Mapping the colonization of a synthetic microbial community inoculum from sugarcan...
| Grant number: | 16/04322-0 |
| Support type: | Scholarships in Brazil - Doctorate (Direct) |
| Effective date (Start): | July 01, 2016 |
| Effective date (End): | August 31, 2018 |
| Field of knowledge: | Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms |
| Principal Investigator: | Paulo Arruda |
| Grantee: | Jaderson Silveira Leite Armanhi |
| Home Institution: | Centro de Biologia Molecular e Engenharia Genética (CBMEG). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil |
Abstract Traditionally, the isolation of a representative set of microorganisms from a given environment requires several rounds of picking and streaking to obtain pure colonies. These methods are costly, time-consuming and may result in the loss of relevant biological information between microorganisms. In many cases the interaction between microbes or even their dependency can restrict access to a reliable environmental sampling. In this work we advance a method to a Community-Based Culture collection (CBC) of microorganisms from a single step of picking, bypassing laborious steps of streaking. We applied these methodology to sampling microbial communities from root and stalks of sugarcane and constructed a CBC harboring around 5 thousands stored cultures in 56 plates of 96 wells. In order to identify these microbes we developed a methodology of sequencing pools of amplicons using barcodes that allowed the identification of microorganisms of each well. The full-length amplicons of microbial 16S rRNA were sequenced with PacBio RS platform. This multi-tagging system allows tracing back individual sequences to their original wells regardless of whether the well contains one of multiple microorganisms. The CBC construction method described here can be used to recover larger fractions of microbiota, cross-referenced to the metagenomics data for further investigation and selection of candidates. (AU) | |