Optimization of Conditions for Callus Induction from BC2F1 Population (Ranbir Basmati x PAU148) through Anther Culture
Mridhu Sharma *
Department of School of Biotechnology, Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, Chatha, Jammu and Kashmir- 180009, India.
A. K. Singh
Department of School of Biotechnology, Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, Chatha, Jammu and Kashmir- 180009, India.
R. K. Salgotra
Department of School of Biotechnology, Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, Chatha, Jammu and Kashmir- 180009, India.
Manmohan Sharma
Department of School of Biotechnology, Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, Chatha, Jammu and Kashmir- 180009, India.
Bupesh Ku. Sharma
Department of Plant Breeding and Genetics, Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, Chatha, Jammu and Kashmir- 180009, India.
S. K. Gupta
Department of Plant Breeding and Genetics, Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, Chatha, Jammu and Kashmir- 180009, India.
Vikas Sharma
Department of Biochemistry, Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, Chatha, Jammu and Kashmir- 180009, India.
*Author to whom correspondence should be addressed.
Abstract
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.
Keywords: Callus induction frequency, anther culture, 2,4- D, cold pre-treatments, amino acids.