Pantnagar Journal of Research

Abstract

Genetic transformation and many other genome engineering processes in wheat still lags behind due to non availability of a better callus induction and regeneration systems. In the present study, an efficient embryogenic based regeneration protocol is using different combinations of plant growth hormones. The combinations of commercial bleach along with mercuric chloride reported for effective sterilization of the explants (mature embryos) used for in vitro culture. The combinations of two auxin viz., 2, 4-D and NAA were taken for an efficient callus culture for callus induction, while cytokinin viz., kinetin and BAP were used to achieve the efficient regeneration capacity of the mature explants in wheat genotypes HD3059. A significant callus induction rate was reported among the callus cultured on MS media supplemented with concentration combinations of 4.0 mg L- 1 2, 4-D, 1.0 mg L-1 NAA and 1.0 mg L-1 ABA. Similarly, the concentration combination of 2.0 mg L-1 BAP; 1.0 mg L-1 NAA gave the maximum regeneration capacity along with maximum number of shoots. Further, MS media supplemented with 1.5 mg L-1 NAA in combination with 1.0 mg L-1 IAA produced better roots in regenerated shoots. The present study would be useful in ameliorating Indian wheat cultivars and would probably get over the problems regarding developing a new plant with a modification of a targeted gene using various recent technologies such CRISPR-Cas9 and gene engineering approaches. Therefore, efficient regeneration system has been achieved in the wheat genotype HD 3059.