MaDREB1F confers cold and drought stress resistance through common regulation of hormone synthesis and protectant metabolite contents in banana

Abstract

Adverse environmental factors severely affect crop productivity. Improving crop resistance to multiple stressors is an important breeding goal. Although CBFs/ DREB1s extensively participate in plant resistance to abiotic stress, the common mechanism underlying CBFs/DREB1s that mediate resistance to multiple stressors remains unclear. Here, we show the common mechanism for MaDREB1F conferring cold and drought stress resistance in banana. MaDREB1F encodes a dehydration-responsive element binding protein (DREB) transcription factor with nuclear localization and transcriptional activity. MaDREB1F expression is significantly induced after cold, osmotic, and salt treatments. MaDREB1F overexpression increases banana resistance to cold and drought stress by common modulation of the protectant metabolite levels of soluble sugar and proline, activating the antioxidant system, and promoting jasmonate and ethylene syntheses. Transcriptomic analysis shows that MaDREB1F activates or alleviates the repression of jasmonate and ethylene biosynthetic genes under cold and drought conditions. Moreover, MaDREB1F directly activates the promoter activities of MaAOC4 and MaACO20 for jasmonate and ethylene syntheses, respectively, under cold and drought conditions. MaDREB1F also targets the MaERF11 promoter to activate MaACO20 expression for ethylene synthesis under drought stress. Together, our findings offer new insight into the common mechanism underlying CBF/DREB1-mediated cold and drought stress resistance, which has substantial implications for engineering cold- and drought-tolerant crops.

Acknowledgements

We thank the National Key Research and Development Program of China (2019YFD1000204), the 2020 Research Program of Sanya Yazhou Bay Science and Technology City (SKJC-2020-02-002), the National Natural Science Foundation of China (32260736), and the Central Public-interest Scientific Institution Basal Research Fund for Chinese Academy of Tropical Agricultural Sciences (1630052020006) for funding resources.

Author contributions

W.H., B.X., and Z.J. conceived the study. Y.X. and W.H. wrote the manuscript. Other authors carried out experiments and analysis.

Data availability

The data that support the findings are available in the paper and the supplementary materials published online.

Conflict of interest

The authors declare no competing interests.

References

[1] Hu W, Huang C, Deng X et al. TaASR1, a transcription factor gene in wheat, confers drought stress tolerance in transgenic tobacco. Plant Cell Environ. 2013; 36: 1449-64.

[2] Fan W, Hai M, Guo Y et al. The ERF transcription factor family in cassava: genome-wide characterization and expression analyses against drought stress. Sci Rep. 2016; 6: 37379.

[3] Lakhwani D, Pandey A, Dhar YV et al. Genome-wide analysis of the AP2/ERF family in Musa species reveals divergence and neofunctionalisation during evolution. Sci Rep. 2016; 6: 18878.

[4] Muller M, Munne-Bosch S . Ethylene response factors: a key regulatory hub in hormone and stress signaling. Plant Physiol. 2015; 169: 32-41.

[5] Huang X, Song X, Chen R et al. Genome-wide analysis of the DREB subfamily in Saccharum spontaneum reveals their functional divergence during cold and drought stresses. Front Genet. 2019; 10: 1326.

[6] Shi Y, Ding Y, Yang S . Molecular regulation of CBF signaling in cold acclimation. Trends Plant Sci. 2018; 23: 623-37.

[7] Stockinger EJ, Gilmour SJ, Thomashow MF . Arabidopsis thaliana CBF1 encodes an AP2 domain-containing transcriptional activator that binds to the C-repeat/DRE, a cis-acting DNA regulatory element that stimulates transcription in response to low temperature and water deficit. Proc Natl Acad Sci USA. 1997; 94: 1035-40.

[8] Novillo F, Alonso JM, Ecker JR et al. CBF2/DREB1C is a negative regulator of CBF1/DREB1B and CBF3/DREB1A expression and plays a central role in stress tolerance in Arabidopsis. Proc Natl Acad Sci USA. 2004; 101: 3985-90.

[9] Zhao C, Zhang Z, Xie S et al. Mutational evidence for the critical role of CBF transcription factors in cold acclimation in Arabidopsis. Plant Physiol. 2016; 171: 2744-59.

[10] Agarwal PK, Agarwal P, Reddy MK et al. Role of DREB transcription factors in abiotic and biotic stress tolerance in plants. Plant Cell Rep. 2006; 25: 1263-74.

[11] Mizoi J, Shinozaki K, Yamaguchi-Shinozaki K . AP2/ERF family transcription factors in plant abiotic stress responses. Biochim Biophys Acta. 2012; 1819: 86-96.

[12] Qin F, Sakuma Y, Li J et al. Cloning and functional analysis of a novel DREB1/CBF transcription factor involved in cold-responsive gene expression in Zea mays L. Plant Cell Physiol. 2004; 45: 1042-52.

[13] Zhang X, Fowler SG, Cheng H et al. Freezing-sensitive tomato has a functional CBF cold response pathway, but a CBF regulon that differs from that of freezing-tolerant Arabidopsis. Plant J. 2004; 39: 905-19.

[14] Savitch LVAA, Allard G, Seki M et al. The effect of overexpression of two Brassica CBF/DREB1-like transcription factors on photosynthetic capacity and freezing tolerance in Brassica napus. Plant Cell Physiol. 2005; 46: 1525-39.

[15] Ito YJIR, Katsura K, Maruyama K et al. Functional analysis of rice DREB1/CBF-type transcription factors involved in cold-responsive gene expression in transgenic rice. Plant Cell Physiol. 2006; 47: 141-53.

[16] Wang Q, Guan Y, Wu Y et al. Overexpression of a rice OsDREB1F gene increases salt, drought, and low temperature tolerance in both Arabidopsis and rice. Plant Mol Biol. 2008; 67: 589-602.

[17] Dubouzet JG, Sakuma Y, Ito Y et al. OsDREB genes in rice, Oryza sativa L., encode transcription activators that function in drought-, high-salt- and cold-responsive gene expression. Plant J. 2003; 33: 751-63.

[18] Haake V, Cook D, Riechmann JL et al. Transcription factor CBF4 is a regulator of drought adaptation in Arabidopsis. Plant Physiol. 2002; 130: 639-48.

[19] Niu X, Luo T, Zhao H et al. Identification of wheat DREB genes and functional characterization of TaDREB3 in response to abiotic stresses. Gene. 2020; 740: 144514.

[20] Wang D, Jin YN, Ding XH et al. Gene regulation and signal transduction in the ICE-CBF-COR signaling pathway during cold stress in plants. Biochemistry (Mosc). 2017; 82: 1103-17.

[21] Liu Y, Dang P, Liu L et al. Cold acclimation by the CBF-COR pathway in a changing climate: lessons from Arabidopsis thaliana. Plant Cell Rep. 2019; 38: 511-9.

[22] Kobayashi F, Takumi S, Nakata M et al. Comparative study of the expression profiles of the Cor/Lea gene family in two wheat cultivars with contrasting levels of freezing tolerance. Physiol Plant. 2004; 120: 585-94.

[23] Tang K, Zhao L, Ren Y et al. The transcription factor ICE1 functions in cold stress response by binding to the promoters of CBF and COR genes. J Integr Plant Biol. 2020; 62: 258-63.

[24] Li H, Ye K, Shi Y et al. BZR1 positively regulates freezing tolerance via CBF-dependent and CBF-independent pathways in Arabidopsis. Mol Plant. 2017; 10: 545-59.

[25] Shi Y, Huang J, Sun T et al. The precise regulation of different COR genes by individual CBF transcription factors in Arabidopsis thaliana. J Integr Plant Biol. 2017; 59: 118-33.

[26] Jia Y, Ding Y, Shi Y et al. The cbfs triple mutants reveal the essential functions of CBFs in cold acclimation and allow the definition of CBF regulons in Arabidopsis. New Phytol. 2016; 212: 345-53.

[27] Liu J, Gao P, Sun X et al. Efficient regeneration and genetic transformation platform applicable to five Musa varieties. Electron J Biotechnol. 2017; 25: 33-8.

[28] Xu Y, Hu W, Liu J et al. An aquaporin gene MaPIP2-7 is involved in tolerance to drought, cold and salt stresses in transgenic banana (Musa acuminata L.). Plant Physiol Biochem. 2020; 147: 66-76.

[29] Sreedharan S, Shekhawat UK, Ganapathi TR . MusaSAP1, a A20/AN1 zinc finger gene from banana functions as a positive regulator in different stress responses. Plant Mol Biol. 2012; 80: 503-17.

[30] Sreedharan S, Shekhawat UK, Ganapathi TR . Transgenic banana plants overexpressing a native plasma membrane aquaporin MusaPIP1;2 display high tolerance levels to different abiotic stresses. Plant Biotechnol J. 2013; 11: 942-52.

[31] Sreedharan S, Shekhawat UK, Ganapathi TR . Constitutive and stress-inducible overexpression of a native aquaporin gene (MusaPIP2;6) in transgenic banana plants signals its pivotal role in salt tolerance. Plant Mol Biol. 2015; 88: 41-52.

[32] Shekhawat UK, Ganapathi TR . MusaWRKY71 overexpression in banana plants leads to altered abiotic and biotic stress responses. PLoS One. 2013; 8: e75506.

[33] Tak H, Negi S, Ganapathi TR . Banana NAC transcription factor MusaNAC042 is positively associated with drought and salinity tolerance. Protoplasma. 2017; 254: 803-16.

[34] Negi S, Tak H, Ganapathi TR . A banana NAC transcription factor (MusaSNAC1) impart drought tolerance by modulating stomatal closure and H₂O₂ content . Plant Mol Biol. 2018; 96: 457-71.

[35] Xu Y, Li J, Song S et al. A novel aquaporin gene MaSIP2-1 confers tolerance to drought and cold stresses in transgenic banana. Mol Breed. 2020; 40: 1380-3743.

[36] Hu Y, Jiang Y, Han X et al. Jasmonate regulates leaf senescence and tolerance to cold stress: crosstalk with other phytohormones. J Exp Bot. 2017; 68: 1361-9.

[37] Moore K, Roberts LJ . Measurement of lipid peroxidation. Free Radic Res. 1998; 28: 659-71.

[38] Xu Y, Hu W, Liu J et al. A banana aquaporin gene, MaPIP1;1, is involved in tolerance to drought and salt stresses. BMC Plant Biol. 2014; 14: 59.

[39] Wang C, Deng P, Chen L et al. A wheat WRKY transcription factor TaWRKY10 confers tolerance to multiple abiotic stresses in transgenic tobacco. PLoS One. 2013; 8: e65120.

[40] Zhu T, Zou L, Li Y et al. Mitochondrial alternative oxidase-dependent autophagy involved in ethylene-mediated drought tolerance in Solanum lycopersicum. Plant Biotechnol J. 2018; 16: 2063-76.

[41] Ullah A, Manghwar H, Shaban M et al. Phytohormones enhanced drought tolerance in plants: a coping strategy. Environ Sci Pollut Res Int. 2018; 25: 33103-18.

[42] Sun X, Zhao T, Gan S et al. Ethylene positively regulates cold tolerance in grapevine by modulating the expression of ETHYLENE RESPONSE FACTOR 057. Sci Rep. 2016; 6: 24066.

[43] Liu W, Wang H, Chen Y et al. Cold stress improves the production of artemisinin depending on the increase in endogenous jasmonate. Biotechnol Appl Biochem. 2017; 64: 305-14.

[44] Ruan J, Zhou Y, Zhou M et al. Jasmonic acid signaling pathway in plants. Int J Mol Sci. 2019; 20: 2479.

[45] Aharoni N. Relationship between leaf water status and endogenous ethylene in detached leaves. Plant Physiol. 1978; 61: 658-62.

[46] Yu J, Jiang M, Guo C . Crop pollen development under drought: from the phenotype to the mechanism. Int J Mol Sci. 2019; 20: 1550.

[47] Arraes FB, Beneventi MA, Lisei de Sa ME et al. Implications of ethylene biosynthesis and signaling in soybean drought stress tolerance. BMC Plant Biol. 2015; 15: 213.

[48] Zhao ML, Wang JN, Shan W et al. Induction of jasmonate signalling regulators MaMYC2s and their physical interactions with MaICE1 in methyl jasmonate-induced chilling tolerance in banana fruit. Plant Cell Environ. 2013; 36: 30-51.

[49] Huang B, Li X, Liu P et al. Transcriptomic analysis of Eruca vesicaria subs. sativa lines with contrasting tolerance to polyethylene glycol-simulated drought stress. BMC Plant Biol. 2019; 19: 419-9.

[50] Zhao R, Xie H, Lv S et al. LeMAPK4 participated in cold-induced ethylene production in tomato fruit. J Sci Food Agric. 2013; 93: 1003-9.

[51] Sun T, Cen G, You C et al. ScAOC1, an allene oxide cyclase gene, confers defense response to biotic and abiotic stresses in sugarcane. Plant Cell Rep. 2020; 39: 1785-801.

[52] Zhao Y, Dong W, Zhang N et al. A wheat allene oxide cyclase gene enhances salinity tolerance via jasmonate signaling. Plant Physiol. 2014; 164: 1068-76.

[53] Hu Y, Jiang L, Wang F et al. Jasmonate regulates the inducer of cbf expression-C-repeat binding factor/DRE binding factor1 cascade and freezing tolerance in Arabidopsis. Plant Cell. 2013; 25: 2907-24.

[54] Houben M, Van de Poel B . 1-Aminocyclopropane-1-carboxylic acid oxidase (ACO): the enzyme that makes the plant hormone ethylene. Front Plant Sci. 2019; 10: 695.

[55] Lelièvre JM, Tichit L, Dao P et al. Effects of chilling on the expression of ethylene biosynthetic genes in Passe-Crassane pear (Pyrus communis L.) fruits. Plant Mol Biol. 1997; 33: 847-55.

[56] Nieuwenhuizen NJ, Chen X, Pellan M et al. Regulation of wound ethylene biosynthesis by NAC transcription factors in kiwifruit. BMC Plant Biol. 2021; 21: 411.

[57] Luo W, Gong Y, Tang Y et al. Glutathione and ethylene biosynthesis reveal that the glume and lemma have better tolerance to water deficit in wheat. Plant Physiol Biochem. 2021; 160: 120-9.

[58] Dalal M, Sahu S, Tiwari S et al. Transcriptome analysis reveals interplay between hormones, ROS metabolism and cell wall biosynthesis for drought-induced root growth in wheat. Plant Physiol Biochem. 2018; 130: 482-92.

[59] Wei H, Xue Y, Chen P et al. Genome-wide identification and functional investigation of 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) genes in cotton. Plants (Basel). 2021; 10: 1699.

[60] Xie Z, Nolan TM, Jiang H et al. AP2/ERF transcription factor regulatory networks in hormone and abiotic stress responses in Arabidopsis. Front Plant Sci. 2019; 10: 228-8.

[61] Xiao Y, Chen JY, Kuang JF et al. Banana ethylene response factors are involved in fruit ripening through their interactions with ethylene biosynthesis genes. J Exp Bot. 2013; 64: 2499-510.

[62] Livak KJ, Schmittgen TD . Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods. 2001; 25: 402-8.

[63] Stewart CN Jr. The utility of green fluorescent protein in transgenic plants. Plant Cell Rep. 2001; 20: 376-82.

[64] Waldo GS, Standish BM, Berendzen J et al. Rapid protein-folding assay using green fluorescent protein. Nat Biotechnol. 1999; 17: 691-5.

[65] Bottin A, Larche L, Villalba F et al. Green fluorescent protein (GFP) as gene expression reporter and vital marker for studying development and microbe-plant interaction in the tobacco pathogen Phytophthora parasitica var. nicotianae. FEMS Microbiol Lett. 1999; 176: 51-6.

[66] Jiang MHKB, Zhang J . Effect of abscisic acid on active oxygen species, antioxidative defence system and oxidative damage in leaves of maize seedlings. Plant Cell Physiol. 2001; 42: 1265-73.

[67] Heath RL, Packer L . Photoperoxidation in isolated chloroplasts. I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys. 1968; 125: 189-98.

[68] Bates LS, Waldren RP, Teare ID . Rapid determination of free proline for water-stress studies. Plant Soil. 1973; 39: 205-7.

[69] Niu Q, Li J, Cai D et al. Dormancy-associated MADS-box genes and microRNAs jointly control dormancy transition in pear (Pyrus pyrifolia white pear group) flower bud. J Exp Bot. 2016; 67: 239-57.