Micropropagation of endemic Muscari latifolium and Muscari coeleste

This study aimed to develop optimized micropropagation protocols for the rapid clonal propagation of the endemic species Muscari latifolium and Muscari coeleste. Seeds were used as the initial explant for M. latifolium, while bulb explants were used for M. coeleste. For M. latifolium, surface sterilization using 20% NaOCl for 30 minutes was effective, achieving 100% decontamination and 100% germination. The maximum bulblet number (21.06 per explant) and bulblet length (0.71 cm) were obtained on MS medium supplemented with 1.0 mg/l BAP. Conversely, MS medium supplemented with 1.0 mg/l BAP in combination with 1.0 mg/l NAA resulted in a reduced bulblet number (5.60 per explant) but produced bulblets with a larger diameter (0.19 cm) and a higher rooting frequency (75%). Root induction was most successful on MS medium containing 2 mg/l IBA, with a rooting rate of 100%. During the acclimatization phase, plantlets grown in a peat–soil mixture exhibited a survival rate of 47%. For M. coeleste, surface sterilization with 80% NaOCl for 40 minutes proved to be the most effective treatment, resulting in complete elimination of contamination (0%) and full explant viability (100%). Shoot induction was most effectively achieved on MS medium supplemented with 4 mg/l BAP in combination with 1–2 mg/l NAA, yielding callus (100%) and shoot formation. During bulblet induction, the highest number of bulblets (5.07±0.27 per explant) was obtained on MS medium containing 4 mg/l BAP + 1 mg/l NAA, whereas the greatest bulblet length (2.42±0.08 cm) was recorded on MS medium supplemented with 4 mg/l BAP + 2 mg/l NAA. Similarly, adventitious bulblet formation followed a comparable trend, with the highest bulblet number (3.47±0.68 per explant) observed on MS medium containing 4 mg/l BAP + 1 mg/l NAA, while the largest bulblet diameter (0.19±0.06 cm) and length (0.77±0.18 cm) were achieved on MS medium supplemented with 4 mg/l BAP + 2 mg/l NAA. Rooting experiments indicated that MS medium supplemented with 2 mg/l IBA produced the most favorable results, and during acclimatization, the peat–soil mixture resulted in the highest survival rate (53%). In conclusion, the developed protocols for both M. latifolium and M. coeleste demonstrated high propagation efficiency, morphological quality, and acclimatization success. The use of seeds as starting material in M. latifolium reduces the need for bulb harvesting from natural populations, contributing to ex situ conservation and sustainable ornamental plant production. It is expected that the protocol developed for M. coeleste, which is used for medicinal purposes, will provide a basis for future pharmacological and medical studies. The obtained findings may also provide a model for the propagation of other rare geophyte species facing similar conservation challenges.