Optimization of Ionic and Chelated Iron and Its Interaction with Disodium Ethylenediaminetetraacetic Acid for Enhancement of Plant Regeneration in Rice (Oryza sativa L)

Abstract

Interactive effects of Fe2(SO4)3 and Na2EDTA on callus induction and plant regeneration in indica rice (Oryza sativa cv Pusa Basmati-1) was investigated. Callus induction and subsequent plant regeneration from seed was obtained on MS medium supplemented with 11.31 µM 2,4-D and 2.68 µM NAA + 8.87 µM BAP respectively. Both the callus induction and the plant regeneration media were supplemented with different levels of Fe2(SO4)3, Na2EDTA and their combinations. Callus induction, its morphogenic potential, average number of regenerated plants as well as the appearance of the regenerants was influenced by the levels of Fe2(SO4)3 and Na2EDTA. Embryogenic callus could be induced in the presence of Fe2(SO4)3 / Na2EDTA. Iron was essential for plant regeneration. Non-chelated iron could induce callus formation as well as plant regeneration, yet the chelated form was more suitable. A higher level of Na2EDTA in the regeneration medium was detrimental. Differential requirement of Fe2(SO4)3 at induction and regeneration level was observed. Method of medium preparation affected the regeneration response. Nearly three fold increase in the number of regenerated plants was achieved with 0.05 mM Fe2(SO4)3 + 0.1 mM Na2EDTA at callus induction and 0.1 mM Fe2(SO4)3+ 0.1 mM Na2EDTA at plant regeneration and standard method of autoclaving.