The invention discloses osa-miR167 mediating differentiation of rice spike-stalks and application thereof. According to the invention, osa-miR167 is specifically expressed in the reproductive organ of rice; an over-expressed two-element expression vector of osa-miR167 is mediated through Agrobacterium, T-DNA is inserted and integrated to the chromosome of the rice, and obtained transgenic rice has abnormal differentiation of spike-stalks and a low fullness rate. Application of osa-miR167 provides an effective method for genetic modification and oriented creation of a novel variety of the rice.