FIELD: agriculture.
SUBSTANCE: invention relates to the field of agricultural biotechnology. Method of producing dihaploid plants of barley from cultured microspores in vitro, comprising: growing donor plants at reduced air temperature of 15-20°C, light regime: 16 h day/8 h night, relative humidity of 60-70%, lighting intensity of 10000-15000 lux, with carrying out phytosanitary treatment, at that growing donor plants is carried out to the stage from opening of the sheath, when the inflorescence is in the flag leaf, - stress treatment of ears at 4°C, in tubes with a poor environment containing KCl - 1.5 g/l, MgSO4×7H2O - 0.25 g/l, CaCl2×2H2O - 0.1 g/l, mannitol - 60 g/l, potassium phosphate buffer - 1 ml/l, pH 7.0, for 7 days for shifting from gametophytic development path of microspore to sporophytic path - microspore isolation from spikelets in sterile conditions, - culturing the isolated microspores in a modified environment for embryogenesis induction, consisting of 6% maltose, 10 sets per 1.5 ml of the culture, and plant growth regulators in amount of 1 mg/l 2,4-D, 0.2 mg/l zeatin, and adding the aforementioned components is carried out before the addition of microspores, - regenerating plants from embryoids by culturing on solid nutrient Murashige and Skoog medium without the adding of plant growth regulators, - processing the barley haploid plants with antimitotic agent N-diacetyl-N-(β,γ-epoxypropyl) aminocolchicine for doubling chromosomes and obtaining dihaploid plants.
EFFECT: invention enables to obtain dihaploid homozygous lines of barley for breeding new varieties and hybrids with improved properties.
1 dwg, 2 tbl, 6 ex
