The object of the invention is to isolate and initiate i n vitro microspore culture using the early to mid-spring barley stage, in particular to produce DH line more efficiently from genotypes/varieties with a high level of albino plant regeneration in the standard method. The method of isolating and initiating in vitro culture the microspore using the early to mid-spring barley stage consists of cutting the stem containing the microspores in the early to mid-to-medium development stage and placing it in water,and then stored at 4.oC temperature until the logs are isolated. The brush is sterilised on the surface with 70-100% ethanol and then the logs are removed and the bone is removed using a sterile brush. The bait shall be taught and stored to provide adequate moisture. Such prepared logs are subjected to controlled cooling. Cold trouble cuts right through you. One millimeter fragments and puts in a cooled blender chamber. The bait blends twice in 0,4 M mannithOil at idle speed. Blinded logs are passed through a 100m sterile filter into a container on ice. The remaining logs are collected from the filter and blended twice into 0,4 M mannitol to release as many microspores as possible and re-filtered through a 100m filter. The suspension is transferred to the test tube and rotates pris 120-130 x g through 8-10 min at 4-oC temperature, supernatant is then removed and microspore sediment is suspended in 0,55 M maltose. The suspension is transferred to a new sterile test tube and 0,4 M mannitol is applied to the surface and rotates in a concentration gradient at 100-110 x g through 8-10 min at 4,C temperature to separate viable microsporeson the edge of the phases, from the non-viable in the pellet. The micropores at the edge of the phase in the ring are collected and rotated to be deposited at 120-130 x g through 8-10-min at 4-oC temperature. The microspore sediment is then dried, and the micro-spores are suspended in a modified KBP inducer medium by adding the hydr
