Endoparasitic root-knot (Meloidogyne spp.) and lesion (Pratylenchus spp.) nematodes cause considerable damage in agriculture. Before they invade roots to complete their life cycle, soil microbes can attach to their cuticle or surface coat and antagonize the nematode directly or by induction of host plant defenses. We investigated whether the nematode-associated microbiome in soil differs between infective stages of Meloidogyne incognita and Pratylenchus penetrans, and whether it is affected by variation in the composition of microbial communities among soils. Nematodes were incubated in suspensions of five organically and two integrated horticultural production soils, recovered by sieving and analyzed for attached bacteria and fungi after washing off loosely adhering microbes. Significant effects of the soil type and nematode species on nematode-associated fungi and bacteria were revealed as analyzed by community profiling using denaturing gradient gel electrophoresis. Attached microbes represented a small specific subset of the soil microbiome. Two organic soils had very similar bacterial and fungal community profiles, but one of them was strongly suppressive towards root-knot nematodes. They were selected for deep amplicon sequencing of bacterial 16S rRNA genes and fungal ITS. Significant differences among the microbiomes associated with the two species in both soils suggested specific surface epitopes. Among the 28 detected bacterial classes, Betaproteobacteria, Bacilli and Actinobacteria were the most abundant. The most frequently detected fungal genera were Malassezia, Aspergillus and Cladosporium. Attached microbiomes did not statistically differ between these two soils. However, Malassezia globosa and four fungal species of the family Plectosphaerellaceae, and the bacterium Neorhizobium galegae were strongly enriched on M. incognita in the suppressive soil. In conclusion, the highly specific attachment of microbes to infective stages of phytonematodes in soil suggested an ecological role of this association and might be involved in soil suppressiveness towards them.

The fluctuation of Meloidogyne population density and the percentage of fungal egg parasitism were determined from July 2011 to July 2013 in two commercial organic vegetable production sites (M10.23 and M10.55) in plastic greenhouses, located in northeastern Spain, in order to know the level of soil suppressiveness. Fungal parasites were identified by molecular methods. In parallel, pot tests characterized the level of soil suppressiveness and the fungal species growing from the eggs. In addition, the egg parasitic ability of ten fungal isolates per site was also assessed. The genetic profiles of fungal and bacterial populations from M10.23 and M10.55 soils were obtained by Denaturing Gradient Gel Electrophoresis (DGGE), and compared with a non-suppressive soil (M10.33). In M10.23, Meloidogyne population in soil decreased progressively throughout the rotation zucchini, tomato, and radish or spinach. The percentage of egg parasitism was 54.7% in zucchini crop, the only one in which eggs were detected. Pochonia chlamydosporia was the only fungal species isolated. In M10.55, nematode densities peaked at the end of the spring-summer crops (tomato, zucchini, and cucumber), but disease severity was lower than expected (0.2 to 6.3). The percentage of fungal egg parasitism ranged from 3 to 84.5% in these crops. The results in pot tests confirmed the suppressiveness of the M10.23 and M10.55 soils against Meloidogyne. The number of eggs per plant and the reproduction factor of the population were reduced (P < 0.05) in both non-sterilized soils compared to the sterilized ones after one nematode generation. Pochonia chlamydosporia was the only fungus isolated from Meloidogyne eggs. In in vitro tests, P. chlamydosporia isolates were able to parasitize Meloidogyne eggs from 50 to 97% irrespective of the site. DGGE fingerprints revealed a high diversity in the microbial populations analyzed. Furthermore, both bacterial and fungal genetic patterns differentiated suppressive from non-suppressive soils, but the former showed a higher degree of similarity between both suppressive soils than the later.

BACKGROUND: The effectiveness of combining resistant tomato with BioAct WG (Purpureocillium lilacinum strain 251, Pl251) against Meloidogyne incognita was assessed in a tomato‐cucumber rotation in a greenhouse over 2 years. Additionally, the enzymatic activity of the fungus, the percentage of fungal egg and juvenile parasitism, cardinal temperatures and the effect of water potential on mycelial growth and the soil receptivity to Pl251 were determined in vitro.RESULTS: Plant resistance was the only factor that suppressed nematode and crop yield losses. Percentage of egg parasitism in plots treated with BioAct WG was less than 2.6%. However, under in vitro conditions, Pl251 showed protease, lipase and chitinase activities and parasitised 94.5% of eggs, but no juveniles. Cardinal temperatures were 14.2, 24–26 and 35.4 °C. The maximum Pl251 mycelial growth was at −0.25 MPa and 25 °C. Soil temperatures and water potential in the greenhouse were in the range of the fungus. However, soil receptivity was lower in greenhouse soil, irrespective of sterilisation, than in sterilised sand.CONCLUSION: Plant resistance was the only factor able to suppress nematode densities, disease severity and yield losses, and to protect the following cucumber crop. Environmental factors involved in soil receptivity could have negatively affected fungus effectiveness. © 2016 Society of Chemical Industry

A survey was conducted to assess the biodiversity and frequency of infection of fungal egg parasites of Meloidogyne spp. and relate results to soil properties in organic and integrated vegetable production in Spain. Forty sites were sampled at the end of the cropping cycle, 30 under integrated and ten under organic production. Fungal egg parasites were isolated from all organically managed sites and from 73 % sites under integrated production. Species richness and Shannon–Wiener index did not differ between production systems but the percentage of fungal egg parasitism did, as well as soil properties. Percentages of egg parasitism higher than 40 % were found in five and three sites under organic and integrated production, respectively. In all these sites, Pochonia chlamydosporia was present alone or co-occurring with other fungal species. The relative frequency of P. chlamydosporia was positively related to the percentage of parasitism in both production systems.

Pot experiments were carried out to characterize the response of two Cucumis metuliferus accessions ( BGV 11135 and BGV 10762) against Mi1.2 gene (a)virulent Meloidogyne arenaria , M. incognita and M. javanica isolates and to determine the compatibility and the effect on physicochemical properties of fruit melons. In addition, histopathological studies were conducted. One week after transplanting, plants were inoculated with one J2 cm ⁻³ of sterilized sand (200 cm ³ pots) and maintained in a growth chamber at 25 °C for 40 days. The susceptible cucumber cv. Dasher II or melon cv. Paloma were included for comparison. The number of egg masses and number of eggs per plant were assessed, and the reproduction index ( RI ) was calculated as the percentage of eggs produced on the C. metuliferus accessions compared to those produced on the susceptible cultivars. The compatibility and fruit quality were assessed by grafting three scions, two of Charentais type and one of type piel de sapo , under commercial greenhouse conditions. The resistance level of both C. metuliferus accessions ranged from highly resistant ( RI < 1%) to resistant (1% ≤ RI ≤ 10%) irrespective of Meloidogyne isolates. Melon plants grafted onto C. metuliferus accession BGV 11135 grew as self‐grafted plants without negatively impacting fruit quality traits. Giant cells induced by Meloidogyne spp. on C. metuliferus were in general poorly developed compared to those on cucumber. Furthermore, necrotic areas surrounding the nematode were observed. Cucumis metuliferus accession BGV 11135 could be a promising melon rootstock to manage Meloidogyne spp., irrespective of their Mi1.2 (a)virulence, without melon fruit quality reduction.