Abstract: Study was carried out on effect of different concentration acetic acid(2,4-D), 6-benzylamino purine(6-BA)and casein hydroly-sate(Hy) under different culture medium on callus induction of three varieties of Poa Pratensis L. with its seeds as explants. The result showed that (1)MN6 culture medium could improve callus induction rate; (2) culture medium of callus induction and regeneration of embryo was MN6 +2,4-D 3mg/L+BA0.2mg/L;(3) effect of different concentration casein hydroly-sate(Hy) was weak on callus induction;(4) callus induction rate of Nuglade was the highest in three varieties of Poa Pratensis L.