Optimization of Conditions for Callus Induction from BC2F1 Population (Ranbir Basmati x PAU148) through Anther Culture

With different culture conditions and concentrations of growth regulators and supplements, Anther culture technique can be easily employed for the production of haploids under in vitro conditions. Aims: The present study was undertaken with the objective to optimize the development of doubled haploids using anthers for in vitro induction of callus on N6 medium. Place and Duration of Study: The samples (BC2F1 seeds) were raised previously in Skuast-J. From total degree program of 3 years, this work related to tissue culture technique was done in one year from January 2018 to January 2019. Methodology: The effect of levels of 2, 4-dichlorophenoxy acetic acid (2, 4-D) i.e. 0 to 3 mg/L in basal N6 media was observed on callus induction frequency (CIF). The effect of duration of cold pre-treatment was observed on callus induction frequency at 2.5 mg/L of 2, 4-D by giving the cold pre-treatment at 4ºC from 8 to 12 days. Also the effect of different amino acids was checked on callus induction frequency. Results: Highest callus induction frequency of 9.39 per cent was observed in N6 medium fortified with 2.5 mg/L 2, 4-D and lowest callus induction frequency of 2.52 per cent at the concentration of 1.0 mg/L. The cold pre-treatment for 10 days gave highest callus induction frequency of 1.44 per cent and lowest callus induction frequency of 0.44 per cent was obtained at cold pre treatment for 8 days. The highest callus induction frequency of 12.55 per cent was observed in case of media supplemented with 25 mg/L tryptophan and 40 mg/L cysteine and lowest callus induction frequency of 7.18 per cent was observed when media was supplemented with 560 mg/L proline. Conclusion: The cold pre-treatment of 10 days at 4ºC on media supplemented with 2.5 mg/L of 2, 4-D and combination of 25 mg/L tryptophan and 40 mg/L cysteine proves to provide best androgenesis conditions for anthers from BC2F1 population.

Identification of high b-Carotene lines of maize in backcross (BC2F1)population by genotyping crtRB1 Allele specific marker

In order to address the global problem of Vitamin A deficiency, biofortification is considered to be a cost effective and sustainable approach. The polymorphism in crtRB1gene is associated with b-carotene levels in maize kernels. The randomly selected 10 seeds of selected maize lines in BC2F1 population cobs when analyzed for b-carotene content through HPLC, showed confirmation of improvement in b-carotene content of 2.37 µg/g. The crtRB1 3’TE polymorphism in BC2F1 population of the cross revealed that the progenies showed segregation distortion(SD). The present study indicated the possiblity to develop a high b-carotene inbred with genetic identity closer to that of the recurrent parent.