Dataset Information

Simultaneous profiling of seed-associated bacteria and fungi reveals antagonistic interactions between microorganisms within a shared epiphytic microbiome on Triticum and Brassica seeds.

ABSTRACT: In order to address the hypothesis that seeds from ecologically and geographically diverse plants harbor characteristic epiphytic microbiota, we characterized the bacterial and fungal microbiota associated with Triticum and Brassica seed surfaces. The total microbial complement was determined by amplification and sequencing of a fragment of chaperonin 60 (cpn60). Specific microorganisms were quantified by qPCR. Bacteria and fungi corresponding to operational taxonomic units (OTU) that were identified in the sequencing study were isolated and their interactions examined. A total of 5477 OTU were observed from seed washes. Neither total epiphytic bacterial load nor community richness/evenness was significantly different between the seed types; 578 OTU were shared among all samples at a variety of abundances. Hierarchical clustering revealed that 203 were significantly different in abundance on Triticum seeds compared with Brassica. Microorganisms isolated from seeds showed 99-100% identity between the cpn60 sequences of the isolates and the OTU sequences from this shared microbiome. Bacterial strains identified as Pantoea agglomerans had antagonistic properties toward one of the fungal isolates (Alternaria sp.), providing a possible explanation for their reciprocal abundances on both Triticum and Brassica seeds. cpn60 enabled the simultaneous profiling of bacterial and fungal microbiota and revealed a core seed-associated microbiota shared between diverse plant genera.

SUBMITTER: Links MG

PROVIDER: S-EPMC4235306 | biostudies-literature | 2014 Apr

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Simultaneous profiling of seed-associated bacteria and fungi reveals antagonistic interactions between microorganisms within a shared epiphytic microbiome on Triticum and Brassica seeds.

Links Matthew G MG Demeke Tigst T Gräfenhan Tom T Hill Janet E JE Hemmingsen Sean M SM Dumonceaux Tim J TJ

The New phytologist 20140121 2

In order to address the hypothesis that seeds from ecologically and geographically diverse plants harbor characteristic epiphytic microbiota, we characterized the bacterial and fungal microbiota associated with Triticum and Brassica seed surfaces. The total microbial complement was determined by amplification and sequencing of a fragment of chaperonin 60 (cpn60). Specific microorganisms were quantified by qPCR. Bacteria and fungi corresponding to operational taxonomic units (OTU) that were ident ...[more]

PMID: 24444052

Similar Datasets

Project description:: Seed-borne fungi in 69 sunflower cultivars were evaluated which comprised 52 confectionery and 17 oilseed types. Seed coats were placed on both NP-10 (Nonylphenol Ethoxylate based surfacant -10) and potato dextrose agar (PDA) media to culture fungi. The rate of contamination among the different varieties was calculated by counting seed coats with fungal colonies. The rate of contamination in the confectionary group (88%) was significantly (p ≤ 0.05) higher than in the oilseed group (71%). Of the 52 confectionery varieties, the dominant fungi recovered were Verticillium dahliae along with Alternaria spp., Fusarium spp., and Rhizopus spp., whereas the oilseed type varieties were contaminated with only V. dahliae. Molecular identification of fungal species via BLAST (Basic Alignment Search Tool) was performed on fungal sequences obtained from PCR (Polymerase Chain Reaction) analysis. The results included five Alternaria spp. that included Alternaria tenuissima, Alternaria alternata, Alternaria helianthiinficiens, Alternaria longipes, and Alternaria tamaricis, three Fusarium spp. such as Fusarium oxysporum, Fusarium incarnatum, and Fusarium proliferatum, and V. dahliae and Cladosporium cladosporioides. These were identified from pure fungal cultures recovered from seed coats. To efficiently control seed-borne fungi, four broad spectrum fungicides (carbendazim, triadimefon, caprio F-500, and flusilazole) were screened against V. dahliae isolate Gn3, which was isolated from a diseased LD 5009 sunflower plant. Flusilazole was selected based on its low half-maximal effective concentration value (EC50), 78.7 µg/mL. Seeds of diseased LD 5009 plants obtained from two different locations treated with formulated flusilazole fungicide at optimum parameters showed a significant (p ≤ 0.05) increase in seed germination and a decrease in contamination rate from 98% to less than 10%. The results affirmed that confectionery cultivars are much more susceptible to fungal contamination than oilseeds, and also that seed pretreatment is a suitable way to prevent the spread of soil- and seed-borne fungi in sunflower production.

Project description:Seed banks were first established to conserve crop genetic diversity, but seed banking has more recently been extended to wild plants, particularly crop wild relatives (CWRs) (e.g., by the Millennium Seed Bank (MSB), Royal Botanic Gardens Kew). CWRs have been recognised as potential reservoirs of beneficial traits for our domesticated crops, and with mounting evidence of the importance of the microbiome to organismal health, it follows that the microbial communities of wild relatives could also be a valuable resource for crop resilience to environmental and pathogenic threats. Endophytic fungi reside asymptomatically inside all plant tissues and have been found to confer advantages to their plant host. Preserving the natural microbial diversity of plants could therefore represent an important secondary conservation role of seed banks. At the same time, species that are reported as endophytes may also be latent pathogens. We explored the potential of the MSB as an incidental fungal endophyte bank by assessing diversity of fungi inside stored seeds. Using banana CWRs in the genus Musa as a case-study, we sequenced an extended ITS-LSU fragment in order to delimit operational taxonomic units (OTUs) and used a similarity and phylogenetics approach for classification. Fungi were successfully detected inside just under one third of the seeds, with a few genera accounting for most of the OTUs-primarily Lasiodiplodia, Fusarium, and Aspergillus-while a large variety of rare OTUs from across the Ascomycota were isolated only once. Fusarium species were notably abundant-of significance in light of Fusarium wilt, a disease threatening global banana crops-and so were targeted for additional sequencing with the marker EF1α in order to delimit species and place them in a phylogeny of the genus. Endophyte community composition, diversity and abundance was significantly different across habitats, and we explored the relationship between community differences and seed germination/viability. Our results show that there is a previously neglected invisible fungal dimension to seed banking that could well have implications for the seed collection and storage procedures, and that collections such as the MSB are indeed a novel source of potentially useful fungal strains.

Project description:BackgroundBrassica napus is of substantial economic value for vegetable oil, biofuel, and animal fodder production. The breeding of yellow-seeded B. napus to improve seed quality with higher oil content, improved oil and meal quality with fewer antinutrients merits attention. Screening the genes related to this phenotype is valuable for future rapeseed breeding.ResultsA total of 85,407 genes, including 4317 novel genes, were identified in the developing seeds of yellow- and black-seeded B. napus, and yellow rapeseed was shown to be an introgression line between black-seeded B. napus and yellow-seeded Sinapis alba. A total of 15,251 differentially expressed genes (DEGs) were identified among all the libraries, and 563 and 397 common DEGs were identified throughout black and yellow seed development, including 80 upregulated and 151 downregulated genes related to seed development and fatty acid accumulation. In addition, 11 up-DEGs and 31 down-DEGs were identified in all developmental stages of yellow rapeseed compared with black seed. Enrichment analysis revealed that many DEGs were involved in biosynthetic processes, pigment metabolism, and oxidation-reduction processes, such as flavonoid and phenylpropanoid biosynthesis, phenylalanine metabolism, flavone and flavonol biosynthesis, and fatty acid biosynthesis and metabolism. We found that more than 77 DEGs were related to flavonoid and lignin biosynthesis, including 4CL, C4H, and PAL, which participated in phenylalanine metabolism, and BAN, CHI/TT5, DFR, F3H, FLS, LDOX, PAP, CHS/TT4, TT5, bHLH/TT8, WD40, MYB, TCP, and CYP, which were involved in flavonoid biosynthesis. Most of these DEGs were downregulated in yellow rapeseed and were consistent with the decreased flavonoid and lignin contents. Both up- and down-DEGs related to fatty acid biosynthesis and metabolism were also analyzed, which could help to explain the improved oil content of yellow rapeseed.ConclusionThis research provided comprehensive transcriptome data for yellow-seeded B. napus with a unique genetic background, and all the DEGs in comparison with the black-seeded counterpart could help to explain seed quality differences, such as lower pigmentation and lignin contents, and higher oil content.

Project description:Given the challenges imposed by climate change and societal challenges, the European Union established ambitious goals as part of its Farm to Fork (F2F) strategy. Focussed on accelerating the transition to systems of sustainable food production, processing and consumption, a key element of F2F is to reduce the use of fertilisers by at least 20% and plant protection products by up to 50% by 2030. In recent years, a substantial body of research has highlighted the potential impact of microbial-based applications to support crop production practices through both biotic/abiotic stresses via maintaining or even improving yields and reducing reliance on intensive chemical inputs. Here, we have characterised the ability of a new soil-borne free-living bacterium strain Ensifer adhaerens OV14 (EaOV14) to significantly enhance crop vigour index by up to 50% for monocot (wheat, Triticum aestivum L., p < 0.0001) and by up to 40% for dicot (oilseed rape, Brassica napus L., p < 0.0001) species under in-vitro conditions (n = 360 seedlings/treatment). The beneficial effect was further studied under controlled glasshouse growing conditions (n = 60 plants/treatment) where EaOV14 induced significantly increased seed yield of spring oilseed rape compared to the controls (p < 0.0001). Moreover, using bespoke rhizoboxes, enhanced root architecture (density, roots orientation, roots thickness etc.) was observed for spring oilseed rape and winter wheat, with the median number of roots 55% and 33% higher for oilseed rape and wheat respectively, following EaOV14 seed treatment compared to the control. In addition, EaOV14 treatment increased root tip formation and root volume, suggesting the formation of a more robust root system architecture post-seed treatment. However, like other microbial formulations, the trade-offs associated with field translation, such as loss or limited functionality due to inoculum formulation or environmental distress, need further investigation. Moreover, the delivery method requires further optimisation to identify the optimal inoculum formulation that will maximise the expected beneficial impact on yield under field growing conditions.

Project description:Brassica napus is an important oilseed crop worldwide. Although seed weight is the main determinant of seed yield, few studies have focused on the molecular mechanisms that regulate seed weight in B. napus. In this study, the immature seeds of G-42 and 7-9, two B. napus doubled haploid (DH) lines with extremely different thousand-seed weight (TSW), were selected for a transcriptome analysis to determine the regulatory mechanisms underlying seed weight at the whole gene expression level and to identify candidate genes related to seed weight. A total of 2,251 new genes and 2,205 differentially expressed genes (DEGs) were obtained via RNA-seq (RNA sequencing). Among these genes, 1,747 (77.61%) new genes and 2020 (91.61%) DEGs were successfully annotated. Of these DEGs, 1,118 were up-regulated and 1,087 were down-regulated in the large-seed line. The Kyoto Encyclopedia of Genes and Genomes (KEGG) database analysis indicated that 15 DEGs were involved in ubiquitin-mediated proteolysis and proteasome pathways, which might participate in regulating seed weight. The Gene Ontology (GO) database indicated that 222 DEGs were associated with the biological process or molecular function categories related to seed weight, such as cell division, cell size and cell cycle regulation, seed development, nutrient reservoir activity, and proteasome-mediated ubiquitin-dependent protein catabolic processes. Moreover, 50 DEGs encoding key enzymes or proteins were identified that likely participate in regulating seed weight. A DEG (GSBRNA2T00037121001) identified by the transcriptome analysis was also previously identified in a quantitative trait locus (QTL) region for seed weight via SLAF-seq (Specific Locus Amplified Fragment sequencing). Finally, the expression of 10 DEGs with putative roles in seed weight and the expression of the DEG GSBRNA2T00037121001 were confirmed by a quantitative real-time reverse transcription PCR (qRT-PCR) analysis, and the results were consistent with the RNA sequencing data. This work has provided new insights on the molecular mechanisms underlying seed weight-related biosynthesis and has laid a solid foundation for further improvements to the seed yield of oil crops.

Dataset Information

Simultaneous profiling of seed-associated bacteria and fungi reveals antagonistic interactions between microorganisms within a shared epiphytic microbiome on Triticum and Brassica seeds.

Publications

Simultaneous profiling of seed-associated bacteria and fungi reveals antagonistic interactions between microorganisms within a shared epiphytic microbiome on Triticum and Brassica seeds.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets