Functional Characterization of Anthocyanin Biosynthesis-Related <i>Dihydroflavonol 4-reductase</i> (<i>DFR</i>) Genes in Blueberries (<i>Vaccinium corymbosum</i>)
<i>Dihydroflavonol 4-reductase</i> (<i>DFR</i>) genes contribute greatly to anthocyanin biosynthesis in plants. Up to now, however, research on the <i>DFR</i> gene family and the key anthocyanin-related <i>DFR</i> members in blueberries (<i>Vacci...
Saved in:
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2025-05-01
|
| Series: | Plants |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2223-7747/14/10/1449 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850126298596245504 |
|---|---|
| author | Yongyan Zhang Sijian Guo Zening Zhang Ruide Li Shitao Du Siyi Hao Chunzhen Cheng |
| author_facet | Yongyan Zhang Sijian Guo Zening Zhang Ruide Li Shitao Du Siyi Hao Chunzhen Cheng |
| author_sort | Yongyan Zhang |
| collection | DOAJ |
| description | <i>Dihydroflavonol 4-reductase</i> (<i>DFR</i>) genes contribute greatly to anthocyanin biosynthesis in plants. Up to now, however, research on the <i>DFR</i> gene family and the key anthocyanin-related <i>DFR</i> members in blueberries (<i>Vaccinium corymbosum</i>) has been limited. In this study, we performed a genome-wide identification of the blueberry <i>DFR</i> gene family, identifying 36 <i>VcDFR</i> genes categorized into five subfamilies. Gene expression analysis showed that three Subfamily III members (<i>VcDFR11/29/34</i>) and four Subfamily V members (<i>VcDFR4/7/30/33</i>) are highly expressed in blueberry fruits, particularly at late ripening stages. Transient overexpression analysis in apple fruits verified the contributions of <i>VcDFR11</i> and <i>VcDFR30</i> to anthocyanin biosynthesis, with <i>VcDFR11</i> showing better promoting effects. Blueberry fruit-based transient overexpression further confirmed the promoting effects of <i>VcDFR11</i> on anthocyanin accumulation and the expression of anthocyanin-related structural genes (especially its downstream <i>anthocyanindin synthase</i> (<i>ANS</i>) and <i>UDP-glucose: flavonoid 3-O-glycosyltransferase</i> (<i>UFGT</i>) genes). The <i>VcDFR11</i> promoter contains binding sites for both bHLH and MYB transcription factors (TFs). Consistently, yeast one-hybrid and dual-luciferase assays confirmed that anthocyanin-related VcMYB-1 and VcbHLHs can bind to and activate the <i>VcDFR11</i> promoter. Furthermore, co-overexpressing <i>VcMYB-1</i>/<i>VcbHLHs</i> with <i>VcDFR11</i> led to much higher anthocyanin accumulation than overexpressing <i>VcDFR11</i> alone, indicating that these TFs positively regulate anthocyanin biosynthesis by upregulating <i>VcDFR11</i>. In summary, our study characterized the blueberry <i>DFR</i> gene family and demonstrated the role of <i>VcDFR11</i> in anthocyanin biosynthesis. |
| format | Article |
| id | doaj-art-0a09dbbd33a04a1c831b45c93ed8f77f |
| institution | OA Journals |
| issn | 2223-7747 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Plants |
| spelling | doaj-art-0a09dbbd33a04a1c831b45c93ed8f77f2025-08-20T02:33:58ZengMDPI AGPlants2223-77472025-05-011410144910.3390/plants14101449Functional Characterization of Anthocyanin Biosynthesis-Related <i>Dihydroflavonol 4-reductase</i> (<i>DFR</i>) Genes in Blueberries (<i>Vaccinium corymbosum</i>)Yongyan Zhang0Sijian Guo1Zening Zhang2Ruide Li3Shitao Du4Siyi Hao5Chunzhen Cheng6College of Horticulture, Shanxi Agricultural University, Taigu 030801, ChinaCollege of Horticulture, Shanxi Agricultural University, Taigu 030801, ChinaCollege of Horticulture, Shanxi Agricultural University, Taigu 030801, ChinaCollege of Horticulture, Shanxi Agricultural University, Taigu 030801, ChinaCollege of Horticulture, Shanxi Agricultural University, Taigu 030801, ChinaCollege of Horticulture, Shanxi Agricultural University, Taigu 030801, ChinaCollege of Horticulture, Shanxi Agricultural University, Taigu 030801, China<i>Dihydroflavonol 4-reductase</i> (<i>DFR</i>) genes contribute greatly to anthocyanin biosynthesis in plants. Up to now, however, research on the <i>DFR</i> gene family and the key anthocyanin-related <i>DFR</i> members in blueberries (<i>Vaccinium corymbosum</i>) has been limited. In this study, we performed a genome-wide identification of the blueberry <i>DFR</i> gene family, identifying 36 <i>VcDFR</i> genes categorized into five subfamilies. Gene expression analysis showed that three Subfamily III members (<i>VcDFR11/29/34</i>) and four Subfamily V members (<i>VcDFR4/7/30/33</i>) are highly expressed in blueberry fruits, particularly at late ripening stages. Transient overexpression analysis in apple fruits verified the contributions of <i>VcDFR11</i> and <i>VcDFR30</i> to anthocyanin biosynthesis, with <i>VcDFR11</i> showing better promoting effects. Blueberry fruit-based transient overexpression further confirmed the promoting effects of <i>VcDFR11</i> on anthocyanin accumulation and the expression of anthocyanin-related structural genes (especially its downstream <i>anthocyanindin synthase</i> (<i>ANS</i>) and <i>UDP-glucose: flavonoid 3-O-glycosyltransferase</i> (<i>UFGT</i>) genes). The <i>VcDFR11</i> promoter contains binding sites for both bHLH and MYB transcription factors (TFs). Consistently, yeast one-hybrid and dual-luciferase assays confirmed that anthocyanin-related VcMYB-1 and VcbHLHs can bind to and activate the <i>VcDFR11</i> promoter. Furthermore, co-overexpressing <i>VcMYB-1</i>/<i>VcbHLHs</i> with <i>VcDFR11</i> led to much higher anthocyanin accumulation than overexpressing <i>VcDFR11</i> alone, indicating that these TFs positively regulate anthocyanin biosynthesis by upregulating <i>VcDFR11</i>. In summary, our study characterized the blueberry <i>DFR</i> gene family and demonstrated the role of <i>VcDFR11</i> in anthocyanin biosynthesis.https://www.mdpi.com/2223-7747/14/10/1449anthocyanin biosynthesisblueberrydihydroflavonol 4-reductasepromoter activationtranscriptional regulation |
| spellingShingle | Yongyan Zhang Sijian Guo Zening Zhang Ruide Li Shitao Du Siyi Hao Chunzhen Cheng Functional Characterization of Anthocyanin Biosynthesis-Related <i>Dihydroflavonol 4-reductase</i> (<i>DFR</i>) Genes in Blueberries (<i>Vaccinium corymbosum</i>) Plants anthocyanin biosynthesis blueberry dihydroflavonol 4-reductase promoter activation transcriptional regulation |
| title | Functional Characterization of Anthocyanin Biosynthesis-Related <i>Dihydroflavonol 4-reductase</i> (<i>DFR</i>) Genes in Blueberries (<i>Vaccinium corymbosum</i>) |
| title_full | Functional Characterization of Anthocyanin Biosynthesis-Related <i>Dihydroflavonol 4-reductase</i> (<i>DFR</i>) Genes in Blueberries (<i>Vaccinium corymbosum</i>) |
| title_fullStr | Functional Characterization of Anthocyanin Biosynthesis-Related <i>Dihydroflavonol 4-reductase</i> (<i>DFR</i>) Genes in Blueberries (<i>Vaccinium corymbosum</i>) |
| title_full_unstemmed | Functional Characterization of Anthocyanin Biosynthesis-Related <i>Dihydroflavonol 4-reductase</i> (<i>DFR</i>) Genes in Blueberries (<i>Vaccinium corymbosum</i>) |
| title_short | Functional Characterization of Anthocyanin Biosynthesis-Related <i>Dihydroflavonol 4-reductase</i> (<i>DFR</i>) Genes in Blueberries (<i>Vaccinium corymbosum</i>) |
| title_sort | functional characterization of anthocyanin biosynthesis related i dihydroflavonol 4 reductase i i dfr i genes in blueberries i vaccinium corymbosum i |
| topic | anthocyanin biosynthesis blueberry dihydroflavonol 4-reductase promoter activation transcriptional regulation |
| url | https://www.mdpi.com/2223-7747/14/10/1449 |
| work_keys_str_mv | AT yongyanzhang functionalcharacterizationofanthocyaninbiosynthesisrelatedidihydroflavonol4reductaseiidfrigenesinblueberriesivacciniumcorymbosumi AT sijianguo functionalcharacterizationofanthocyaninbiosynthesisrelatedidihydroflavonol4reductaseiidfrigenesinblueberriesivacciniumcorymbosumi AT zeningzhang functionalcharacterizationofanthocyaninbiosynthesisrelatedidihydroflavonol4reductaseiidfrigenesinblueberriesivacciniumcorymbosumi AT ruideli functionalcharacterizationofanthocyaninbiosynthesisrelatedidihydroflavonol4reductaseiidfrigenesinblueberriesivacciniumcorymbosumi AT shitaodu functionalcharacterizationofanthocyaninbiosynthesisrelatedidihydroflavonol4reductaseiidfrigenesinblueberriesivacciniumcorymbosumi AT siyihao functionalcharacterizationofanthocyaninbiosynthesisrelatedidihydroflavonol4reductaseiidfrigenesinblueberriesivacciniumcorymbosumi AT chunzhencheng functionalcharacterizationofanthocyaninbiosynthesisrelatedidihydroflavonol4reductaseiidfrigenesinblueberriesivacciniumcorymbosumi |