In vitrotissue culture protocol of ancient einkorn (Triticum monococcumssp.monococum) wheat via indirect shoot regeneration

Einkorn indirect shoot regeneration was obtained from leaf, coleoptile, and root explants cultured on full, 1/2, and 1/4 strength Murashige and Skoog (MS) medium containing 0.5 to 10 mg L(-1)2,4-dichlorophenoxyacetic acid (2,4-D). Calluses derived from coleoptile and root explants were cultured on full-strength MS medium supplemented with 0.5 to 5 mg L(-1)thidiazuron (TDZ) for 6 wk to induce plant regeneration. Only callus cultures derived from coleoptile explants regenerated shoots. For root induction, 1.0 to 10 mg L(-1)indole-3-acetic acid (IAA) was used for 4 wk. The highest rate of callus formation (93%) was from root explants cultured on full-strength MS medium with 4 mg L(-1 )2,4-D. Coleoptile explants had the highest callus formation (100%) after culture on full-strength MS medium with 3 to 6 mg L(-1)of 2,4-D. The highest indirect shoot regeneration (7.0 +/- 1.1 shoots produced per callus with a 67% shoot formation frequency) was from calluses derived from coleoptile explant cultured on full MS medium supplemented with 5 mg L(-1)of TDZ. Of the different IAA concentrations investigated for rooting, the greatest number of roots per explant (7.7 +/- 0.088 roots produced per regenerated shoot with a 100% root formation frequency) was observed on MS medium supplemented with 6 mg L(-1)IAA. This indirect shoot regeneration protocol for coleoptile-derived einkorn wheat callus will be useful for future wheat genetic and improvement studies.