The class B heat shock factor HSFB1 regulates heat tolerance in grapevine

Abstract

Grape is a widely cultivated crop with high economic value. Most cultivars derived from mild or cooler climates may not withstand increasing heat stress. Therefore, dissecting the mechanisms of heat tolerance in grapes is of particular significance. Here, we performed comparative transcriptome analysis of Vitis davidii ‘Tangwei’ (heat tolerant) and Vitis vinifera ‘Jingxiu’ (heat sensitive) grapevines after exposure to 25℃, 40℃, or 45℃ for 2 h. More differentially expressed genes (DEGs) were detected in ‘Tangwei’ than in ‘Jingxiu’ in response to heat stress, and the number of DEGs increased with increasing treatment temperatures. We identified a class B Heat Shock Factor, HSFB1, which was significantly upregulated in ‘Tangwei’, but not in ‘Jingxiu’, at high temperature. VdHSFB1 from ‘Tangwei’ and VvHSFB1 from ‘Jingxiu’ differ in only one amino acid, and both showed similar transcriptional repression activities. Overexpression and RNA interference of HSFB1 in grape indicated that HSFB1 positively regulates the heat tolerance. Moreover, the heat tolerance of HSFB1- overexpressing plants was positively correlated to HSFB1 expression level. The activity of the VdHSFB1 promoter is higher than that of VvHSFB1 under both normal and high temperatures. Promoter analysis showed that more TATA-box and AT∼TATA-box cis-elements are present in the VdHSFB1 promoter than the VvHSFB1 promoter. The promoter sequence variations between VdHSFB1 and VvHSFB1 likely determine the HSFB1 expression levels that influence heat tolerance of the two grape germplasms with contrasting thermotolerance. Collectively, we validated the role of HSFB1 in heat tolerance, and the knowledge gained will advance our ability to breed heat-tolerant grape cultivars.

Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grant no. U21A20227), the National Key Research and Development Program of China (Grant no. 2018YFD1000300), and the Strategic Priority Research Program of the Chinese Academy of Sciences (Grant no. XDA23080602). Research conducted as part of the LIA INNOGRAPE International Associated Laboratory.

Author contributions

H.C. performed the work and initiated the draft. X.L., S.L., and H.M. participated in the plant material screening, treatment, and collection. Y.W. participated in data analysis. Y.L. helped to do the transformation of grape plants and cells. L.Y., W.D., P.F., and Z.L. helped to design the experiment. L.W. obtained funding and designed experiments. L.Y. and L.W. did the final editing.

Data availability

The raw data of RNA-sequencing for this article have been deposited in National Genomics Data Center (https://ngdc.cncb. ac.cn/). The BioProject accession is PRJCA009515. And the data used to support the findings of this study are available from the corresponding authors upon reasonable request.

Conflict of interest

The authors have no conflicts of interest to declare.

References

[1] Kumar RR, Goswami S, Singh K et al. Memorizing stress: elucidating the role of Rubisco activase in modulating the carbon assimlatory process and defense related network of Wheat (Triticum aestivum) under heat stress . In: AFHABEC-2016. 2016.

[2] Allakhverdiev SI, Kreslavski VD, Klimov VV et al. Heat stress: an overview of molecular responses in photosynthesis. Photosynth Res 2008; 98: 541-50.

[3] Sharkey TD, Zhang R . High temperature effects on electron and proton circuits of photosynthesis. J Integr Plant Biol. 2010; 52: 712-22.

[4] Wahid A, Gelani S, Ashraf M et al. Heat tolerance in plants: an overview. Environ Exp Bot. 2007; 61: 199-223.

[5] Hasanuzzaman M, Nahar K, Alam MM et al. Physiological, biochemical, and molecular mechanisms of heat stress tolerance in plants. Int J Mol Sci. 2013; 14: 9643-84.

[6] Li B, Gao K, Ren H et al. Molecular mechanisms governing plant responses to high temperatures. J Integr Plant Biol. 2018; 60: 757-79.

[7] Zhang X, Wu Y, Zhang Y et al. Identification of potential key genes in resveratrol biosynthesis via transcriptional analyses of berry development in grapevine (Vitis spp.) genotypes varying in trans-resveratrol content . Fruit Res. 2022; 2: 6.

[8] Ren C, Lin Y, Liang Z . CRISPR/Cas genome editing in grapevine: recent advances, challenges and future prospects. Fruit Res. 2022; 2: 7.

[9] Wu W, Fu P, Lu J . Grapevine WRKY transcription factors. Fruit Res. 2022; 2: 10.

[10] Venios X, Korkas E, Nisiotou A et al. Grapevine responses to heat stress and global warming. Plan Theory. 2020; 9: 1754.

[11] Pereira HS, Delgado M, Avo AP et al. Pollen grain development is highly sensitive to temperature stress in Vitis vinifera. Aust J Grape Wine Res. 2015; 20: 474-84.

[12] Lecourieux F, Kappel C, Pieri P et al. Dissecting the biochemical and transcriptomic effects of a locally applied heat treatment on developing cabernet sauvignon grape berries. Front Plant Sci. 2017; 8: 53.

[13] Movahed N, Pastore C, Cellini A et al. The grapevine VviPrx31 peroxidase as a candidate gene involved in anthocyanin degradation in ripening berries under high temperature. J Plant Res. 2016; 129: 513-26.

[14] Torregrosa L, Bigard A, Doligez A et al. Developmental, molecular and genetic studies on grapevine response to temperature open breeding strategies for adaptation to warming. OENO One. 2017; 51: 155-65.

[15] Liu G, Wang J, Cramer G et al. Transcriptomic analysis of grape (Vitis vinifera L.) leaves during and after recovery from heat stress . BMC Plant Biol. 2012; 12: 174.

[16] Liu G, Ma L, Duan W et al. Differential proteomic analysis of grapevine leaves by iTRAQ reveals responses to heat stress and subsequent recovery. BMC Plant Biol. 2014; 14: 110.

[17] Jiang J, Liu X, Liu C et al. Integrating omics and alternative splicing reveals insights into grape response to high temperature. Plant Physiol. 2017; 173: 1502-18.

[18] Liu G, Jiang J, Liu X et al. New insights into the heat responses of grape leaves via combined phosphoproteomic and acetylproteomic analyses. Hortic Res. 2019; 6: 100.

[19] Lecourieux D, Kappel C, Claverol S et al. Proteomic and metabolomic profiling underlines the stage- and time-dependent effects of high temperature on grape berry metabolism. J Integr Plant Biol. 2020; 62: 1132-58.

[20] Seth R, Maritim TK, Parmar R et al. Underpinning the molecular programming attributing heat stress associated thermotolerance in tea (Camellia sinensis (L.) O. Kuntze). Hortic Res. 2021; 8: 99.

[21] Scharf KD, Berberich T, Ebersberger I et al. The plant heat stress transcription factor (HSF) family: structure, function and evolution. Biochim Biophys Acta. 2012; 1819: 104-19.

[22] Nover L, Bharti K, Döring P et al. Arabidopsis and the heat stress transcription factor world: how many heat stress transcription factors do we need? Cell Stress Chaperones. 2001; 6: 177-89.

[23] Kotak S, Port M, Ganguli A et al. Characterization of C-terminal domains of Arabidopsis heat stress transcription factors (HSFs) and identification of a new signature combination of plant class a HSFs with AHA and NES motifs essential for activator function and intracellular localization. Plant J. 2004; 39: 98-112.

[24] Mishra SK, Tripp J, Winkelhaus S et al. In the complex family of heat stress transcription factors, HSFA1 has a unique role as master regulator of thermotolerance in tomato. Genes Dev. 2002; 16: 1555-67.

[25] Yoshida T, Ohama N, Nakajima J et al. Arabidopsis HSFA1 transcription factors function as the main positive regulators in heat shock-responsive gene expression. Mol Gen Genomics. 2011; 286: 321-32.

[26] Liu H, Liao H, Charng Y . The role of class A1 heat shock factors (HSFA1s) in response to heat and other stresses in Arabidopsis. Plant Cell Environ. 2011; 34: 738-51.

[27] Charng Y, Liu H, Liu N et al. A heat-inducible transcription factor, HSFA2, is required for extension of acquired thermotolerance in Arabidopsis. Plant Physiol. 2007; 143: 251-62.

[28] Yoshida T, Sakuma Y, Todaka D et al. Functional analysis of an Arabidopsis heat-shock transcription factor HSFA3 in the transcriptional cascade downstream of the DREB2A stress-regulatory system. Biochem Biophys Res Commun. 2008; 368: 515-21.

[29] Schramm F, Larkindale J, Kiehlmann E et al. A cascade of transcription factor DREB2A and heat stress transcription factor HSFA3 regulates the heat stress response of Arabidopsis. Plant J. 2008; 53: 264-74.

[30] Fragkostefanakis S, Simm S, El-Shershaby A et al. The repressor and co-activator HSFB1 regulates the major heat stress transcription factors in tomato. Plant Cell Environ. 2019; 42: 874-90.

[31] Hu Y, Han Y, Zhang K et al. Identification and expression analysis of heat shock transcription factors in the wild Chinese grapevine (Vitis pseudoreticulata). Plant Physiol Biochem. 2016; 99: 1-10.

[32] Liu G, Chai F, Wang Y et al. Genome-wide identification and classification of HSF family in grape, and their transcriptional analysis under heat acclimation and heat stress. Hortic Plant J. 2018; 4: 133-43.

[33] Giorno F, Guerriero G, Baric S et al. Heat shock transcriptional factors in Malus domestica: identification, classification and expression analysis . BMC Genomics. 2012; 13: 639.

[34] Lin Q, Jiang Q, Lin J et al. Heat shock transcription factors expression during fruit development and under hot air stress in Ponkan (Citrus reticulata Blanco cv. Ponkan) fruit . Gene. 2015; 559: 129-36.

[35] Hu Y, Han Y, Wei W et al. Identification, isolation, and expression analysis of heat shock transcription factors in the diploid woodland strawberry Fragaria vesca. Front Plant Sci. 2015; 6: 736.

[36] Peng S, Zhu Z, Zhao K et al. A novel heat shock transcription factor, VpHSF1, from Chinese wild Vitis pseudoreticulata is involved in biotic and abiotic stresses . Plant Mol Biol Report. 2013; 31: 240-7.

[37] Liu X, Chen H, Li S et al. Natural variations of HSFA2 enhance thermotolerance in grapevine. Hortic Res. 2022; 10: uhac250.

[38] Ashraf M, Harris P . Photosynthesis under stressful environments: an overview. Photosynthetica. 2013; 51: 163-90.

[39] Xu H, Liu G, Liu G et al. Comparison of investigation methods of heat injury in grapevine (Vitis) and assessment to heat tolerance in different cultivars and species . BMC Plant Biol. 2014; 14: 156.

[40] Ashraf MF, Yang S, Wu R et al. Capsicum annuum HSFB2a positively regulates the response to Ralstonia solanacearum infection or high temperature and high humidity forming transcriptional cascade with CaWRKY6 and CaWRKY40. Plant Cell Physiol. 2018; 59: 2608-23.

[41] Ikeda M, Mitsuda N, Ohme-Takagi M . Arabidopsis HSFB1 and HSFB2b act as repressors of the expression of heat-inducible HSFs but positively regulate the acquired thermotolerance. Plant Physiol. 2011; 157: 1243-54.

[42] Czarnecka-Verner E, Pan S, Salem T et al. Plant class B HSFs inhibit transcription and exhibit affinity for TFIIB and TBP. Plant Mol Biol. 2004; 56: 57-75.

[43] Ren C, Liu X, Zhang Z et al. CRISPR/Cas9-mediated efficient targeted mutagenesis in chardonnay (Vitis vinifera L.). Sci Rep. 2016; 6: 32289.

[44] Ilík P, Špundová M, Šicner M et al. Estimating heat tolerance of plants by ion leakage: a new method based on gradual heating. New Phytol. 2018; 218: 1278-87.

[45] Lescot M, Déhais P, Thijs G et al. PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences. Nucleic Acids Res. 2002; 30: 325-7.

[46] Zhang M, Lv Y, Wang Y et al. TATA box insertion provides a selection mechanism underpinning adaptations to Fe deficiency. Plant Physiol. 2017; 173: 715-27.

[47] Iglesias A, Garrote L, Flores F et al. Challenges to manage the risk of water scarcity and climate change in the Mediterranean. Water Resour Manag. 2007; 21: 775-88.

[48] White MA, Diffenbaugh NS, Jones GV et al. Extreme heat reduces and shifts United States premium wine production in the 21st century. Proc Natl Acad Sci U S A. 2006; 103: 11217-22.

[49] Schultz HR, Stoll M . Some critical issues in environmental physiology of grapevines: future challenges and current limitations. Aust J Grape Wine Res. 2010; 16: 4-24.

[50] Banilas G, Korkas E, Kaldis P et al. Olive and Grapevine Biodiversity in Greece and Cyprus-A Review. Netherlands: Springer; 2009.

[51] Wolkovich EM, García de Cortázar-Atauri I, Morales-Castilla I et al. From pinot to Xinomavro in the world’s future wine-growing regions. Nat Clim Chang. 2018; 8: 29-37.

[52] Banilas G, Korkas E, Englezos V et al. Genome-wide analysis of the heat shock protein 90 gene family in grapevine (Vitis vinifera L.). Aust J Grape Wine Res. 2012; 18: 29-38.

[53] Guo M, Liu J, Ma X et al. The plant heat stress transcription factors (HSFs): structure, regulation, and function in response to abiotic stresses. Front Plant Sci. 2016; 7: 114.

[54] Ikeda M, Ohme-Takagi M . A novel group of transcriptional repressors in Arabidopsis. Plant Cell Physiol. 2009; 50: 970-5.

[55] Bharti K, Von Koskull-Döring P, Bharti S et al. Tomato heat stress transcription factor HSFB1 represents a novel type of general transcription coactivator with a histone-like motif interacting with the plant CREB binding protein ortholog HAC1. Plant Cell. 2004; 16: 1521-35.

[56] Ma H, Wang C, Yang B et al. CarHSFB2, a class B heat shock transcription factor, is involved in different developmental processes and various stress responses in chickpea (Cicer Arietinum L.). Plant Mol Biol Rep. 2016; 34: 1-14.

[57] Zhang S, Xu Z, Li P et al. Overexpression of TaHSF3 in transgenic Arabidopsis enhances tolerance to extreme temperatures . Plant Mol Biol Rep. 2013; 31: 688-97.

[58] Xiang J, Ran J, Zou J et al. Heat shock factor OsHSFB2b negatively regulates drought and salt tolerance in rice. Plant Cell Rep. 2013; 32: 1795-806.

[59] Zhang Z, Li J, Pan Y et al. Natural variation in CTB4a enhances rice adaptation to cold habitats . Nat Commun. 2017; 8: 14788.

[60] Xu Y, Zhang L, Ou S et al. Natural variations of SLG1 confer high-temperature tolerance in indica rice . Nat Commun. 2020; 11: 5441.

[61] Basi G, Schmid E, Maundrell K . TATA box mutations in the Schizosaccharomyces pombe nmt1 promoter affect transcription efficiency but not the transcription start point or thiamine repressibility. Gene. 1993; 123: 131-6.

[62] Coutos-Thevenot P, Maes O, Jouenne T et al. Extracellular protein patterns of grapevine cell suspensions in embryogenic and non-embryogenic situations. Plant Sci. 1992; 86: 137-45.

[63] Wang Z, Wong DCJ, Wang Y et al. GRAS-domain transcription factor PAT1 regulates jasmonic acid biosynthesis in grape cold stress response. Plant Physiol. 2021; 186: 1660-78.

[64] Xu M, Tong Q, Wang Y et al. Transcriptomic analysis of grapevine LEA gene family in response to osmotic and cold stress, and functional analyses of VamDHN3 gene . Plant Cell Physiol. 2020; 61: 775-86.

[65] Bolger AM, Lohse M, Usadel B . Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics. 2014; 30: 2114-20.

[66] Jaillon O, Aury JM, Noel B et al. The grapevine genome sequence suggests ancestral hexaploidization in major angiosperm phyla. Nature. 2007; 449: 463-7.

[67] Kim D, Pertea G, Trapnell C et al. TopHat2: accurate alignment of transcriptomes in the presence of insertions, deletions and gene fusions. Genome Biol. 2013; 14: R36.

[68] Trapnell C, Roberts A, Goff L et al. Differential gene and transcript expression analysis of RNA-seq experiments with TopHat and cufflinks. Nat Protoc. 2012; 7: 562-78.

[69] Mao X, Cai T, Olyarchuk JG et al. Automated genome annotation and pathway identification using the KEGG Orthology (KO) as a controlled vocabulary. Bioinformatics. 2005; 21: 3787-93.

[70] Kerschen A, Napoli CA, Jorgensen RA et al. Effectiveness of RNA interference in transgenic plantlets. FEBS Lett. 2004; 566: 223-8.

[71] Mu H, Li Y, Yuan L et al. MYB30 and MYB14 form a repressor-activator module with WRKY8 that controls stilbene biosynthesis in grapevine. Plant Cell. 2023; 35: 552-73.