In vitro tissue culture protocol of ancient einkorn (Triticum monococcum ssp. monococum) wheat via indirect shoot regeneration

• Published in: In vitro cellular & developmental biology. Plant 2021-02, Vol.57 (1), p.143-151
• Main Authors: Örgeç, Mehmet, Verma, Sandeep Kumar, Şahin, Günce, Zencirci, Nusret, Gürel, Ekrem
• Format: Article
• Language: English
• Subjects: 2,4-D; Acetic acid; Agricultural production; Biomedical and Life Sciences; Callus; Cell Biology; Coleoptile; Developmental Biology; Dichlorophenoxyacetic acid; Embryos; Explants; Indoleacetic acid; Life Sciences; Plant Breeding/Biotechnology; Plant Genetics and Genomics; Plant Sciences; PLANT TISSUE CULTURE; Regeneration; Rooting; Roots; Seeds; Shoots; Strength; Thidiazuron; Tissue culture; Triticum monococcum; Variance analysis; Wheat
• ISSN: 1054-5476; 1475-2689
• DOI: 10.1007/s11627-020-10122-8

Einkorn indirect shoot regeneration was obtained from leaf, coleoptile, and root expiants cultured on full, ½, and ¼ 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 expiants 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 expiants 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 expiants 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 expiant 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 expiant (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.