The invention belongs to the technical field of edible mushrooms, and particularly relates to a white needle mushroom JK01 strain and application thereof. A needle mushroom provided by the invention is named JK01, is obtained by hybridizing a Taichangbai strain and a hybridized strain 19 serving as parents through systemic breeding, belongs to needle mushroom (Flammulina velutipes), and is collected in the Institute of Microbiology, Chinese Academy of Sciences (address: No. 3, 1# Yard, Beichen West Road, Chaoyang District, Beijing) with a collection number CGMCCNo.13877 on June 14, 2017. Compared with the prior art, the white needle mushroom JK01 strain provided by the invention has the advantages of short culturing period, consistent growth, high stress resistance, high yield, suitability for industrial cultivation, thick stems, superior shape characteristic, crispy mouthfeel, contribution to splitting and cooking of products, high comprehensive performance, and wide market prospect.