Recipient Organization
UNIVERSITY OF CALIFORNIA, DAVIS
410 MRAK HALL
DAVIS,CA 95616-8671
Performing Department
Microbiology & Molecular Genetics
Non Technical Summary
This project supports the mission of the Agricultural Experiment Station by addressing the Hatch Act area(s) of: biotechnology.Alternative pest control management approaches are important to prevent the rampant use of toxic chemical compounds from contaminating ecologically sensitive areas. This study will provide information as to how MYXOCOCCUS XANTHUS can be used to control many microbial agricultural pests. This study will also examine how M.XANTHUS interacts with other microbes in natural habitats, including agricultural pathogens and non-pathogens in agricultural soils. We will also learn about the effectiveness of M. XANTHUS as a potential biological control organism, providing alternatives to chemical pesticides. Finally, this work will provide information on how antibiotics produced by M. XANTHUS to attack agricultural pests, are regulated and produced, potentially leading to the development of alternative antimicrobial agents.
Animal Health Component
50%
Research Effort Categories
Basic
50%
Applied
50%
Developmental
0%
Goals / Objectives
For the past several years we have been working under the hypothesis that myxobacteria growth and development both influence and are influenced by the local microbial environment. Myxobacteria are known microbial predators, which includes a variety of plant pathogens in the soil and rhizosphere. The interaction between myxobacteria and plant pathogens, which can lead to an alterations in the progression of the disease to the plants. The specific objectives of this research are to develop tools to rapidly quantify and identify myxobacteria from agricultural soils and to understand the influence of plants and plant pathogens on the cell cycle and gene expression in myxobacteria. We propose to use these tools to define the nature of the predator-prey relationship between myxobacteria and soil prey bacteria. The information and tools developed as a result of this research are needed for our long-term goal of manipulating the interactions between myxobacteria and plant pathogens to mitigate plant disease. (1) Identify genes and their products involved in predation behaviors, using standard molecular approaches and RNA HiSeq methodologies(2) Define the role of secondary metabolites and developmental regulators in biocontrol and/or predation of plant pathogens in agricultural soils using RNA HiSeq methodologies.
Project Methods
We are taking a two-prong approach to look at the use of Myxococcus xanthus as a possible biological control agent. First, we have developed two independent predation assays to date, the streak assay and the plate assay, (21) which will be used to determine the effectiveness of M. XANTHUs as a biological control agent on variety soil bacteria. Second, we have previously identified candidate genes that we predicted are involved in the production of secondary metabolites and/or secreted digestive enzymes, such as lipases, proteases, cellulases and chitinases. We are making in-frame null mutations in each of these genes and will determine their phenotype with respect to predation, vegetative growth and development using standard laboratory protocols and the predation assays that we developed.Mutants with predation and/or developmental phenotypes will be further analyzed for regulatory phenotypes using RNAseq protocols. RNAseq will also be used for the identification of novel predatory pathways.The phenotypic analysis of these mutants, specifically with respect to predation, will allow us to quantify the importance of each identified gene product on predation. In addition, we will be able to determine which gene sets are required for which type of pathogen, bacterial and/or fungal. Finally, we propose to determine the expression patterns for each of these genes, using RNAseq and quantitative PCR. These data will allow us to define their regulatory circuitry in order to better engineer more efficient bio-control agents.Standard Myxobacterial isolation procedures are being used on a variety of soil isolates form the Salinas Valley