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. 2021 Jun 19;40(9):1631–1646. doi: 10.1007/s00299-021-02707-3

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© The Author(s) 2021

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — Gene expression of DlP5βR1 and DlP5βR2 in Digitalis lanata WT and RNAi-mediated knockdown shoots. Relative RNA expression levels were calculated using the qPCR method by applying the 2^−ΔΔCT method with actin as the reference gene. The y axis denotes the normalized relative transcript accumulation of the genes indicated in the x axis (mean ± SEM, n = 3; Tukey’s test, *P > 0.01). (a) Relative transcripts of P5βRs (DlP5βR1 and DlP5βR2) in D. lanata WT; (b) relative expression of DlP5βR1 and DlP5βR2 in a Dl P5βR1-RNAi knockdown line 1 (L1) and line 2 (L2) compared to WT shoots; c relative expression of DlP5βR1 and DlP5βR2 in a Dl P5βR2-RNAi knock down line 1 (L1) and line 2 (L2) compared to WT shoots; (d) quantification of digoxigenin and digitoxigenin in D. lanata WT shoots and transgenic shoots with reduced expression of P5βRs. Shoots transformed with 679p935s-GusIo-rbs (VC) were used as control to exclude artificial effects created by the transformation process (mean ± SEM; n = 9; Tukey’s test, *P > 0.05)