An Optimized Protocol for Seed Sterilization and Shoot Regeneration from Mature Embryo in Wheat (Triticum Aestivum L.) var. HD2967

Abstract

In the present study, an efficient protocol for seed sterilization was developed. This protocol used a 1.0 % bavistin solution, which provides a protective response against fungal contamination. It was observed that there was no significant difference among the effective sterilization treatments in comparison to seed germination and seedling length under in vitro conditions. Furthermore, the embryo excised from bavistin-treated seeds showed improvement in callus induction and shot regeneration in var. HD2967. 0.1% HgCl2-SDS solution used for seed sterilization was also optimized with 2.5 to 3.0 minutes as the best-suited duration, providing a completely sterilized environment for the seeds for healthy and fast growth. In addition, plant growth regulators (PGRs) were further used to screen the most effective concentration for callus to shoot formation using mature embryos. Auxin concentration of 2.0 mg/l in MS agar media was quite effective for callus induction from embryo explant. For shoot regeneration (SI-7) combination is suitable for a maximum number of shoots per callus. This combination of 2, 4-D, Zeatin, CuSO4, and TDZ showed good regeneration in the wheat variety HD2967. The optimized protocol displayed 100% callus induction and up to 80-85% regeneration in HD2967, with a high reproducibility rate paving the way for regenerating plantlets from mature wheat embryos.