Callus induction and efficient plant regeneration in wheat (Triticum aestivum L.) through mature embryo culture

Abstract

Mature embryos of wheat were taken for callus development and efficient regeneration system through in vitro culture. Mature embryos in wheat have an added advantage that these are easily available round the year. In the present investigation we developed a protocol for sterilization of wheat seeds, callusing and regeneration. The technique developed used as a novel tool to assist plant breeders in crop improvements. Standardization of sterilization process was achieved successfully with ethanol (70%) for 30 second + HgCl2 (0.1%) for 1 min + NaOCl (1.0%) for 5 minute in both the genotypes viz. HD 2967 and C 306 selected for this study.. The different concentrations and combinations of plant growth regulators (PGRs) such as ABA, 2,4-D and NAA were used for callusing and BAP, NAA and TDZ were used for regeneration of plants from calluses. In the present study we observed that 1.0 mg/l ABA + 1.0 mg/l 2-4D + 2.0 mg/l NAA was optimum for callus development in genotype C306 and 70.00 percent callus induction achieved with no precocious germination. Genotype HD2967 showed 76.6 percent callus induction with low precocious germinations with the same concentration combinations of PGRs. The highest regeneration percentage 93.33% was recorded with the concentration combinations of PGRs (1.0 mg/l BAP + 2.0 mg/l TDZ + 0.5 mg/l NAA) in both the genotypes. The concentration of 0.5 mg/L NAA + 0.1 mg/L IBA produced better roots in regenerated shoots in genotype HD2967 and for genotype C 306 the rooting media gave good rooting with PGRs (0.1 mg/L NAA + 0.5 mg/L IBA).