How to make haploid Arabidopsis thaliana

The haploid inducer is a cenh3-1 null mutant in the Col-0 ecotype that is complemented by a CENH3 transgene called GFP-tailswap (cenh3-1 is embryo lethal). It has a vegetative phenotype that varies in intensity depending on growth conditions. Rosette leaves are slightly curled, and it also has shorter internodes after bolting. The mutant is mostly male sterile, but has reasonable female fertility (about 60-70% of wild type in our hands). It can be maintained as a homozygote because it yields a few hundred seed per plant if treated carefully. Later siliques have higher fertility. Importantly, a majority (90%) of the offspring are diploids similar to the parent plant. We do observe a low frequency of aneuploid offspring which can be easily distinguished from diploid siblings by their distinctive phenotypes. Seed collected from cenh3-1/+ GFP-tailswap/ GFP-tailswap plants are available from ABRC and from NASC (stock number CS66982). 1/4 will be the desired cenh3-1 GFP-tailswap.