Efficient Callus Induction and Regeneration in Selected Indica Rice

Abstract

An efficient callus induction and in vitro regeneration were developed using plant growth regulators, carbon sources, and basal media for three selected Malaysian wetland rice varieties (MR220, MR220-CL2, and MR232) and one upland variety (Bario). Effect of plant growth regulator (PGR) was carried out using four different concentrations (1–4 mg/L) of 2,4-D (2, 4-dichlorophenoxyacetic acid), and NAA (1-naphthalene acetic acid) (2.5, 5.0, 7.5, and 10 mg/L) with optimized 2,4-D. Effects of carbon sources (maltose and sorbitol), and basal media (MS, N6, and LS) were also studied with optimized PGR to maximize the induction of regenerable calli. This study found that all four varieties exhibited high frequency of callus induction on MS (Murashige and Skoog) medium that was supplemented with 3 mg/L 2,4-D and 30 g/L maltose. Callus induction frequencies in the cases of MR220, MR220-CL2, MR232, and Bario were found to be 76%, 94%, 85%, and 42% respectively. Morphological analysis through scanning electron microscopy (SEM) and histological analysis revealed the embryogenicity of the induced callus. In the regeneration study, it was observed that combination of 2 mg/L BAP (6-benzylaminopurine), 2 mg/L Kin (Kinetin) and 0.5 mg/L NAA supplemented MS medium has the potential to promote regeneration of selected indica rice varieties with higher regeneration percentage, i.e., 82% (MR220-CL2), 68% (both in MR220 and MR232), and 40% (Bario). The optimized conditions for callus formation and regeneration can be useful for biotechnological practices for the genetic improvement of Malaysian indica rice.