Location: Crop Bioprotection Research
Project Number: 5010-22410-024-000-D
Project Type: In-House Appropriated
Start Date: Mar 2, 2022
End Date: Mar 1, 2027
Objective:
Objective 1: Discover and optimize the use of bioactive metabolites associated with beneficial microbes.
Sub-objective 1A: Genome sequencing of Bacillus microbial resources.
Sub-objective 1B: Heterologous expression of biosynthetic gene clusters.
Sub-objective 1C: Creation of lipopeptide producer strains and evaluation of synergy and efficacy.
Objective 2:Evaluate the application of microbes, such as seed coatings, for their interaction with plant pathogens and their role in biocontrol efficacy.
Sub-objecitve 2A: Evaluation of seed coatings and biocontrol agent genotype.
Sub-objective 2B: Development of genetic modification protocols and functional genomics to understand the determinants of biocontrol efficacy.
Approach:
Our approach will be to apply technologies allied with the fields of fermentation science, microbial physiology, metabolomics, genomics, and proteomics for two purposes: to enhance the efficacy and shelf-life of the antagonist biomass manufactured and to produce gnotobiotic (i.e., all of a limited number of organisms in a culture are known) or axenic cultures of nutritionally fastidious plant pathogens. More specifically, the shelf-life and efficacy of biocontrol strains will be improved by isolating efficacious stress tolerant variants of a yeast biocontrol agent and then testing the more promising strains isolated in small pilot tests against Fusarium head blight of wheat. Other studies will strive to discover cell production methodologies that promote the production of compounds that enhance cell stress tolerance. Strain transcriptional response to culture conditions will be determined to facilitate optimizing these cell production studies. This will include studies to elucidate the transcriptional response of a yeast biocontrol strain to cold-adaptation that improves cell survival and biocontrol efficacy. Gnotobiotic culturing studies will include establishing a selection of host plants in sterile tissue culture boxes or as callus cell cultures and evaluating methods for infecting these host tissues with axenic propagules of an obligate pathogen. The transcriptional response of gnotobiotic host cell tissue to infection by an obligate plant pathogen will then be determined as a prelude to attempting to grow one or more obligate plant pathogens in axenic culture.