Recipient Organization
UNIVERSITY OF FLORIDA
G022 MCCARTY HALL
GAINESVILLE,FL 32611
Performing Department
Microbiology and Cell Science
Non Technical Summary
Beneficial soil bacteria are important determinants of crop health, yield, and disease resistance. This proposal aims to understand the molecular details of how the beneficial soil bacterium Bacillus subtilis responds to extreme changes in its environment. Better understanding of how soil microbes respond to environmental change may lead to develpment of microbial strains with improved plant growth-promoting and disease-resistance conferring properties.
Animal Health Component
0%
Research Effort Categories
Basic
100%
Applied
0%
Developmental
0%
Goals / Objectives
Because beneficial soil bacteria are essential for crop health, it is of high importance to understand how they respond to changes in the soil environment. Soils are subjected to: (i) physical extremes of temperature and water availability; (ii) chemical extremes such as nutrient availability and application of pesticides, and (iii) biological extremes such as predation and niche competition. The overall goal of this project is to understand how a common beneficial soil bacterium, Bacillus subtilis, behaves in response to such environmental extremes. We have shown in previous work that chronic exposure to environmental stresses results in the evolution of B. subtilis strains with enhanced abilities to withstand such stresses. Using classical genetics and molecular biology, coupled with various -omics technologies, (genomics, transcriptomics, and phenomics), our goal is to: (i) identify mutations associated with evolution of enhanced growth and/or tolerance to stress; (ii) measure the global patterns of gene expression to stress, and; (iii) elucidate the regulatory circuitry underlying stress adaptation.
Project Methods
Evolution experiments will be performed in our lab using our routine published protocols. Whole-genome sequencing will be performed using the PacBio platform at UF Interdisciplinary Center for Biotechnology Research (ICBR) and mutations will be identified by standard bioinformatics software at the UF High Performance Computing center (HPC). Total RNA will be purified from cells using the Ribopure™ Bacterial RNA kit, quantified by Qubit™ analysis, and RNA Integrity Number (RIN) derived by Bioanalyzer 2100™ analysis. RNA-seq will be performed by the Hudson Alpha Institute and data will be analyzed using UF HPC software. Phenotype Microarrays will be performed in the Omnilog™ instrument housed in our laboratory using Phenotype Microarray (PM) plates 1-20. Competition experiments will be performed in the lab using our standard protocols. All molecular biology and genetic techniques used are routine in our lab.