Abstract: In this study, with the young spikes （2-3cm） of hybrid F1 of Elymus scibiricus × E.purpuraristatus as explants and MS as basic medium, the effects of 4 kinds of hormones （2,4-D,6- BA,NAA and IBA） on callus induction, plantlet differentiation and rooting were studied. The optimal medium for callus induction of hybrid F1 was MS＋2,4-D 2.0mg/L, with the induction rate of 63.3%;The optimal medium for plantlets differentiation was MS＋6-BA 2.0mg/L＋NAA 0.4mg/L,with the induction rate of 75%. The optimal medium for rooting was MS＋IBA 0.5 mg/L, with the induction rate of 90%;and the regenerated plant was obtained after acclimatization and transplanting. This regeneration system provides conditions for the following colchicin-induced chromosome doubhng, and ultimately overcoming its sterility to create new grass germplasm resource.