The invention discloses an endophytic penicillium strain of Xiangyang ophiopogon japonicus and an application of the endophytic penicillium strain. The endophytic penicillium strain of Xiangyang ophiopogon japonicus is penicillium sp. strain EF010, wherein the preservation number is CCTCC NO: M2018778, and the preservation time is November 12, 2018. The penicillium sp. strain EF010 provided by theinvention is obtained by separating and screening fresh tuberous roots of Xiangyang ophiopogon japonicus plants through steps of culture, screening, purification and the like, and the strain can be metabolized by itself to generate Hubei ophiopogon japonicus polysaccharide, so the strain can be applied to preparation of Hubei ophiopogon japonicus polysaccharide by microbial liquid fermentation. The liquid fermentation mode is less interfered by external environment, and the yield of fermentation production is high, so that Hubei ophiopogon polysaccharide can be rapidly, stably and massively produced through liquid fermentation of the penicillium strain EF010. In addition, compared with a conventional chemical extraction process, the liquid fermentation process has little pollution to theenvironment, so that production of Hubei ophiopogon polysaccharide is more environmentally friendly.