The molecular mechanism underlying anthocyanin metabolism in apple using the MdMYB16 and MdbHLH33 genes
| Autor: | Jingxuan Liu, Ninglin Lu, Xuesen Chen, Wang Yicheng, Zongying Zhang, Jiang Shenghui, Changzhi Qu, Wang Nan, Deyun Wang, Xu Haifeng |
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| Rok vydání: | 2017 |
| Předmět: |
0106 biological sciences
0301 basic medicine Malus Repressor Plant Science 01 natural sciences Anthocyanins Gene Knockout Techniques 03 medical and health sciences chemistry.chemical_compound Bimolecular fluorescence complementation Gene Expression Regulation Plant Genetics Cloning Molecular Gene Transcription factor Plant Proteins biology fungi food and beverages Promoter General Medicine biology.organism_classification 030104 developmental biology Biochemistry chemistry Fruit Callus Anthocyanin Phyllachorales Agronomy and Crop Science Transcription Factors 010606 plant biology & botany |
| Zdroj: | Plant Molecular Biology. 94:149-165 |
| ISSN: | 1573-5028 0167-4412 |
| Popis: | MdMYB16 forms homodimers and directly inhibits anthocyanin synthesis via its C-terminal EAR repressor. It weakened the inhibitory effect of MdMYB16 on anthocyanin synthesis when overexpressing MdbHLH33 in callus overexpressing MdMYB16. MdMYB16 could interact with MdbHLH33. Anthocyanins are strong antioxidants that play a key role in the prevention of cardiovascular disease, cancer, and diabetes. The germplasm of Malus sieversii f. neidzwetzkyana is important for the study of anthocyanin metabolism. To date, only limited studies have examined the negative regulatory mechanisms underlying anthocyanin synthesis in apple. Here, we analyzed the relationship between anthocyanin levels and MdMYB16 expression in mature Red Crisp 1-5 apple (M. domestica) fruit, generated an evolutionary tree, and identified an EAR suppression sequence and a bHLH binding motif of the MdMYB16 protein using protein sequence analyses. Overexpression of MdMYB16 or MdMYB16 without bHLH binding sequence (LBSMdMYB16) in red-fleshed callus inhibited MdUFGT and MdANS expression and anthocyanin synthesis. However, overexpression of MdMYB16 without the EAR sequence (LESMdMYB16) in red-fleshed callus had no inhibitory effect on anthocyanin. The yeast one-hybrid assay showed that MdMYB16 and LESMdMYB16 interacted the promoters of MdANS and MdUFGT, respectively. Yeast two-hybrid, pull-down, and bimolecular fluorescence complementation assays showed that MdMYB16 formed homodimers and interacted with MdbHLH33, however, the LBSMdMYB16 could not interact with MdbHLH33. We overexpressed MdbHLH33 in callus overexpressing MdMYB16 and found that it weakened the inhibitory effect of MdMYB16 on anthocyanin synthesis. Together, these results suggested that MdMYB16 and MdbHLH33 may be important part of the regulatory network controlling the anthocyanin biosynthetic pathway. |
| Databáze: | OpenAIRE |
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