Host Suitability of Lettuce and Bean Germplasm for Meloidogyne incognita and M. javanica Isolates from Spain.

Meloidogyne spp. are an important threat to horticulture and cause substantial yield losses. Plant resistance is an alternative control method for chemical nematicides. This study highlights the host suitability of the lettuces cultivars Grand Rapids and Salinas 88 and the beans cultivars Aporé, Cornell 49242, Macarrão Atibaia and Ouro Negro to four Meloidogyne incognita and seven M. javanica isolates from Spain in a pot experiment. Moreover, the response of these cultivars to increasing M. incognita densities (Pi) was assessed in a plastic greenhouse. The lettuce cultivar Regina 71 and the bean cultivar Bolinha were included as susceptible standards for comparison. It was found that Grand Rapids and Salinas 88 lettuces were resistant to the most nematode isolates in the pot experiment but were classified as slightly and moderately resistant, respectively, in the plastic greenhouse at increasing Pi. Regarding the beans, Aporé was resistant to the majority of the Meloidogyne isolates whereas Macarrão Atibaia and Ouro Negro were slightly resistant and Cornell 49242 was susceptible in the pot experiment. In the plastic greenhouse, Aporé was the only cultivar able to effectively suppress the nematode reproduction irrespective of Pi, while Ouro Negro became less resistant as Pi increased. These results play an important role in enhancing the effective and ecofriendly Meloidogyne management strategies.

Crop rotation with Meloidogyne-resistant germplasm is useful to manage and revert the (a)virulent populations of Mi1.2 gene and reduce yield losses.

A rotation sequence of ungrafted and grafted tomato-melon-pepper-watermelon on resistant rootstocks 'Brigeor', Cucumis metuliferus, 'Oscos' and Citrullus amarus, respectively, was carried out in a plastic greenhouse, ending with a susceptible or resistant tomato crop. The rotation was conducted in plots infested by an avirulent (Avi) or a partially virulent (Vi) Meloidogyne incognita population to the Mi1.2 gene. At the beginning of the study, the reproduction index (RI, relative reproduction in the resistant respect susceptible tomato) of Avi and Vi populations was 1.3% and 21.6%, respectively. Soil nematode density at transplanting (Pi) and at the end (Pf) of each crop, disease severity and crop yield were determined. Moreover, the putative virulence selection and fitness cost were determined at the end of each crop in pot tests. In addition, a histopathological study was carried out 15 days after nematode inoculation in pot test. The volume and number of nuclei per giant cell (GC) and the number of GC, their volume and the number of nuclei per feeding site in susceptible watermelon and pepper were compared with C. amarus and resistant pepper. At the beginning of the study, the Pi of Avi and Vi plots did not differ between susceptible and resistant germplasm. At the end of the rotation, the Pf of Avi was 1.2 the Pi in susceptible and 0.06 in resistant, the cumulative yield of grafted crops was 1.82 times higher than that of the ungrafted susceptible ones, and the RI in resistant tomato less than 10% irrespective of the rotation sequence. Concerning the Vi, Pf was below the detection level at the end of the rotation in resistant and 3 times Pi in the susceptible. The cumulative yield of grafted crops was 2.83 times higher than that of the ungrafted and the RI in resistant tomato was 7.6%, losing the population's virulence. In the histopathological study, no differences in number of GC per feeding site were observed in watermelon compared to C. amarus, but they were more voluminous and contained higher number of nuclei per GC and per feeding site. Regarding pepper, Avi population did not penetrate resistant rootstock.

Tomato and Melon Meloidogyne Resistant Rootstocks Improve Crop Yield but Melon Fruit Quality Is Influenced by the Cropping Season.

Four rotation sequences consisting of ungrafted tomato cv. Durinta - melon cv. Paloma or tomato grafted onto the resistant rootstock 'Aligator' - melon grafted onto the resistant Cucumis metuliferus accession BGV11135, and in reverse order, were conducted from 2015 to 2017 in a plastic greenhouse infested or not with Meloidogyne incognita to determine the plant tolerance (T), the minimum relative crop yield (m) and fruit quality. The relationship between M. incognita densities in soil at transplanting (Pi) of each crop and the crop yield was assessed and T and m were estimated by the Seinhorst's damage model. In addition, the volume and the number of nuclei of single giant cells and the number of giant cells, its volume and the number of nuclei per feeding site in susceptible tomato and melon were compared to those in the resistant tomato and C. metuliferus 15 days after nematode inoculation in pot test. The relationship between the Pi and the relative crop yield fitted the Seinhorst's damage model in both ungrafted and grafted tomato and melon, but not for all years and cropping seasons. The estimated T for ungrafted and grafted tomato did not differ but m was lower in the former (34%) than the latter (67%). Sodium concentration in fruits from ungrafted but not from grafted tomato increased with nematode densities in spring 2015 and 2016. The estimated ungrafted melon T did not differ from the grafted melon cultivated in spring, but it did when it was cultivated in summer. The relative crop yield of ungrafted melon was lower (2%) than the grafted cultivated in spring (62%) and summer (20%). Sodium concentration in melon fruits from ungrafted plants increased with nematode densities. No variations in fruit quality from grafted melon cultivated in spring were found, although less dry matter and soluble solid content at highest nematode densities were registered when it was cultivated in summer. Lower number of giant cells per feeding site was observed in both susceptible tomato germplasms compared to the resistant ones but they were more voluminous and held higher number of nuclei per giant cell and per feeding site.

Cucumis metuliferus reduces Meloidogyne incognita virulence against the Mi1.2 resistance gene in a tomato-melon rotation sequence.

BACKGROUND: Susceptible tomato cv. Durinta, ungrafted or grafted onto cv. Aligator resistant rootstock, both followed by the susceptible melon cv. Paloma, ungrafted or grafted onto Cucumis metuliferus BGV11135, and in the reverse order, were cultivated from 2015 to 2017 in the same plots in a plastic greenhouse, infested or not with Meloidogyne incognita. For each crop, soil nematode densities, galling index, number of eggs per plant and crop yield were determined. Virulence selection was evaluated in pot experiments. RESULTS: In the tomato-melon rotation, nematode densities increased progressively for the grafted tomato, being higher than for ungrafted plants at the end of the study; this was not the case in the melon-tomato rotation. Grafted crops yielded more than ungrafted crops in the infested plots. Virulence against the Mi1.2 gene was detected, but not against C. metuliferus. Reproduction of M. incognita on the resistant tomato was ∼ 120% that on the susceptible cultivar after the first grafted tomato crop, but this decreased to just 25% at the end of the experiment. CONCLUSION: Alternating different resistant plant species suppresses nematode population growth rate and yield losses. Although this strategy does not prevent virulence selection, the level was reduced. © 2018 Society of Chemical Industry.

Effect of plant resistance and BioAct WG (Purpureocillium lilacinum strain 251) on Meloidogyne incognita in a tomato-cucumber rotation in a greenhouse.

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.

Microbiomes associated with infective stages of root-knot and lesion nematodes in soil.

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.

Characterization of Soil Suppressiveness to Root-Knot Nematodes in Organic Horticulture in Plastic Greenhouse.

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 10 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 (0.2-6.3). 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. P. 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.