Dataset Information

Developing tools for evaluating inoculation methods of biocontrol Streptomyces sp. strains into grapevine plants.

ABSTRACT: The endophytic Streptomyces sp. VV/E1, and rhizosphere Streptomyces sp. VV/R4 strains, isolated from grapevine plants were shown in a previous work to reduce the infection rate of fungal pathogens involved in young grapevine decline. In this study we cloned fragments from randomly amplified polymorphic DNA (RAPD), and developed two stably diagnostic sequence-characterized amplified region (SCAR) markers of 182 and 160 bp for the VV/E1 and VV/R4 strains, respectively. The SCAR markers were not found in another 50 actinobacterial strains isolated from grapevine plants. Quantitative real-time PCR protocols based on the amplification of these SCAR markers were used for the detection and quantification of both strains in plant material. These strains were applied on young potted plants using two methods: perforation of the rootstock followed by injection of the microorganisms or soaking the root system in a bacterial suspension. Both methods were combined with a booster treatment by direct addition of a bacterial suspension to the soil near the root system. Analysis of uprooted plants showed that those inoculated by injection exhibited the highest rate of colonization. In contrast, direct addition of either strain to the soil did not lead to reliable colonization. This study has developed molecular tools for analyzing different methods for inoculating grapevine plants with selected Streptomyces sp. strains which protect them from fungal infections that enter through their root system. These tools are of great applied interest since they could easily be established in nurseries to produce grafted grapevine plants that are protected against fungal pathogens. Finally, this methodology might also be applied to other vascular plants for their colonization with beneficial biological control agents.

SUBMITTER: Gonzalez-Garcia S

PROVIDER: S-EPMC6345443 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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Developing tools for evaluating inoculation methods of biocontrol Streptomyces sp. strains into grapevine plants.

González-García Sandra S Álvarez-Pérez Jose Manuel JM Sáenz de Miera Luis E LE Cobos Rebeca R Ibañez Ana A Díez-Galán Alba A Garzón-Jimeno Enrique E Coque Juan José R JJR

PloS one 20190124 1

The endophytic Streptomyces sp. VV/E1, and rhizosphere Streptomyces sp. VV/R4 strains, isolated from grapevine plants were shown in a previous work to reduce the infection rate of fungal pathogens involved in young grapevine decline. In this study we cloned fragments from randomly amplified polymorphic DNA (RAPD), and developed two stably diagnostic sequence-characterized amplified region (SCAR) markers of 182 and 160 bp for the VV/E1 and VV/R4 strains, respectively. The SCAR markers were not fo ...[more]

PMID: 30677098

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Project description:Microorganisms related to plant roots are vital for plant growth and health and considered to be the second genome of the plant. When the plant is attacked by plant pathogens, the diversity and community structure of plant-associated microbes might be changed. The goal of this study is to characterize differences in root-associated endophytic actinobacterial community composition and antifungal activity between Fusarium wilt diseased and healthy cucumber and screen actinobacteria for potential biological control of Fusarium wilt of cucumber. In the present research, three healthy plants (also termed "islands") and three obviously diseased plants (naturally infected by F. oxysporum f. sp. cucumerinum) nearby the islands collected from the cucumber continuous cropping greenhouse were chosen as samples. Results of culture-independent and culture-dependent analysis demonstrated that actinomycetes in the healthy roots were significantly more abundant than those of diseased roots. Moreover, there were seven strains with antifungal activity against F. oxysporum f. sp. cucumerinum in healthy cucumber roots, but only one strain in diseased cucumber roots. Out of these eight strains, the isolate HAAG3-15 was found to be best as it had the strongest antifungal activity against F. oxysporum f. sp. cucumerinum, and also exhibited broad-spectrum antifungal activity. Thus, strain HAAG3-15 was selected for studying its biocontrol efficacy under greenhouse conditions. The results suggested that the disease incidence and disease severity indices of cucumber Fusarium wilt greatly decreased (p < 0.05) while the height and shoot fresh weight of cucumber significantly increased (p < 0.05) after inoculating strain HAAG3-15. On the basis of morphological characteristics, physiological and biochemical properties and 100% 16S ribosomal RNA (rRNA) gene sequence similarity with Streptomyces sporoclivatus NBRC 100767T, the isolate was assigned to the genus Streptomyces. Moreover, azalomycin B was isolated and identified as the bioactive compound of strain HAAG3-15 based on analysis of spectra using a bioactivity-guided method. The stronger antifungal activity against F. oxysporum f. sp. cucumerinum, the obvious effect on disease prevention and growth promotion on cucumber seedlings in the greenhouse assay, and the excellent broad-spectrum antifungal activities suggest that strain HAAG3-15 could be developed as a potential biocontrol agent against F. oxysporum f. sp. cucumerinum used in organic agriculture. These results suggested that the healthy root nearby the infected plant is a good source for isolating biocontrol and plant growth-promoting endophytes.

Dataset Information

Developing tools for evaluating inoculation methods of biocontrol Streptomyces sp. strains into grapevine plants.

Publications

Developing tools for evaluating inoculation methods of biocontrol Streptomyces sp. strains into grapevine plants.

Similar Datasets

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