Location: Natural Products Utilization Research
Project Number: 6060-21410-012-005-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Jun 1, 2023
End Date: May 31, 2026
Objective:
Discovery of the mechanisms of action for newly discovered and existing bioactive phytotoxins using molecular genetics approaches.
Approach:
The identification of naturally produced compounds with phytotoxic activity enables researchers to explore their potential to use as herbicides. However, to make better use of these potential herbicides, we should identify their mechanisms of action, i.e., molecular target sites. The strategies for discovery of molecular target sites of phytotoxins are complex, due to the multitude of potential molecular targets in plants. Few established protocols or standard methods are available to conduct such experiments for natural products such as phytotoxins for that matter. One direct approach for determining the potential molecular targets, which are directly related to the biological functions of genes in an organism, could be to produce mutants with altered responses to the phytotoxins. Arabidopsis has been widely used as a model organism for the identification of genes that are involved in many aspects of herbicide resistance. In this project, Arabidopsis mutants will be identified against newly discovered phytotoxins. Specifically, ethyl methane sulfonate (EMS)-mutagenized Arabidopsis seeds or T-DNA tagged overexpression lines will be used to screen for mutants that are resistant to the phytotoxins. The loci that lead to resistance will be identified and characterized using genetic mapping, genome sequencing or other available methods. The isolated mutants will be subjected to gene cloning for functional characterization. More focused studies will be conducted to provide a definitive identification of the molecular target site.
