Location: Natural Products Utilization Research
Project Number: 6060-21410-012-004-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Aug 1, 2022
End Date: Aug 1, 2024
Objective:
1. To develop plant allelochemicals as plant incorporated pesticides for crop species.
2. To identify components of the S. bicolor transport apparatus required for the extracellular transport of the allelochemical sorgoleone from root hair cells.
Approach:
The demonstrated efficacy of sorgoleone as a potent broad-spectrum plant growth inhibitor active against many agronomically important monocotyledonous and dicotyledonous weed species has led to substantial interest in its development as a plant-incorporated pesticide. RNAseq analyses will be performed using a panel of S. bicolor vegetative and reproductive tissues, including root hairs isolated from both wild-type and previously-generated sorgoleone-deficient RNAi knockout lines. The RNA-seq work will provide datasets of genes preferentially expressed in root hairs, as well as genes responsive to perturbations in sorgoleone biosynthesis. These datasets will be mined for differentially expressed transporter sequences, and candidate sequences will be subjected to CRISPR/Cas-mediated gene editing for generating knockout lines. Additionally, in vitro assays will be performed with isolated S. bicolor embryos to evaluate the efficiency of various guide RNA sequences prior to their use in stable transformation studies. The ability of the resulting CRISPER/Cas knockouts to transport and secrete sorgoleone will then be evaluated, ideally leading to the identification of key cellular components required for the transport of sorgoleone from its site of synthesis in root hairs to the root system-soil interface.
