Transcription factors LvBBX24 and LvbZIP44 coordinated anthocyanin accumulation in response to light in lily petals

Abstract

Lily (Lilium spp . ), a horticultural crop serving both ornamental and edible functions, derives its coloration primarily from anthocyanins. However, limited studies have been conducted on the accumulation of anthocyanins within lilies. In this study, we cloned a light-induced transcription factor named as LvBBX24 in lilies. Through genetic and biochemical analysis, we determined that LvBBX24 could upregulate the transcription of LvMYB5 and facilitate anthocyanin synthesis. Moreover, we identified that darkness promoted the degradation of LvBBX24 protein. Through screening a yeast library, we identified LvbZIP44 acts as its interacting partner. Genetic testing confirmed that LvbZIP44 also plays a role in promoting lily anthocyanin synthesis. This indicates a potential synergistic regulatory effect between LvBBX24 and LvbZIP44. Our study indicates that LvBBX24 and LvbZIP44 cooperate to regulate anthocyanin accumulation in lily petals. These findings provide compelling evidence supporting the idea that LvBBX24 and LvbZIP44 may form a looped helix surrounding the LvMYB5 promoter region to regulate anthocyanin biosynthesis.

Acknowledgements

This work was financially supported by Funds for the China Agriculture Research System (CARS-23) and National Key R & D Program of China (Grant No. 2019YFD1001002).

Author contributions

L.C. developed the research concept and design, Z.G. and Z.Z. carried out the experiment and wrote the draft for the first time with the help of Y.S. N.N. and M.N. helped to review and edit the manuscript. S.S., W.D., and M.I. have provided experimental materials and valuable opinions. L.C. and L.Z. had overall responsibility for this project, including project ideas, guidance on experimental design, data analysis, paper writing and revision. L.C. and Y.S. took part in the project administration and funding acquisition. All authors have read and approved the final manuscript.

Data availability

All relevant data in this study are provided in the article and its supplementary figure files.

Conflict of interest statement

The authors declare no competing interests.

Supplementary data

Supplementary data is available at Horticulture Research online.

References

[1] Rana MS, Samant SS. Population biology of Lilium polyphyllum D. Don ex Royle—A critically endangered medicinal plant in a protected area of Northwestern Himalaya. J Nat Conserv. 2011; 19 : 137-42

[2] Yu L, Sun Y, Zhang X. et al. ROS1 promotes low temperature-induced anthocyanin accumulation in apple by demethylating the promoter of anthocyanin-associated genes. Hortic Res. 2022; 9 :uhac007

[3] Shin DY, Ryu CH, Lee WS. et al. Induction of apoptosis and inhibition of invasion in human hepatoma cells by anthocyanins from meoru. Ann N Y Acad Sci. 2009; 1171 :137-48

[4] Chen J, Xu B, Sun J. et al. Anthocyanin supplement as a dietary strategy in cancer prevention and management: A comprehensive review. Crit Rev Food Sci Nutr. 2022; 62 :7242-54

[5] Yamagishi M. How genes paint lily flowers: regulation of colouration and pigmentation patterning. Sci Hortic. 2013; 163 : 27-36

[6] Jiang W, Zhou X. Hydrolysis of radish anthocyanins to enhance the antioxidant and antiproliferative capacities. Food Chem. 2019; 294 :477-85

[7] Liu Y, Tikunov Y, Schouten RE. et al. Anthocyanin biosynthesis and degradation mechanisms in solanaceous vegetables: a review. Front Chem. 2018; 6 :52

[8] Li P, Chen B, Zhang G. et al. Regulation of anthocyanin and proanthocyanidin biosynthesis by Medicago truncatula bHLH transcription factor MtTT8. New Phytol. 2016; 210 :905-21

[9] Zhou H, Lin-Wang K, Wang F. et al. Activator-type R2R3-MYB genes induce a repressor-type R2R3-MYB gene to balance anthocyanin and proanthocyanidin accumulation. New Phytol. 2019; 221 :1919-34

[10] Yang X, Wang J, Xia X. et al. OsTTG1, a WD40 repeat gene, regulates anthocyanin biosynthesis in rice. Plant J. 2021; 107 : 198-214

[11] McNellis TW, von Arnim AG, Araki T. et al. Genetic and molecular analysis of an allelic series of cop 1 mutants suggests functional roles for the multiple protein domains. Plant Cell. 1994; 6 : 487-500

[12] Lau OS, Deng XW. The photomorphogenic repressors COP1 and DET1: 20 years later. Trends Plant Sci. 2012; 17 :584-93

[13] Dong H, Liu X, Zhang C. et al. Expression of Tomato UVR8 in Arabidopsis reveals conserved photoreceptor function. Plant Sci. 2021; 303 :110766

[14] Ramsay NA, Glover BJ. MYB-bHLH-WD40 protein complex and the evolution of cellular diversity. Trends Plant Sci. 2005; 10 : 63-70

[15] Yamagishi M. Involvement of a LhMYB18 transcription factor in large anthocyanin spot formation on the flower tepals of the Asiatic hybrid lily (Lilium spp.) cultivar “Grand Cru”. Mol Breeding. 2018; 38 :60

[16] Yin X, Zhang Y, Zhang L. et al. Regulation of MYB transcription factors of anthocyanin synthesis in lily flowers. Front Plant Sci. 2021; 12 :761668

[17] Yan Y, Zhao J, Lin S. et al. Light-mediated anthocyanin biosynthesis in rose petals involves a balanced regulatory module comprising transcription factors RhHY5, RhMYB114a, and RhMYB3b. J Exp Bot. 2023; 74 :5783-804

[18] Tirumalai V, Swetha C, Nair A. et al. miR828 and miR858 regulate VvMYB114 to promote anthocyanin and flavonol accumulation in grapes. J Exp Bot. 2019; 70 :4775-92

[19] Jiang S, Wang N, Chen M. et al. Methylation of MdMYB1 locus mediated by RdDM pathway regulates anthocyanin biosynthesis in apple. Plant Biotechnol J. 2020; 18 :1736-48

[20] Cai H, Zhang M, Chai M. et al. Epigenetic regulation of anthocyanin biosynthesis by an antagonistic interaction between H2A.Z and H3K4me3. New Phytol. 2019; 221 :295-308

[21] Rehman RNU, You Y, Zhang L. et al. High temperature induced anthocyanin inhibition and active degradation in Malus profusion. Front Plant Sci. 2017; 8 :1401

[22] Wang N, Qu C, Jiang S. et al. The proanthocyanidin-specific transcription factor MdMYBPA1 initiates anthocyanin synthesis under low-temperature conditions in red-fleshed apples. Plant J. 2018; 96 :39-55

[23] Wang S, Zhang X, Li B. et al. Genome-wide identification and characterization of bZIP gene family and cloning of candidate genes for anthocyanin biosynthesis in pomegranate (Punica granatum ). BMC Plant Biol. 2022; 22 :170

[24] Nakatsuka A, Yamagishi M, Nakano M. et al. Light-induced expression of basic helix-loop-helix genes involved in anthocyanin biosynthesis in flowers and leaves of Asiatic hybrid lily. Sci Hortic. 2009; 121 :84-91

[25] Zheng T, Li Y, Lei W. et al. SUMO E3 Ligase SIZ1 stabilizes MYB75 to regulate anthocyanin accumulation under high light conditions in Arabidopsis. Plant Sci. 2020; 292 :110355

[26] Li S, Dong Y, Li D. et al. Eggplant transcription factor SmMYB5 integrates jasmonate and light signaling during anthocyanin biosynthesis. Plant Physiol. 2023; 194 :1139-65

[27] Reymond A, Meroni G, Fantozzi A. et al. The tripartite motif family identifies cell compartments. EMBO J. 2001; 20 :2140-51

[28] Putterill J, Robson F, Lee K. et al. The CONSTANS gene of Arabidopsis promotes flowering and encodes a protein showing similarities to zinc finger transcription factors. Cell. 1995; 80 : 847-57

[29] Xu DB, Gao SQ, Ma YN. et al. The G-protein β subunit AGB1 promotes hypocotyl elongation through inhibiting transcription activation function of BBX21 in Arabidopsis. Mol Plant. 2017; 10 : 1206-23

[30] Zhang X, Huai J, Shang F. et al. A PIF1/PIF3-HY5-BBX23 transcription factor cascade affects photomorphogenesis. Plant Physiol. 2017; 174 :2487-500

[31] Luo D, Xiong C, Lin A. et al. SlBBX20 interacts with the COP 9 signalosome subunit SlCSN5-2 to regulate anthocyanin biosynthesis by activating SlDFR expression in tomato. Hortic Res. 2021; 8 :163

[32] An JP, Wang XF, Zhang XW. et al. Apple B-box protein BBX37 regulates jasmonic acid mediated cold tolerance through the JAZ-BBX37-ICE1-CBF pathway and undergoes MIEL1-mediated ubiquitination and degradation. New Phytol. 2021; 229 :2707-29

[33] Bai S, Tao R, Yin L. et al. Two B-box proteins, PpBBX18 and PpBBX21, antagonistically regulate anthocyanin biosynthesis via competitive association with Pyrus pyrifolia ELONGATED HYPOCOTYL 5 in the peel of pear fruit. Plant J. 2019; 100 :1208-23

[34] Gangappa SN, Holm M, Botto JF. Molecular interactions of BBX24 and BBX25 with HYH, HY 5 HOMOLOG, to modulate Arabidopsis seedling development. Plant Signal Behav. 2013; 8 :e25208

[35] Zhou LJ, Wang Y, Wang Y. et al. Transcription factor CmbHLH16 regulates petal anthocyanin homeostasis under different lights in Chrysanthemum. Plant Physiol. 2022; 190 :1134-52

[36] Xing Y, Sun W, Sun Y. et al. MPK6-mediated HY5 phosphorylation regulates light-induced anthocyanin accumulation in apple fruit. Plant Biotechnol J. 2023; 21 :283-301

[37] Liu Y, Tang L, Wang Y. et al. The blue light signal transduction module FaCRY1-FaCOP1-FaHY5 regulates anthocyanin accumulation in cultivated strawberry. Front Plant Sci. 2023; 14 :1144273

[38] Jiang L, Yue M, Liu Y. et al. A novel R2R3-MYB transcription factor FaMYB5 positively regulates anthocyanin and proanthocyanidin biosynthesis in cultivated strawberries (Fragaria × ananassa). Plant Biotechnol J. 2023; 21 :1140-58

[39] Zhang Y, Zheng S, Liu Z. et al. Both HY5 and HYH are necessary regulators for low temperature-induced anthocyanin accumulation in Arabidopsis seedlings. J Plant Physiol. 2011; 168 : 367-74

[40] Huang Y, Xiong H, Xie Y. et al. BBX24 interacts with DELLA to regulate UV-B-induced photomorphogenesis in Arabidopsis thaliana. Int J Mol Sci. 2022; 23 :7386

[41] Jiang L, Wang Y, Li QF. et al. Arabidopsis STO/BBX24 negatively regulates UV-B signaling by interacting with COP1 and repressing HY5 transcriptional activity. Cell Res. 2012; 22 : 1046-57

[42] Gangappa SN, Crocco CD, Johansson H. et al. The Arabidopsis B-BOX protein BBX25 interacts with HY5, negatively regulating BBX22 expression to suppress seedling photomorphogenesis. Plant Cell. 2013; 25 :1243-57

[43] Chang CS, Maloof JN, Wu SH. COP1-mediated degradation of BBX22/LZF1 optimizes seedling development in Arabidopsis. Plant Physiol. 2011; 156 :228-39

[44] Wang Y, Xiao Y, Sun Y. et al. Two B-box proteins, PavBBX6/9, positively regulate light-induced anthocyanin accumulation in sweet cherry. Plant Physiol. 2023; 192 :2030-48

[45] Bursch K, Toledo-Ortiz G, Pireyre M. et al. Identification of BBX proteins as rate-limiting cofactors of HY5. Nat Plants. 2020; 6 : 921-8

[46] An JP, Zhang XW, Liu YJ. et al. ABI 5 regulates ABA-induced anthocyanin biosynthesis by modulating the MYB1-bHLH3 complex in apple. J Exp Bot. 2021; 72 :1460-72

[47] Liu W, Mei Z, Yu L. et al. The ABA-induced NAC transcription factor MdNAC1 interacts with a bZIP-type transcription factor to promote anthocyanin synthesis in red-fleshed apples. Hortic Res. 2023; 10 :uhad049

[48] Xu H, Wang S, Larkin RM. et al. The transcription factors DcHB30 and DcWRKY75 antagonistically regulate ethylene-induced petal senescence in carnation (Dianthus caryophyllus ). J Exp Bot. 2022; 73 :7326-43

[49] Morgunova E, Taipale J. Structural perspective of cooperative transcription factor binding. Curr Opin Struct Biol. 2017; 47 : 1-8

[50] Chen L, Glover JN, Hogan PG. et al. Structure of the DNA-binding domains from NFAT, Fos and Jun bound specifically to DNA. Nature. 1998; 392 :42-8

[51] Sicoli G, Vezin H, Ledolter K. et al. Conformational tuning of a DNA-bound transcription factor. Nucleic Acids Res. 2019; 47 : 5429-35

[52] Tomăž S, Petek M, Lukan T. et al. A mini-TGA protein modulates gene expression through heterogeneous association with transcription factors. Plant Physiol. 2023; 191 :1934-52

[53] Kumar S, Zavaliev R, Wu Q. et al. Structural basis of NPR1 in activating plant immunity. Nature. 2022; 605 :561-6

[54] Chen K, Liu H, Lou Q. et al. Ectopic expression of the grape hyacinth (Muscari armeniacum) R2R3-MYB transcription factor gene, MaAN2, induces anthocyanin accumulation in Tobacco. Front Plant Sci. 2017; 8 :965