## 4 Conclusions and Discussion
Furthermore, the dual-luciferase assay confirmed that the promoter of _FaMADS1_ could be significantly inhibited by FaTRAB1, whereas FaABIS-5 and FaABIS had little influence on the promoter of _FaMADS1_. These results suggested that the ABA signal was transmitted to the _FaMADS1_ promoter with the interaction of FaTRAB1. _FaMADS1_ regulated the transcriptional level of the downstream anthocyanin-related genes, softening-related genes, and aroma-related genes, eventually initiating fruit ripening. In conclusion, _FaMADS1_ expression negatively depended on ABA to regulate strawberry fruit ripening. The yeast one-hybrid analysis and dual-luciferase assay suggested thatthe _FaMADS1_ promoter was inhibited by the interaction with FaTRAB1, resulting in the blockage of _FaMADS1_ translation. ABA application and _FaMADS1_-RNAi data reflected a negative regulation of _FaMADS1_ to the downstream anthocyanin-related genes (_PAL6_, _C4H_, _4 CL_, _DFR_, and _UFGT_), softening-related genes (_PL_ and _XTH_), and aroma-related genes (_QR_ and _AAT2_), as well as the enzyme activities (Figure 10). This observation enhanced our understanding of the molecular mechanism of ABA signaling during the non-climacter fruit ripening. **Supplementary Materials:** The following supporting information can be downloaded at: [[https://www.mdpi.com/article/10.3390/foods12091802/s1](https://www.mdpi.com/article/10.3390/foods12091802/s1)]([https://www.mdpi.com/article/10.3390/foods12091802/s1](https://www.mdpi.com/article/10.3390/foods12091802/s1)), Table S1: All primers used in the study for PCR analysis; Table S2: Bioinformatics analysis of the _FaMADS1_ gene promoter. **Author Contributions:** Conceptualization, L.M. and W.L. ; methodology, W.L. ; software, X.W. ; validation, X.W., X.H. and R.C. ; formal analysis, W.L. ; investigation, X.H. ; resources, X.W. ; data curation, W.L. ; original draft preparation, W.L. ; review and editing, X.Z. and L.M. ; visualization, W.L. ; supervision, L.M. ; project administration, C.X. and X.W. ; funding acquisition, L.M. All authors have read and agreed to the published version of the manuscript. **Funding:** This research was funded by the National Natural Science Foundation of China (31972468). Figure 10: A model for ABA receptors-mediated signaling pathway connected to _FaMADS1_ regulating strawberry fruit ripening. In the presence of ABA, ABA signal activated ABI and TRAB transcription factors, and then _FaMADS1_ was inhibited. Depression of _FaMADS1_ induced the expression of anthocyanin-related _FaPAL_, _FaC4H_, _FaC1_, and _FaUFIGT_, softening-related _FaPL_ and _FaXTH_, and aroma-related _FaQR_ and _FaMAT_. Abbreviations: AAT2, alcohol acyltransferase; ABA, abscisic acid; 4 CL, \(\rho\)-cumarate ligase; C4H, cinnamicacid-4-hydroxylase; PAL6, phenylalanine ammonia lyase; PL, pectate lyase; QR, quinone oxidoreductase; SnRK2s, phosphorylation of ABA-activated sucrose non-fermenting 1-related protein kinases subfamily 2; TRAB, transcription factor responsible for ABA regulation; UFGT, flavonoid 3-O-glucosyltransferase; XTH, xyloglucan endotransglycosylation-lase/hydrolase. The solid arrows indicate direct action and the dashed arrow indicates indirect action.