High-efficiency micropropagation of Rhodiola species with concurrent bioactive compound validation

Rhodiola species are highly valuable medicinal resources that hold significant importance in both traditional medicine and modern health fields. However, existing breeding techniques suffer from pain points such as low breeding efficiency and unstable active ingredients, which seriously restrict the development and utilization of their medicinal value. To develop a rapid tissue culture propagation system for endangered medicinal plants ( Rhodiola spp.) in plateau regions, and to determine the optimal conditions for seed germination, proliferation, seedling strengthening, rooting induction, and transplanting substrates across stages. L₉(3 3 ) orthogonal design and Box-Behnken response surface methodology were employed to explore the optimal tissue culture conditions. Results showed that the highest seed germination 92.67% was realized with 75% ethanol (30 s) pre-treatment, and then 1.0% NaClO (10 min). This study established a rapid tissue culture system for R. crenulate and R. rosea , achieving a proliferation coefficient of 7.38, 89% rooting rate, and enhanced active compound production. The most suitable transplanting substrate was a 2:1:1 mixture of vermiculite, sphagnum peat, and river sand, resulting in a survival rate of 68.54%. Post-transplantation, R. crenulate rhizomes accumulates higher salidroside and gallic acid, while R. rosea L. shows superior rosavin and tyrosol accumulation, revealing distinct interspecific patterns in active compound production. In conclusion, a tissue culture system for Rhodiola species was established providing a solid foundation for their sustainable resource utilization.