Response of tomato rootstocks carrying the <Emphasis Type="Italic">Mi</Emphasis>-resistance gene to populations of <Emphasis Type="Italic">Meloidogyne arenaria,M. incognita and M. javanica</Emphasis>

The response of four Mi-resistance gene tomato rootstocks to seven populations of Meloidogyne was determined in pot tests conducted in a glasshouse. Rootstocks PG76 (Solanum lycopersicum × Solanum sp.) and Brigeor (S. lycopersicum × S. habrochaites) and resistant cv. Monika (S. lycopersicum) were assessed against one population of M. arenaria, three of M. incognita, and three of M. javanica. Rootstocks Beaufort and Maxifort were assessed against one population of M. arenaria, two of M. incognita and two of M. javanica. Rootstock PG76 was highly resistant (reproduction index <10%) to all the populations, whereas rootstock Brigeor and cv. Monika were highly to moderate resistant. Rootstocks Beaufort and Maxifort showed reduced resistance or inability to suppress nematode reproduction, and their responses varied according to the population tested. Beaufort and Maxifort were susceptible to the two populations of M. javanica as Maxifort was to one of M. incognita. The reproduction index of the nematode was higher (P < 0.05) on Maxifort than Beaufort for all root-knot nematode populations.     

Identification of Pathotypes of the Potato Cyst Nematode1

Since the discovery of pathotypes of the potato cyst nematode (Heterodera rostochiensis Wollenweber, 1923), surveys on the distribution of these pathotypes have been carried out in different European countries. Test varieties of potato were selected according to findings resulting in the present situation of varying distinctive host-plants and of diverging programs for breeding potatoes resistant to the potato cyst nematode. With respect to rational selection and use of resistant potato varieties, there is a great need for standardization in pathotype identification (nomenclature), maintenance and supply of appropriate genotypes of Solanum species and methods for testing resistance in new potato varieties. Anticipating a cross-experiment with pathotypes and test varieties from Great Britain, the Federal Republic of Germany and the Netherlands in 1974, a preliminary test with all recognized or presumed pathotypes from these countries, but with only part of the potential differential host-plants was carried out by the Plant Protection Service at Wageningen. The results of this experiment are discussed.     

Selection of nematodes by resistant plants has implications for local adaptation and cross‐virulence

The variability of resistance durability in different potato genotypes harbouring the same resistance QTL but differing by their genetic background was explored. The indirect consequences of the resistance adaptation in terms of local (i.e. genotype‐specific) adaptation and cross‐virulence was also investigated. Following the virulence of the potato cyst nematode Globodera pallida in a long‐term experimental evolution protocol, the results showed that nematode populations were able to adapt to the resistance of four potato genotypes carrying the QTL GpaV from Solanum vernei, and that the plant genetic background has an impact upon the durability of resistance. The pattern of local adaptation observed here indicates that divergent selection has occurred during the experimental evolution performed from the same initial nematode population, and revealed a trade‐off between the adaptation to a resistant potato genotype and the adaptation to another resistant genotype differing in its genetic background. In terms of cross‐virulence between potato genotypes derived from different resistance sources (S. sparsipilum and S. spegazzinii), this study shows that the adaptation to resistance QTL GpaV_vrn does not necessarily allow the adaptation to collinear GpaV loci. The results presented here could be useful for predicting evolution of nematode populations in natural agro‐ecosystems and identifying durable strategies for resistance deployment.     

Selection forMeloidogyne incognita virulence against resistance genes from tomato and pepper and specificity of the virulence/resistance determinants

Experiments were designed to analyze the relationships between the root-knot nematodeMeloidogyne incognita and resistant tomato and pepper genotypes. From a natural avirulent isolate, near-isogenic nematode lineages were selected with virulence either against the tomatoMi resistance gene or the pepperMe3 resistance gene. Despite the drastic selection pressure used, nematodes appeared unable to overcome the pepperMe1 gene, therefore suggesting some differences in the resistance conferred byMe1 andMe3 in this species. Nematodes virulent onMi-resistant tomatoes were not able to reproduce onMe1-resistant nor onMe3-resistant peppers, and nematodes virulent onMe3-resistant peppers were not able to reproduce onMi-resistant tomatoes nor onMe1-resistant peppers. These results clearly demonstrate the specificity ofM. incognita virulence against resistance genes from both tomato and pepper, and indirectly suggest that gene-for-gene relationships could occur between these two solanaceous crops and the nematode.     

Interactions Between the Potato Cyst Nematode Globodera rostochiensis and Diseases Caused by Rhizoctonia solani AG3 in Potatoes Under Field Conditions

Findings from 2 years of field experiments investigating the relationship between Globodera rostochiensis and Rhizoctonia solani on unique field sites are reported. In 2000, a field experiment was positioned on land that had previously been used for experimental work investigating integrated potato cyst nematode (PCN) management methods. This study had produced an ‘untypical’ mosaic of PCN population densities ranging from 5 to 221 eggs g⁻¹ soil. In 2001, the field experiment was conducted on a different field site and overlaid on a focus of G. rostochiensis population densities ranging from 11 to 108 eggs g⁻¹ soil. In each experiment, potatoes (cv. Désirée) were grown in plots with similar population densities of G. rostochiensis that were either uninoculated or inoculated with R. solani. A series of potato plant harvests were undertaken to investigate the effects of nematode infestation on the incidence and severity of R. solani diseases and the associated development of plants. In both experiments, a clear relationship was found between the density of G. rostochiensis juveniles present in potato roots and the incidence of stolons infected by R. solani, 6 weeks after planting. For the first time this interaction has been determined under field conditions. The results of the study suggest that the interaction between nematode and fungus is indirect and possible mechanisms are discussed.     

Identification and characterization of RB-orthologous genes from the late blight resistant wild potato species Solanum verrucosum

Late blight, caused by the oomycete pathogen, Phytophthora infestans, is a devastating disease of potatoes and tomatoes. A key long-term management strategy for combating this disease is to develop potato cultivars with high levels of durable resistance through identification and integration of major resistance genes. The RBgene, cloned from the Mexican diploid potato species Solanum bulbocastanum, confers broad-spectrum resistance to potato late blight. Here, we have determined the late blight resistance phenotypes of eight accessions of Solanum verrucosum, another wild diploid potato species, using greenhouse inoculations and discovered variability among the accessions. While most accessions were resistant, one accession was notably more susceptible. Transcribed orthologs of the RB gene from the eight S. verrucosum accessions were cloned using a homology-based PCR approach. Sequence analysis revealed that the RB^ver orthologs share up to 83.5% nucleotide identity with RB^blb. Stable introduction of the RB ortholog from late blight resistant S. verrucosum PI275260 into susceptible S. tuberosum confers resistance to P. infestans. Interestingly, this functional RB^ver ortholog contains an insertion of a complete leucine rich repeat when compared to RB^blb, and differs from the RB^ver ortholog from a susceptible accession at only four amino acid residues.     

Identification of the Beet Cyst Nematode Heterodera schachtii by PCR

PCR-RFLPs of ITS-rDNA and PCR with species-specific primers were developed for identification of cysts and juveniles of the beet cyst nematode Heterodera schachtii. Restrictions of PCR product by MvaI or ScrFI distinguish H. schachtii, H. betae, H. trifolii and H. medicaginis. RFLP profiles with eight restriction enzymes for these four nematode species are presented. Based on Internal Transcribed Spacer sequences of populations from several Schachtii group species, a specific primer for H. schachtii was designed, permitting amplification of the target sequence from juveniles and cysts of the beet cyst nematode. A duplex PCR protocol tested with a wide range of nematode samples is described.     

Identification and management of virulence genes in potato cyst nematodes

Efficient and accurate diagnostic assays are essential for the design and evaluation of control measures of the potato cyst nematodesGlobodera rostochiensis andG. pallida by means of resistance. The hybridoma technology and the polymerase chain reaction (PCR) offer in potential various possibilities to design such diagnostic tests for routine purposes. We set out to devise a refined advisory system based on biochemical assays by using the following stepwise approach.In the early 80's a research program was started to develop an immunoassay to differentiate the two sibling species of potato cyst nematodes. Species specific monoclonal antibodies were raised against nematode proteins which are thermostable, abundant and homologous, and which enable reliable species identification using single eggs.     

Selection of virulence in Meloidogyne chitwoodi to resistance in the wild potato Solanum fendleri

The possibilities of selecting virulence from a virtually avirulent root-knot nematode population towards resistance in wild potato have been investigated. Single egg masses of Meloidogyne chitwoodi, which had occasionally been produced on roots of resistant Solanum fendleri gave rise to eight lines after one generation on tomato. Five lines were able to circumvent completely the resistance of S. fendleri 93-114-12 resulting in a susceptible response similar to that of the control potato cv Nicola. Subsequently, a resistance test with other resistant genotypes of S. fendleri, S. bulbocastanum, S. stoloniferum and S. hougasii revealed that the virulent lines were also able to break through the resistance in most other species, but clear differences were noticed between the virulent lines. The results suggest a simple inheritance of virulence in M. chitwoodi towards resistance in S. fendleri. However, more virulence factors are involved to explain the differences on the other Solanum species between the virulent lines. The implications of the ease to select virulence with respect to the practical use of resistance in potato breeding and growing are discussed.     

The influence of cyst nematodes and drought on potato growth. 1. Effects on plant growth under semi-controlled conditions

Potatoes were grown under a permanent rain shelter in mobile containers in soil with or without potato cyst nematodes (Globodera pallida). The plants were subjected to an early drought stress period starting at planting until 43 days after planting, to a late drought stress period starting at 43 days until senescence at 92 days and to a drought control. Dry matter weight and characteristics of leaves, stems, stolons and roots were determined at periodic harvests. The early drought stress and nematode infection affected many plant organ characteristics in similar ways. Numbers of leaves, specific leaf area, plant height, specific stem weight, leaf area ratio, mean tuber weight and harvest index were reduced by both stress factors at early stages of growth.Later on, interactions between both stress factors which influence the development rate of the plants led to more diverse plant reactions. Plants of all treatments rapidly senesced at about 90 days after planting. Uninfected plants had then depleted the soil nutrient supply whereas the plants grown in the inoculated soil senesced as a result of the potato cyst nematode infection.     

Increased virulence of Globodera pallida during repeated rearing on different resistant potato cultivars explained by a simple model

Selection for virulence of Globodera pallida on potato cultivars was studied for four generations under controlled conditions. The reproduction rate (P_f/P_i) of a mixed Pa2/3 population increased by a factor of 61 during rearing on the partially resistant potato cv. Darwina compared to rearing on the susceptible cv. Irene. This was a result of selection for virulence on cv. Darwina, and achieving the Hardy–Weinberg equilibrium on cv. Irene. Increased virulence also significantly raised the reproduction rate on several other Solanum genotypes. These changes could be explained reasonably well by the monogenic inheritance of a virulence factor breaking the Grp1 locus. The virulence changes were probably mainly evoked by this gene only, inherited from S. vernei 1-3 or S. vernei 24/20. The Grp1 locus has probably provided the differential S. vernei hybrid (VTⁿ)² 62-33-3 with its resistance to the Pa2 group and not to the Pa3 group. Alternation of cultivars did not halt selection if the cultivars highly differentiated between the Pa2 and Pa3 populations. Only when alternation was with cultivars that harboured a different resistance gene against Pa3 was selection for virulence delayed. Differences in virulence levels (i.e. reproduction rates) within the nematode population determined the rate of selection, not the resistance level itself. Selection of a Pa3 population for three generations on cv. Karakter not only increased the reproduction rate on cv. Karakter itself by a factor 4.2, but also raised the reproduction on other potato genotypes. A simple monogenic model could explain these changes in virulence.     

Co-evolution of potato cyst nematodes and their hosts: implications for pathotypes and resistance1

The co-evolutionary process is believed to have resulted, through interaction of wild populations of potato cyst nematodes and their hosts in geological time, in the resistant hosts now utilized in plant breeding programmes and in nematode populations with genes for resistance-breaking or virulence. It is argued that all such interactions between highly adapted, truly parasitic plant nematodes and their hosts are likely to be governed by gene-for-gene interrelationships. Practical implications of this hypothesis are that only pathotypes (resistance-breaking races) defined against identified resistance genes are scientifically sound and of practical value; that, in the case of potato cyst nematodes, other pathotypes (Ro2, Ro3, Ro5 and Pa2 and Pa3) should be abandoned; and that oligogenically based resistance to potato cyst nematodes, especially important in providing resistance to Globodera pallida, is non-durable. Working definitions of the terms ‘pathotype’, ‘host-race’ and ‘virulence’ are provided.     

Potato cyst nematodes: a critical review of the current pathotyping scheme1

The pathotype scheme proposed by Kort et al. (1977) is critically reviewed. It is proposed that many of their pathotypes are artifacts resulting from their using an arbitrary Pf/Pi value of ± 1 and differential clones with quantitatively based polygenic resistance to separate pathotypes. It is concluded that most European populations of potato cyst nematodes are heterogeneous for virulence genes and that Pf/Pi measurements are influenced too readily by several factors, including initial population density, the vigour of plant growth, and the viability of the nematode, to be used as either a measure of nematode virulence or plant resistance. Instead, reproduction should be related to that on appropriate non-resistant or partially resistant controls and the aim should be to quantify the proportion of virulent individuals in the population. For greatest accuracy it will be necessary to use procedures which minimize genotype × environmental interactions.     

The influence of cyst nematodes and drought on potato growth. 2. Effects on plant water relations under semi-controlled conditions

Potatoes were grown under a permanent rain shelter in mobile containers in soil with and without potato cyst nematodes (Globodera pallida). The plants were either subjected to an early drought stress period from planting until 43 days after planting, to a late drought stress period during tuber bulking or to a drought control. Leaf water potentials, stomatal diffusion resistances for water vapour, transpiration rates, dry matter accumulation and water use efficiencies of the plants were determined periodically. Both drought and nematodes decreased leaf water potential and increased stomatal resistance.Drought led to a higher water use efficiency, cyst nematode infection, however, reduced the water use efficiency at early stages of growth, and increased it at later stages. It is concluded that at least two main growth reducing mechanisms exist of which the relative importance varies with time. Firstly, reduced apparent assimilation rates, which are unrelated to a change in the water balance caused by the initial attack by the cyst nematodes. Secondly, a reduced dry matter accumulation resulting from a decrease of water uptake. Effects of drought and cyst nematode infection on plant growth and water relations were not always additive mainly because infected plants used up less water leading to less drought stress.     

Genetic Structure of the Population of Phytophthora infestans Attacking Solanum ochranthum in the Highlands of Ecuador

Thirty-nine isolates of Phytophthora infestans were collected from the wild host Solanum ochranthum in the highland tropics of Ecuador and characterized with a set of phenotypic and molecular markers (mating type, metalaxyl sensitivity, the allozyme loci Gpi, and Pep, mitochondrial DNA haplotype, RFLP, and SSR), as well as for pathogenicity on various hosts. Three groups of isolates (A, B, and C) were identified based on their multilocus genotypes and variable abilities to cause disease on different hosts. Group A had a combination of alleles for the Gpi (86/100), Pep (96/100) and mtDNA (Ia) loci, as well as an RFLP fingerprint, that have not been reported for P. infestans in Ecuador, or elsewhere. Group B shares many marker characteristics with the US-1 lineage described in Ecuador on tomato, pear melon (S. muricatum), and S. caripense, but has SSR alleles not present in typical US-1 isolates. Group C for all markers tested is identical to the EC-1 lineage described on cultivated and wild potatoes in Ecuador. All isolates from S. ochranthum were able to re-infect their host of origin in the detached leaf assay; however, we did not draw clear conclusions as to the relative aggressiveness of the three groups on this host. Isolates of group A were the most specialized and were generally non-pathogenic or weakly pathogenic on all hosts other than S. ochranthum. Groups B and C infected tuber-bearing hosts, including the cultivated potato but were generally non-pathogenic on other non-tuber bearing hosts. Solanum ochranthum was infected by isolates coming from tuber-bearing Solanum hosts (i.e., the EC-1 lineage of P. infestans) and some US-1 isolates from non-tuber bearing hosts. Thus, in nature this species might be a potential reservoir of inoculum of different pathogen populations able to infect the cultivated hosts potato, tomato and pear melon (S.␣muricatum). Unlike potato and tomato in Ecuador, each of which is primarily attacked by a highly specialized pathogen population, S. ochranthum appears to harbour at least three pathogen groups of␣different genetic make-up. The unresolved issue of potential host specificity in isolates found on S.␣ochranthum could complicate efforts to use this species in tomato improvement.     

Expression of resistance to the root-knot nematodes,Meloidogyne hapla andM. fallax,in wildSolanum spp. under field conditions

In 1995 two fields in the Netherlands, naturally infested withMeloidogyne hapla (Wageningen) andM. fallax (Baexem), were used to evaluate resistant and susceptibleSolanum genotypes under natural conditions. In April, genotypes were planted in circular microplots. Soil samples were taken and analyzed for the occurrence of second-stage juveniles every six weeks. From August onwards, large differences between resistant and susceptible genotypes in numbers of juveniles were found in the soil. For all resistant wildSolanum genotypes the level of infection in soil at the end of the growing season in October was equal to or lower than at the beginning. Glasshouse experiments were performed with the same genotypes and nematode populations (i.e. originally derived from these fields) and the results were comparable with the observations from the field. It is concluded that resistance, as selected in glasshouse trials, corresponds well with resistant behaviour in the field and that it is worthwhile to transfer the resistance from theseSolanum sources to commercial potato cultivars for successful control of root-knot nematodes.     

Behaviour of Solanum spp. on inoculation with different isolates of Fusarium oxysporum f. sp. melongenae*

Twelve wild Solanum accessions were tested in a glasshouse at the seedling stage for resistance to Fusarium oxysporum f. sp. melongenae, the causal agent of fusarium wilt of aubergine. Four isolates of the fungus (three Turkish and one Italian) were used. Solanum incanum and S. linneanum were highly susceptible, whereas S. sisymbrifolium, S. torvum and S. aethiopicum Gilo group (one accession) were resistant. In Solanum aethiopicum Aculeatum (two accessions), S. aethiopicum Gilo, S. viarum and S. macrocarpon there were both resistant and susceptible individuals. The sources of resistance found in these wild Solanum spp. could be conveniently used to breed aubergine cultivars resistant to fusarium wilt.     

New genes for disease resistances through somatic hybridization

Somatic hybridization, a process of combining protoplasts from different plants, can provide new sources of disease resistances for plants. In the case of wild and cultivatedSolanum species, the hybrids express resistances from each partner in the fusion and can often be crossed with cultivars to improve agronomic characteristics of the tubers. Restriction fragment length polymorphism (RFLP) analyses can provide a means for determining that the plants being investigated are actually hybrids as well as a means for following the introgression of DNA into progeny lines. These points are addressed in this paper with specific reference to somatic hybrids betweenSolanum brevidens and potato.     

Partial stem and leaf resistance against the fungal pathogen <Emphasis Type="Italic">Botrytis cinerea</Emphasis> in wild relatives of tomato

Tomato (Solanum lycopersicum) is one of many greenhouse crops that can be infected by the necrotrophic ascomycete Botrytis cinerea. Commercial cultivation of tomato is hampered by the lack of resistance. Quantitative resistance has been reported in wild tomato relatives, mostly based on leaf assays. We aimed to identify wild tomato relatives with resistance to B. cinerea based on quantitative assays both on leaves and stem segments, monitoring infection frequency and disease expansion rate as parameters. A quantitative tomato stem segment assay was developed. This stem assay and a previously described leaf assay were used to screen a collection of 22 Solanum accessions. Significant differences in disease parameters were observed among accessions. Resistance to B. cinerea was observed in a number of wild Solanum species, including accessions of S. chilense, S. habrochaites and S. neorickii, both in the leaf assay and the stem segment assay. A number of resistant and susceptible accessions were evaluated as adult plants under greenhouse conditions. The data obtained in greenhouse assays confirmed the leaf and stem disease data. The expression of several defence-related genes was studied in a subset of accessions. There was no apparent correlation between the expression levels of the genes tested and the quantitative resistance level to B. cinerea. Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users.