Genetic and physical localization of the root-knot nematode resistance locus Mi in tomato

As part of a map-based cloning strategy designed to isolate the root-knot nematode resistance gene Mi, tomato F2 populations were analyzed in order to identify recombination points close to this economically important gene. A total of 21 089 F2 progeny plants were screened using morphological markers. An additional 1887 F2 were screened using PCR-based flanking markers. Fine-structure mapping of recombinants with newly developed AFLP markers, and RFLP markers derived from physically mapped cosmid subclones, localized Mi to a genomic region of about 550 kb. The low frequency of recombinants indicated that recombination was generally suppressed in these crosses and that crossovers were restricted to particular regions. To circumvent this problem, a population of Lycopersicon peruvianum, the species from which Mi was originally introgressed, that was segregating for resistance was developed. Screening of this population with PCR, RFLP and AFLP markers identified several plants with crossovers near Mi. Recombination frequency was approximately eight-fold higher in the Mi region of the L. peruvianum cross. However, even within the wild species cross, recombination sites were not uniformly distributed in the region. By combining data from the L. esculentum and L. peruvianum recombinant analyses, it was possible to localize Mi to a region of the genome spanning less than 65 kb. Received: 15 July 1997 / Accepted: 1 October 1997     

Genetic and physical localization of the root-knot nematode resistance locus Mi in tomato

As part of a map-based cloning strategy designed to isolate the root-knot nematode resistance gene Mi, tomato F2 populations were analyzed in order to identify recombination points close to this economically important gene. A total of 21?089 F2 progeny plants were screened using morphological markers. An additional 1887 F2 were screened using PCR-based flanking markers. Fine-structure mapping of recombinants with newly developed AFLP markers, and RFLP markers derived from physically mapped cosmid subclones, localized Mi to a genomic region of about 550?kb. The low frequency of recombinants indicated that recombination was generally suppressed in these crosses and that crossovers were restricted to particular regions. To circumvent this problem, a population of Lycopersicon peruvianum, the species from which Mi was originally introgressed, that was segregating for resistance was developed. Screening of this population with PCR, RFLP and AFLP markers identified several plants with crossovers near Mi. Recombination frequency was approximately eight-fold higher in the Mi region of the L. peruvianum cross. However, even within the wild species cross, recombination sites were not uniformly distributed in the region. By combining data from the L.?esculentum and L. peruvianum recombinant analyses, it was possible to localize Mi to a region of the genome spanning less than 65?kb.     

High resolution RFLP map around the root knot nematode resistance gene (Mi) in tomato

Summary In the 1940's the root-knot nematode resistance gene (Mi) was introgressed into the cultivated tomato from the wild species, L. peruvianum, and today it provides the only form of genetic resistance against this pathogen. We report here the construction of a high resolution RFLP map around the Mi gene that may aid in the future cloning of this gene via chromosome walking. The map covers the most distal nine map units of chromosome 6 and contains the Mi gene, nine RFLP markers, and one isozyme marker (Aps-1). Based on the analysis of more than 1,000 F₂ plants from four crosses, we were able to pinpoint the Mi gene to the interval between two of these markers — GP79 and Aps-1. In crosses containing the Mi gene, this interval is suppressed in recombination and is estimated to be 0.4 cM in length. In contrast, for a cross not containing Mi, the estimated map distance is approximately 5 times greater (ca. 2 cM).Using RFLP markers around Mi as probes, it was possible to classify nematode resistant tomato varieties into three types based on the amount of linked peruvianum DNA still present. Two of these types (representing the majority of the varieties tested) were found to still contain more than 5 cM of peruvianum chromosome — a result that may explain some of the negative effects (e.g. fruit cracking) associated with nematode resistance. The third type (represented by a single variety) is predicted to carry a very small segment of peruvianum DNA (<2 cM) and may be useful in the identification of additional markers close to Mi and in the orientation of clones during a chromosome walk to clone the gene.     

Tomato chromosome 1: high resolution genetic and physical mapping of the short arm in an interspecific Lycopersicon esculentum × L. peruvianum cross

 A detailed map of part of the short arm of chromosome 1 proximal to the Cf-4/Cf-9 gene cluster was generated by using an F₂ population of 314 plants obtained from the cross between the remotely related species Lycopersicon esculentum and L. peruvianum. Six markers that cosegregate in an L. esculentum×L. pennellii F₂ population showed high recombination frequencies in the present interspecific population, spanning an interval of approximately 13 cM. Physical distances between RFLP markers were estimated by pulsed field gel electrophoresis of high-molecular-weight DNA and by identifying YACs that recognized more than one RFLP marker. In this region 1 cM corresponded to 55–110 kb. In comparsion with the value of 730 kb per cM averaged over the entire genome, this reflects the remarkably high recombination frequencies in this region in the hybrid L. esculentum×L. peruvianum progeny population. The present data underline the fact that recombination is not a process that occurs randomly over the entire genome, but can vary dramatically in intensity between chromosomal regions and among populations. Received: 20 May 1996 / Accepted: 10 September 1996     

The nematode-resistance gene,<Emphasis Type="Italic"> Mi-1</Emphasis>, is associated with an inverted chromosomal segment in susceptible compared to resistant tomato

The gene Mi-1 confers effective resistance in tomato (Lycopersicon esculentum) against root-knot nematodes and some isolates of potato aphid. This locus was introgressed from L. peruvianum into the corresponding region on chromosome 6 in tomato. In nematode-resistant tomato, Mi-1 and six homologs are grouped into two clusters separated by 300 kb. Analysis of BAC clones revealed that the Mi-1 locus from susceptible tomato carried the same number and distribution of Mi-1 homologs, as did the resistant locus. Molecular markers flanking the resistant and susceptible loci were in the same relative orientation, but markers between the two clusters were in an inverse orientation. The simplest explanation for these observations is that there is an inversion between the two clusters of homologs when comparing the Mi-1 loci from L. esculentum and L. peruvianum. Such an inversion may explain previous observations of severe recombination suppression in the region. Two Mi-1 homologs identified from the BAC library derived from susceptible tomato are not linked to the chromosome 6 locus, but map to chromosome 5 in regions known to contain resistance gene loci in other solanaceous species.Communicated by J.S. Heslop-Harrison     

RFLP markers linked to the root knot nematode resistance gene Mi in tomato

Summary The Mi gene originating from the wild tomato species Lycopersicon peruvianum confers resistance to all major root knot nematodes (Meloidogyne spp.). This single dominant gene is located on chromosome 6 and is very closely linked to the acid phosphatase-1 (Aps-1) locus. Resistance to nematodes has been introgressed into various cultivars of the cultivated tomato (L. esculentum), in many cultivars along with the linked L. peruvianum Aps-1 ¹ allele. By using a pair of nearly isogenic lines differing in a small chromosomal region containing the Mi and Aps-1 loci, we have identified two RFLP markers, GP79 and H6A2c2, which are located in the introgressed L. peruvianum region. Analysis of a test panel of 51 L. esculentum genotypes of various origins indicated that GP79 is very tightly linked to the Mi gene and allows both homozygous and heterozygous nematode-resistant genotypes to be distinguished from susceptible genotypes, irrespective of their Aps-1 alleles. Marker H6A2c2 is linked to the Aps-1 locus and is capable of discriminating between the L. peruvianum Aps-1 ¹ allele and the L. esculentum Aps-1 ³ and Aps-1 ⁺ alleles. In combination, these RFLP markers may provide a powerful tool in breeding tomatoes for nematode resistance.     

Mapping a new nematode resistance locus in Lycopersicon peruvianum

Accessions of the wild tomato species L. peruvianum were screened with a root-knot nematode population (557R) which infects tomato plants carrying the nematode resistance gene Mi. Several accessions were found to carry resistance to 557R. A L. peruvianum backcross population segregating for resistance to 557R was produced. The segregation ratio of resistant to susceptible plants suggested that a single, dominant gene was a major factor in the new resistance. This gene, which we have designated Mi-3, confers resistance against nematode strains that can infect plants carrying Mi. Mi-3, or a closely linked gene, also confers resistance to nematodes at 32°C, a temperature at which Mi is not effective. Bulked-segregant analysis with resistant and susceptible DNA pools was employed to identify RAPD markers linked to this gene. Five-hundred-and-twenty oligonucleotide primers were screened and two markers linked to the new resistance gene were identified. One of the linked markers (NR14) was mapped to chromosome 12 of tomato in an L. esculentum/L. pennellii mapping population. Linkage of NR14 and Mi-3 with RFLP markers known to map on the short arm of chromosome 12 was confirmed by Southern analysis in the population segregating for Mi-3. We have positioned Mi-3 near RFLP marker TG180 which maps to the telomeric region of the short arm of chromosome 12 in tomato.     

Resistance in Lycopersicon peruvianum to Isolates of Mi Gene-Compatible Meloidogyne Populations

Root-knot nematode resistance of F₁ progeny of an intraspecific hybrid (Lycopersicon peruvianum var. glandulosum Acc. No. 126443 x L. peruvianum Acc. No. 270435), L. esculentum cv. Piersol (possessing resistance gene Mi), and L. esculentum cv. St. Pierre (susceptible) was compared. Resistance to 1) isolates of two Meloidogyne incognita populations artificially selected for parasitism on tomato plants possessing the Mi gene, 2) the wild type parent populations, 3) four naturally occurring resistance (Mi gene)-breaking populations of M. incognita, M. arenaria, and two undesignated Meloidogyne spp., and 4) a population of M. hapla was indexed by numbers of egg masses produced on root systems in a greenhouse experiment. Artificially selected M. incognita isolates reproduced abundantly on Piersol, but not (P = 0.01) on resistant F₁ hybrids. Thus, the gene(s) for resistance in the F₁ hybrid differs from the Mi gene in Piersol. Four naturally occurring resistance-breaking populations reproduced extensively on Piersol and on the F₁ hybrid, demonstrating ability to circumvent both types of resistance. Meloidogyne hapla reproduced on F₁ hybrid plants, but at significantly (P = 0.01) lower levels than on Piersol.     

Introgression and DNA marker analysis of Lycopersicon peruvianum, a wild relative of the cultivated tomato, into Lycopersicon esculentum, followed through three successive backcross generations

 Segregation of the Lycopersicon peruvianum genome was followed through three generations of backcrossing to the cultivated tomato L. esculentum cv ‘E6203’ using molecular markers. Thirteen BC₁ plants were genotyped with 113 markers, 67 BC₂ plants with 84 markers, and finally 241 BC₃ plants were genotyped with 177 markers covering the entire genome and a BC₃ map constructed. Several segments of the genome, including parts of chromosomes 3, 4, 6, and 10, quickly became fixed for esculentum alleles, possibly due to sterility problems encountered in the BC₁. Observed overall heterozygosity and chromosome segment lengths at each generation were very near the expected theoretical values. Markers located near the top telomeric region of chromosome 9 showed segregation highly skewed towards the wild allele through all generations, suggesting the presence of a gamete promoter gene. One markers, TG9, mapped to a new position on chromosome 9, implying an intrachromosomal translocation event. Despite the great genetic distance between the two parents, overall recombination was only 25% less than that observed in a previous tomato cross, indicating that L. peruvianum genes may be more readily introgressed into cultivated germplasm than originally believed. Received: 9 April 1997 / Accepted : 20 May 1997     

Tomato chromosome 1: high resolution genetic and physical mapping of the short arm in an interspecific Lycopersicon esculentum×L. peruvianum cross

A detailed map of part of the short arm of chromosome 1 proximal to the Cf-4/Cf-9 gene cluster was generated by using an F₂ population of 314 plants obtained from the cross between the remotely related species Lycopersicon esculentum and L. peruvianum. Six markers that cosegregate in an L. esculentum×L. pennellii F₂ population showed high recombination frequencies in the present interspecific population, spanning an interval of approximately 13?cM. Physical distances between RFLP markers were estimated by pulsed field gel electrophoresis of high-molecular-weight DNA and by identifying YACs that recognized more than one RFLP marker. In this region 1?cM corresponded to 55–110?kb. In comparsion with the value of 730?kb per cM averaged over the entire genome, this reflects the remarkably high recombination frequencies in this region in the hybrid L. esculentum×L. peruvianum progeny population. The present data underline the fact that recombination is not a process that occurs randomly over the entire genome, but can vary dramatically in intensity between chromosomal regions and among populations.     

Unilateral incongruity in crosses involvingLycopersicon pennellii andL. esculentum is distinct from self-incompatibility in expression,timing and location

Both interspecific and intraspecific mechanisms restrict the exchange of genes between plants. Much research has focused on self incompatibility (SI), an intraspecific barrier, but research on interspecific barriers lags behind. We are using crosses betweenLycopersicon esculentum andL. pennellii as a model with which to study interspecific crossing barriers. The crossL. esculentum×L. pennellii is successful, but the reciprocal cross fails. Since the cross can be successfully made in one direction but not the other, gross genomic imbalance or chromosomal abnormality are precluded as causes. We showed that the lack of seed set observed in the crossL. pennellii×L. esculentum is due to the inability of pollen tubes to grow more than 2–3 mm into the style, whereas S1 crosses show continued slow pollen tube growth but, also, fail to set seed. These results indicate that the unilateral response is a barrier distinct from SI, differing from SI in the timing and location of expression in the style. We therefore suggest that this unilateral response in theL. pennellii×L. esculentum cross is more accurately referred to as unilateral incongruity (UI) rather than interspecific incompatibility. Periclinal chimeras were used to determine the tissues involved in UI. The results of crosses with the available chimeras indicate that the female parent must beL. pennellii at either LI (layer 1) or both LI and LII (layer 2) and the male parent must beL. esculentum at either LII or both LI and LII to observe UI similar to that seen in theL. pennellii×L. esculentum cross. Pollinations with a mixture of pollen fromL. pennellii and from transgenicL. esculentum plants harboring a pollen-specific GUS reporter gene marker were used to ascertain whether the growth of the pollen tubes of either species was modified as a possible means of overcoming UI. We found no evidence of communication between the two types of pollen tubes to either enhance or restrict all pollen tube growth.     

An RFLP linkage map of Lycopersicon peruvianum

In order to map genes determining resistance to bacterial canker in tomato, backcrosses were made between a resistant and a susceptible Lycopersicon peruvianum accession. The linkage study with RFLP markers yielded a genetic map of L. Peruvianum. This map was compared to that derived from a L. esculentum x L. pennellii F₂ population, based on 70 shared RFLP markers. The maps showed a good resemblance in both the order of markers and the length of the chromosomes, with the exception of just one relocated marker on chromosome 9. Because backcrosses were made with the F₁, either as the pollen parent or as the pistil parent, linkage maps from male and female meioses could be estimated. It was concluded that recombination at male meiosis was reduced, and that gametophytic selection for parental genotypes at more than one locus per chromosome might be partly responsible for the reduction of the estimated male map length.     

The development of bridge lines for interspecific gene transfer between Lycopersicon esculentum and L. peruvianum

Summary Using a modified embryo callus culture technique, hybrids between Lycopersicon esculentum and L. peruvianum were developed and their usefulness as bridge lines for facilitating interspecific gene transfer was evaluated. Four of these lines showed a high level of sexual compatibility with several other L. peruvianum var. typicum accessions, as well as with accessions of L. peruvianum var. humifusum and L. peruvianum var. glandulosum and L. esculentum. These bridge line x L. peruvianum hybrids could be crossed with L. esculentum to introgress genes from L. peruvianum into L. esculentum.     

Homoeologous pairing and recombination in backcross derivatives of tomato somatic hybrids [Lycopersicon esculentum (+) L. peruvianum]

 Genomic in situ hybridization (GISH) was used to examine genome interactions in two allohexa ploid (2n=6x=72) Lycopersicon esculentum (+) L. peruvianum somatic hybrids and their seed progenies originated from subsequent backcrosses to L. esculentum. The ability of GISH to distinguish between chromatin derived from two closely related species, L. esculentum and L. peruvianum (both 2n=2x=24), allowed the precise chromosomal constitution of somatic hybrids and their backcross progenies to be unequivocally established. This enabled the interaction of species genomes to be observed at meiosis, providing clear evidence of strictly regular homoeologous pairing and the high degree of homoeologous recombination in allodiploid plants (2n=2x=24) of the BC₁ generation. In segmental allodiploids of the BC₂ and BC₃ generations, the recombinant chromosomes continued to pair with a homoeologous partner (in the absence of a homologous one), and therefore could be stably incorporated into gametes. Chiasmata were found almost exclusively in more distal, rather subterminal, chromosome segments. A considerable proportion of meiotic recombination was detected in subterminal heterochromatic regions, often involving distal euchromatin, located in close proximity. GISH also supplied information on the extent of the overall sequence homology between the genomes of L. esculentum and L. peruvianum, indicating that despite their different breeding systems, these species may not be differentiated to a high degree genetically. The present study has demonstrated that somatic hybridization between two such closely related, but sexually incompatible or difficult to cross species, provides a way of transferring genes, via homoloeogous crossing-over and recombination, across the incompatibility barriers. Indeed, such hybrids may offer the preferred route for gene transfer, which subsequently results in more stable gene introgression than other methods. Received: 22 July 1996 / Accepted: 23 August 1996     

Differentiation of Meloidogyne incognita and M. arenaria novel resistance phenotypes in Lycopersicon peruvianum and derived bridge-lines

Lycopersicon peruvianum PI 270435 clone 2R2 and PI 126443 clone 1MH were crossed reciprocally with three L. esculentum-L. peruvianum bridge-lines. The incongruity barrier between the two plant species was overcome; F₁ progeny were obtained from crosses between four parental combinations without embryo-rescue culture. Hybridity was confirmed by leaf and flower morphology and by the production of nematode-resistant F₁ progeny on homozygous susceptible parents. Clones of the five F₁ bridgeline hybrids were highly resistant to Mi-avirulent root-knot nematode (Meloidogyne incognita) at both 25°C and 30°C soil temperatures. However, only clones from PI 270435-3MH and PI 126443-1MH, and hybrids from PI 126443-1MH, were resistant to Mi-virulent M. incognita isolates at high soil temperature. Clones and hybrids from PI 270435-2R2 were not resistant to two Mi-virulent M. incognita isolates at high soil temperature. A source of heat-stable resistance was identified in bridge-line EPP-2, and was found to be derived from L. peruvianum LA 1708. Accessions of the L. peruvianum Maranon races, LA 1708 and LA 2172, and bridge-line EPP-2, segregated for heat-stable resistance to Mi-avirulent M. incognita, but were susceptible to Mi-virulent M. incognita isolates. Clone LA 1708-I conferred heat-stable resistance to M. arenaria isolate W, which is virulent to heat-stable resistance genes in L. peruvianum PI 270435-2R2, PI 270435-3MH, and PI 126443-1MH. Clone LA 1708-I has a distinct heat-stable factor for resistance to Mi-avirulent M. arenaria isolate W, for which the gene symbol Mi-4 is proposed. A Mi-virulent M. arenaria isolate Le Grau du Roi was virulent on all Lycopersicon spp. accessions tested, including those with novel resistance genes.     

Mapping of ripening-related or -specific cDNA clones of tomato (Lycopersicon esculentum)

Summary Nineteen ripening-related or -specific clones from Lycopersicon esculentum were mapped via RFLP analysis using an F₂ population from the cross L. esculentum x L. pennellii and cDNA or genomic clones of known map location. The map produced using cDNA and genomic clones of known map location corresponded well with previously published maps of tomato. The number of loci detected for each ripening-related or-specific clone varied from one to seven. These loci were located on all 12 chromosomes of the tomato genome. There was no significant clustering of ripening-related or-specific genes. Regions of very low recombination were observed. The clone for polygalacturonase (TOM6) mapped to a single region on chromosome 10, the same chromosome as the nor and alc ripening mutants. To fine map this chromosome, two backcross populations were produced from the cross of L. esculentum x L. pimpenillifolium, in which the esculentum parents used were homozygous for either the alc or the nor. The coding region for polygalacturonase is functionally unlinked to either of these two ripening mutants.     

Endosperm dosage relationships among Lycopersicon species

Summary Accessions of eight Lycopersicon species and five yellow-flowered Solanum species were used as males in crosses with 2x and 4x L. esculentum to observe seed set and progeny ploidy. Species which failed in crosses to L. esculentum were crossed as males to 2x and 4x L. peruvianum. In cases of low seed set, chromosome counts were undertaken to establish the nature of the progeny. Endosperm Balance Number (EBN) relationships were determined for the crossability groups. Results support the basic concept of an L. esculentum crossability complex and an L. peruvianum crossability complex. Within the L. esculentum complex, all EBNs appear identical with a value of 2. Within the L. peruvianum complex, more variability appears to exist. The EBN values of this group are higher, and may be approximately double those of the L. esculentum complex. The EBN of L. peruvianum var humifusum appears to be somewhat lower than other L. peruvianum types. The EBN values of S. lycopersicoides, S. rickii, S. ochranthum and S. juglandtfolium could not be determined experimentally. Differential aspects of Lycopersicon and tuber-bearing Solanum evolution may be interpreted on the basis of endosperm compatibility.Co-operative investigation of the Vegetable Crops Research Unit, U.S. Department of Agriculture, Agricultural Research Service, and the Wisconsin Agricultural Experiment Station     

Fine mapping of the nematode resistance gene <Emphasis Type="Italic">Mi-3</Emphasis> in <Emphasis Type="Italic">Solanum peruvianum</Emphasis> and construction of a <Emphasis Type="Italic">S. lycopersicum</Emphasis> DNA contig spanning the locus

Currently, the only genetic resistance against root-knot nematodes in the cultivated tomato Solanum lycopersicum (Lycopersicon esculentum) is due to the gene Mi-1. Another resistance gene, Mi-3, identified in the related wild species Solanum peruvianum (Lycopersicon peruvianum) confers resistance to nematodes that are virulent on tomato lines that carry Mi-1, and is effective at temperatures at which Mi-1 is not effective (above 30°C). Two S. peruvianum populations segregating for Mi-3 were used to develop a high-resolution map of the Mi-3 region of chromosome 12. S. lycopersicum BACs carrying flanking markers were identified and used to construct a contig spanning the Mi-3 region. Markers generated from BAC-end sequences were mapped in S. peruvianum plants in which recombination events had occurred near Mi-3. Comparison of the S. peruvianum genetic map with the physical map of S. lycopersicum indicated that marker order is conserved between S. lycopersicum and S. peruvianum. The 600 kb contig between Mi-3-flanking markers TG180 and NR18 corresponds to a genetic distance of about 7.2 cM in S. peruvianum. We have identified a marker that completely cosegregates with Mi-3, as well as flanking markers within 0.25 cM of the gene. These markers can be used to introduce Mi-3 into cultivated tomato, either by conventional breeding or cloning strategies.     

Characterization and mapping of a gene controlling shoot regeneration in tomato

The superior regeneration capacity of Lycopersicon peruvianum was introduced into the cultivated tomato Lycopersicon esculentum by backcrossing hybrid material with the tomato genotype VF11. In segregating material derived from these backcrosses, the ability to regenerate shoots on root explants cultured on a zeatin-containing medium, was highly correlated with the ability to regenerate shoots on established callus cultures. The efficient shoot-regenerating root explant system permitted us to study the genetics of this trait and to locate the genes involved, using a set of morphological markers defining all 12 tomato chromosomes. Depending on the tomato genotype, mono, -di- or trigenic ratios were observed. It is concluded that a dominant L. peruvianum allele at a locus (Rg-1) near the middle of chromosome 3 determines efficient shoot regeneration on root explants in tomato in combination with dominant alleles at one or two other loci of either L. peruvianum or L. esculentum origin. The map location of the Rg-1 locus was refined further using a number of chromosome-3-specific RFLPs. The addition of new classical and RFLP linkage data to existing literature data and subsequent processing resulted in a revised and integrated map of tomato chromosome 3. From a morphological and physiological analysis of genotypes differing in Rg phenotype, it is concluded that the genetic component associated with regeneration determines the maintenance of morphogenetic competence and not the sensitivity to hormones.     

Resistance to the potato cyst-nematode, Heterodera rostochiensis, in the plant genus Lycopersicon

Forty-eight lines of Lycopersicon and four lines of Solanum were screened for resistance to twelve Heterodera rostochiensis populations of known patho-type(s). Plant lines were assessed for resistance first by examining the outside of the root ball for cysts and later by washing the root ball to extract all cysts. Possible resistant plant selections were re-tested against three eelworm populations, including the one to which they were first shown resistant. Resistance was discovered in two lines of Lycopersicon pimpinelli-folium, two L. esculentum L. pimpinellifolium crosses, L. esculentum var. cerasiforme, six lines of L. peruvianum, in L. peruvianum var. humifusum, L. hirsutum var. glabratum, and in Solanum indicum. Because resistance was found most commonly in L. peruvianum and because it has already been used as a resistant parent in breeding programmes to incorporate resistance to root-knot nematode (Meloidogyne spp.) in tomato, L. peruvianum seems to be the best source of resistance among plants tested so far. The host-parasite relationships of resistant L. hirsutum var. glabratum (B 6013) were compared with those of a commercial, susceptible tomato, L. esculentum‘Ailsa Craig’. Plants were inoculated with three eelworm isolates; the extent of eelworm invasion, plant reaction and eelworm development were studied. Larvae invaded and penetrated roots of the resistant plant as freely and in as large numbers as they penetrated roots of the susceptible tomato. In the latter, numerous larvae matured while, in contrast, few larvae matured in the roots of L. hirsutum var. glabratum. L. hirsutum var. glabratum was shown to possess a root diffusate as active in hatching larvae of Heterodera rostochiensis as that of L. esculentum‘Ailsa Craig’. The existence of pathotypes of H. rostochiensis, identifiable by their differing abilities to increase on resistant tomato lines, was not clearly revealed in the experiments.