" In situ similis " Culturing of Plant Microbiota: A Novel Simulated Environmental Method Based on Plant Leaf Blades as Nutritional Pads.

Autor:	Nemr RA; Environmental Studies and Research Unit, Department of Microbiology, Faculty of Agriculture, Cairo University, Giza, Egypt., Khalil M; Environmental Studies and Research Unit, Department of Microbiology, Faculty of Agriculture, Cairo University, Giza, Egypt., Sarhan MS; Environmental Studies and Research Unit, Department of Microbiology, Faculty of Agriculture, Cairo University, Giza, Egypt., Abbas M; Department of Microbiology, Faculty of Agriculture and Natural Resources, Aswan University, Aswan, Egypt., Elsawey H; Environmental Studies and Research Unit, Department of Microbiology, Faculty of Agriculture, Cairo University, Giza, Egypt., Youssef HH; Environmental Studies and Research Unit, Department of Microbiology, Faculty of Agriculture, Cairo University, Giza, Egypt., Hamza MA; Environmental Studies and Research Unit, Department of Microbiology, Faculty of Agriculture, Cairo University, Giza, Egypt., Morsi AT; Environmental Studies and Research Unit, Department of Microbiology, Faculty of Agriculture, Cairo University, Giza, Egypt., El-Tahan M; Regional Center for Food and Feed, Agricultural Research Center, Giza, Egypt., Fayez M; Environmental Studies and Research Unit, Department of Microbiology, Faculty of Agriculture, Cairo University, Giza, Egypt., Patz S; Algorithms in Bioinformatics, Center for Bioinformatics, University of Tübingen, Tübingen, Germany., Witzel K; Department of Plant Microbe Systems, Leibniz Institute of Vegetable and Ornamental Crops, Großbeeren, Germany., Ruppel S; Department of Plant Microbe Systems, Leibniz Institute of Vegetable and Ornamental Crops, Großbeeren, Germany., El-Sahhar KF; Department of Botany, Faculty of Agriculture, Cairo University, Giza, Egypt., Hegazi NA; Environmental Studies and Research Unit, Department of Microbiology, Faculty of Agriculture, Cairo University, Giza, Egypt.
Jazyk:	angličtina
Zdroj:	Frontiers in microbiology [Front Microbiol] 2020 Apr 07; Vol. 11, pp. 454. Date of Electronic Publication: 2020 Apr 07 (Print Publication: 2020).
DOI:	10.3389/fmicb.2020.00454
Abstrakt:	High-throughput cultivation methods have recently been developed to accelerate the recovery of microorganisms reluctant to cultivation. They simulate in situ environmental conditions for the isolation of environmental microbiota through the exchange of growth substrates during cultivation. Here, we introduce leaf-based culture media adopting the concept of the plant being the master architect of the composition of its microbial community. Pre-physical treatments of sunflower plant leaves, namely punching, freezing, and/or autoclavation, allowed the diffusion of electrolytes and other nutrients to configure the leaf surface as a natural pad, i.e., creating an " in situ similis " environment suitable for the growth of rarely isolated microbiota. We used surface inoculation and membrane-filtration methods to assess the culturability of endophytic bacteria from the sunflower phyllosphere and rhizosphere. Both methods supported excellent colony-forming unit (CFU) development when compared to standard R2A medium, with a special affinity to support better growth of epiphytic and endophytic populations of the phyllosphere compared with the rhizosphere. A 16S rRNA gene analysis of >122 representative isolates indicated the cultivation of a diverse set of microorganisms by application of the new methods. It indicated the predominance of 13 genera of >30 potential species, belonging to Firmicutes, Proteobacteria, and Actinobacteria, and especially genera not commonly reported for sunflower, e.g., Rhizobium , Aureimonas , Sphingomonas , Paracoccus , Stenotrophomonas , Pantoea , Kosakonia , and Erwinia . The strategy successfully extended diversity and richness in the endophyllosphere compared to the endorhizosphere, while CFUs grown on the standard R2A medium mainly pertain to Firmicutes, especially Bacillus spp. MALDI-TOF MS analysis clustered the isolates according to their niche and potential functions, where the majority of isolates of the endorhizosphere were clustered away from those of the endophyllosphere. Isolates identified as Gammaproteobacteria and Alphaproteobacteria were distinguishably sub-clustered, which was in contrast to the heterogeneous isolates of Firmicutes ( Bacillus spp.). In conclusion, leaf in situ similis cultivation is an effective strategy to support the future application of culturomics of plant microbiota. This is an effort to access novel isolates that are more adapted and competitive in their natural environments, especially those subjected to abiotic stresses like those prevailing in arid/semi-arid zones, and, consequently, to support the application of agro-biotechnologies, among other technologies, to improving agriculture in such zones. (Copyright © 2020 Nemr, Khalil, Sarhan, Abbas, Elsawey, Youssef, Hamza, Morsi, El-Tahan, Fayez, Patz, Witzel, Ruppel, El-Sahhar and Hegazi.)
Databáze:	MEDLINE
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