Establishment of a regeneration system with anther tissue culture of Medicago falcata ‘Hulunbeier’ and haploid identification

The establishment of a regeneration system using the anther tissue culture method is an important approach in haploid breeding. In the present study, the anther tissue culture of Medicago falcata ‘Hulunbeier’ was investigated and the regeneration system of anther tissue culture established. The results showed that liquid suspension medium was more suitable than solid medium for M. falcata in the induction of anther callus. Mixtures of B5+0.5 mg·L-1 2,4-D+0.25 mg·L-1 6-BA+0.4 mg·L-1 NAA+3.0 mg·L-1 KT, MS+1 mg·L-1 2,4-D+0.5 mg·L-1 6-BA+sucrose 2%+ager 0.7%, and 1/2 MS+0.1 mg·L-1 NAA+sucrose 2%+ager 0.7% were suitable for callus induction, callus differentiation, and rooting culture, respectively. The percentage of haploids of regenerated plants was measured by flow cytometry as being 27% for M. falcata.