Microgenomics 2021 webinar :  Single haploid nucleus genotyping in barley
Channel: The International Symposium on Microgenomics 2023
Date: 2022-02-03

Stefan Heckmann (Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) Gatersleben, Germany. Invited by our GOLD sponsor Stilla): Single haploid nucleus genotyping in barley – measuring meiotic recombination rates in pollen nuclei

Meiotic recombination assures novel allelic combinations that are harnessed during breeding. Assessing meiotic recombination outcome in gametes prior fertilization enables measuring large sample numbers from single individuals without the need of growing large segregating populations. So far, methods to genotype or sequence a single barley pollen nucleus rely on whole-genome amplification (WGA) of individual single pollen nuclei, thus restricting the number of analyzable samples.

Using Crystal Digital PCRTM (naica, STILLA Technologies) we developed a single pollen nucleus genotyping assay to measure meiotic recombination rates within defined chromosomal intervals in high-throughput without WGA. Measured rates of meiotic recombination in hybrid pollen nuclei corresponded to that in segregating populations. Initial data acquired using the 6-color naicaTM system suggest that the throughput can be further increased by multiplexing our assays.

Nuclei from several crop plants with different nuclear sizes and amounts of DNA are compatible with the set-up, suggesting a broad application of Crystal Digital PCRTM-based single nuclei genotyping assays.