Establishment of efficient regeneration protocol from leaf explants of Iris pumila shoots cultured in vitro

An efficient plant regeneration protocol via somatic embyogenesis by leaf base culture of in vitro grown Iris pumila shoots was developed. Induction of embryogenic calli was achieved on MS media supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D), kinetin (4.5 μM, each) and some additives (L-proline, casein hydrolysate, adenine sulphate and tyrosine). Further differentiation of embryogenic calli was achieved on MS hormone-free media, and on media supplemented with either BAP (4.5 μM) or BAP + zeatin (4.5 and 0.2 μM, respectively), which allowed somatic embryos, as well as shoot-like structures to form. Fully developed somatic embryos germinated on an MS hormone-free medium. An anatomical study confirmed that shoot-like structures represented early germinating stages of somatic embryos. Acclimatization of plants derived from somatic embryos was 64% after 1 year and no morphological variation was observed.