Moussous, AsmaMoussous, WalidKhelifi, Lakhdar2026-06-082026-06-082026-05-132784-0379https://repository.iuls.ro/handle/20.500.12811/6539This study evaluated the growth kinetics and hyoscyamine biosynthesis of transgenic hairy roots from Datura innoxia (DI), Datura stramonium var. stramonium (DS) and Datura stramonium var. tatula (DT) induced by Agrobacterium rhizogenes strain A4 and cultured in vitro for five years after their induction using liquid chromatography–tandem mass spectrometry for alkaloid analysis. The lines were elicited with AgNO3 for 24 and 48 h to determine its effects on the biomass, hyoscyamine and scopolamine yield of hairy roots. DT grew better, resulting in 0.295 g of dry mass after 21 d of culture on B5 medium in the first year, compared to 0.229 g for DI and 0.205 g for DS. Over five years, DT grew better than DS and DI. Furthermore, the hyoscyamine yield was higher in DT than in DI and DS with the best yield occurring in the first year of culture (8.575 mg g−1 DW). After chemical elicitation, the biomass, expressed as the dry weight of DS and DT, decreased under all AgNO3 treatments. However, the DI biomass increased after AgNO3 treatment for 24 and 48 h (74%) of the control. Only two treatments promoted hyoscyamine biosynthesis, and the best yield was obtained with 7.89 mg g−1 DW of hyoscyamine after 48 h of elicitation. However, the scopolamine yield was improved by AgNO3 elicitation for 24 h with the best result obtained for DT hairy roots (0.539 mg g−1 DW).EnglishAttribution 4.0 International (CC BY 4.0)http://creativecommons.org/licenses/by/4.0alkaloidsbiomassDaturahairy rootsin vitroArticle2784-0360https://doi.org/10.46909/alse-591203