Nematode quantitative resistance conferred by the pepper genetic background presents additive effects and is stable against different isolates of Meloidogyne incognita

Root‐knot nematodes (RKNs), Meloidogyne spp., are a major disease problem in solanaceous crops worldwide, including pepper (Capsicum spp.). Genetic control provides an economically and environmentally sustainable protection alternative to soil fumigants. In pepper, resistance to the main RKN species (M. incognita, M. javanica and M. arenaria) is conferred by the major genes (R genes) Me1, Me3 and N. However, RKNs are able to develop virulence, thus endangering the efficiency of R genes. Quantitative resistance (QR) against Meloidogyne spp. is expected to provide an alternative to R genes, or to be combined with R genes, to increase the resistance efficiency and durability in pepper. In order to explore the ability of QR to protect pepper against RKNs, five pepper inbred lines, differing in their QR level, were tested directly, or after combination with the Me1 and Me3 genes, for their resistance to a panel of M. arenaria, M. javanica and M. incognita isolates. The M. arenaria and M. javanica isolates showed low pathogenicity to pepper, unlike the M. incognita isolates. The QR, controlled by the pepper genetic background, displayed a high resistance level with a broad spectrum of action, protecting pepper against Me3‐virulent as well as avirulent M. incognita isolates. The QR was also expressed when combined with the Me1 and Me3 genes, but presented additive genetic effects so that heterozygous F₁ hybrids proved less resistant than homozygous inbred lines. The discovery of this QR is expected to provide promising applications for preserving the efficiency and durability of nematode resistance.     

Host suitability of Vitis rootstocks to root‐knot nematodes (Meloidogyne spp.) and the dagger nematode Xiphinema index,and plant damage caused by infections

The host suitability of commercial Vitis rootstocks commonly used in Spain (161‐49C, 41B, 1103P, 110R, 140Ru and SO4) to root‐knot nematodes (Meloidogyne arenaria, M. incognita, M. javanica) and Xiphinema index, and damage caused by nematode infection were determined under controlled conditions. The three root‐knot nematodes reproduced with a rate higher than one in all rootstocks, indicating that they are suitable hosts for these nematodes. Growth of rootstocks infected with the root‐knot nematodes was less vigorous than that of nematode‐uninfected controls in the majority of the rootstocks studied. Root infection resulted in moderate to severe root galling in all rootstocks. The shoot and main stem diameters appeared to be the most sensitive variables of damage caused by infection by Meloidogyne spp., with reduction rates from 36% and 53% in 161‐49C to 57% and 66% in 140Ru, respectively. The shoot height was not significantly affected by the root‐knot nematodes and the root fresh weight generally increased as a consequence of intensive galling. The nematode X. index caused significant root damage with a reproduction factor higher than one in all rootstocks. However, reproduction factor was significantly influenced by the rootstock and significantly decreased by about 12‐fold (5·7 to 18·1‐fold) with the increase in inoculum density from 100 to 1000 nematodes per plant. The root dry weight was reduced by X. index infections, and was the plant growth variable most affected by the nematode infection in all rootstocks at both inoculum densities. Meloidogyne arenaria, M. incognita, M. javanica and X. index, prevalent in many world vineyards, are all shown to have a damaging effect on the six tested rootstocks.     

Characterisation of Meloidogyne species from China using Isozyme Phenotypes and Amplified Mitochondrial DNA Restriction Fragment Length Polymorphism

Forty-six Meloidogyne populations from 14 provinces of China were characterised in terms of malate dehydrogenase, esterase phenotypes and HinfI restriction profiles of amplification products from the mitochondrial DNA (mtDNA) region between the COII and lrRNA genes. Isozyme phenotyping revealed that 29 of the populations were M. incognita, six were M. javanica, six were M. arenaria, three were M. hapla and two were M. enterolobii. HinfI restriction patterns of the COII-lrRNA region correlated with nematode isozyme phenotypes and enabled reliable differentiation and identification of the five root-knot nematodes occurring in China. The size and sequence of the mtDNA amplification product were determined for the first time for M. enteroloii, a potentially economically important crop pathogen. Sequence comparison showed that the sequence of the intergenic region between the COII and lrRNA genes for M. enterolobii was identical to that reported for M. mayaguensis. Together with published observations on morphology, host range and esterase phenotype of the two nominal species, the mtDNA sequence evidence suggests that M. mayaguensis could be conspecific with M. enterolobii.     

The quarantine root-knot nematode Meloidogyne enterolobii – a potential threat to Portugal and Europe

In 2017, during a survey on subsistence farms and gardens in Coimbra region, Portugal, 40 infected root samples were collected and 47 root-knot nematode (RKN) isolates identified, based on esterase phenotype. The phenotypes A2, H1, Hi2/Hi4, I1/I2/I3 and J3 associated to five Meloidogyne species (M. arenaria, M. hapla, M. hispanica, M. incognita and M. javanica) were found in 43 RKN isolates. The esterase phenotype En2/En4/En5, corresponding to M. enterolobii (=M. mayaguensis), was detected in four RKN isolates from Cereus hildmannianus (Cactaceae), Lampranthus sp. (Aizoaceae), Physalis peruviana (Solanaceae) and Callistemon sp. (Myrtaceae) infected roots. In order to validate the biochemical identification of the M. enterolobii isolates, molecular studies performed with species-specific primers yielded the expected fragment of c.520 bp, and the amplification of cytochrome oxidase subunits I and II regions of 800 bp. The DNA sequences of one of the isolates were compared with available Meloidogyne species sequences in databases. The Portuguese isolate grouped with 99–100% bootstrap support with all M. enterolobii sequences included for comparison, confirming the presence of this RKN species in Portugal. In the EPPO region, M. enterolobii has been reported in France and Switzerland and intercepted in the Netherlands, Germany and the UK associated with plant material from Asia, South America and Africa. Taking into account the pathogen aggressiveness and its distribution, there is a high probability of its spread not only in the Mediterranean region but also in Europe, and of it becoming a threat to the agricultural economy, where there are no effective strategies for its control.     

<Emphasis Type="Italic">Solanum sisymbriifolium</Emphasis> - a new approach for the management of plant-parasitic nematodes

Plant-parasitic nematodes are serious pests causing important crop losses worldwide. After extensive screening of non-tuber-bearing Solanaceae, a resistant trap crop, Solanum sisymbriifolium, with a high production level of hatching agents, seemed an ideal control method for potato cyst nematodes (PCN), Globodera spp. Recently, root-knot nematodes (RKN), Meloidogyne spp., were found coexisting with PCN. Therefore, it is important to find alternative methods to control both nematode genera. The chemical properties of S. sisymbriifolium turns this plant into an excellent candidate for further nematicidal studies and to develop new crop production models. Studies concerning the effects of this plant on plant-parasitic nematodes are presented. Pathogenicity studies with four S. sisymbriifolium cvs (Domino, Pion, Sis 4004 and Sharp) and five Meloidogyne species showed that all cultivars of S. sisymbriifolium studied were resistant to M. chitwoodi and hypersusceptible to M. arenaria and M. hapla. For M. hispanica only cv Pion was susceptible. M. javanica induced different responses: cvs Pion and Sharp were susceptible; cv Domino resistant and Sis 4004 hypersusceptible. The studies of the hatching effects of root exudates from these cvs showed that they had an influence on the hatching inhibition of second stage juveniles of the five Meloidogyne species tested.     

Screening of common bean (Phaseolus vulgaris) for resistance against temperate root-knot nematodes (Meloidogyne spp.)

BACKGROUND: An important part of the production area of common bean (Phaseolus vulgaris L.) in Belgium is located on the sandy soils of the provinces of Antwerp and Limburg where Meloidogyne chitwoodi (Golden), M. fallax (Karssen) and M. hapla (Chitwood) are present. The host plant status of ten bean cultivars for root‐knot nematodes was determined by evaluating penetration, development and egg mass formation after inoculation with second‐stage juveniles. RESULTS: The tested cultivars were poor to good hosts for M. chitwoodi, non‐hosts or bad hosts for M. fallax and excellent hosts for M. hapla. Significantly fewer M. fallax were found in the roots, and their development was delayed. Penetration of M. hapla took place over a longer period than that of M. chitwoodi and M. fallax. The number of mature females of M. chitwoodi in cv. Polder 6 weeks after inoculation was no different from that in other cultivars, although fewer egg masses were found on this cultivar in the screening test. There was no influence of M. chitwoodi on vegetative growth of cv. Polder. CONCLUSION: The differences found in host plant status of bean cultivars stress the importance of a correct diagnosis of the Meloidogyne species in agricultural fields. Cultivar Polder showed potential as a trap crop for M. chitwoodi. Copyright © 2011 Society of Chemical Industry     

Reaction of a heterozygous tomato hybrid bearing the Mi-1.2 gene to 15 Meloidogyne species

Many root-knot nematode (RKN) species (Meloidogyne spp.) are polyphagous and cultivated tomato (Solanum lycopersicum) is one of their preferential hosts, leading to significant losses. It is known that the dominant Mi-1.2 gene in tomato confers resistance to the three most important RKN species—M. incognita, M. javanica, and M. arenaria, and minor species—M. ethiopica, M. hispanica, and M. luci. However, little information is available about resistance of tomatoes carrying this gene to other tomato-infecting RKN species. In this study, resistance conferred by the Mi-1.2 gene/locus was evaluated against populations of 15 Meloidogyne species, employing tomato cultivars Santa Clara (homozygous recessive mi-1.2/mi-1.2, susceptible) and Debora Plus (heterozygous Mi-1.2/mi-1.2, resistant). Debora Plus was susceptible only to M. enterolobii and M. hapla, and was resistant to the other Brazilian populations of M. arenaria, M. ethiopica, M. exigua, M. hispanica, M. incognita, M. inornata, M. izalcoensis M. javanica, M. konaensis, M. luci, M. morocciensis, M. paranaensis, and M. petuniae. Mi-1.2 is located on tomato chromosome 6 within a cluster of seven homologous genes of the nucleotide-binding site leucine-rich repeat (NBS-LRR) family; further research is required to confirm if this multiple Meloidogyne spp. resistance phenotype is controlled exclusively by Mi-1.2 or by combined action of other closely linked genes. This evaluation of resistance of the Debora Plus cultivar to several Meloidogyne species suggests that the Mi-1.2 gene/locus may reduce losses induced by a wide range of Meloidogyne spp. Further studies using additional resistant cultivars and other populations of Meloidogyne spp. are needed to confirm these results.     

Screening Tomato Cultivars for Resistance to Meloidogyne Species

Abstract

Screening tests on 162 tomato cultivars and hybrids conducted at the Division of Horticulture, Indian Agricultural Research Institute, New Delhi during 1967-8 indicated a high degree of resistance to root-knot nematodes in Nematex, VFN-8, 65N215-1, 65N255-1 and S1-120, under field and laboratory conditions. The degree of resistance varied with the species of Meloidogyne. Nematex was immune to M. javanica, M. incognita and M. arenaria. VFN-8, 65N215-1 and 65N255-1 were immune to M. incognita and M. arenaria and resistant to M. javanica. S1-120, a commercial cultivar, showed a high degree of tolerance to M. javanica, M. incognita and M. arenaria. The degree of resistance varied with nematode population density.     

Nematicidal activity of Ochradenus baccatus against the root‐knot nematode Meloidogyne javanica

Ochradenus baccatus is a widely distributed shrub in desert regions of the Middle East and North Africa. This plant's nematicidal activity against the root‐knot nematode Meloidogyne javanica was evaluated because it has been found to contain exceptionally high levels of glucosinolates. In in vitro assays with aqueous extracts of the plant, 100% of second‐stage juveniles were immobilized after exposure to 4% root‐core extract for 48 h; 8% root‐core extract suppressed their hatching by 87%, whereas stem, flower and root bark showed lower activity. Incorporation of root core or bark into the soil, as fresh or dry powder at 1 and 0·5% (w/w), respectively, reduced the number of nematodes recovered from the soil by 95–100%, whereas the flower and stem were much less effective. Results from further pot experiments indicated that only the root bark consistently contains nematicidal compounds which are effective in soil, whereas the nematicidal activity of the root core in soil was inconsistent. The presence of non‐volatile lipophilic and lipophobic nematicidal compounds in the root bark was suggested by extraction with different polar solvents, but these compounds do not seem to be isothiocyanates – glucosinolate‐hydrolysed compounds with nematicidal activity. Very poor host status of Ochradenus baccatus to M. javanica, M. incognita and M. hapla, but with root‐penetration rates of juveniles similar to those in tomato roots, suggest that this plant may be used as a cover plant or trap plant to reduce nematode populations in the soil.     

Occurrence of virulent root-knot nematode populations on tomatoes bearing the <Emphasis Type="Italic">Mi</Emphasis> gene in protected vegetable-growing areas of Turkey

Root-knot nematodes (Meloidogyne spp.; RKN) are one of the most important pathogens of vegetables in Turkey. Assessing the existing virulent RKN populations is of importance for pathogen mapping in the west Mediterranean region of Turkey. Therefore, 95 populations of RKN were collected from different protected vegetable-growing locations in the region. Pure cultures were obtained and identified by means of species-specific primers. Virulence of the populations against the Mi-1 gene conferring resistance to Meloidogyne incognita, M. javanica and M. arenaria was determined according to their egg masses and gall rating on resistant and susceptible tomato varieties. Results showed that seven populations of M. incognita and six populations of M. javanica were able to overcome the resistance controlled by the Mi-1 gene. The frequency of virulent populations of M. incognita and M. javanica collected from different protected-grown vegetables was 11.7% and 21.4%, respectively. To our knowledge, this is the first report of populations of RKN virulent to the Mi-1 gene in Turkey.     

Detection of Meloidogyne enterolobii in potatoes in South Africa and phylogenetic analysis based on intergenic region and the mitochondrial DNA sequences

Root-knot nematodes (Meloidogyne spp.) are a major problem facing crop production globally including potatoes. During the 2011/2012 potato growing season, root-knot nematode infected potato tubers were obtained from different potato growing regions in South Africa for identification of Meloidogyne spp. Using the intergenic region of the ribosomal DNA (IGS-rDNA) together with the region between the cytochrome oxidase small subunit II (COII) and the 16S rRNA gene in the mitochondrial DNA (mtDNA), five of the 78 composite samples received produced amplicon sizes of 705 bp for COII and 780 bp for IGS typical of M. enterolobii. These five samples were from the KwaZulu-Natal potato producing region. Nucleotide sequencing and phylogenetic analysis of the COII and IGS fragment showed that the five Meloidogyne populations were 100 % similar and they clustered closely with those of M. enterolobii in the GenBank database. The high damage potential of resistance-breaking populations of Meloidogyne species is a threat to profitable potato production and will require effective pest management programmes to be put in place.     

Host suitability of Solanum torvum cultivars to Meloidogyne incognita and M. javanica and population dynamics

Several experiments were carried out to assess the performance of commercial Solanum torvum cultivars against the root knot nematodes Meloidogyne incognita and M. javanica in Spain. The response of S. torvum rootstock cultivars Brutus, Espina, Salutamu and Torpedo against M. incognita and Mi-1.2 (a)virulent M. javanica isolates was determined in pot experiments, and of ‘Brutus’ to an N-virulent isolate of M. incognita, compared with that of the eggplant S. melongena ‘Cristal’. The relationship between the initial and final population densities of M. javanica on ungrafted and grafted ‘Cristal’ onto the S. torvum ‘Brutus’ was assessed, together with the effect on dry shoot biomass. Finally, the population growth rate and the resistance level of the four S. torvum cultivars against M. incognita was assessed under plastic greenhouse conditions in two cropping seasons. All S. torvum rootstocks responded as resistant to the M. incognita isolates and from highly resistant to susceptible against M. javanica isolates. The maximum multiplication rates of M. javanica on the ungrafted or grafted eggplant were 270 and 49, respectively, and the equilibrium densities were 1318 and 2056 eggs and J2 per 100 cm³ soil, respectively. The tolerance of the ungrafted eggplant was 10.9 J2 per 100 cm³ soil, and the minimum relative dry shoot biomass was 0.76. The population growth rate of M. incognita on eggplant cv. Cristal differed from that of the S. torvum cultivars in both cropping seasons. These results suggest that S. torvum is a valuable rootstock for managing the two Meloidogyne species irrespective of the (a)virulence status.     

Suitability of weed species prevailing in Spanish vineyards as hosts for root-knot nematodes

Commercial vineyards in southern Spain were surveyed and sampled during October to December 2004 to determine the extent to which common weeds present were suitable hosts of root-knot nematodes infesting soils of those vineyards. Seven weed species commonly growing in grapevine soils in southern Spain were found infected by either Meloidogyne incognita or M. javanica: Amaranthus retroflexus (redroot pigweed), Anchusa azurea (ox-tongue), Chenopodium album (goosefoot), Erodium moschatum (musk stork’s bill), Malva rotundifolia (low mallow), Sinapis alba (white mustard), and Solanum nigrum (black nightshade). The host suitability of the weeds to root-knot nematodes was evaluated on the basis of root galling severity and nematode population densities in soil and roots. Also, the host–parasite relationship in these naturally Meloidogyne-infected weeds was examined. All the weed species in the study were considered suitable hosts for M. incognita and M. javanica because: (a) high Meloidogyne spp. populations occurred in roots and surrounding soil of the weed species; (b) the severity of root galling was high, and (c) well-established permanent feeding sites were observed in the histopathological studies of infected root tissues. In addition, this study presents the first reports of S. alba and A. azurea as hosts for M. incognita, and of E. moschatum as a new host for M. javanica, thus increasing the list of reported weed hosts for Meloidogyne spp. These results indicate that noticeable population densities of M. incognita and M. javanica can be maintained or increased in these weeds, at population levels higher than those previously reported for the same nematodes infecting grapevine roots. The weeds infesting vineyards thus represent an important source of inoculum of Meloidogyne spp., and furthermore may act as reservoirs of these nematodes which can be disseminated within or among vineyards by agricultural operations.     

Species-specific DNA markers for identification of two root-knot nematodes of coffee: Meloidogyne arabicida and M. izalcoensis

Seven root-knot nematodes (RKN), including Meloidogyne exigua, M. incognita, M. paranaensis, M. enterolobii, M. arabicida, M. izalcoensis and M. arenaria are major pathogens of coffee crop in the Americas. Species-specific primers for their identification have been developed for five of them and constitute a fast and reliable method of identification. Here we report a PCR-based assay for specific detection of M. arabicida and M. izalcoensis. Random Amplified Polymorphic DNA fragments specific for these two species were converted into sequence characterized amplified region (SCAR) markers. PCR amplification using the SCAR primers produced a specific fragment of 300 bp and 670 bp for M. arabicida and M. izalcoensis, respectively, which were absent in other coffee-associated Meloidogyne spp. tested. SCAR primers also allowed successful amplification of DNA from single second-stage juveniles (J2), males and females. In addition, these primers were able to unambiguously detect the target species in nematode suspensions extracted from soil and roots samples, in different isolates of the same species or when used in multiplex PCR reactions containing mixtures of species. These results demonstrated the effectiveness of these SCAR markers and their multiplex use with those previously developed for M. exigua, M. incognita, M. paranaensis, M. enterolobii and M. arenaria constitute an essential detection tool. This diagnostic kit will contribute for specific J2 identification of the major RKN infecting coffee from field samples in the Americas.     

Thermal requirements and population dynamics of root‐knot nematodes on cucumber and yield losses under protected cultivation

Several studies were carried out to determine (i) thermal requirements for development, egg production and emergence of juveniles, and completion of the life cycle of Meloidogyne incognita and Meloidogyne javanica on cucumber, (ii) the maximum multiplication rate and the equilibrium density of root‐knot nematodes on cucumber and yield losses in pot and plastic greenhouse experiments, and (iii) the relationships between relative leaf chlorophyll content (RLCC) and relative cucumber dry top weight biomass (RDTWB) in relation to increasing nematode densities at planting (P_i) in pot experiments. Thermal requirements of M. incognita and M. javanica on cucumber did not differ, irrespective of the biological stage. In the pot experiments, M. javanica completed one generation. The maximum multiplication rate (a) was 833, and the equilibrium density (E) varied according to the effective inoculum densities. The relationship between RDTWB and P_i fitted the Seinhorst damage function model. The RLCC value at 40 or 50 days post‐inoculation also fitted the damage model and was related to RDTWB. In greenhouse experiments, conducted from 2009 to 2012, M. incognita completed three generations. The values for a and E were 1147 and 625 second stage juveniles (J2) per 250 cm³ soil, respectively. The tolerance limit was below zero, and the minimum relative yield ranged from 0·12 to 0·34.     

Genetic diversity of the root‐knot nematode Meloidogyne enterolobii and development of a SCAR marker for this guava‐damaging species

The objectives of this work were to evaluate the genetic variability of Meloidogyne enterolobii by molecular markers, and develop species‐specific molecular markers for application in detection. Sixteen M. enterolobii isolates from different geographical regions (Brazil and other countries) and hosts were used in this study. The identification and purification of the populations were carried out based on isoenzyme phenotype. The DNA amplification of the intergenic region (IGS) of the rDNA and of the region between the cytochrome oxidase subunit II (COII) and 16S rRNA genes (mtDNA) produced specific fragments of the expected size for this nematode, i.e. 780 and 705 bp, respectively. Intraspecific variability among the isolates was evaluated with three different neutral molecular markers: AFLP, ISSR and RAPD. The results showed a low level of diversity among the isolates tested, indicating that M. enterolobii is a genetically homogeneous root‐knot nematode species. The RAPD method allowed the identification of a species‐specific RAPD fragment for M. enterolobii. This fragment was cloned and sequenced, and from the sequence obtained, a set of primers was designed and tested. The amplification of a 520‐bp‐long fragment occurred only for the 16 isolates of M. enterolobii and not for the 10 other Meloidogyne species tested. In addition, positive detection was achieved in a single individual female, egg‐mass and second stage juvenile of this nematode. This SCAR species‐specific marker for M. enterolobii represents a new molecular tool to be used in the detection of this nematode from field samples and as a routine diagnostic test for quarantine devices .     

Meloidogyne luci,a new root‐knot nematode parasitizing potato in Portugal

In 2013, during a field survey conducted in Portugal on potato, Solanum tuberosum, an unusual esterase (EST) phenotype was detected in a root‐knot nematode (RKN) from potato roots collected in Coimbra. This Portuguese isolate was purified and maintained on tomato, S. lycopersicum, and morphological, biochemical and molecular characteristics were studied. Perineal pattern morphology was highly variable, similar to Meloidogyne ethiopica and not useful for identification. The EST phenotype, from young egg‐laying females, displayed three bands similar to the Brazilian M. luci (L3) and distinct from M. ethiopica (E3). Phylogenetic analyses of mitochondrial cytochrome oxidase subunit I and the mitochondrial DNA region between COII and 16S rRNA genes revealed that the Portuguese isolate grouped with M. luci isolates close to M. ethiopica isolates. However, considering the ITS1‐5.8S‐ITS2 region, the Portuguese isolate grouped with isolates of M. luci, M. ethiopica and M. hispanica, which limits the confidence of this region for M. luci diagnosis, and its differentiation from other species with morphological similarities. The M. luci pathogenicity to potato was also assessed in 16 commercial cultivars and compared with M. chitwoodi, considered to be a quarantine RKN species by EPPO. All potato cultivars were susceptible to both Meloidogyne species with gall indices of 5 and higher reproduction factor values ranging from 12.5 to 122.3, which suggests that M. luci may constitute a potential threat to potato production. In the present study, M. luci is reported for the first time attacking potato in Portugal.     

Comparison of two short DNA barcoding loci (COI and COII) and two longer ribosomal DNA genes (SSU & LSU rRNA) for specimen identification among quarantine root-knot nematodes (Meloidogyne spp.) and their close relatives

Root-knot nematodes (Meloidogyne spp.) are important pests of numerous crops worldwide. Some members of this genus have a quarantine status, and accurate species identification is required to prevent further spreading. DNA barcoding is a method for organism identification in non-complex DNA backgrounds based on informative motifs in short DNA stretches (≈600 bp). As part of the EU 7th Framework project QBOL, 15 Meloidogyne species were chosen to compare the resolutions offered by two typical DNA barcoding loci, COI and COII, with the distinguishing signals produced by two ribosomal DNA genes (small and large subunit rDNA; SSU?≈?1,700 and LSU?≈?3,400 bp). None of the four markers distinguished between the tropical species Meloidogyne incognita, M. javanica and M. arenaria. Taking P ID (Liberal) values ≥0.93 as a measure for species delimitation, the four mtDNA and rDNA markers performed well for the tropical Meloidogyne species complex, M. enterolobii, M. hapla, and M. maritima. Within cluster III A (Holterman et al. Phytopathology, 99, 227–235, 2009), SSU rDNA did not offer resolution at species level. Both mtDNA loci COI and COII did, whereas for LSU rDNA a longer fragment (≥700 bp) is required. The high level of mitochondrial heteroplasmy recently reported for M. chitwoodi (Humphreys-Pereira and Elling Nematology, 15, 315–327, 2013) was not found in the populations under investigation, suggesting this could be a regional phenomenon. For identification of RKNs, we suggest the combined use of SSU rDNA with one of three other markers presented here.     

Differential reproduction of Meloidogyne incognita and M. javanica in watermelon cultivars and cucurbit rootstocks

The suitability of watermelon cultivars and cucurbit rootstocks as hosts to Meloidogyne incognita and M. javanica was determined in pot and field experiments. Meloidogyne incognita showed higher reproduction than did M. javanica on watermelon and cucurbit rootstocks. The watermelon cultivars did not differ in host status when challenged with these two species and supported lower nematode reproduction than the cucurbit rootstocks. Rootstocks Lagenaria siceraria cv. Pelops and Cucurbita pepo AK15 supported lower reproduction than did the squash hybrid rootstocks (C. maxima × C. moschata). Egg production increased (P < 0·05) with a rising initial inoculum level (Pi) in the non‐grafted Sugar Baby but the reproduction factor Rf (eggs per plant/Pi) was similar at two Pi levels. The total egg production in the plants grafted onto squash hybrids RS841 and Titan was greater (P < 0·05) at the higher Pi, but the Rf values were lower. The development of field‐grown non‐grafted watermelon plants was significantly stunted in plots where nematodes were detected at planting. However, no differences were observed in plots with grafted plants. In plots with nematodes, non‐grafted and Titan‐grafted plants had similar yields that were higher than that of RS841‐grafted plants. In the commercial plastic houses with grafted watermelon, the average Rf value was 42‐fold, confirming the high susceptibility of squash hybrids as rootstocks for grafted watermelon. The Titan–Sugar Baby combination was tolerant to M. javanica.     

Interaction of Root-knot Nematodes (RKN) and the Bacterium Agrobacterium Tumefaciens in Roots of Prunus Cerasifera: Evidence of the Protective Effect of the Ma RKN Resistance Genes Against Expression of Crown Gall Symptoms

Agrobacterium tumefaciens (AT) is the causal agent of crown gall, a major problem in the family Rosaceae and particularly for Prunus spp. Crown gall symptoms result from the bacterial infection of the cells damaged mechanically at the collar or by root parasitic nematodes. Myrobalan plum (P. cerasifera) is susceptible to AT and is not a host for the root-knot nematode (RKN), M. hapla. Some clones of this plum carry single Ma resistance genes that control M. arenaria, M. incognita and M. javanica. The four above mentioned RKN and Myrobalan progenies segregating for Ma were used in experiments aimed at obtaining a better knowledge of the interaction between AT and RKN in relation to the RKN resistance genes. Prunus rooted cuttings, naturally infected with the bacterium were repotted, grown and inoculated individually with RKN. In a first experiment, Prunus plants were (i) either inoculated with 10,000 juveniles (J2s) of M. arenaria to provide a short inoculum pressure (SIP) or (ii) inoculated by association with one M. arenaria-galled tomato root system that produced a high and durable inoculum pressure of the same nematode species. Four months after RKN inoculation, plants were rated for nematode and bacterial root galling symptoms. RKN and AT galls were more numerous and more homogenous under DIP than under SIP. Nevertheless, for both inoculum regimes, AT galls were present in the RKN-susceptible clones (= carrying none of the Ma genes) and absent in the RKN-resistant clones. Subsequent experiments, conducted under DIP with M. arenaria, M. incognita, M. javanica and M. hapla, also showed, for the three first species, the presence of AT galls only in RKN-susceptible clones whereas Prunus plants inoculated with M. hapla and nematode-free controls were free of AT galls. Consequently RKN act as a wound agent in the AT infection process of Myrobalan plum only when the plant develops a compatible reaction (i.e. when it lacks the Ma resistance genes). Considering that J2s do penetrate the roots of resistant plants, the absence of crown gall symptoms on this material even under durable inoculum pressure strengthens the hypothesis that this nematode stage has a very weak effect on plant cells during the infection process. This is the first evidence of the protective effect of a RKN resistance gene against the expression of root crown gall consecutive to RKN infection. The protective effect of Ma and presumably of other RKN resistance genes against AT is a strong argument for their introgression into Prunus and other Rosaceae or bacterium-susceptible crops.