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      Influence of vintage, geographic location and cultivar on the structure of microbial communities associated with the grapevine rhizosphere in vineyards of San Juan Province, Argentina

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          Abstract

          Soil microbiomes, as a primary reservoir for plant colonizing fungi and bacteria, play a major role in determining plant productivity and preventing invasion by pathogenic microorganisms. The use of 16S rRNA and ITS high-throughput amplicon sequencing for analysis of complex microbial communities have increased dramatically in recent years, establishing links between wine specificity and, environmental and viticultural factors, which are framed into the elusive terroir concept. Given the diverse and complex role these factors play on microbial soil structuring of agricultural crops, the main aim of this study is to evaluate how external factors, such as vintage, vineyard location, cultivar and soil characteristics, may affect the diversity of the microbial communities present. Additionally, we aim to compare the influence these factors have on the structuring of bacterial and fungal populations associated with Malbec grapevine rhizosphere with that of the more widespread Cabernet Sauvignon grapevine cultivar. Samples were taken from Malbec and Cabernet Sauvignon cultivars from two different vineyards in the San Juan Province of Argentina. Total DNA extracts from the rhizosphere soil samples were sequenced using Illumina’s Miseq technology, targeting the V3-V4 hypervariable 16S rRNA region in prokaryotes and the ITS1 region in yeasts. The major bacterial taxa identified were Proteobacteria, Bacteroidetes and Firmicutes, while the major fungal taxa were Ascomycetes, Basidiomycetes, Mortierellomycetes and a low percentage of Glomeromycetes. Significant differences in microbial community composition were found between vintages and vineyard locations, whose soils showed variances in pH, organic matter, and content of carbon, nitrogen, and absorbable phosphorus.

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          QIIME allows analysis of high-throughput community sequencing data.

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            UPARSE: highly accurate OTU sequences from microbial amplicon reads.

            Amplified marker-gene sequences can be used to understand microbial community structure, but they suffer from a high level of sequencing and amplification artifacts. The UPARSE pipeline reports operational taxonomic unit (OTU) sequences with ≤1% incorrect bases in artificial microbial community tests, compared with >3% incorrect bases commonly reported by other methods. The improved accuracy results in far fewer OTUs, consistently closer to the expected number of species in a community.
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              Search and clustering orders of magnitude faster than BLAST.

              Biological sequence data is accumulating rapidly, motivating the development of improved high-throughput methods for sequence classification. UBLAST and USEARCH are new algorithms enabling sensitive local and global search of large sequence databases at exceptionally high speeds. They are often orders of magnitude faster than BLAST in practical applications, though sensitivity to distant protein relationships is lower. UCLUST is a new clustering method that exploits USEARCH to assign sequences to clusters. UCLUST offers several advantages over the widely used program CD-HIT, including higher speed, lower memory use, improved sensitivity, clustering at lower identities and classification of much larger datasets. Binaries are available at no charge for non-commercial use at http://www.drive5.com/usearch.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: SoftwareRole: Writing – original draftRole: Writing – review & editing
                Role: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: ResourcesRole: SoftwareRole: Writing – original draftRole: Writing – review & editing
                Role: Data curationRole: Formal analysisRole: MethodologyRole: SoftwareRole: Writing – review & editing
                Role: MethodologyRole: ResourcesRole: ValidationRole: Writing – review & editing
                Role: MethodologyRole: Project administrationRole: SupervisionRole: Writing – review & editing
                Role: ConceptualizationRole: SupervisionRole: Writing – review & editing
                Role: ConceptualizationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: SupervisionRole: Writing – original draftRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS One
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                14 December 2020
                2020
                : 15
                : 12
                : e0243848
                Affiliations
                [1 ] Instituto de Biotecnología y Biología Molecular (IBBM), CCT-La Plata, CONICET, Dto de Cs. Biológicas, Fac. Cs. Exactas, UNLP, La Plata, Argentina
                [2 ] Section of Microbial Ecology and Biotechnology, Department of Plant and Environmental Sciences, University of Copenhagen, Frederiksberg, Denmark
                [3 ] Department of Environmental Science, Aarhus University, Roskilde, Denmark
                [4 ] Laboratorio de Microbiología Molecular, Departamento de Ciencia y Tecnología, Instituto de Microbiología Básica y Aplicada, Universidad de Nacional de Quilmes (UNQ), Bernal, Buenos Aires, Argentina
                Institute for Sustainable Plant Protection, C.N.R., ITALY
                Author notes

                Competing Interests: The authors declare no competing interests.

                Author information
                https://orcid.org/0000-0003-3541-4732
                https://orcid.org/0000-0002-7235-9026
                https://orcid.org/0000-0001-7931-8721
                Article
                PONE-D-20-24593
                10.1371/journal.pone.0243848
                7735631
                33315910
                7e0ed427-7134-45e2-b5ef-3fa05a14fc7e
                © 2020 Oyuela Aguilar et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 6 August 2020
                : 29 November 2020
                Page count
                Figures: 4, Tables: 0, Pages: 19
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/100010661, Horizon 2020 Framework Programme;
                Award ID: 643063
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100006668, Fondo para la Investigación Científica y Tecnológica;
                Award ID: PICT-2017-2833
                Award Recipient :
                This study was funded by the Horizon 2020 Programme of the European Commission within the Marie Skłodowska-Curie Innovative Training Network “MicroWine” (grant number 643063), and by Agencia Nacional de Promoción Científica y Técnica PICT-2017-2833 to MP. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Ecology
                Plant Ecology
                Plant-Environment Interactions
                Rhizosphere
                Ecology and Environmental Sciences
                Ecology
                Plant Ecology
                Plant-Environment Interactions
                Rhizosphere
                Biology and Life Sciences
                Plant Science
                Plant Ecology
                Plant-Environment Interactions
                Rhizosphere
                Biology and Life Sciences
                Organisms
                Bacteria
                Biology and Life Sciences
                Agriculture
                Agricultural Soil Science
                Earth Sciences
                Soil Science
                Agricultural Soil Science
                Biology and Life Sciences
                Organisms
                Eukaryota
                Plants
                Flowering Plants
                Grapevine
                Biology and Life Sciences
                Microbiology
                Medical Microbiology
                Microbiome
                Biology and Life Sciences
                Genetics
                Genomics
                Microbial Genomics
                Microbiome
                Biology and Life Sciences
                Microbiology
                Microbial Genomics
                Microbiome
                Biology and life sciences
                Biochemistry
                Nucleic acids
                RNA
                Non-coding RNA
                Ribosomal RNA
                Biology and life sciences
                Biochemistry
                Ribosomes
                Ribosomal RNA
                Biology and life sciences
                Cell biology
                Cellular structures and organelles
                Ribosomes
                Ribosomal RNA
                Biology and Life Sciences
                Mycology
                Fungal Structure
                Biology and Life Sciences
                Nutrition
                Diet
                Beverages
                Alcoholic Beverages
                Wine
                Medicine and Health Sciences
                Nutrition
                Diet
                Beverages
                Alcoholic Beverages
                Wine
                Custom metadata
                The authors confirm that the sequencing data have been deposited in the Sequence Read Archive (SRA) ( https://www.ncbi.nlm.nih.gov/sra) and are accessible through the study accession number PRJNA640285.

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                Uncategorized

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