葡萄果实MYBA1与UFGT、DFR的作用机制

doi:10.3864/j.issn.0578-1752.2018.12.013

Abstract

Abstract: 【Objective】MYBA1 transcription factor plays an important role in the biosynthesis of anthocyanins. To further explore the synergistic regulatory mechanism of anthocyanins, we analyzedthe mutual mechanism and expression pattern of VvMYBA1, VvUFGT, VvDFR and content of anthocyanins during grape berry development.【Method】The real-time PCR, spectrophotometer and SAS8.0 were used to analyze the expression patterns of VvMYBA1, VvUFGT, VvDFR content of anthocyanins and correlation in different time periods, respectively. The transcriptional activity of VvMYBA1 and the interactions between the VvMYBA1 and VvUFGT, VvDFR were determined by yeast hybrid system. 【Result】The expression pattern analysis showed that the expression of VvMYBA1, VvUFGT and VvDFR was increased until veraison (60-80 d after full bloom) and then declined. The content of anthocyanins was fist increased until veraison (80 d after full bloom) and then stabilization. Correlation analysis showed that the expression of MYBA1 with expression ofthe DFR, UFGT had positive correlation, and the content of anthocyanins with the expression of MYBA1, DFR, UFGT had positive correlation. Yeast hybrid assay indicated that VvMYBA1 had transcriptional activation functions which could specifically combine promoter of VvUFGT, VvDFR and had no interaction with VvUFGT and VvDFR encoded proteins.【Conclusion】The content of anthocyanins was significantly correlated with expression of VvMYBA1, VvDFR, and VvUFGT. The VvMYBA1 had transcriptional activation functions. All results indicated that VvMYBA1 could regulate the accumulation of anthocyanins in grape berry development by regulating VvUFGT and VvDFR expression combined with VvUFGT and VvDFR promoter specifically.

Key words: grape, VvMYBA1; VvUFGT, VvDFR, yeast hybrid system

NIU TieQuan, DONG YanMei, LIU HaiXia, ZHANG XiaoJun, GAO Yan, ZHANG PengFei, LIANG ChangMei, WEN PengFei. The Regulations of the MYBA1 in UFGT and DFR from the Grape Berries[J].Scientia Agricultura Sinica, 2018, 51(12): 2368-2377.

References

[1] 孙欣, 韩键, 房经贵, 上官凌飞, 王西成, 宋长年, 李晓颖. 葡萄浆果着色分子机理的重要研究进展. 植物生理学报, 2012, 48(4): 333-342.

Sun X, Han J, Fang J G, Shangguan L F, Wang X C, Song C N, Li X Y. Important research progress of coloring molecular mechanisms in grape berry. Plant Physiology Journal, 2012, 48(4): 333-342. (in Chinese)

[2] Mizuno H, Okamoto G, Hirano K. Effect of anthocyanin composition in grape skin on anthocyanic vacuolar inclusion development and skin coloration. Vitis Journal of Grapevine Research, 2006, 45(4): 173-177.

[3] LIU W, XU J, LIU Y, YU X, TANG X, WANG Z, LI X. Anthocyanins potentiate the activity of trastuzumab in human epidermal growth factor receptor 2?positive breast cancer cells in vitro and in vivo. Molecular Medicine Reports, 2014, 10(4): 1921-1926.

[4] PEIFFER D S, WANG L S, ZIMMERMAN N P, RANSON B W, CARMELLA S G, KUO C T, CHEN J H, OSHIMA K, HUANG Y W, HECHT S S, STONER G D. Dietary consumption of black raspberries or their anthocyanin constituents alters innate immune cell trafficking in esophageal cancer. Cancer Immunology Research, 2016, 4(1): 72-82.

[5] REIS J F, MONTEIRO V V S, GOMES R D S, CARNO, M M D, COSTA G V D, RIBERA P C. Action mechanism and cardiovascular effect of anthocyanins: a systematic review of animal and human studies. Journal of Translational Medicine, 2016, 14(1): 315.

[6] ZHAO Q, DUAN C Q, WANG J. Anthocyanins profile of grape berries of Vitis amurensis, its hybrids and their wines. International Journal of Molecular Sciences, 2010, 11(5): 2212-2228.

[7] CZEMMEL S, HEPPEL S C, BOGS J. R2R3 MYB transcription factors: key regulators of the flavonoid biosyntlietic patliway ingrapevine. Protoplasma, 2012, 249: 109-118.

[8] AZUMA A, YAKUSHIJI H, KOSHITA Y, KOBAYASHI S. Flavonoid biosynthesis-related genes in grape skin are differentially regulated by temperature and light conditions. Planta, 2012, 236(4): 1067-1080.

[9] 张龙, 李卫华, 姜淑梅, 朱根发, 王碧青, 李洪清. 花色素苷生物合成与分子调控研究进展. 园艺学报, 2008, 35(6): 909-916.

ZHANG L, LI W H, JIANG S M, ZHU G F, WANG B Q, LI H Q. Progress of molecular basis of biosynthesis and transcriptional regulation of anthocyanins. Acta Horticulturae Sinica, 2008, 35(6): 909-916. (in Chinese)

[10] CHEN S M, LI C H, ZHU X R, SUN W, WANG L S, CHEN F D. The identification of flavonoids and the expression of genes of anthocyanin biosynthes is in the chrysanthemum flowers. Biotechnology Plantarum, 2012, 56(3): 458-464.

[11] Viljoen C D, Snyman M C, Spies J J. Identification and expression analysis of chalcone synthase, and dihydroflavonol 4-reductase, in Clivia miniata. South African Journal of Botany, 2013, 87(1): 18-21.

[12] Yildiz M, Willis D K, Cavagnaro P F, Iorizzo M, Abak K, Simon P W. Expression and mapping of anthocyanin biosynthesis genes in carrot. Theoretical & Applied Genetics, 2013, 126(7): 1689-1702.

[13] 文习成, 姜卫兵, 韩键, 翁忙玲, 房经贵. 环境因子和外源化学物质对果树UFGT基因的影响. 植物生理学报, 2012, 48(2): 129-134.

Wen X C, Jiang W B, Han J, Weng M L, Fang J G. The effects of environmental factors and exogenous chemicals on the fruit UFGT gene. Plant Physiology Journal, 2012, 48(2): 129-134. (in Chinese)

[14] Kobayashi S, Ishimaru M, Ding C K, Yakushiji H, Goto N. Comparison of UDP-glucose: flavonoid 3-O-glucosyltransferase (UFGT) gene sequences between white grapes (Vitis vinifera) and their sports with red skin. Plant Science An International Journal of Experimental Plant Biology, 2001, 160(3): 543.

[15] 冯立娟, 苑兆和, 尹燕雷, 招雪晴, 许鑫科, 徐榕, 李自峰. 槭属2品种叶变色期花青苷含量与相关酶活性的变化. 林业科学, 2009, 45(8): 56-60.

Feng L J, Yuan Z H, Yin Y L, Zhao X Q, Xu X K, Xu R, Li Z F. Anthocyanin content and the relevant enzymes activities during leaf color changing of two Acer species. Scientla Silvae Sinicae, 2009, 45(8): 56-60. (in Chinese)

[16] 刘金, 魏景立, 刘美艳, 宋杨, 冯守千, 王传增, 陈学森. 早熟苹果花青苷积累与其相关酶活性及乙烯生成之间的关系. 园艺学报, 2012, 39(7): 1235-1242.

Liu J, Wei J L, Liu M Y, Song Y, Feng S Q, Wang C Z, Chen X S. The relationships between the enzyme activityof anthocyanin. Acta Horticulturae Sinica, 2012, 39(7): 1235-1242. (in Chinese)

[17] 冯守千, 陈学森, 张春雨, 刘晓静, 刘遵春, 王海波, 王延玲, 周朝华. 砂梨品种‘满天红’及其芽变品系‘奥冠’花青苷合成与相关酶活性研究. 中国农业科学, 2008, 41(10): 3184-3190.

Feng S Q, Chen X S, Zhang C Y, Liu X J, Liu Z C, Wang H B, Wang Y L, Zhou C H. A study of the relationship between anthocyanin biosynthesis and related enzymes activity inpyrus pyrifolia ‘mantianhong’ and its bud sports ‘aoguan’. Scientia Agricultura Sinica, 2008, 41(10): 3184-3190. (in Chinese)

[18] Ali M B, Howard S, Chen S,Wang Y, Yu O, Kovacs L G, Qiu W. Berry skin development in Norton grape: distinct patterns of transcriptional regulation and flavonoid biosynthesis. BMC Plant Biology, 2011, 11(1): 7.

[19] Boss P K, Davies C, Robinson S P. Expression of anthocyanin biosynthesis pathway genes in red and white grapes. Plant Molecular Biology, 1996, 32(3): 565-569.

[20] Kobayashi S, Ishimaru M, Hiraoka K, Honda C. Myb-related genes of the Kyoho grape (Vitis labruscana) regulate anthocyanin biosynthesis. Planta, 2002, 215(6): 924.

[21] 任国慧, 陶然, 王晨, 孙欣, 房经贵. 葡萄浆果着色与UFGT和MYBA基因表达量的关系研究. 南京农业大学学报, 2013, 36(4): 30-36.

Ren G H, Tao R, Wang C, Sun X, Fang J G. The research of the relationship between coloring and UFGT and MYBA gene expression level of the grape berry. Journal of Nanjing Agricultural University, 2013, 36(4): 30-36. (in Chinese)

[22] Kobayashi S, Goto-Yamamoto N, Hirochika H. Retrotransposon-induced mutations in grape skin color. Science, 2004, 304(5673): 982-982.

[23] Vashisth T, Johnson L K, Malladi A. An efficient RNA isolation procedure and identification of reference genes for normalization of gene expression in blueberry. Plant Cell Reports, 2011, 30(12): 2167-2176.

[24] 田莉. 葡萄果实查耳酮合成酶研究—基因克隆、表达、纯化、抗体的制备以及UV、糖对葡萄果实查耳酮合成酶的调节[D]. 北京: 中国农业大学, 2007.

Tian L. Study on chalcone synthase of grape berry-cloning, expression, purification, antibody, preparation and regulation of chalcone synthase by UV and sugars in grape berry [D]. Beijing: China Agricultural University, 2007. (in Chinese)

[25] 刘晓静, 冯宝春, 冯守千, 王海波, 石俊, 王娜, 陈为一, 陈学森. ‘国光’苹果及其红色芽变花青苷合成与相关酶活性的研究. 园艺学报, 2009, 36(09): 1249-1254.

Liu X J, Feng B C, Feng S Q, Wang H B, Shi J, Wang N, Chen W Y, Chen X S. Studies on anthocyanin biosynthesis and activities of related enzymes of ‘Ralls’ and its bud mutation. Acta Horticulturae Sinica, 2009, 36(9): 1249-1254. (in Chinese)

[26] Cutandaperez M C, Ageorges A, Gomez C, Vialet S, Terrier N, Romieu C, Torregrosa L. Ectopic expression of VlmybA1 in grapevine activates a narrow set of genes involved in anthocyanin synthesis and transport. Plant Molecular Biology, 2009, 69(6): 633-648.

[27] 朱婷婷, 梁东, 夏惠. R2R3-MYB调控果实花色苷合成的研究进展. 基因组学与应用生物学, 2016, 35(4): 985-991.

Zhu T T, Liang D, Xia H. Research progress of R2R3-MYB regulating fruit anthocyanin synthesis. Genomics and Applied Biology, 2016, 35(4): 985-991. (in Chinese)

[28] Rinaldo A, Cavallini E, Jia Y, Moss S M, McDavid D A, Hooper L C, Walker A R. A grapevine anthocyanin acyltransferase, transcriptionally regulated by VvMYBA, can produce most acylated anthocyanins present in grape skins. Plant Physiologic, 2015, 169: 1897-1916.

[29] Kobayashi S, Ishimaru M, Hiraoka K, Honda C. Myb-related genes of the Kyoho grape (Vitis labruscana) regulate anthocyanins biosynthesis. Planta, 2002, 215: 924-933.

[30] Holton T A, Cornish E C. Genetics and biochemistry of anthocyanins biosynthesis. Plant Cell, 1997, 7: 1071-1083.

[31] Kim S H, Lee J R, Hong S T, Yoo Y K, An G, Kim S R. Molecular cloning and analysis of anthocyanin biosynthesis genes preferentially expressed in apple skin. Plant Science, 2003, 165(2): 403-413.

[32] 刘金, 魏景立, 刘美艳, 宋杨, 冯守千, 王传增, 陈学森. 早熟苹果花青苷积累与其相关酶活性及乙烯生成之间的关系. 园艺学报, 2012, 39(7): 1235-1242.

Liu J, Wei J L, Liu M.Y, Song Y, Feng S Q, Wang C Z, Chen X S. The relationships between the enzyme activity of anthocyan in biosynthesis，ethylene release and anthocyanin accumulation in fruits of precocious apple cultivars. Acta Horticulturae Sinica, 2012, 39(7): 1235-1242 . (in Chinese)

[33] 王海竹, 曲红云, 周婷婷, 徐启江. 茄萼花色苷合成相关基因DFR和MYB克隆及表达分析. 中国农业科学, 2017, 50(14): 2781-2792.

Wang H z, Qu H Y, Zhou T T, Xu Q J. Cloning and expression analysis of DFR and MYB genes related to anthocyanin synthesis in eggplant. Scientia Agricultura Sinica, 2017, 50(14): 2781-2792. (in Chinese)

[34] Nakatsuka T, Haruta K S, Pitaksutheepong C, Abe Y, Kakizaki Y, Yamamoto K, Shimada N, Yamamura S, Nishihara M. Identification and characterization of R2R3- MYB and bHLH transcription factors regulating anthocyanin biosynthesis in gentian flowers. Plant & Cell Physiology, 2008, 49(12): 1818.

[35] Takos A M, Jaffã© F W, Jacob S R, Boqs J, Robinson S P, Walker A R. Light-induced expression of a MYB gene regulates anthocyanin biosynthesis in red apples. Plant Physiology, 2006, 142(3): 1216-1232.

[36] 张传义, 曹秋芬, 孟玉平, 周慧. 一个调控葡萄花和果实色素形成的基因家族—MYB相关基因家族. 分子植物育种, 2007, 5(S6): 85-88.

Zhang C Y, Cao Q f, Meng Y P, Zhou H. A gene family of genes related to the formation of vines and fruit pigments- MYB-related gene family. Molecular Plant Breeding, 2007, 5(S6): 85-88. (in Chinese)

[37] 许玲, 卫培培, 张大勇, 徐照龙, 何晓兰, 黄益洪, 马鸿翔, 邵宏波. 大豆转录因子基因GmMYB111的克隆及功能分析. 中国农业科学, 2015, 48(15): 3079-3089.

Xu L, WeI P P, Zhang D Y, Xu Z L, He X L, Huang Y H, Ma H X, Shao H B. Expression and function analysis of the transcription factor Gm MYB111 in soybean.Scientia Agricultura Sinica, 2015, 48(15): 3079-3089. (in Chinese)

[38] Kim C Y, Lee S H, Park H C, Bae C G, Cheong Y H, Chol Y J, Han C, Lee S Y, Lim C O, Cho M J. Identification of rice blast fungal elicitor-responsive genes by differential display analysis. Molecular Plant-Microbe Interactions, 2000, 13(4): 470-474.

[39] Abe H K, Shinozaki Y,Urao T, Iwasaki T, Hosokawa D, Shinozaki K. Role of Arabidopsis MYC and MYB homologs in drought-and abscisic acid-regulated gene expression. Plant Cell, 1997, 9: 1859.

[40] Villalobos M A, Bartels D, Iturriaga G. Stress tolerance and glucose insensitive phenotypes in Arabidopsis over expressing the CpMYB10 transcriptional factor gene. Plant Physiology, 2004, 135: 309-324.

[41] Legay S, Lacombe E, Goicoechea M, Briere C, Seguin A, Mackay J. Molecular characterization of EgMYB1, a putative transcriptional repressor of the lignin biosynthetic pathway. Plant Science, 2007, 173: 542-549.

Related Articles 15

[1]	FENG WeiQing, NI YuanQian, FEI Teng, LI YouMei, XIE ZhaoSen. Differences in Vascular Bundle Morphological Structure, Distribution, and Water Transport Function in Grape Fruits of Different Shapes [J]. Scientia Agricultura Sinica, 2026, 59(1): 161-178.
[2]	WANG SiQi, ZOU LiRen, BAI RuiWen, YAN Ke, WANG SiYang, QI XiaoGuang, SHEN HaiLin, WEN JingHui. Screening of Key Genes Related to Gibberellic Acid Regulation of Rachis Hardening in Honey Grapes [J]. Scientia Agricultura Sinica, 2026, 59(1): 179-189.
[3]	TAN XiBei, LAN XuYing, LIU ChongHuai, FAN XiuCai, JIANG JianFu, SUN Lei, LI Peng, YU ShuXin, ZHANG Ying. Changes of Secondary Metabolites in Grapes with Different Resistance Levels in Response to White Rot Infection [J]. Scientia Agricultura Sinica, 2025, 58(9): 1767-1778.
[4]	TANG XueShen, DANG ShiZhuo, ZHOU Juan, LI JiaHao, LI MeiHua, HU Hao, ZHANG YaHong. Analysis of VvBES1-1 Involvement in Flower Bud Differentiation of Red Globe Grape Based on Red and Blue Light Regulation [J]. Scientia Agricultura Sinica, 2025, 58(8): 1650-1662.
[5]	YANG CaiLi, LI YongZhou, HE LiangLiang, SONG YinHua, ZHANG Peng, LIU ZhaoXian, LI PengHui, LIU SanJun. Genome-Wide Identification and Analysis of TPS Gene Family and Functional Verification of VvTPS4 in the Formation of Monoterpenes in Grape [J]. Scientia Agricultura Sinica, 2025, 58(7): 1397-1417.
[6]	GUO AoLin, LIN JunXuan, LAI GongTi, HE LiYuan, CHE JianMei, PAN Ruo, YANG FangXue, HUANG YuJi, CHEN GuiXin, LAI ChengChun. Effect of VdF3′5′H2 Overexpression on the Accumulation of Anthocyanin Composition in Spine Grape Cells [J]. Scientia Agricultura Sinica, 2025, 58(4): 802-818.
[7]	ZHANG XiangKun, LI JiaYing, QIAO RuMeng, HE JingLei, WANG Li, SHI XiaoXin, DU GuoQiang. Effects of GFabV Under Different Zn Levels on Photosynthetic Efficiency and Photosynthesis-Related Gene Expression of ‘Shine Muscat’ Grapevine [J]. Scientia Agricultura Sinica, 2025, 58(24): 5190-5200.
[8]	WANG Di, HAN ShouAn, ZHANG Wen, WANG Min, SHI HuiDong, ZHU XueHui, BAI ShiJian, LIU XuPeng, TIAN Jia, XIE Hui. Effects of Different Plant Growth Regulators on Fruit and Raisin Quality of Thompson Seedless Grapes [J]. Scientia Agricultura Sinica, 2025, 58(18): 3767-3782.
[9]	YU Huan, LIN Ling, GUO RongRong, CAO XiongJun, WANG Bo, FANG JingGui, XIE ShuYu, HUANG XiaoYun, HAN JiaYu, BAI XianJin. Investigation and Evaluation of Inflorescence Attachment and Quality of Grape Germplasm Resources in The Hot Zone Guangxi [J]. Scientia Agricultura Sinica, 2025, 58(17): 3503-3515.
[10]	WANG HuiLing, ZHANG YingYing, YAN AiLing, WANG XiaoYue, LIU ZhenHua, REN JianCheng, XU HaiYing, SUN Lei. Multi-Omics Analysis Reveals the Changes of Monoterpenes and Anthocyanins Accumulation During Veraison in Red Muscat-Type Grape [J]. Scientia Agricultura Sinica, 2025, 58(13): 2645-2662.
[11]	CAO XiongJun, WANG Bo, HAN JiaYu, LIAO YongFeng, XIE ShuYu, BAI Yang, HUANG XiaoYun, LU Li, HUANG QiuMi, JIANG ChunFen, PAN FengPing, BAI XianJin. Research and Practice on High Photosynthetic Efficiency Breeding of Grapes in Hot Climate Regions [J]. Scientia Agricultura Sinica, 2025, 58(10): 1994-2007.
[12]	DONG Jie, ZHANG Peng, LI WangZe, LI HeFang, ZHOU GuoChao, CHEN KeQin, FANG YuLin, ZHANG KeKun. Effects of Seedlessness and Swelling Treatments Based on GA₃ and CPPU on the Fruit Quality of 'Shine Muscat' Grapes [J]. Scientia Agricultura Sinica, 2025, 58(10): 2008-2021.
[13]	GE Yi, ZHENG QiuLing, CHEN MengXia, XIA JiaXin, FANG Xiang, TANG MeiLing, FANG JingGui, SHANGGUAN LingFei. Cloning and Functional Analysis of the Autophagy Gene ATG8f in the Grapevine [J]. Scientia Agricultura Sinica, 2025, 58(1): 156-169.
[14]	FENG Fan, JIANG XingRui, WANG LingYun, ZHANG YongGang, LI AiHua, TAO YongSheng. The Stabilization of Aroma and Color During Hutai-8 Rose Winemaking by Gallic Acid Treatment [J]. Scientia Agricultura Sinica, 2024, 57(8): 1592-1605.
[15]	YUAN Miao, ZHOU Juan, DANG ShiZhuo, TANG XueShen, ZHANG YaHong. Functional Analysis of VvARF18 Gene in Red Globe Grape [J]. Scientia Agricultura Sinica, 2024, 57(7): 1363-1376.

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed

Comments

Recommended 0

No Suggested Reading articles found!

The Regulations of the MYBA1 in UFGT and DFR from the Grape Berries

RichHTML

PDF