Artificial moss biocrusts (moss-dominating biological soil crusts) have attracted attention as a new type of windproof and sand fixation method in recent years. In order to further optimize the existing breeding and propagation technology, this study used a 4 factor and 2 level orthogonal test design to discuss the effects of microorganism (Actinomyces bovis, Bacillus megaterium), algae (Chlorella, Microcolusvaginatus), artemisia gum (yes, no), and substrate type (sand soil matrix, formulation matrix) on the indoor propagation of sand-land moss biocrusts. The results showed that: (1) Among the four factors, only the substrate type significantly affected the coverage and plant density of moss biocrusts (P<0.01), which increased to 61.04% and 17.48 strains/cm², were higher than that of the formulation substrate treatment. (2) Algae, microorganism, and substrate types significantly influenced the physiological characteristics of moss biocrusts (P<0.01), while the effect of artemisiagum was not significant. The sandy substrates and the addition of actinomycetes and chlorella significantly increased the peroxidase (POD) activity and soluble protein content and reduced the content of malondialdehyde (MDA) of moss biocrusts. (3)The optimal treatment for propagation was sandy substrate + actinomycete (1 g/kg substrate) + chlorella (2 g/m²). Under the conditions of the incubator (set illumination, photoperiod, temperature, air humidity as 6 000 lx, 12 h/d, 20 ℃, 75%), we propagated 1 m² moss biocrusts (collected from the field and fragmented into provenance) to 4 m² moss biocrusts with a coverage of 97.14%, which achieved 4 times multiplication. This research would provide technical support for large-scale industrialization and efficient propagation of moss biocrusts in the future.