THE OPTIMIZATION OF CULTIVATION CONDITIONS <i>IN VITRO </i>OF ALFALFA (MEDICAGO SATIVA L.)
In the current work we optimized the conditions of cultivation in vitro of alfalfa (Medicago sativa L.). To obtain callus and regenerants were used the medium with contain the macro-and microelements in Gamborg, Shenk and Hildebrandt, with the addition of 3mg/l BAP(6-benzylaminopurine) and 0.1 mg/l NAA (NAA--naphthaleneacetic acid) and with different contents of vitamins: from 1-10 mg/l of thiamine, 0.5-1 mg/l pyridoxine, 0.5-5 mg/l of nicotinic acid,100-100 mg/l of meso-inositol and sucrose (from 10-30g/l). Of the eight tested medium the medium Schenk- Hildebrandt with 5 mg / l thiamine, 0.5 mg / l pyridoxine, 5 mg/l of nicotinic acid and 10 g / l sucrose was chosen. The morphogenic callus frequency ranged from 68 to 94% and the regenerants frequency ranged from 55-75%. Since this medium was used in all stages of cell cultivation of alfalfa, it is possible to simplify the procedure for forming regenerated plants and reduce the duration of their production to 2-3 months.