Lycium barbarum is a potential high-quality woody forage whose crude protein concentration is up to 30% and it is rich in numerous secondary metabolites. This study aimed to investigate the effects of L. barbarum included in alfalfa-tall fescue mixture on in vitro dry matter digestibility (IVDMD), total gas production (TGP), fermentation characteristics, productions of carbon dioxide (CO2), methane (CH4), and nitrous oxide (N2O), and global warming potential (GWP). Three cultivars of L. barbarum, ‘Ningqicai (NQC) No.1’, ‘Ningqi (NQ) No.9’, ‘Qixin (QX) No.2’, were included at 4%, 8%, and 12% levels of substrate with 7 replicates. After 72 h of incubation, the highest IVDMD of 3 cultivars were observed at 4% compared to the control. Although TGP and GWP of 3 cultivars at 4% were similar to the control (P > 0.05), they increased quadratically (P < 0.05) with increasing substitution levels for ‘NQC No.1’ and reduced quadratically (P < 0.05) for ‘NQ No.9’, corresponding with the productions of CO2 and CH4, and ammonia nitrogen (AN) and violate fatty acids (VFA). The productions of CO2 and CH4, acetate to propionate ratio, and GWP of ‘QX No.2’ were significantly lower than the control at 12% level (P < 0.05). L. barbarum inclusion accelerated N2O production for all treatments (P < 0.05). Overall, 4%, 8% and 12% were recommended for ‘NQC No.1’, ‘NQ No.9’ and ‘QX No.2’, respectively, which increased IVDMD and fermentation characteristics, and reduced CO2, CH4 and N2O productions. These findings suggest that L. barbarum can be a kind of new potential woody forage for ruminant production with high degradability and low GWP at optimal inclusion levels.