DgbZIP3 interacts with DgbZIP2 to increase the expression of DgPOD for cold stress tolerance in chrysanthemum

Abstract

The bZIP transcription factor plays a very important role in abiotic stresses, e.g. drought, salt, and low-temperature stress, but the mechanism of action at low temperature is still unclear. In this study, overexpression of DgbZIP3 led to increased tolerance of chrysanthemum (Chrysanthemum morifolium Ramat.) to cold stress, whereas antisense suppression of DgbZIP3 resulted in decreased tolerance. Electrophoretic mobility shift assay (EMSA), chromatin immunoprecipitation (ChIP), luciferase complementary imaging analysis (LCI), and dual-luciferase reporter gene detection (DLA) experiments indicated that DgbZIP3 directly bound to the promoter of DgPOD and activated its expression. DgbZIP2 was identified as a DgbZIP3-interacting protein using yeast two-hybrid, co-immunoprecipitation, LCI, and bimolecular fluorescence complementation assays. Overexpression of DgbZIP2 led to increased tolerance of chrysanthemum to cold stress, whereas antisense suppression of DgbZIP2 resulted in decreased tolerance. A ChIP–qPCR experiment showed that DgbZIP2 was highly enriched in the promoter of DgPOD, while DLA, EMSA, and LCI experiments further showed that DgbZIP2 could not directly regulate the expression of DgPOD. The above results show that DgbZIP3 interacts with DgbZIP2 to regulate the expression of DgPOD to promote an increase in peroxidase activity, thereby regulating the balance of reactive oxygen species and improving the tolerance of chrysanthemum to low-temperature stress.

References

[1] Knight M-R, Knight H . Low-temperature perception leading to gene expression and cold tolerance in higher plants. New Phytol. 2012; 195: 737-51.

[2] Bohnert H-J. Adaptation to environmental stresses. Plant Cell. 1995; 7: 1099.

[3] Chinnusamy V, Zhu J, Zhu J-K . Cold stress regulation of gene expression in plants. Trends Plant Sci. 2007; 12: 444-51.

[4] Rohit J, Wani S-H, Balwant S et al. Transcription factors and plants response to drought stress: current understanding and future directions. Front Plant Sci. 2016; 7: 1029.

[5] Jia J-T, Zhao P-C, Cheng L-Q et al. MADS-box family genes in sheepgrass and their involvement in abiotic stress responses. BMC Plant Biol. 2018; 18: 42.

[6] Corrêa L, Riaño-Pachón D-M, Schrago C-G et al. The role of bZIP transcription factors in green plant evolution: adaptive features emerging from four founder genes. PLoS One. 2008; 3: e2944.

[7] Nijhawan A, Jain M, Tyagi A-K et al. Genomic survey and gene expression analysis of the basic leucine zipper transcription factor family in rice. Plant Physiol. 2008; 146: 333-50.

[8] Wang D, Guo Y, Wu C et al. Genome-wide analysis of CCCH zinc finger family in Arabidopsis and rice. BMC Genomics. 2008; 9: 44.

[9] Liao Y, Zou H-F, Wei W et al. Soybean GmbZIP44, GmbZIP62 and GmbZIP78 genes function as negative regulator of ABA signaling and confer salt and freezing tolerance in transgenic Arabidopsis . Planta. 2008; 228: 225-40.

[10] Ji X-Y, Liu G-F, Liu Y-J et al. The bZIP protein from Tamarix hispida, ThbZIP1, is ACGT elements binding factor that enhances abiotic stress signaling in transgenic Arabidopsis . BMC Plant Biol. 2013; 13: 151.

[11] Liu C, Mao B, Ou S et al. OsbZIP71, a bZIP transcription factor, confers salinity and drought tolerance in rice. Plant Mol Biol. 2014; 84: 19-36.

[12] Sun X, Yong L, Hua C et al. The Arabidopsis AtbZIP1 transcription factor is a positive regulator of plant tolerance to salt, osmotic and drought stresses. J Plant Res. 2012; 125: 429-38.

[13] Li Y-H, Wang C-L, Li Y et al. Cloning and expression analysis of the fatty acid desaturase gene CmFAD7 in chrysanthemum. J Hortic. 2015; 42: 65-74.

[14] Chen L, Chen Y, Jiang J-F et al. The constitutive expression of Chrysanthemum dichrum ICE1 in Chrysanthemum grandiflorum improves the level of low temperature, salinity and drought tolerance . Plant Cell Rep. 2012; 31: 1747-58.

[15] Huang Q-X, Liao X-Q, Yang X-H et al. Lysine crotonylation of DgTIL1 at K72 modulates cold tolerance by enhancing DgnsLTP stability in chrysanthemum. Plant Biotechnol J. 2021; 19: 1125-40.

[16] Yang X-H, Luo Y-C, Bai H-R et al. DgMYB2 improves cold resistance in chrysanthemum by directly targeting DgGPX1. Hortic Res. 2022, 9, uhab028.

[17] Yang X-H, Lin P, Luo Y-C et al. Lysine decrotonylation of glutathione peroxidase at lysine 220 site increases glutathione peroxidase activity to resist cold stress in chrysanthemum. Ecotoxicol Environ Saf. 2022; 232: 113295.

[18] Bai H-R, Lin P, Li X et al. DgC3H1, a CCCH zinc finger protein gene, confers cold tolerance in transgenic chrysanthemum. Sci Hortic. 2021; 281: 109901.

[19] Izawa T, Foster R, Chua N-H . Plant bZIP protein DNA binding specificity. J Mol Biol. 1993; 230: 1131-44.

[20] Jakoby M, Weisshaar B, Dröge-Laser W et al. bZIP transcription factors in Arabidopsis . Trends Plant Sci. 2002; 7: 106-11.

[21] Verona UDSD, Adami L, Vicentini D et al. Genome-wide analysis and expression profile of the bZIP transcription factor gene family in grapevine (Vitis vinifera) . BMC Genomics. 2014; 15: 1-18.

[22] Jin J-P, Tian F, Yang D-C et al. PlantTFDB 4.0: toward a central hub for transcription factors and regulatory interactions in plants. Nucleic Acids Res. 2017; 45: D1040-5.

[23] Li D-Y, Fu F-Y, Zhang H-J et al. Genome-wide systematic characterization of the bZIP transcriptional factor family in tomato (Solanum lycopersicum L.) . BMC Genomics. 2015; 16: 771.

[24] Wei K-F, Chen J, Wang Y-M et al. Genome-wide analysis of bZIP-encoding genes in maize. DNA Res. 2012; 19: 463-76.

[25] Wang J-Z, Zhou J-X, Zhang B-L et al. Genome-wide expansion and expression divergence of the basic leucine zipper transcription factors in higher plants with an emphasis on sorghum. J Integr Plant Biol. 2011; 53: 212-31.

[26] Zhao J, Guo R-R, Guo C-L et al. Evolutionary and expression analyses of the apple basic leucine zipper transcription factor family. Front Plant Sci. 2016; 7: 376.

[27] Agarwal P, Baranwal VK, Khurana P . Genome-wide analysis of bZIP transcription factors in wheat and functional characterization of a TabZIP under abiotic stress. Sci Rep.(2019) 9: 4608.

[28] Liu C-T, Wu Y-B, Wang X-P . bZIP transcription factor OsbZIP52/RISBZ5: a potential negative regulator of cold and drought stress response in rice. Planta. 2012; 235: 1157-69.

[29] Zhang Y, Zhang G, Xia N et al. Cloning and characterization of a bZIP transcription factor gene in wheat and its expression in response to stripe rust pathogen infection and abiotic stresses. Physiol Mol Plant Pathol. 2008; 73: 88-94.

[30] Kobayashi F, Maeta E, Terashima A et al. Development of abiotic stress tolerance via bZIP-type transcription factor LIP19 in common wheat. J Exp Bot. 2008; 59: 891-905.

[31] Wang L, Cao H-L, Qian W-J et al. Identification of a novel bZIP transcription factor in Camellia sinensis as a negative regulator of freezing tolerance in transgenic Arabidopsis . Ann Bot. 2017; 119: 1195-209.

[32] Zhang L-N, Zhang L-C, Xia C et al. A novel wheat C-bZIP gene, TabZIP14-B, participates in salt and freezing tolerance in transgenic plants. Front Plant Sci. 2017.00710. https://doi.org/10.3389//fpls.2017.00710.

[33] Zhang L, Zhang L, Xia C et al. A novel wheat bZIP transcription factor, TabZIP60, confers multiple abiotic stress tolerances in transgenic Arabidopsis . Physiol Plant. 2015; 153: 538.

[34] Pompelli M-F, Barata-Luis R, Vitorino H-S et al. Photosynthesis, photoprotection and antioxidant activity of purging nut under drought deficit and recovery. Biomass Bioenergy. 2010; 34: 1207-15.

[35] Tang L-L, Cai H, Ji W et al. Overexpression of GsZFP1 enhances salt and drought tolerance in transgenic alfalfa (Medicago sativa L.) . Plant Physiol Biochem. 2013; 71: 22-30.

[36] Suzuki N, Koussevitzky S, Mittler R et al. ROS and redox signalling in the response of plants to abiotic stress. Plant Cell Environ. 2012; 35: 259-70.

[37] Farmer E-E, Mueller M-J . ROS-mediated lipid peroxidation and RES-activated signaling. Annu Rev Plant Biol. 2013; 64: 429-450.

[38] Navrot N, Rouhier N, Gelhaye E et al. Reactive oxygen species generation and antioxidant systems in plant mitochondria. Physiol Plant. 2007; 129: 185-95.

[39] Passardi F, Longet D, Penel C et al. The class III peroxidase multigenic family in rice and its evolution in land plants. Phytochemistry. 2004; 65: 1879-93.

[40] Gao C-Q, Wang Y-C, Liu G-F et al. Cloning of ten peroxidase (POD) genes from Tamarix hispida and characterization of their responses to abiotic stress. Plant Mol Biol Rep. 2010; 28: 77-89.

[41] Liu C, Ou S-J, Mao B-G et al. Early selection of bZIP73 facilitated adaptation of japonica rice to cold climates. Nat Commun. 2018; 9: 3302.

[42] Wang J-Y, Qian L, Mao X-G et al. Wheat transcription factor TaAREB3 participates in drought and freezing tolerances in Arabidopsis . Int J Biol Sci. 2016; 12: 257-69.

[43] Kusano T, Berberich T, Harada M et al. A maize DNA-binding factor with a bZIP motif is induced by low temperature. Mol Gen Genet. 1995; 248: 507-17.

[44] An J-P, Yao J-F, Wang X-N et al. MdHY5 positively regulates cold tolerance via CBF-dependent and CBF-independent pathways in apple. J Plant Physiol. 2017; 218: 275-81.

[45] Bi C-X, Yu Y-H, Dong C-H et al. The bZIP transcription factor TabZIP15 improves salt stress tolerance in wheat. Plant Biotechnol J. 2021; 19: 209-11.

[46] Cui L-G, Shan JX, Min S et al. DCA1 acts as a transcriptional co-activator of DST and contributes to drought and salt tolerance in rice. PLoS Genet. 2015; 11: e1005617.

[47] Liu C-T, Schläppi M-R, Mao B-G et al. The bZIP73 transcription factor controls rice cold tolerance at the reproductive stage. Plant Biotechnol J. 2019; 17: 1834-1849.

[48] Hidekazu S, Kazuhito S, Thomas B et al. LIP19, a basic region leucine zipper protein, is a Fos-like molecular switch in the cold signaling of rice plants. Plant Cell Physiol. 2005; 46: 1623-34.

[49] Wang K, Bai Z-Y, Liang Q-Y et al. Transcriptome analysis of chrysanthemum (Dendranthema grandiflorum) in response to low temperature stress . BMC Genomics. 2018; 19: 319.

[50] An G, Watson B-D, Chiang C-C . Transformation of tobacco, tomato, potato, and Arabidopsis thaliana using a binary Ti vector system. Plant Physiol. 1986; 81: 301-5.

[51] Xue J-P, Miao Y, Zhang A-M . Studies on callus induced from leaves and plantlets regeneration of the traditional Chinese medicine Chrysanthemum morifolium . Zhongguo Zhong Yao Za Zhi. 2003; 28: 213-6.

[52] An J-P, Qu F-J, Yao J-F et al. The bZIP transcription factor MdHY5 regulates anthocyanin accumulation and nitrate assimilation in apple. Hortic Res. 2017; 4: 17023.

[53] Luo D-L, Ba L-J, Chen J-Y et al. Banana MaTIFY1 transcription factor characteristics and analysis of gene expression during maturation. J Hortic. 2017; 44: 10.

[54] Ba L-J, Kuang J-F, Chen J-Y et al. MaJAZ1 attenuates the MaLBD5-mediated transcriptional activation of jasmonate biosynthesis gene MaAOC2 in regulating cold tolerance of banana fruit. J Agric Food Chem. 2016; 64: 738-45.

[55] An J-P, Li R, Qu F-J et al. R2R3-MYB transcription factor MdMYB23 is involved in the cold tolerance and proanthocyanidin accumulation in apple. Plant J. 2018; 96: 562-77.

[56] Wang K, Zhong M, Wu Y-H et al. Overexpression of a chrysanthemum transcription factor gene DgNAC1 improves the salinity tolerance in chrysanthemum. Plant Cell Rep. 2017; 36: 571-81.

[57] Gilmour S-J, Zarka D-G, Stockinger E-J et al. Low temperature regulation of the Arabidopsis CBF family of AP2 transcriptional activators as an early step in cold-induced COR gene expression. Plant J. 2010; 16: 433-42.

[58] Kjellsen T-D, Shiryaeva L, Schröder W-P et al. Proteomics of extreme freezing tolerance in Siberian spruce (Picea obovata) . J Proteome. 2010; 73: 965-75.