Abstract: Pennisetum sp. is monocotyledon plant and widely planted in China and Africa as it can be used as forage and culture substrates of edible fungi. However, the lack of cold tolerance limits its popularization. Genetic technology or somatic hybridization has potential to improve the cold tolerance or quality of Pennisetum sp. which require the establishment of Pennisetum sp. tissue culture system. In the present stydy, the effects of different disinfection ways and different growth hormone such as 2,4D, IAA, NAA and cytokinins (6BA and KT) on callus induction were studied using caulicle of Pennisetum sp. as explants. The results showed that the ideal disinfection method was soaking in 2% sodium hypochlorite solution for 10 min after wiping stripping leaf sheath with 75% alcohol. All of the medium with the different concentrations of 2,4D ranged from 1.0 mgL-1 to 4.0 mgL-1 can induce callus without significant difference. The medium with 0.2 mgL-1 IAA can induce more callus. The medium with different cytokinins can not induce more callus. Based on these observation, the optimal callus induction medium of Pennisetum sp. was MS + 2,4D (3.0 mgL-1) + IAA (0.2 mgL-1).