## 2 Results and Discussion
### Secoriridoids-Related Biosynthetic Genes Expression Profiles in Response to Wounding
erythraea_ leaf transcriptome database [59], we identified nucleotide sequences of several transcription factors candidates (_CeMYC2_, _CeBIS1_, _CeJAZ1_, _CeCO11_, _CeJAM2_, and _CeJAM3_) for which we expected to be included in wounding-induced regulation of SGs biosynthesis. On the other hand, no putative genes for the ORCA2 and ORCA3 transcriptional factors were found. In addition for BIS2, which has previously been identified in _C. roseus_[31], no candidate genes were retrieved in the _C. erythraea_ leaves transcriptome database. It is possible that this functional homologue of BIS1 is specific for the regulation of MIA-related genes downstream of secologanin in _C. roseus_, and it has no homologue in _C. erythraea_. _CeJAZ1_ expression was induced 2 HAW, while minor increase in _CeJAM3_ and _CeCO11_ expression was noted 2/4 HAW and 48 HAW, respectively, according to the qPCR results (Figure 3). The enhanced expression of _CeBIS1_ was detected already 2 HAW, and the max transcript amounts were recorded 24 HAW (Figure 3). Interestingly, factorial ANOVA revealed differences between factors that influence the expression of each of the analyzed _C. erythraea_ TFs. To be precise, time was the only factor that has a significant impact on the expression of _CeCO11_, _CeBIS1_, _CeJAM2_, and _CeJAM3_ upon wounding (_p_ < 0.05), as it was for all genes encoding for enzymes that are involved in SGs biosynthesis. The type of leaves (local or systemic) was the factor that influenced _CeMYC2_ expression, while expression of CeJAZ1 was influenced by time, by the type of leaves, as well as by the interaction of these two factors (Figure 3). The most important difference in the expression of _CeJAZ1_ was observed 2 HAW in LL (Figure 3). Similarly, [PERSON] et al. [30] reported an increase of _JAZ1_ expression upon wounding in leaves, but not in stems of _C. roseus_. JAZs themselves are wound inducible, and _JAZ_ genes exhibit different transcription patterns, which indicate that specific JAZ proteins might regulate different TFs and downstream responses to environmental stresses in specific development stages, tissues, or cell types [58; 60]. The expression of _CeBIS1_ in leaves of _C. erythraea_ upon wounding followed the pattern of _CeSLS_ expression, both in terms of time course and intensity. According to the obtained results (Figure 3), _CeBIS1_ might be indicated as the TF that is positively involved in wounding-induced regulation of SGs biosynthesis. This is in accordance with the previous finding of [PERSON] et al. [30], which suggests that overexpression of _BIS1_ caused a dramatic increase of loganic acid and of downstream (seco)-iridoid (secologanin) and MIAs in _C. roseus_ (strictosidine, ajanicine, serpentine, and tabersonine). Expression of _BIS2_ showed analogous pattern of changes as _BIS1_[31]. Similarly, [PERSON] et al. [52] correlated detected low iridoid and MIA content with the lowered expression of _BIS1_/_BIS2_ and of several secologanin-related biosynthetic genes that are expressed in IPAP cells. Figure 3: Relative gene expression patterns of selected transcription factors in _C. erythraea_ leaves at determined time points (h) after wounding (HAW). The relative expressions of the genes were normalized against _CeEF1_ gene as an internal control; unwounded leaves at 0 HAW were set as a calibrator. Since factorial ANOVA revealed that leaf type does not significantly contribute to the expression of _CeCO11_, _CeBIS1_, _CeJAM2_, and _CeJAM3_ transcription factors, only the effect of time was  
post hoc tested. In these cases, bars with different letters are significantly different (\(p<0.05\)) according to post hoc [PERSON]'s test. Leaf type (SL or LL) was the factor that significantly contributed to gene expression of _CeMYC2_. Factorial ANOVA indicated that time, leaf type, and interaction time x condition significantly contribute to the _CeJAZ1_ gene expression. In this case each bar with different letters is significantly different (\(p<0.05\)) according to post hoc [PERSON]'s test.