Dataset Information

Specific microbial attachment to root knot nematodes in suppressive soil.

ABSTRACT: Understanding the interactions of plant-parasitic nematodes with antagonistic soil microbes could provide opportunities for novel crop protection strategies. Three arable soils were investigated for their suppressiveness against the root knot nematode Meloidogyne hapla. For all three soils, M. hapla developed significantly fewer galls, egg masses, and eggs on tomato plants in unsterilized than in sterilized infested soil. Egg numbers were reduced by up to 93%. This suggested suppression by soil microbial communities. The soils significantly differed in the composition of microbial communities and in the suppressiveness to M. hapla. To identify microorganisms interacting with M. hapla in soil, second-stage juveniles (J2) baited in the test soil were cultivation independently analyzed for attached microbes. PCR-denaturing gradient gel electrophoresis of fungal ITS or 16S rRNA genes of bacteria and bacterial groups from nematode and soil samples was performed, and DNA sequences from J2-associated bands were determined. The fingerprints showed many species that were abundant on J2 but not in the surrounding soil, especially in fungal profiles. Fungi associated with J2 from all three soils were related to the genera Davidiella and Rhizophydium, while the genera Eurotium, Ganoderma, and Cylindrocarpon were specific for the most suppressive soil. Among the 20 highly abundant operational taxonomic units of bacteria specific for J2 in suppressive soil, six were closely related to infectious species such as Shigella spp., whereas the most abundant were Malikia spinosa and Rothia amarae, as determined by 16S rRNA amplicon pyrosequencing. In conclusion, a diverse microflora specifically adhered to J2 of M. hapla in soil and presumably affected female fecundity.

SUBMITTER: Adam M

PROVIDER: S-EPMC3993313 | biostudies-literature | 2014 May

REPOSITORIES: biostudies-literature

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Specific microbial attachment to root knot nematodes in suppressive soil.

Adam Mohamed M Westphal Andreas A Hallmann Johannes J Heuer Holger H

Applied and environmental microbiology 20140214 9

Understanding the interactions of plant-parasitic nematodes with antagonistic soil microbes could provide opportunities for novel crop protection strategies. Three arable soils were investigated for their suppressiveness against the root knot nematode Meloidogyne hapla. For all three soils, M. hapla developed significantly fewer galls, egg masses, and eggs on tomato plants in unsterilized than in sterilized infested soil. Egg numbers were reduced by up to 93%. This suggested suppression by soil ...[more]

PMID: 24532076

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Project description: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.

Project description: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.

Project description:The utilization of Trichoderma longibrachiatum filtrate as a safe biocontrol method for producing zinc nanoparticles is a promising approach for managing pests and diseases in agricultural crops. The identification of Trichoderma sp. was achieved through PCR amplification and sequencing of 18s as ON203115, while the synthesis of ZnO-NPs was accomplished by employing Trichoderma filtration. The presence of ZnO-NPs was confirmed by observing a color change to dark green, along with the use of visible and UV spectrophotometers, and the formation and chemical structure of ZnO-NPs were examined. Direct exposure to ZnO-NPs exhibited a significant inhibitory effect on the growth of Fusarium oxysporum at 80.73% compared with control. Also, the percent mortality of Meloidogyne incognita second juveniles stage (J2s) results showed 11.82%, 37.63%, 40.86%, and 89.65% after 6, 12, 24, and 72 h, respectively in vitro. Disease resistance was assessed in the greenhouse against M. incognita and F. oxysporum using the drench application of ZnO-NPs. The application of ZnO-NPs significantly reduced the disease severity of F. oxysporum and improved the quality and quantity of sweet pepper yield. In addition, the application of ZnO-NPs to M. incognita resulted in a significant reduction in the number of nematode galls, egg masses per root, eggs/egg mass, and females by 98%, 99%, 99.9%, and 95.5% respectively.Furthermore, it was observed that the application of ZnO-NPs to pepper plants not only inhibited the growth of F. oxysporum and M. incognita, but also promoted the recovery of pepper plants as indicated by improvements in stem length by 106%, root length 102%, fresh weight 112%, root fresh weight 107%, and leaf area 118% compared to healthy control plants. Additionally, real-time PCR application and DD-PCR technique revealed that the application of ZnO-NPs stimulated the secretion of certain enzymes. These findings suggest that the biosynthesized ZnO-NPs possess anti-nematode and antifungal properties, making them effective for protecting plants against M. incognita and F. oxysporum invasion in soil. This study significantly contributes to our understanding of the nematicidal and fungicidal activities of ZnO-NPs in suppressing soil-borne diseases.

Dataset Information

Specific microbial attachment to root knot nematodes in suppressive soil.

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Specific microbial attachment to root knot nematodes in suppressive soil.

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