Abstract

An efficient regeneration protocol from sugarcane callus has been developed through inclusion of amino acids in regeneration medium. Sugarcane (Saccharum officinarum L.) SP-241 callus was induced from meristematic explants cultured on Murashige and Skoog medium supplemented with B5 vitamins containing 13.6 µM 2-4, dichlorophenoxyacetic acid, 0.05% (w/v) casein hydrolysate, 10% (v/v) coconut water and 3% glucose. Five levels (0.1, 0.25, 0.5, 0.75, 1.0 mM) of five different amino acids (glutamine, asparagine, glycine, cysteine and arginine) were tested on the same medium containing 6.8 µM 2, 4-D to compare their ability to induce somatic embryogenesis and shoot regeneration from six week old callus. Among the tested amino acids, glycine (0.75 mM), arginine (0.5 mM) and cysteine (0.25 mM) showed significant effect on  smatic embryogeensis (94%) and shoot production as compared to non-amino acid medium. Of the evaluated amino acids, glycine was most effective to promote somatic embryogeensis and maximum shoot regeneration. Except low callus growth index (G.I) amino acid treatment resulted in high percentages of somatic embryogenesis and shoot regeneration as compared to non-amino acid medium. Regenerated shoots when transferred to same medium supplemented with 19.7 µM IBA, grew normal and developed roots.These results indicate the efficacy of amino acids in stimulating sugarcane plant regeneration from non embryogenic callus, and may be suitable for future use in genetic transformation studies to enhance regeneration of transgenic sugarcane plants.

Key words: Saccharum officinarum L, amino acids, embryogenic calli, plant regeneration, 2, 4 dichlorophenoxyacetic acid.