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Bioprospecting and Structure of Fungal Endophyte Communities Found in the Brazilian Biomes, Pantanal, and Cerrado.

ABSTRACT: Medicinal plants have been recognized as hosts of high diverse endophytic microorganisms, including fungi that produce secondary metabolites with biological activity. Two biomes in Brazil, Pantanal (wetland), and Cerrado (savannah), are known as biodiversity hotspots, and despite their importance as a reservoir for several species, knowledge about the fungal biodiversity in these biomes is very limited. Fungal endophytic communities associated with leaves and petioles of the medicinal plants Vochysia divergens (from Pantanal) and Stryphnodendron adstringens (from Cerrado) were analyzed and studied for their antimicrobial activity against human and plant pathogens. A total of 1,146 isolates of endophytic fungi were obtained from plants collected in January and June of 2016 and grouped into 124 morphotypes. One isolate of each morphotype was identified by sequencing of internal transcribed spacer (ITS) region of the rDNA gene, which revealed the presence of 24 genera, including 3 possible new genera, and 48 taxa. Differences in the endophytic community according to the biomes were observed concerning the analyzed morphotypes. However, when we analyzed the diversity of genera and richness, they were similar for both plants, with Diaporthe, Phyllosticta, and Neofusicoccum as dominant genera. In addition, the community composition of V. divergens differs according to the analyzed plant tissues (petiole and leaf). These data suggested that both, the plant species and plant tissues play a role in the composition of endophytic community. As regards the biotechnological potential, 5 isolates showed activity against the phytopathogens Phyllosticta citricarpa, Colletotrichum abscissum, and Fusarium verticilioides, and 8 isolates showed high activity against clinical pathogens and were selected for the production of crude extract in different culture media. Extract from cultivation of Diaporthe sp. LGMF1548 and LGMF1583 and Neofusicoccum brasiliense LGMF1535 showed activity against methicillin-resistant Staphylococcus aureus, Klebssiella pneumonia, and Candida albicans. In addition, extracts of Diaporthe cf. heveae LGMF1631 inhibited 90% of the mycelial growth of the P. citricarpa and 70% of C. abscissum and may represent an alternative to be used in the biological control of these phytopathogens. Future research will focus on the chemical characterization and structural elucidation of these bioactive compounds.

SUBMITTER: Noriler SA

PROVIDER: S-EPMC6066559 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Plants harbor complex and highly diverse fungal endophyte communities (FECs), making it difficult to evaluate the functional role of individual taxa, subsets of the community, or the FEC as a whole. To reduce the complexity of this system, we aimed to produce fungi-null wheat (Triticum aestivum) plants. To this end, we treated seeds with heat and fungicides and generated plants from rescued embryos and callus tissue. A culture-based approach and reverse transcription PCR analysis were negative, indicating that all treatments produced plants apparently free of fungi. However, the analysis of DNA using digital droplet PCR and next-generation sequencing revealed that tissues from all treatments retained low levels but diversity-rich FECs. While the FECs varied in composition across treatments and tissues, they all included core taxa of the mycobiome. The reduced fungal biomass, along with the changes in FEC composition, negatively affected plant development, supporting a FEC contribution to proper plant development and fitness. Our discovery that a large part of the FEC cannot be separated from plants and can be transmitted through seeds and tissue culture calls for reevaluation of particular microbiome paradigms, such as core taxa concepts, transmission modes, and functional species.IMPORTANCEThe native microbiome in a given plant must be considered when evaluating the effect of a single taxon or synthetic community. The pre-existing microbiome can interact with artificially added microbial cargo, which affects the final outcome. Such issues can be at least partially solved by the use of endophyte-free plants, which provide a clean background that should be useful in determining the effect of a single taxon, taxa combinations, or the entire microbiome on plant performance. Previous reports regarded plants as endophyte-free or axenic by the lack of fungal growth on culture media or the generation of plants from tissue cultures. We showed here that while fungi could not be isolated from fungicide-treated or tissue culture-regenerated plants, nevertheless, all plants contained rich fungal endophyte communities; namely, it was impossible to create fungi-free wheat plants. Our results call for rethinking fundamental microbiome-related concepts, such as core taxa, transmission mode, and functional species.

Project description:Studies of fungi in upland cotton (Gossypium hirsutum) cultivated in the United States have largely focused on monitoring and controlling plant pathogens. Given increasing interest in asymptomatic fungal endophytes as potential biological control agents, surveys are needed to better characterize their diversity, distribution patterns and possible applications in integrated pest management. We sampled multiple varieties of cotton in Texas, USA and tested for temporal and spatial variation in fungal endophyte diversity and community composition, as well as for differences associated with organic and conventional farming practices. Fungal isolates were identified by morphological and DNA identification methods. We found members of the genera Alternaria, Colletotrichum and Phomopsis, previously isolated as endophytes from other plant species. Other recovered species such as Drechslerella dactyloides (formerly Arthrobotrys dactyloides) and Exserohilum rostratum have not, to our knowledge, been previously reported as endophytes in cotton. We also isolated many latent pathogens, but some species such as Alternaria tennuissima, Epicoccum nigrum, Acremonium alternatum, Cladosporium cladosporioides, Chaetomium globosum and Paecilomyces sp., are known to be antagonists against plant pathogens, insects and nematode pests. We found no differences in endophyte species richness or diversity among different cotton varieties, but did detect differences over time and in different plant tissues. No consistent patterns of community similarity associated with variety, region, farming practice, time of the season or tissue type were observed regardless of the ecological community similarity measurements used. Results indicated that local fungal endophyte communities may be affected by both time of the year and plant tissue, but the specific community composition varies across sites. In addition to providing insights into fungal endophyte community structure, our survey provides candidates for further evaluation as potential management tools against a variety of pests and diseases when present as endophytes in cotton and other plants.

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Bioprospecting and Structure of Fungal Endophyte Communities Found in the Brazilian Biomes, Pantanal, and Cerrado.

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