The “green revolution” is responsible for the unprecedented expansion in agricultural productivity. Advances were delivered by improved varieties, and changes to agricultural practices e.g. fertilisers and better irrigation. These increases in productivity are unsustainable with land and water usage, climate change and resource intensive systems dependent on chemical inputs from fossil fuels. The outputs from the green revolution have been distributed unevenly between the developing and developed world.
PROSTRIG will address food insecurity in a durable and environmentally sustainable way by generating the tools and resources needed to develop new productive, resilient and low input crops. The focus will be the emerging class of plant growth regulators, Strigolactones.
- An expression construct for SaCas9, the specific nuclease mediating the genome editing process was designed and cloned into an appropriate vector.
- Promoter sequences in target genes CCD7 and CCD8, the two genes controlling the formation of the specific strigolactone molecules we aim to modulate to achieve the desired phenotype and outcomes, were identified.
- SaCas9 activity was validated by digestion of cloned CCD7 and CCD8 promoter sequences in the presence of the appropriate gRNAs (sequences which guide the nuclease to its target site in the maize genome).
- Transformation experiments to generate maize plants containing SaCas9 and gRNAs were initiated but were severely disrupted during the protracted Covid-19 lock-down. A small number of transgenic maize plants containing Cas9 and gRNAs were recovered. Some of these plants only contain Cas9, providing a back-up strategy through breeding with plants containing gRNAs to screen for the target mutations.
- We established all analytical methods for mutant plants and created a biochemical network onto which perturbation can be painted over to represent changes.
- Quality and general overview of data was revised using principal component analysis, pair-wise comparisons of genotypes per time-point using statistical packages.
- Results were painted over bespoken pathways constructed from dedicated databases and literature references.