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Haploid maize cells as target for the regeneration of homozygous genome edited maize lines (HAGEMA)
Project
Project code: 031B0543
Contract period: 01.07.2018
- 30.06.2020
Budget: 237,866 Euro
Purpose of research: Experimental development
Keywords: haploid techniques, microspores, corn, recalcitrance, Impedance Flow Cytometry
Haploid cells are desirable targets for the delivery of genes or genome-editing compounds, since they would allow the integration into a completely homozygous plant. As a unicellular system, isolated microspores would avoid the formation of chimeric regenerants. In maize, many efforts have been made to take advantage of such a single cell based microspore system. The feasibility of DNA delivery into haploid maize cells has already been successfully demonstrated in microspores, haploid callus cells and protoplasts derived thereof. However, the strong genotype dependency accompanied by a poor regeneration ability impeded extensive application possibilities so far. The HAGEMA research project is aiming to improve the regeneration ability of maize microspores by taking into account latest knowledge in cell biology. Recently, fundamental studies described the role of HDAC proteins in repressing totipotency during pollen development. In microspores it was demonstrated that HDAC inhibitor TSA can maintain totipotency. That finding may be a key factor to overcome recalcitrance in maize microspore culture. Impedance Flow Cytometry (IFC) is a novel technique to estimate plant cell viability, especially in pollen. It was also demonstrated that the electric characterization of maize pollen allows a visualization of the developmental nuclear stage. Both information generated by IFC will allow to increase the quality of the starting material for subsequent microspore culture. The working plan of the project proposes the screening of a set of genotypes with regard to microspore quality and susceptibility to TSA application. Embryogenic microspore cultures will be the basis for the establishment of callus suspension cultures, subsequently protoplasts will be derived thereof. The project goal is reached, when a robust single cell based regeneration system has been established as target for genome-editing approaches to end up with homozygous edited double haploid maize plants.
Section overview
Subjects
- Plant Breeding
- Process engineering
