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

Autor: Sandra González-García, José Manuel Álvarez-Pérez, Rebeca Cobos, Juan José R. Coque, Ana B. Ibáñez, Enrique Garzón-Jimeno, Alba Diez-Galán, Luis E. Sáenz de Miera
Rok vydání: 2019
Předmět:
0301 basic medicine
Artificial Gene Amplification and Extension
Plant Science
Pathology and Laboratory Medicine
Polymerase Chain Reaction
Plant Roots
Medicine and Health Sciences
Endophytes
Methods
Vitis
Colonization
Flowering Plants
Fungal Pathogens
Rhizosphere
Multidisciplinary
Ecology
Plant Anatomy
Eukaryota
Plants
Wood
Streptomyces
Random Amplified Polymorphic DNA Technique
RAPD
Biological Control Agents
Medical Microbiology
Medicine
Grapevine
Pathogens
Rootstock
Research Article
Science
030106 microbiology
Perforation (oil well)
Mycology
Biology
Research and Analysis Methods
Biomolecular isolation
Microbiology
03 medical and health sciences
Plant-Environment Interactions
Molecular Biology Techniques
Microbial Pathogens
Molecular Biology
Plant Diseases
Bacteria
Inoculation
Plant Ecology
Ecology and Environmental Sciences
Organisms
Biology and Life Sciences
Agricultural Inoculants
biology.organism_classification
DNA isolation
030104 developmental biology
Zdroj: PLoS ONE, Vol 14, Iss 1, p e0211225 (2019)
PLoS ONE
ISSN: 1932-6203
Popis: 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.
Databáze: OpenAIRE
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