Modifying recombination in plants

About the Project

In eukaryotes, chromosomes recombine during meiosis to create novel combinations of alleles that may be selected for in the next generation. Bringing together the best traits is central to plant breeding, but recombination is limited both in number and position in the majority of crops (Desjardins et al. 2022, Nature Comms. doi: 10.1038/s41467-022-31438-6). Therefore, understanding and implementing ways to increase recombination is important scientifically and economically. We have recently identified an uncharacterised E3 SUMO ligase that alters recombination in Arabidopsis and now aim to transfer this knowledge to barley. The project will involve training a doctoral student in Oxford Nanopore DNA sequencing to analyse recombining barley genomes as well as super-resolution immunofluorescence to monitor proteins that control recombination at the cellular level. Experiments will also include yeast-2-hybrid to identify interacting proteins of the E3 SUMO ligase and CRISPR/Cas to target specific SUMO sites for site directed mutagenesis, followed up by recombination analysis. This will provide you with the opportunity to develop a modern set of skills for analysing eukaryotes as well as perform world-leading research.

Objective 1: Perform super-resolution immunofluorescence on wild type barley and E3 SUMO ligase mutant with a panel of antibodies that bind to recombination proteins

Objective 2. Cross the barley E3 SUMO ligase mutant to generate a mapping population that will be sequenced in the progeny to determine recombination pattern

Objective 3. Design CRISPR guide RNAs and mutate SUMO sites in proteins that control recombination and then analyse with molecular and cytological techniques

Techniques that will be undertaken during the project

Super-resolution fluorescence microscopy, DNA extraction, cloning, PCR, genomic DNA sequencing, yeast-2-hybrid analysis, bioinformatics.

Funding Notes

This project is only available on a self funded basis or if you have your own sponsorship.