies published during the last 15 years showed the occurrence of insecticide resistance in Triatoma infestans populations. The different toxicological profiles and mechanisms of resistance to insecticides is due to a genetic base and environmental factors, being the insecticide selective pressure the best studied among the last factors. The studies on insecticide resistance on T. infestans did not consider the effect of environmental factors that may influence the distribution of resistance to pyrethroid insecticides. To fill this knowledge gap, the present study aims at studying the association between the spatial distribution of pyrethroid resistant populations of T. infestans and environmental variables.Conclusions/SignificanceThe occurrence of these two groups concentrated over a particular region that coincides with the area where populations of the intermediate cytogenetic group were found might reflect the spatial heterogeneity of the genetic variability of T. infestans, that seems to be the cause of the insecticide resistance in the area, even on sylvatic populations of T. infestans, never before exposed to pyrethroid insecticides, representing natural and wild toxicological phenotypes. The strong linear relationship found between LD₅₀ and RR₅₀ suggest RR₅₀ might not be the best indicator of insecticide resistance in triatomines.     

Plant integrity: An important factor in plant-pathogen interactions

The effect of plant integrity and of aboveground-belowground defense signaling on plant resistance against pathogens and herbivores is emerging as a subject of scientific research. There is increasing evidence that plant defense responses to pathogen infection differ between whole intact plants and detached leaves. Studies have revealed the importance of aboveground-belowground defense signaling for plant defenses against herbivores, while our studies have uncovered that the roots as well as the plant integrity are important for the resistance of the potato cultivar Sarpo Mira against the hemibiotrophic oomycete pathogen Phytophthora infestans. Furthermore, in the Sarpo Mira–P. infestans interactions, the plant’s meristems, the stalks or both, seem to be associated with the development of the hypersensitive response and both the plant’s roots and shoots contain antimicrobial compounds when the aerial parts of the plants are infected. Here, we present a short overview of the evidence indicating the importance of plant integrity on plant defense responses.     

Multiple factors control the level of resistance induced in tomato by Fusarium oxysporum f. sp. cucumerinum

Fourteen wild type and three UV-irradiated isolates of Fusarium oxysporum f. sp. cucumerinum (Foc) were evaluated as to the level of resistance they could induce in tomato to late blight caused by Phytophthora infestans. Tomato plants were induced by applying a suspension of Foc microconidia directly to the surface of the potting media without disturbing the tomato roots. Upper leaves of tomato plants were inoculated with P. infestans, and a reduction in lesion expansion was used as an index of induced resistance. All fourteen wild type isolates of Foc significantly reduced expansion of late blight lesions. One of the wild type isolates produced a significantly weaker resistance response than the other isolates. None of the UV-irradiated isolates induced significant resistance. The same Foc isolates were compared as to their virulence and their pigment production in culture, and considerable variation among them was revealed for both characteristics. Positive correlations existed both between the level of induced resistance and virulence, and between the level of induced resistance and pigmentation. The gradual increment in the level of induced resistance and the exceptions to the correlations between induced resistance and the two characteristics investigated suggest that multiple factors contribute to the induction of resistance by Foc.     

Down-regulation of <Emphasis Type="Italic">Arabidopsis</Emphasis><Emphasis Type="Italic">DND1</Emphasis> orthologs in potato and tomato leads to broad-spectrum resistance to late blight and powdery mildew

Multiple susceptibility genes (S), identified in Arabidopsis, have been shown to be functionally conserved in crop plants. Mutations in these S genes result in resistance to different pathogens, opening a new way to achieve plant disease resistance. The aim of this study was to investigate the role of Defense No Death 1 (DND1) in susceptibility of tomato and potato to late blight (Phytophthora infestans). In Arabidopsis, the dnd1 mutant has broad-spectrum resistance against several fungal, bacterial, and viral pathogens. However this mutation is also associated with a dwarfed phenotype. Using an RNAi approach, we silenced AtDND1 orthologs in potato and tomato. Our results showed that silencing of the DND1 ortholog in both crops resulted in resistance to the pathogenic oomycete P. infestans and to two powdery mildew species, Oidium neolycopersici and Golovinomyces orontii. The resistance to P. infestans in potato was effective to four different isolates although the level of resistance (complete or partial) was dependent on the aggressiveness of the isolate. In tomato, DND1-silenced plants showed a severe dwarf phenotype and autonecrosis, whereas DND1-silenced potato plants were not dwarfed and showed a less pronounced autonecrosis. Our results indicate that S gene function of DND1 is conserved in tomato and potato. We discuss the possibilities of using RNAi silencing or loss-of-function mutations of DND1 orthologs, as well as additional S gene orthologs from Arabidopsis, to breed for resistance to pathogens in crop plants.     

Paranoid potato: Phytophthora-resistant genotype shows constitutively activated defense

Phytophthora is the most devastating pathogen of dicot plants. There is a need for resistance sources with different modes of action to counteract the fast evolution of this pathogen. In order to better understand mechanisms of defense against P. infestans, we analyzed several clones of potato. Two of the genotypes tested, Sarpo Mira and SW93-1015, exhibited strong resistance against P. infestans in field trials, whole plant assays and detached leaf assays. The resistant genotypes developed different sizes of hypersensitive response (HR)-related lesions. HR lesions in SW93-1015 were restricted to very small areas, whereas those in Sarpo Mira were similar to those in Solanum demissum, the main source of classical resistance genes. SW93-1015 can be characterized as a cpr (constitutive expressor of PR genes) genotype without spontaneous microscopic or macroscopic HR lesions. This is indicated by constitutive hydrogen peroxide (H₂O₂) production and PR1 (pathogenesis-related protein 1) secretion. SW93-1015 is one of the first plants identified as having classical protein-based induced defense expressed constitutively without any obvious metabolic costs or spontaneous cell death lesions.     

Sequence Evolution and Expression Regulation of Stress-Responsive Genes in Natural Populations of Wild Tomato

The wild tomato species Solanum chilense and S. peruvianum are a valuable non-model system for studying plant adaptation since they grow in diverse environments facing many abiotic constraints. Here we investigate the sequence evolution of regulatory regions of drought and cold responsive genes and their expression regulation. The coding regions of these genes were previously shown to exhibit signatures of positive selection. Expression profiles and sequence evolution of regulatory regions of members of the Asr (ABA/water stress/ripening induced) gene family and the dehydrin gene pLC30-15 were analyzed in wild tomato populations from contrasting environments. For S. chilense, we found that Asr4 and pLC30-15 appear to respond much faster to drought conditions in accessions from very dry environments than accessions from more mesic locations. Sequence analysis suggests that the promoter of Asr2 and the downstream region of pLC30-15 are under positive selection in some local populations of S. chilense. By investigating gene expression differences at the population level we provide further support of our previous conclusions that Asr2, Asr4, and pLC30-15 are promising candidates for functional studies of adaptation. Our analysis also demonstrates the power of the candidate gene approach in evolutionary biology research and highlights the importance of wild Solanum species as a genetic resource for their cultivated relatives.     

Genetics,Genomics and Breeding of Late Blight and Early Blight Resistance in Tomato

Late blight (LB), caused by the oomycete Phytophthora infestans, and early blight (EB), caused by the fungi Alternaria solani and A. tomatophila, are two common and destructive foliar diseases of the cultivated tomato (Solanum lycopersicum) and potato (Solanum tuberosum) in the United States and elsewhere in the world. While LB can infect and devastate tomato plants at any developmental stages, EB infection is usually associated with plant physiological maturity and fruit load where older senescing plants exhibit greater susceptibility and a heavy fruit set enhances the disease. At present, cultural practices and heavy use of fungicides are the most common measures for controlling LB and EB. Genetic resources for resistance have been identified for both diseases, largely within the tomato wild species, in particular the red-fruited species S. pimpinellifolium and the green-fruited species S. habrochaites. A few race-specific major resistance genes (e.g., Ph-1, Ph-2 and Ph-3) and several race-nonspecific resistance QTLs have been reported for LB. Ph-3 is a strong resistance gene and has been incorporated into many breeding lines of fresh market and processing tomato. However, new P. infestans isolates have been identified which overcome Ph-3 resistance. Recently, a new resistance gene (Ph-5) has been identified, which confers resistance to several pathogen isolates including those overcoming the previous resistance genes. Advanced breeding lines including Ph-5 alone and in combinations with Ph-2 and Ph-3 are being developed. Genetic controls of EB resistance have been studied and advanced breeding lines and cultivars with improved resistance have been developed through traditional breeding. Additionally, QTLs for EB resistance have been identified, which can be utilized for marker-assisted resistance breeding. Currently, new inbred lines and cultivars of tomato with good levels of EB resistance and competitive yield performance are being developed at the Pennsylvania State University. This review will focus on the current knowledge of both LB and EB with respect to the causal pathogens, host resistances, and genetics and breeding progresses.     

Comparative next-generation mapping of the Phytophthora infestans resistance gene Rpi-dlc2 in a European accession of Solanum dulcamara

Phytophthora infestans, the causal agent of late blight, remains the main threat to potato production worldwide. Screening of 19 accessions of Solanum dulcamara with P. infestans isolate Ipo82001 in detached leaf assays revealed strong resistance in an individual belonging to accession A54750069-1. This plant was crossed with a susceptible genotype, and an F₁ population consisting of 63 individuals was obtained. This population segregated for resistance in 1:1 ratio, both in detached leaf assays and in an open-field experiment. Presence of the formerly mapped Rpi-dlc1 gene as the cause of the observed segregating resistance could be excluded. Subsequently, AFLP analyses using 128 primer combinations enabled identification of five markers linked to a novel resistance gene named Rpi-dlc2. AFLP markers did not show sequence similarity to the tomato and potato genomes, hampering comparative genetic positioning of the gene. For this reason we used next-generation mapping (NGM), an approach that exploits direct sequencing of DNA (in our case: cDNA) pools from bulked segregants to calculate the genetic distance between SNPs and the locus of interest. Plotting of these genetic distances on the tomato and potato genetic map and subsequent PCR-based marker analysis positioned the gene on chromosome 10, in a region overlapping with the Rpi-ber/ber1 and -ber2 loci from S. berthaultii. Pyramiding of Rpi-dlc2 and Rpi-dlc1 significantly increased resistance to P. infestans, compared with individuals containing only one of the genes, showing the usefulness of this strategy to enhance resistance against Phytophthora.     

Mapping and introgression of a tomato yellow leaf curl virus tolerance gene,TY-1

The whitefly-transmitted tomato yellow-leaf curl gemini-virus (TYLCV) is a major pathogen of tomatoes. The wild tomato species Lycopersicon chilense, which is resistant to the virus, was crossed to the cultivated tomato, L. esculentum. The backcross-1 selfed (BC1S1) generation was inoculated and a symptomless plant was selected. This plant was analyzed using 61 molecular markers, which span the tomato genome, to determine which L. chilense chromosome segments were introgressed. A BC2S1 population was cage-inoculated with viroliferous whiteflies (Bemisia tabaci), the natural insect vector of the virus, and subjected to RFLP analysis. Markers on chromosomes 3 and 6 were significantly associated with the level of tolerance; the association of chromosome-6 markers was further substantiated in two additional BC2S1 populations. A tolerant BC2S1 plant which was homozygous for L. chilense introgressions in chromosomes 3, 6 and 7 was crossed to generate a BC3S1 population which was planted in an infested field. A TYLCV-tolerance gene with partial dominance, TY-1, was mapped to chromosome 6; two modifier genes were mapped to chromosomes 3 and 7. Field and whitefly-mediated cage inoculations of nearly-isogenic lines in BC3S3 supported our conclusion that TY-1 is the major TYLCV-tolerance locus.     

Evidence for the Clustering of Antibacterial Resistance Phenotypes of Enterococci Within Integrated Poultry Companies

From July to December 2006, a panel of 401 enterococci was isolated from carcass rinse samples collected in five poultry processing plants in New Zealand. Agar diffusion assays for nine antibacterial drugs were used to obtain a resistance phenotype for each isolate. Hierarchical clustering techniques and diversity indices showed a high diversity of resistance phenotypes within each plant, with populations of Enterococcus faecalis showing greater heterogeneity than Enterococcus faecium. Bayesian modelling identified three clusters of phenotype patterns within the panel: the E. faecium isolates showed a high probability of containing two distinct clusters, whilst the E. faecalis isolates all grouped together to form the third cluster. The validity of these three clusters was examined using pairwise fixation indices and analysis of variance. Comparing the three clusters to the structure of the participating companies showed that resistance phenotypes for E. faecium isolated from processing plants that were geographically separated but were operated by the same integrated poultry company were more similar than E. faecium isolated from unconnected companies. Company-level management factors, such as the routine use of antibacterial drugs and the genetic line of birds reared, mirrored the structure of these clusters, thus indicating that company-level factors were the dominant selective pressures upon resistance phenotypes across all operating units within these integrated poultry companies.     

Genotyping selection for resistance against tomato yellow leaf curl virus (TYLCV) conferred by Ty-1 and Ty-3 genes in tomato

The tomato yellow leaf curl virus (TYLCV), transmitted by whitefly, causes major disease losses to tomato crops in tropical and subtropical regions of the world. Several genes conferring resistance to TYLCV, mainly Ty-1 and Ty-3 genes, have been introgressed to cultivated tomato (Solanum lycopersicum) from the wild relative species Solanum chilense. By combining bulked segregant analysis and amplified fragment length polymorphisms (AFLP), several AFLP markers closely linked to Ty-1 and Ty-3 genes were identified from the resistant breeding line TZ841-4. Cloning and sequencing of the selected AFLP fragments allowed us to develop codominant cleaved amplified polymorphic sequence and dominant sequence characterized amplified region markers closely linked to Ty-1. In addition, Ty-3-linked allelic-specific markers have been discriminated by a quantitative real-time PCR protocol. Taken together, these markers constitute useful tools for marker-assisted selection breeding programs to improve genetic resistance to TYLCV, and also to initiate map-based cloning approaches to isolate the resistance genes.     

Tolerance and overcompensation to infection by Phytophthora infestans in the wild perennial climber Solanum dulcamara

Studies of infection by Phytophthora infestans—the causal agent of potato late blight—in wild species can provide novel insights into plant defense responses, and indicate how wild plants might be influenced by recurrent epidemics in agricultural fields. In the present study, our aim was to investigate if different clones of Solanum dulcamara (a relative of potato) collected in the wild differ in resistance and tolerance to infection by a common European isolate of P. infestans. We performed infection experiments with six S. dulcamara genotypes (clones) both in the laboratory and in the field and measured the degree of infection and plant performance traits. In the laboratory, the six evaluated genotypes varied from resistant to susceptible, as measured by degree of infection 20 days post infection. Two of the four genotypes susceptible to infection showed a quadratic (concave downward) relationship between the degree of infection and shoot length, with maximum shoot length at intermediate values of infection. This result suggests overcompensation, that is, an increase in growth in infected individuals. The number of leaves decreased with increasing degree of infection, but at different rates in the four susceptible genotypes, indicating genetic variation for tolerance. In the field, the inoculated genotypes did not show any disease symptoms, but plant biomass at the end of the growing season was higher for inoculated plants than for controls, in‐line with the overcompensation detected in the laboratory. We conclude that in S. dulcamara there are indications of genetic variation for both resistance and tolerance to P. infestans infection. Moreover, some genotypes displayed overcompensation. Learning about plant tolerance and overcompensation to infection by pathogens can help broaden our understanding of plant defense in natural populations and help develop more sustainable plant protection strategies for economically important crop diseases.