Vegetative propagation of mosses under controlled conditions: A reproducible, low-cost method for sustainable moss cultivation

This protocol presents a reproducible, sustainable method for vegetative propagation of mosses under controlled but non-sterile conditions. It enables scalable cultivation of mosses for green infrastructure while reducing dependence on wild collections. The method is based on a fundamental and unavoidable trade-off: maintaining high humidity, which is essential for moss growth, while simultaneously limiting fungal and microbial contamination. This balance determines success, restricts species selection, and defines facility and management requirements. All stages of cultivation and validation must therefore consider this constant challenge: promoting moss vitality while preventing overgrowth of competing microorganisms. Compared to traditional sterile cultivation, this open-system method lowers costs and technical complexity without compromising growth efficiency. Four moss species - <i>Dicranum scoparium, Plagiomnium affine, Pleurozium schreberi,</i> and <i>Hypnum cupressiforme</i> - were successfully propagated on hemp mats under controlled light, humidity, and temperature regime. After four months, moss coverage exceeded 80% on most 60 × 60 × 0.4 cm tiles, demonstrating the method's reproducibility and efficiency. 1. The protocol provides a cost-effective, replicable method for moss cultivation under controlled conditions in non-sterile, open environments. 2. The method minimizes ecological impact by reducing wild moss harvesting. 3 The method supports sustainable applications of mosses in green infrastructure and environmental biotechnology.