Gaolejeunea was previously known as a monospecific genus endemic to China. Specimens of an undescribed species belonging to the genus were recently found in Singapore. Critical assessments of morphological characters and phylogenetic analyses based on genetic sequence data of three regions (trnL-F, rbcL, nrITS) unequivocally confirmed and supported its recognition as a new and second species of Gaolejeunea. Gaolejeunea hoi sp. nov. is here fully described and illustrated. The new species is characterized by its faintly Allorgella-type marginal denticulations, the ental hyaline papilla attached to the apical margin, the bracteoles present only at the base of each androecium, gynoecia with innovation(s), perianth with wing-shaped lateral keels.