The in vitro shoot propagation of Cannabis sativa L. is an emerging research area for large-scale plant material production. However, how in vitro conditions influence the genetic stability of maintained material, as well as whether changes in the concentration and composition of secondary metabolites can be expected are aspects that need to be better understood. These features are essential for the standardised production of medicinal cannabis. This work aimed to find out whether the presence of the auxin antagonist α-(2-oxo-2-phenylethyl)-1H-indole-3-acetic acid (PEO-IAA) in the culture media influenced the relative gene expression (RGE) of the genes of interest (OAC, CBCA, CBDA, THCA) and the concentrations of studied cannabinoids (CBCA, CBDA, CBC, ∆9-THCA, and ∆9-THC). Two C. sativa cultivars, 'USO-31' and 'Tatanka Pure CBD', were cultivated by in vitro conditions with PEO-IAA presence and then analysed. The RT-qPCR results indicated that even though some changes in the RGE profiles could be observed, no differences were statistically significant compared with the control variant. The results of the phytochemical analyses demonstrate that although there were some differences from the control variant, only the cultivar 'Tatanka Pure CBD' showed a statistically significant increase (at a statistical significance level α = 0.05) in the concentration of the cannabinoid CBDA. In conclusion, it would appear that using PEO-IAA in the culture medium is a suitable approach to improve in vitro cannabis multiplication.

Centre of the Region Haná for Biotechnological and Agricultural Research Department of Genetic Resources for Vegetables Medicinal and Special Plants Crop Research Institute 78371 Olomouc Czech Republic

Czech Advanced Technology and Research Institute Palacký University Olomouc 78371 Olomouc Czech Republic

Department of Biochemistry Faculty of Science Palacký University 78371 Olomouc Czech Republic

Department of Botany Faculty of Science Palacký University Olomouc 78371 Olomouc Czech Republic

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