In vitro Production of Double Haploid in Maize (Zea mays L.) Through Pollen Culture

Abstract

Abstract Maize (Zea mays L.) has a distinct genetic system and is recalcitrant to generate haploid and subsequently double haploid plants from pollen grains in aseptic conditions. It is a technique with lot of potential for producing pure lines for crop improvement and introducing the desired variability through modification in biochemical and applied genetics. The experiment was conducted using different doses of 2,4-Dichlorophenoxyacetic acid (2, 4-D), Indole-3-acetic acid (IAA), 6-Benzylaminopurine(BAP), Zeatin(ZEA), Kinetin and Naphthalene Acetic Acid(NAA) ranging from 0.5 mg/L to 5 mg/L for the callus induction and plantlet regeneration. The callus was sub-cultured for double haploid induction with doses of 100–500 mg/l colchicine for 7h, 12h and 7 days. Microscopic examination and morphological identification were proposed to check ploidy level. The results exhibited the callus formation at 1, 2 and 3 mg/l concentrations of 2, 4-D and IAA respectively. Based on the time required for the 2, 4-D, callus induction at the concentration of 2mg/l exhibited (47 days) and at the same concentration of IAA exhibited the minimum days of callus induction (34 days). The colchicine treated pollen derived callus regenerated into roots and shoots on IAA and BAP with 3mg/l and 2mg/l respectively. In contrast to pollen derived callus, colchicine treated seed callus gave better results. Double haploid plant produced successfully through in vitro culture of pollen. Current study will be helpful for the achievement of homozygosity in relatively less time and varietal improvement for desired traits.