Somatic hybridization represents a powerful tool for generating novel chemotypes with enhanced biosynthetic capabilities. This study provides the first comprehensive phytochemical characterization of interspecific somatic hybrids between Gentiana cruciata L. and Gentiana tibetica King ex Hook.f., two medicinally important yet regionally rare gentians. A total of 107 compounds were detected using UHPLC-DAD-ESI-MS³, of which 31 were identified as metabolites across eight phytochemical classes. Comparative profiling revealed that all hybrids retained a conserved core of iridoids and secoiridoids while integrating lineage-specific compounds and producing hybrid-specific metabolites not detected in either parent. Despite inheriting plastids from G. tibetica, hierarchical clustering showed that the phytochemical profiles of hybrid lines were more similar to G. cruciata, the donor of the nuclear genome. Quantitative analysis of the major secoiridoids, such as gentiopicroside, swertiamarin, and sweroside, demonstrated that several hybrid lines, particularly F30A-5 and F30A-6, matched or surpassed the biosynthetic output of G. tibetica, the more productive parent. These lines also exhibited elevated antioxidant capacity, underscoring their phytochemical and functional potential. Altogether, our findings show that somatic hybridization not only preserves but may amplify the secondary metabolite capacity of the parental genotypes, offering a viable platform for sustainable in vitro production of pharmacologically relevant secoiridoids.

Helichrysum arenarium (L.) Moench is a European native species typically associated with dry, sunny, sandy, and nutrient-poor habitats. In European herbal practice, preparations from H. arenarium inflorescences are traditionally used for the symptomatic relief of digestive complaints. Polyphenolic constituents, particularly flavonoid and chalcone derivatives, are among the most characteristic compounds detected in the inflorescences. Intensive harvesting and degradation of natural habitats have increased pressure on wild populations. Since H. arenarium is legally protected in many countries, biotechnological approaches for plant biomass production represent a promising alternative. In the present study, shoot and callus cultures were established, and detailed extract profiling was performed using ultra-high-performance liquid chromatography coupled with diode-array detection and electrospray ionization multistage mass spectrometry (UHPLC-DAD-ESI-MS³). Cytotoxic, antioxidant, anti-inflammatory, wound-healing, anti-collagenase, and anti-hyaluronidase activities were also evaluated. In vitro-derived materials were compared with inflorescences and shoots of the maternal plant. The analyzed materials showed pronounced tissue-dependent metabolic differentiation. The highest callus growth rate was observed for the Ha-C-1 line, which also accumulated the highest levels of chlorogenic acid (25.80 ± 3.09 mg/g dry extract) and isochlorogenic acid A (55.83 ± 13.78 mg/g dry extract). Apigenin was detected in the inflorescence extract, reaching 3.55 ± 1.42 mg/g dry extract. The extracts did not markedly affect HaCaT viability at 15.63–500 µg/mL, while the strongest bioactivities were observed for inflorescence and maternal plant extracts.