干旱区生态环境知识资源中心(Arid): In situ similis Culturing of Plant Microbiota: A Novel Simulated Environmental Method Based on Plant Leaf Blades as Nutritional Pads

Arid

DOI	10.3389/fmicb.2020.00454
	In situ similis Culturing of Plant Microbiota: A Novel Simulated Environmental Method Based on Plant Leaf Blades as Nutritional Pads
	Nemr, Rahma A.1; Khalil, Mohab 1; Sarhan, Mohamed S.1,7; Abbas, Mohamed 2; Elsawey, Hend 1; Youssef, Hanan H.1; Hamza, Mervat A.1; Morsi, Ahmed T.1; El-Tahan, Mahmoud 3; Fayez, Mohamed 1; Patz, Sascha 4; Witzel, Katja 5; Ruppel, Silke 5; El-Sahhar, Kassem F.6; Hegazi, Nabil A.1
通讯作者	Hegazi, Nabil A.
来源期刊	FRONTIERS IN MICROBIOLOGY
ISSN	1664-302X
出版年	2020
卷号	11
英文摘要	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.
英文关键词	plant microbiota plant-based culture media endophyllosphere endorhizosphere MALDI-TOF-MS culturomics arid semi-arid zones
类型	Article
语种	英语
国家	Egypt ; Germany ; Italy
开放获取类型	gold, Green Published
收录类别	SCI-E
WOS记录号	WOS:000529235700001
WOS关键词	TOF MASS-SPECTROMETRY ; ELECTROLYTE LEAKAGE ; ENDOPHYTIC BACTERIA ; CULTIVATION ; SUNFLOWER ; RHIZOSPHERE ; DIVERSITY ; GROWTH ; TOOL ; MICROORGANISMS
WOS类目	Microbiology
WOS研究方向	Microbiology
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/318573
作者单位	1.Cairo Univ, Fac Agr, Dept Microbiol, Environm Studies & Res Unit, Giza, Egypt; 2.Aswan Univ, Fac Agr & Nat Resources, Dept Microbiol, Aswan, Egypt; 3.Agr Res Ctr, Reg Ctr Food & Feed, Giza, Egypt; 4.Univ Tubingen, Ctr Bioinformat, Algorithms Bioinformat, Tubingen, Germany; 5.Leibniz Inst Vegetable & Ornamental Crops, Dept Plant Microbe Syst, Grossbeeren, Germany; 6.Cairo Univ, Fac Agr, Dept Bot, Giza, Egypt; 7.Eurac Res Inst Mummy Studies, Bolzano, Italy
推荐引用方式 GB/T 7714	Nemr, Rahma A.,Khalil, Mohab,Sarhan, Mohamed S.,et al. In situ similis Culturing of Plant Microbiota: A Novel Simulated Environmental Method Based on Plant Leaf Blades as Nutritional Pads[J],2020,11.
APA	Nemr, Rahma A..,Khalil, Mohab.,Sarhan, Mohamed S..,Abbas, Mohamed.,Elsawey, Hend.,...&Hegazi, Nabil A..(2020).In situ similis Culturing of Plant Microbiota: A Novel Simulated Environmental Method Based on Plant Leaf Blades as Nutritional Pads.FRONTIERS IN MICROBIOLOGY,11.
MLA	Nemr, Rahma A.,et al."In situ similis Culturing of Plant Microbiota: A Novel Simulated Environmental Method Based on Plant Leaf Blades as Nutritional Pads".FRONTIERS IN MICROBIOLOGY 11(2020).