Dahliae Race Research Articles

Tissues and mechanisms associated with Verticillium wilt resistance in tomato using bi-grafted near-isogenic lines.

Host resistance is the primary means to control Verticillium dahliae, a soil-borne pathogen causing major losses on a broad range of plants, including tomato. The tissues and mechanisms responsible for resistance remain obscure. In the field, resistant tomato used as rootstocks does not confer resistance. Here, we created bi-grafted plants with near-isogenic lines (NILs) exhibiting (Ve1) or lacking (ve1) resistance to V. dahliae race 1. Ten days after inoculation, scion and rootstock tissues were subjected to differential gene expression and co-expression network analyses. Symptoms only developed in susceptible scions regardless of the rootstock. Infection caused more dramatic alteration of tomato gene expression in susceptible compared to resistant tissues, including pathogen receptor, signaling pathway, PR protein, and cell wall modification genes. Differences were observed between scions and rootstocks, primarily related to physiological processes in these tissues. Gene expression in scions was influenced by the rootstock genotype. A few genes were associated with the Ve1 genotype, which was independent of infection or tissue type. Several were physically clustered, some near the Ve1 locus on chromosome 9. Transcripts mapped to V. dahliae were dominated by secreted candidate effector proteins. These findings advance knowledge of molecular mechanisms underlying the tomato-V. dahliae interaction.

Mapping Quantitative Trait Loci for Lettuce Resistance to Verticillium dahliae Race 3, Plant Development, and Leaf Color Using an Ultra-High-Density Bin Map Constructed from F2 Progeny

Verticillium wilt is one of the most devastating soilborne diseases in lettuce, and the use of host resistance is the most optimal choice for its management. This study focused on identifying and mapping the genetic loci for resistance against Verticillium dahliae race 3 in a mapping population of 200 F2:3 families developed from a cross between moderately resistant red-leaf lettuce ‘Sentry’ and susceptible green-leaf lettuce ‘La Brillante’. The population was genotyped using the tunable genotyping-by-sequencing (tGBS) approach. An ultra-high-density genetic linkage map containing 34,838 single nucleotide polymorphism markers grouped into 1,734 bins was constructed using F2 progeny and a sliding window approach. Three quantitative trait loci (QTLs) for resistance to V. dahliae race 3 were located on linkage groups (LGs) LG 2 ( qVR3-2.1) and LG 4 ( qVR3-4.1 and qVR3-4.2). Each of these QTLs explained up to ∼10% of the total phenotypic variation for the trait. At each locus, the resistance alleles were derived from cultivar Sentry that is partially resistant to the pathogen. Additional loci resistant to the disease are expected in this population, and transgressive segregation indicates that some of those loci could originate from the susceptible cultivar La Brillante. In addition, two QTLs for plant development were identified on LG 2 ( qIPD-2.1) and LG 7 ( qIPD-7.1), although no relationship was detected between resistance in these genotypes and the rate of plant growth. A major effect of QTL for red leaf color was detected on LG 9 ( qRLC-9.1). Candidate genes linked to some of the QTLs for V. dahliae race 3 resistance, plant development, and leaf color were identified. The QTLs for resistance identified in Sentry could diversify the resistance gene pool and provide an alternative tool to manage a newly emerged V. dahliae race 3. [Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Dynamics of Verticillium dahliae race 1 population under managed agricultural ecosystems

BackgroundPlant pathogens and their hosts undergo adaptive changes in managed agricultural ecosystems, by overcoming host resistance, but the underlying genetic adaptations are difficult to determine in natural settings. Verticillium dahliae is a fungal pathogen that causes Verticillium wilt on many economically important crops including lettuce. We assessed the dynamics of changes in the V. dahliae genome under selection in a long-term field experiment.ResultsIn this study, a field was fumigated before the Verticillium dahliae race 1 strain (VdLs.16) was introduced. A derivative 145-strain population was collected over a 6-year period from this field in which a seggregating population of lettuce derived from Vr1/vr1 parents were evaluated. We de novo sequenced the parental genome of VdLs.16 strain and resequenced the derivative strains to analyze the genetic variations that accumulate over time in the field cropped with lettuce. Population genomics analyses identified 2769 single-nucleotide polymorphisms (SNPs) and 750 insertion/deletions (In-Dels) in the 145 isolates compared with the parental genome. Sequence divergence was identified in the coding sequence regions of 378 genes and in the putative promoter regions of 604 genes. Five-hundred and nine SNPs/In-Dels were identified as fixed. The SNPs and In-Dels were significantly enriched in the transposon-rich, gene-sparse regions, and in those genes with functional roles in signaling and transcriptional regulation.ConclusionsUnder the managed ecosystem continuously cropped to lettuce, the local adaptation of V. dahliae evolves at a whole genome scale to accumulate SNPs/In-Dels nonrandomly in hypervariable regions that encode components of signal transduction and transcriptional regulation.

Genetics of Partial Resistance Against Verticillium dahliae Race 2 in Wild and Cultivated Lettuce.

Lettuce (Lactuca sativa) is one of the most economically important vegetables in the United States, with approximately 50% of the domestic production concentrated in the Salinas Valley of California. Verticillium wilt, caused by races 1 and 2 of the fungal pathogen Verticillium dahliae, poses a major threat to lettuce production in this area. Although resistance governed by a single dominant gene against race 1 has previously been identified and is currently being incorporated into commercial cultivars, identification of resistance against race 2 has been challenging and no lines with complete resistance have been identified. In this study, we screened germplasm for resistance and investigated the genetics of partial resistance against race 2 using three mapping populations derived from crosses involving L. sativa × L. sativa and L. serriola × L. sativa. The inheritance of resistance in Lactuca species against race 2 is complex but a common quantitative trait locus (QTL) on linkage group 6, designated qVERT6.1 (quantitative Verticillium dahliae resistance on LG 6, first QTL), was detected in multiple populations. Additional race 2 resistance QTLs located in several linkage groups were detected in individual populations and environments. Because resistance in lettuce against race 2 is polygenic with a large genotype by environment interaction, breeding programs to incorporate these resistance genes should be aware of this complexity as they implement strategies to control race 2.

First report of Verticillium dahliae race 2 on scarlet eggplant in Brazil

First report of Verticillium dahliae race 2 on scarlet eggplant in Brazil

Identification of Tomato Ve1 Homologous Proteins in Flax and Assessment for Race-Specific Resistance in Two Fiber FlaxCultivars against Verticillium dahliae Race 1.

In the last decade, the soil borne fungal pathogen Verticillium dahliae has had an increasingly strong effect on fiber flax (Linum usitatissimum L.), thus causing important yield losses in Normandy, France. Race-specific resistance against V. dahliae race 1 is determined by tomato Ve1, a leucine-rich repeat (LRR) receptor-like protein (RLP). Furthermore, homologous proteins have been found in various plant families. Herein, four homologs of tomato Ve1 were identified in the flax proteome database. The selected proteins were named LuVe11, LuVe12, LuVe13 and LuVe14 and were compared to other Ve1. Sequence alignments and phylogenic analysis were conducted and detected a high similarity in the content of amino acids and that of the Verticillium spp. race 1 resistance protein cluster. Annotations on the primary structure of these homologs reveal several features of tomato Ve1, including numerous copies of a 28 amino acids consensus motif [XXIXNLXXLXXLXLSXNXLSGXIP] in the LRR domain. An in vivo assay was performed using V. dahliae race 1 on susceptible and tolerant fiber flax cultivars. Despite the presence of homologous genes and the stronger expression of LuVe11 compared to controls, both cultivars exhibited symptoms and the pathogen was observed within the stem. Amino acid substitutions within the segments of the LRR domain could likely affect the ligand binding and thus the race-specific resistance. The results of this study indicate that complex approaches including pathogenicity tests, microscopic observations and gene expression should be implemented for assessing race-specific resistance mediated by Ve1 within the large collection of flax genotypes.

The LsVe1L allele provides a molecular marker for resistance to Verticillium dahliae race 1 in lettuce.

BackgroundVerticillium wilt caused by the fungus Verticillium dahliae race 1 is among the top disease concerns for lettuce in the Salinas and Pajaro Valleys of coastal central California. Resistance of lettuce against V. dahliae race 1 was previously mapped to the single dominant Verticillium resistance 1 (Vr1) locus. Lines of tomato resistant to race 1 are known to contain the closely linked Ve1 and Ve2 genes that encode receptor-like proteins with extracellular leucine-rich repeats; the Ve1 and Ve2 proteins act antagonistically to provide resistance against V. dahliae race 1. The Vr1 locus in lettuce contains a cluster of several genes with sequence similarity to the tomato Ve genes. We used genome sequencing and/or PCR screening along with pathogenicity assays of 152 accessions of lettuce to investigate allelic diversity and its relationship to race 1 resistance in lettuce.ResultsThis approach identified a total of four Ve genes: LsVe1, LsVe2, LsVe3, and LsVe4. The majority of accessions, however, contained a combination of only three of these LsVe genes clustered on chromosomal linkage group 9 (within ~ 25 kb in the resistant cultivar La Brillante and within ~ 127 kb in the susceptible cultivar Salinas).ConclusionsA single allele, LsVe1L, was present in all resistant accessions and absent in all susceptible accessions. This allele can be used as a molecular marker for V. dahliae race 1 resistance in lettuce. A PCR assay for rapid detection of race 1 resistance in lettuce was designed based on nucleotide polymorphisms. Application of this assay allows identification of resistant genotypes in early stages of plant development or at seed-level without time- and labor-intensive testing in the field.

The island cotton NBS-LRR gene GbaNA1 confers resistance to the non-race 1 Verticillium dahliae isolate Vd991.

Wilt caused by Verticillium dahliae significantly reduces cotton yields, as host resistance in commercially cultivated Gossypium species is lacking. Understanding the molecular basis of disease resistance in non-commercial Gossypium species could galvanize the development of Verticillium wilt resistance in cultivated species. Nucleotide-binding site leucine-rich repeat (NBS-LRR) proteins play a central role in plant defence against pathogens. In this study, we focused on the relationship between a locus enriched with eight NBS-LRR genes and Verticillium wilt resistance in G. barbadense. Independent virus-induced gene silencing of each of the eight NBS-LRR genes in G. barbadense cultivar Hai 7124 revealed that silencing of GbaNA1 alone compromised the resistance of G. barbadense to V. dahliae isolate Vd991. In cultivar Hai 7124, GbaNA1 could be induced by V. dahliae isolate Vd991 and by ethylene, jasmonic acid and salicylic acid. Nuclear protein localization of GbaNA1 was demonstrated by transient expression. Sequencing of the GbaNA1 orthologue in nine G.hirsutum accessions revealed that all carried a non-functional allele, caused by a premature peptide truncation. In addition, all 10 G. barbadense and nine G. hirsutum accessions tested carried a full-length (∼1140 amino acids) homologue of the V. dahliae race 1 resistance gene Gbve1, although some sequence polymorphisms were observed. Verticillium dahliae Vd991 is a non-race 1 isolate that lacks the Ave1 gene. Thus, the resistance imparted by GbaNA1 appears to be mediated by a mechanism distinct from recognition of the fungal effector Ave1.

Short-Term Host Selection Pressure Has Little Effect on the Evolution of a Monoclonal Population of Verticillium dahliae Race 1.

Understanding pathogen evolution over time is vital for plant breeding and deployment of host resistance. In the context of a soilborne pathogen, the potential of host-directed evolution of a Verticillium dahliae race 1 isolate and genotypic variation of V. dahliae associated with two major hosts (lettuce and tomato) were determined. In total, 427 isolates were recovered over 6 years from a resistance screening nursery infested with a single V. dahliae race 1 isolate. In a separate study, an additional 206 isolates representing 163 and 43 isolates from commercial lettuce and tomato fields, respectively, were collected. Analyses of isolates recovered from the screening nursery over 6 years revealed no changes in the race and mating type composition but did uncover seven simple sequence repeat (SSR) variant genotypes. No significant genotypic variation in V. dahliae was observed between or within fields of either lettuce or tomato but pathogen populations were significantly differentiated between these two hosts. Replicated virulence assays of variant SSR genotypes on lettuce differential cultivars suggested no significant difference in virulence from the wild-type race 1 isolate introduced into the field. This suggests that deployed race 1 host resistance will be robust against the widespread race 1 populations in lettuce-growing regions at least for 6 years unless novel pathogen genotypes or races are introduced into the system.

Genetics of resistance in lettuce to races 1 and 2 of Verticillium dahliae from different host species

Race 1 resistance against Verticillium dahliae in lettuce was originally shown in the cultivar La Brillante to be conditioned by a single dominant gene (Verticillium resistance 1, Vr1). Multiple, morphologically diverse sources of germplasm have been identified as resistant to race 1. In this study, allelism tests indicated that resistance in these different lettuce cultivars is closely linked or allelic to the Vr1 gene. The Vr1 gene is defeated by race 2 isolates of V. dahliae. Only partial resistance to race 2 isolates is available in a few plant introductions (PIs). Greenhouse and field experiments conducted with these PIs demonstrated partial resistance to V. dahliae race 1 as well as race 2 isolates from lettuce. Cultivars resistant to race 1 and PIs with partial resistance to race 2 were challenged with several race 1 and 2 isolates originating from hosts other than lettuce. This indicated that cultivars resistant to race 1 and the breeding lines derived from them would also be resistant to race 1 isolates from other hosts; similarly, the partial resistance would be effective against race 1 and 2 isolates from hosts other than lettuce. Nevertheless, there were specific interactions that warrant further study. Although race 1 currently predominates in the major lettuce production area of the Salinas Valley, CA, breeding lettuce for resistance to V. dahliae should take both races into account.

Variation of pathotypes and races and their correlations with clonal lineages in Verticillium dahliae

Understanding pathogenic variation in plant pathogen populations is key for the development and use of host resistance for managing verticillium wilt diseases. A highly virulent defoliating (D) pathotype in Verticillium dahliae has previously been shown to occur only in one clonal lineage (lineage 1A). By contrast, no clear association has yet been shown for race 1 with clonal lineages. Race 1 carries the effector gene Ave1 and is avirulent on hosts that carry resistance gene Ve1 or its homologues. The hypothesis tested was that race 1 arose once in a single clonal lineage, which might be expected if V. dahliae acquired Ave1 by horizontal gene transfer from plants, as hypothesized previously. In a diverse sample of 195 V. dahliae isolates from nine clonal lineages, all race 1 isolates were present only in lineage 2A. Conversely, all lineage 2A isolates displayed the race 1 phenotype. Moreover, 900‐bp nucleotide sequences from Ave1 were identical among 27 lineage 2A isolates and identical to sequences from other V. dahliae race 1 isolates in GenBank. The finding of race 1 in a single clonal lineage, with identical Ave1 sequences, is consistent with the hypothesis that race 1 arose once in V. dahliae. Molecular markers and virulence assays also confirmed the well‐established finding that the D pathotype is found only in lineage 1A. Pathogenicity assays indicated that cotton and olive isolates of the D pathotype (lineage 1A) were highly virulent on cotton and olive, but had low virulence on tomato.

Broad taxonomic characterization of Verticillium wilt resistance genes reveals an ancient origin of the tomato Ve1 immune receptor.

Plant-pathogenic microbes secrete effector molecules to establish themselves on their hosts, whereas plants use immune receptors to try and intercept such effectors in order to prevent pathogen colonization. The tomato cell surface-localized receptor Ve1 confers race-specific resistance against race 1 strains of the soil-borne vascular wilt fungus Verticillium dahliae which secrete the Ave1 effector. Here, we describe the cloning and characterization of Ve1 homologues from tobacco (Nicotiana glutinosa), potato (Solanum tuberosum), wild eggplant (Solanum torvum) and hop (Humulus lupulus), and demonstrate that particular Ve1 homologues govern resistance against V. dahliae race 1 strains through the recognition of the Ave1 effector. Phylogenetic analysis shows that Ve1 homologues are widely distributed in land plants. Thus, our study suggests an ancient origin of the Ve1 immune receptor in the plant kingdom.

Host Range of Verticillium isaacii and Verticillium klebahnii from Artichoke, Spinach, and Lettuce.

Verticillium is a genus that includes major vascular wilt pathogens. Recently, multilocus phylogenetic analyses of the genus identified five new species, including Verticillium isaacii and V. klebahnii, both of which occur in agricultural soils in coastal California and have been isolated from asymptomatic and diseased spinach and lettuce plants. Little data are available regarding their pathogenicity and virulence on a broader range of crops important to the region. Four isolates each of V. isaacii and V. klebahnii along with two reference isolates of V. dahliae races 1 and 2 were inoculated on eight crops (artichoke, cauliflower, eggplant, lettuce, pepper, tomato, spinach, and strawberry) in a greenhouse experiment. After 8 weeks, plants were assessed for disease severity to determine the relative host ranges of Verticillium isolates. Additionally, 13 lettuce lines resistant to race 1 and partially resistant to race 2 of V. dahliae were screened against V. isaacii and V. klebahnii to evaluate their responses. Three of four V. isaacii and four of four V. klebahnii isolates tested were nonpathogenic on all crops tested except those indicated below. One V. isaacii isolate caused wilt on artichoke and 'Salinas' lettuce and most isolates of both species caused varying degrees of Verticillium wilt on strawberry. Lettuce lines resistant to V. dahliae race 1 and partially resistant to V. dahliae race 2 also exhibited resistance to all of the isolates of V. isaacii and V. klebahnii. Thus, at least some isolates in the populations of V. isaacii and V. klebahnii have the potential to become significant pathogens of coastal California crops. However, resistance developed against V. dahliae also offers resistance to the pathogenic isolates of both species, at least in lettuce.

Fifteen Years of Verticillium Wilt of Lettuce in America's Salad Bowl: A Tale of Immigration, Subjugation, and Abatement.

Fifteen Years of Verticillium Wilt of Lettuce in America's Salad Bowl: A Tale of Immigration, Subjugation, and Abatement.

The inheritance of resistance to Verticillium wilt caused by race 1 isolates of Verticillium dahliae in the lettuce cultivar La Brillante.

Verticillium wilt of lettuce caused by Verticillium dahliae can cause severe economic damage to lettuce producers. Complete resistance to race 1 isolates is available in Lactuca sativa cultivar (cv.) La Brillante and understanding the genetic basis of this resistance will aid development of new resistant cultivars. F(1) and F(2) families from crosses between La Brillante and three iceberg cultivars as well as a recombinant inbred line population derived from L. sativa cv. Salinas 88 × La Brillante were evaluated for disease incidence and disease severity in replicated greenhouse and field experiments. One hundred and six molecular markers were used to generate a genetic map from Salinas 88 × La Brillante and for detection of quantitative trait loci. Segregation was consistent with a single dominant gene of major effect which we are naming Verticillium resistance 1 (Vr1). The gene described large portions of the phenotypic variance (R(2) = 0.49-0.68) and was mapped to linkage group 9 coincident with an expressed sequence tag marker (QGD8I16.yg.ab1) that has sequence similarity with the Ve gene that confers resistance to V. dahliae race 1 in tomato. The simple inheritance of resistance indicates that breeding procedures designed for single genes will be applicable for developing resistant cultivars. QGD8I16.yg.ab1 is a good candidate for functional analysis and development of markers suitable for marker-assisted selection.

Fontes de resistência em acessos de Solanum (secção Lycopersicon) a Verticillium dahliae raças 1 e 2

A murcha-de-verticílio (Verticillium dahliae) é uma das doenças mais severas do tomateiro (Solanum lycopersicum L.). Duas raças fisiológicas de V. dahliae foram caracterizadas infectando o tomateiro e ambas estão presentes no Brasil. A estratégia de controle mais efetiva para isolados da raça 1 tem sido a utilização de cultivares resistentes em associação com práticas culturais. Entretanto, não existem cultivares comerciais de tomate resistentes à raça 2 do patógeno. Neste contexto, torna-se importante a identificação de novas fontes de resistência que sejam efetivas contra as duas raças de V. dahliae. Buscou-se fontes de resistência a isolados de V. dahliae (raças 1 e 2). Cem acessos de uma coleção de germoplasma contendo espécies cultivadas e selvagens de Solanum (secção Lycopersicon) foram inicialmente inoculados pelo método de imersão de raízes com um isolado de V. dahliae raça 1 (5 mL de uma suspensão de 10(6) conídios/mL) em condições de casa de vegetação. A avaliação foi feita aos 30 dias após a inoculação, usando uma escala de notas variando de 1 (planta sadia) a 5 (planta morta). Foram considerados resistentes os acessos que apresentaram nota média entre 1 e 2. Um grupo composto por 38 acessos classificados como resistentes neste primeiro ensaio foi reavaliado para resistência a outros quatro isolados de V. dahliae pertencentes às raças 1 e 2. As cultivares Ponderosa e Floradade foram utilizadas como controles. O ensaio foi conduzido em casa de vegetação em delineamento experimental de blocos ao acaso em arranjo fatorial 5 x 40, com três repetições (vasos com quatro plantas). A avaliação foi feita baseada nos parâmetros epidemiológicos: período de incubação e índice de doença. Foram identificados acessos com resistência raça-específica e também com resistência a ambas as raças. Estes acessos podem ser indicados para futuros programas de melhoramento genético visando incorporar resistência ampla à doença.

First report of Verticillium dahliae race 2 in Tunisia

Vascular wilt diseases have recently become a serious problem on tomato (Lycopersicon esculentum) throughout the Tunisian Sahel. Since 2002, plants of cv. Colibri with symptoms characteristic of Verticillium infection (wilting, yellowing, stunting and dark brown vascular discoloration) have been seen in many greenhouses in the Chott Mariem region. Isolates from diseased plants were identified as Verticillium dahliae on the basis of microsclerotium production (Hawksworth & Talboys, 1970). Cultivar Colibri carries a single dominant gene (Ve) conferring resistance to race 1 of V. dahliae (Schaible et al., 1951). This resistance has been effective for over 20 years. Wilt caused by V. dahliae has also been confirmed in several other resistant cultivars (e.g. Amel, Cenkara, Rio Grande). Fifty-one Tunisian isolates were tested to determine their race, with Canadian and Israeli race 1 and 2 isolates (one each) included as controls. Conidial suspensions of 107 conidia mL−1 were used to inoculate plants of the cvs Ventura and Sun 6200 (susceptible to both races), and Rio Grande, Colibri and Naya (resistant to race 1), at the two-leaf stage by root dipping. Seedlings were replanted into a sterile 2:1 mixture of peat/perlite (v/v) and maintained in a growth chamber at 23 ± 3°C. Equal numbers of plants of each cultivar were dipped into water as controls. The experimental design was a completely randomized type with ten replicates (pots). Tests were conducted twice. Susceptible plants developed typical symptoms of wilt as above, starting two to three weeks after inoculation, followed by plant death. Race 1 isolates gave either no or only very mild symptoms in the resistant cultivars. Control plants showed no symptoms. In cultivars where infection occurred, most plants developed typical symptoms. V. dahliae was recovered by in vitro isolation and incubation from all plants with symptoms. Based on the differential reactions of the five cultivars, 10 isolates were classified as race 1 and 41 as race 2. The appearance of V. dahliae race 2 in tomatoes in Tunisia poses a major threat to this crop, particularly in protected cultivation where no rotations are used and the absence of soil disinfection favours the build-up of soil-borne inoculum. Race 2 has been reported from southern Europe, North and South America and North and South Africa (Pegg & Brady, 2002), but this is the first report from Tunisia. The authors would like to thank Aymen Youssef for his excellent technical assistance.

A method of subtraction hybridization mediated by oligo(dT) particles

A subtraction hybridization procedure using magnetic oligo(dT) particles originally designed for mRNA isolation was developed. Magnetic particles are used to isolate mRNA, prime cDNA synthesis, and eliminate transcripts common to control and experimental samples. The protocol was utilized to isolate 2 tomato cDNAs upregulated during infection withVerticillium dahliae race II.

Mapping of Ve in tomato: a gene conferring resistance to the broad-spectrum pathogen, Verticillium dahliae race 1

The soil-borne fungi Verticillium spp. cause vascular wilt disease in a wide range of crop plants. In tomato, resistance to Verticillium dahliae race 1 is conferred by a single dominant gene, Ve. Previous efforts to map Ve in tomato have yielded confusing results, locating it on different chromosomes, which subsequently raised the possibility that Verticillium resistance may be controlled by a number of loci. We used three different mapping populations to obtain an unambiguous map location of Ve: a recombinant inbred (RI) line population; an F2 population segregating for Verticillium resistance; and a population of 50 introgression lines (IL). In all of the mapping populations Ve was positioned on the short arm of chromosome 9 tightly linked to the RFLP marker GP39. This linkage was confirmed by screening for GP39 in different breeding lines with known resistance or susceptibility to Verticillium. A perfect match was found between GP39 and the Verticillium response of the lines, indicating the potential of GP39 in the rapid detection of Verticillium resistance and as a starting point for map-based cloning of Ve. This approach is particularly relevant for Verticillium dahliae race 1, since in the present work we also show that the isolate that infects tomato is responsible for wilt disease in other important crop plants.

DNA fingerprinting and vegetative compatibility analysis indicate multiple origins for Verticillium dahliae race 2 tomato isolates from Ontario, Canada

RFLP, DNA fingerprinting and VCG methods were used to characterize fourteen Verticillium dahliae isolates collected from southwestern Ontario, Canada. The isolates were typed as not pathogenic to tomato (NP), race 1 (avirulent on cvs carrying the Ve resistance gene) or race 2 (virulent on Ve cvs). On the basis of RFLPs, RAPDs, and DNA fingerprints detected by hybridization to a dispersed, repetitive genomic DNA probe, the isolates were classified into five DNA types. Type I included two NP isolates. Type II included four race 2, and three NP isolates. Types III and V were represented by single race 2 and race 1 isolates, respectively. Type IV included one race 2, and two race 1 isolates. Vegetative compatibility was determined for selected NP, race 1, and race 2 isolates of each race type/DNA type combination. Isolates of the same DNA type were compatible, as were type II and III isolates (VCG 4B), and type IV (VCG 2A) and V isolates (VCG 2B). These data show a level of genetic diversity not previously identified in the V. dahliae tomato pathogen population, and suggest multiple origins for the Ontario race 2 pathotype.