Shuwen He

As a Chinese national key protected medicinal fern naturally occurring in forest understories, Cibotium barometz faces severe threats of wild population degradation, while standardized large-scale artificial breeding technology for conservation purposes remains immature. To establish an efficient spore-based conservation propagation system for this endangered forest fern, this study quantified the independent and interactive effects of spore storage temperature, storage duration and sowing density on spore germination, gametophyte growth and sporophyte seedling establishment. Spores were preserved under four gradient temperature treatments with sequential sampling at multiple storage durations, followed by sowing trials with a series of density gradients; germination rate, seedling establishment rate and gametophyte–sporophyte conversion rate were dynamically recorded and statistically analyzed. The results demonstrated that appropriately extended storage significantly shortened the germination phase and simultaneously elevated both spore germination and sporophyte seedling formation rates. Among all temperature treatments, storage at −4 °C achieved the maximum germination and seedling establishment capacity, whereas ultra-low-temperature cryopreservation at −196 °C greatly promoted gametophyte–sporophyte conversion rate. The optimal sowing density balancing growth space and survival rate was determined to be 30 spores per cm². The complete dynamic developmental traits covering the full spore propagation life cycle of C. barometz were systematically summarized in this work. Our findings supply reliable technical parameters to standardize spore breeding protocols, and offer critical support for ex situ conservation, wild forest population restoration and sustainable resource utilization of C. barometz.