Optimization of regeneration protocol and prospecting spectinomycin resistance in barley (Hordeum vulgare L.) cv Haider-93

Abstract

Immature zygotic embryos from spring barley cv. Haider-93 were used to induce somatic embryogenesis. The type of the explant, the level of 2,4-dichlorophenoxyacetic acid (2,4-D) and handling of calli during subculture are critical factors to obtain maximum number of regenerants. Different concentrations of 2,4-D (0, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0 mg/L) and Kinetin (0.5, 1.0, 1.5, 2.0 mg/L) were used for callus induction and shoot initiation, respectively. Use of immature embryos having damaged axis as explants revealed a pronounced gradient of callus formation on Murashige and Skoog (MS) medium supplemented with 2.5 mg/L of 2,4-D and maximum regeneration response at 1 mg/L kinetin. Regenerated shoots were rooted on MS medium supplemented with 1 mg/L IAA. A kill curve was developed to find out the sensitivity level of barley cells to spectinomycin, a broad-spectrum antibiotic. This study fulfils an antique prerequisite of a reproducible regeneration system required for the improvement of barley via genetic engineering and also falls under, “Establish Good Health and Well-Being” Sustainable Development Goals of United Nations Organization.   Key words: 2,4-Dichlorophenoxyacetic acid, indole acetic acid, kinetin, antibiotic resistance, spectinomycin, Barley 53.