The invention provides a method for improving rapid seedling of barley tissue culture. The method for improving rapid seedling of barley tissue culture comprises the following steps of: (1) selectingbarley immature embryos with a diameter of 1-3 cm, and sterilizing the surfaces of the barley immature embryos with alcohol and sodium hypochlorite so as to reduce the rate of bacterial contamination;(2) separating coleoptiles from the immature embryos accurately, conducting secondary treatment on scutellum, and inducing rapid germination of callus tissue; (3) adjusting the time of pre-culture and agrobacterium infection according to the size of the immature embryos so as to prevent overdose of bacteria liquid; (4) adopting a weak-light shielding method to improve the differentiation rate ofgreen spots and the percentage of seedling emergency; and (5) adjusting the antibiotics ratio and usage time of a callus induction medium, a differentiation medium and a rooting medium, and inducing rapid seedling of the callus tissue. The method for improving rapid seedling of the barley tissue culture is a method for the tissue culture of the barley immature embryos, the germination of the callus tissue is rapid, and the rate of embryonic callus tissue is high; besides, the operability is high, the differentiation rate of the green spots and the percentage of seedling emergency are high, andno restriction of material genotypes exists; an established transformation regeneration system of the barley immature embryos can provide technical support for the research on genetic improvement andcell engineering seed breeding of the barley.