**5. Discussion**

This study reviewed the effects of CE and COO on OPB in times of pandemics utilizing the TPB in a developing country. It is essential to point out that the study results align with the TPB and confirm the theory's strength within the e-commerce environment. Although CE, COO, and TPB have been studied before, this study addresses two critical issues in international business and marketing: evaluating the three in the e-commerce environment and their significance in times of health, economic, and social crisis.


*2.4. Comparison of Frequency of Binding in the DOT Region and Other Residues of a Protein*

To estimate the difference between binding in DOT regions and other part of proteins, we calculated the binding frequency of amino acids in these regions, as shown in Figure 6 and Figure S5. Amino acids significantly differ in their binding with RNA in DOT regions and in the complete protein (*p*-value for the mean is less than 0.01). In non-ribosomal proteins, when the 3.5 Å cut-off is considered, nonpolar and aromatic residues mostly have high frequency values in the DOT regions than in the overall protein. All the frequencies are observed to be significant when statistical analysis is performed for the bootstrapped sample of the frequencies (*p*-value is less than 0.01). Residues such as His, Phe, and Leu are found to have a more than 3-fold increase in the frequency of binding in the DOT regions than in other parts of the proteins. A similar trend is observed in the NR6 dataset (Figure S5).

**Figure 6.** Binding frequency of each amino acid in the DOT region and in the overall protein for non-ribosomal complexes using the 3.5 Å cut-off.


*2.7. Identification of Transcription Factors Regulating Petal Color Formation*

Transcription factors (TFs) are important regulators that play key roles in regulating the expression of the genes during anthocyanin biosynthesis in flowering plants [30]. According to the annotation in Plant TF Database v4.0, a total of 302 TFs from 36 families were identified in the *R. pulchrum*, of which 162, 139, and 179 TFs were found in the comparisons of D vs. A, C vs. A, and B vs. A (Figure 7a and Table S5). Most of the genes encoding TFs were down-regulated in the A sample relative to the B, C, and D samples. In the D vs. A comparison, 101 TFs were up-regulated and 61TFs were down-regulated. In the C vs. A comparison, 95 TFs were up-regulated and 44 TFs were down-regulated. In the B vs. A comparison, 96 TFs were up-regulated and 83 TFs were down-regulated. Moreover, 11 TF families containing more than 10 DEGs were identified: ERF (42), MYB (29), *bHLH* (24), WRKY (20), MYB related (16), FAR1 (15), NAC (14), LBD (12), C2H2 (12), bZIP (12), and TCP (11) (Figure 7b). Notably, the ERF, MYB, *bHLH*, and WRKY families were relatively large TF families in the three pairwise groups (Table S5).

**Figure 7.** Differentially expressed TFs in the three pairwise comparisons. (**a**) Distribution of the TF families that include more than 10 DEGs. (**b**) Venn diagram of differentially expressed TFs in D vs. A, C vs. A, B vs. A comparisons.

Furthermore, we investigated ERF, MYB, and *bHLH* TFs to enhance our understanding of their involvement in regulating color formation in *R*. *pulchrum*. The overlaps among the three pairwise comparisons were calculated, and 56 TFs were found in the overlapping regions (Table S5). In our present study, five DEGs (*Rhsim02G0040100*, *Rhsim03G0176600*, *Rhsim07G0072500*, *Rhsim08G0214100*, *Rhsim12G0192100*) encoding ERF and three DEGs (*Rhsim03G0180500*, *Rhsim06G0163100*, *Rhsim09G0042000*) encoding MYB were up-regulated in the D vs. A, C vs. A, and B vs. A comparisons. Moreover, the expression level of one gene (*Rhsim08G0230000*) linked to *bHLH* was up-regulated in the A sample, as compared to the B, C, and D samples (Table S5).


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