As. Pac. J. Mol. Biol. & Biotech., Jan 2014 Vol. 1, 127-136
Application of cold pretreatment and optimisation of media for enhancement of anther culture response in two barley (Hordeum vulgare L.) genotypes derived from Bangladesh
Mozidul Haque and S. M. Shahinul Islam*
Plant Genetic Engineering Laboratory, Institute of Biological Sciences, University of Rajshahi, Rajshahi-6205, Bangladesh.
* Author for correspondence: Dr. S. M. Shahinul Islam
Associate Professor, Plant Genetic Engineering Lab., Institute of Biological Sciences, University of Rajshahi, Rajshahi-6205, Bangladesh.
Rapid development of embryoids and the efficiency of regeneration by anther culture was observed in two barley genotypes. Five semi-solid media viz. MS, B5, N6, FHG and AMS3 were used for embryo induction as well as plant regeneration. Harvested spikes were pretreated with cold for 2, 4, 6, 8, 10, 12, 14 or 16 days at 4°C in the dark. Cold pretreatment for 8-12 days produced the highest frequency of embryoid induction on FHG medium. The FHG medium was modified by MS that was supplemented with 2,4-D (2.0 mg/l), kinetin (0.5 mg/l) and an amino acid (730 mg/l L-glutamine). Out of two genotypes, BB-6 performed better in terms of embryo formation and green plant regeneration in MSR medium. The highest level of embryoid induction (14.6%) was observed at T5 (10 days), producing a total of 13.8% regenerated plantlets and 10.72% green plants in BARI barley-6. Analysis of variance (ANOVA) showed significant differences in embryoids induction rate depending on the duration of cold pretreatment, culture media and between the genotypes. It was observed that the interactions of medium and cold pretreatment, and genotype and medium, were significant in determining the rate of embryoid induction. Cold pretreatment for 10 days of excised anthers from spikes produced optimum results in both genotypes, and the FHG medium was the best out of the five tested for embryoid induction and plant regeneration.