A high-throughput regeneration protocol for recalcitrant tropical Indian maize (Zea mays L) inbreds

Abstract

Immature embryos from five select recalcitrant maize (Zea mays L) inbreds used as explants were evaluated for their ability to form callus, somatic embryos and subsequent regeneration into plants. The embryos were placed on N6 basal media with varying levels of 2,4-D (0.5, 1.0, 1.5, 2.0, and 2.5 mg l-1) and were regenerated on MS me¬dium supplemented with BAP (2 - 10 mg l-1), 2,4-D (0.25 mg l-1) and silver nitrate (0.85 mg l-1). Explants cultured on N6 medium supplemented with 2,4-D (2.0 mg l-1) were associated with the highest frequency of embryogenic calli and that of UMI 29 were highly embryogenic (78.67%). When synergism between dicamba and 2,4-D on Type II callus production in UMI 29 was sought to be investigated using 2,4-D (1 or 2 mg l-1) individually and in combina¬tion with dicamba (3.7 mg l-1) production of Type II callus with the greatest frequency of 83.33% was observed on N6 medium containing 3.7 mg l-1 dicamba + 1 mg l-1 2,4-D. The greatest percentage of shoot induction (82.67%) was observed on MS medium supplemented with BAP (10 mg l-1). Among the five genotypes tested, UMI 29 was associated with the highest percentage of callus initiation, shoot induction and mean number of developed shoots. The protocol described in this study can reliably be used to transform tropical maize inbreds as a routine.