Callus Induction and Regeneration Response of Mustard (Brassica spp.)

Abstract

Mustard (Brassica spp.) is the third most important edible oil crop belonging to the family Brassicaceae (Cruciferae) and the genus Brassica. Due to the growing world population and increasing industrialization, the demand for edible oil and biofuels is increasing worldwide. Stress-tolerant mustard variety creation by transformation or genome editing procedures, as well as varietal in vitro regeneration responses, are critical challenges. The objective of the study was to design a tissue culture protocol for better use the explant by using hypocotyls and cotyledon explants to observe the in vitro response to callus induction and regeneration of three mustard types, namely BARI Sarisha-14, Binasarisha-4, and Binasarisha-9.Explants were cultured on Murashige and Skoog medium (MS) supplemented with different concentrations and combinations of NAA(10 mg mL-1), BAP (10 mg mL-1 for callus induction and shoot initiation medium , 2.5 mg mL-1 for shoot outgrowth medium), AgNO3(50 mg mL-1), GA3(2 mg mL-1), and IBA (10 mg mL-1)for callus induction, shoot initiation, shoot outgrowth, and root initiation. Binasarisha-4 showed the best performance with hypocotyls76.67% callus induction, 70.03% shoot initiation, 92.23% shoot outgrowth, and 40% root initiation followed by BARI Sarisha-14 (71.67%, 58.23%, 73.33% and 23.33% respectively) and Binasarisha-9 (54.17%, 38.87%, 45.83% and 11.11% respectively). It is recommended that in vitro hypocotyls culture of Binasarisha-4 can be used for frequent and successful genetic transformation in Mustard.