Effective callus induction and plant regeneration in callus and protoplast cultures of Nigella damascena L.

• DOI: 10.1007/s11240-020-01953-9
• Publication Date: 2020-10-21T10:03:16Z
• Authors: Magdalena Klimek-...; Dariusz Kadluczka; Aneta Lukasiewicz; Aneta Malec-Pala; Rafal Baranski; Ewa Grzebelus
• Keywords: Callus; Protoplast; Kinetin; Cotyledon; Rosa × damascena

Abstract In this study we report the development of effective in vitro systems for a medicinal plant Nigella damascena L. comprising: (1) callus induction, (2) somatic embryogenesis cultures with subsequent regeneration, and (3) isolation regeneration callus-derived protoplasts. Callus was achieved on 83–100% hypocotyl cotyledon explants, whereby Murashige Skoog medium (MS) supplemented 3 mg L −1 6-benzylaminopurine 0.5 α -naphthaleneacetic acid (NAA; BN medium) more advantageous than MS kinetin NAA (KN medium). Histological observations calli revealed presence embryogenic zones from which embryos developed hormone-free medium. Plant observed 76–95% calli. A high capacity to form maintained long-lasting cultures, i.e. even 2 year old callus. The obtained also good source tissue protoplast isolation. By applying mixture cellulase pectolyase, acceptable yield viable protoplasts achieved, especially hypocotyl-derived Protoplasts embedded an alginate matrix cultured modified Kao Michayluk media re-constructed their cell wall re-entered mitotic divisions. About 30% small aggregates formed microcalli, which, after release alginate, proliferated continuously KN media, irrespective variant used as source. Somatic embryo formation were successful media. An system N. has been is being reported first time.