METHOD FOR OBTAINING DOUBLED HAPLOIDS OF CARROTS IN CULTURE OF ISOLATED MICROSPORES IN VITRO

摘要

FIELD: biotechnology.;SUBSTANCE: invention relates to the field of biotechnology. The invention is a method for producing doubled haploids of carrots in the culture of isolated microspores in vitro. In the method providing for the isolation of microspores, their cultivation on a nutrient medium, the surface sterilization of the umbrellas is carried out in 2% sodium hypochlorite with the addition of Tween 20 for 15 minutes, followed by three rinsing in sterile water, the isolation of microspores is carried out by crushing the umbrellas in a liquid nutrient medium B5 with the addition of 130 g/l of sucrose and the additional inclusion of 75 g/l of mannitol, the density of the resulting suspension is optimized to 5×10⁴ microspores/ml and cultured on NLN nutrient medium with the addition of 130 g/L sucrose, 0.2 mg/L 2,4-D, 0.2 mg/L NAA, 200 mg/L activated coal and 400 mg/L of casein hydrolyzate at a temperature of 33°C for 5 days, followed by cultivation at 25°C in the dark until the formation of embryoids.;EFFECT: invention allows increasingthe yield of doubled haploids in the culture of isolated microspheres.;1 cl, 1 tbl