Natural variations of HSFA2 enhance thermotolerance in grapevine

Abstract

Heat stress limits growth and development of crops including grapevine which is a popular fruit in the world. Genetic variability in crops thermotolerance is not well understood. We identified and characterized heat stress transcription factor HSFA2 in heat sensitive Vitis vinifera ‘Jingxiu’ (named as VvHSFA2) and heat tolerant Vitis davidii ‘Tangwei’ (named as VdHSFA2). The transcriptional activation activities of VdHSFA2 are higher than VvHSFA2, the variation of single amino acid (Thr315Ile) in AHA1 motif leads to the difference of transcription activities between VdHSFA2 and VvHSFA2. Based on 41 Vitis germplasms, we found that HSFA2 is differentiated at coding region among heat sensitive V. vinifera, and heat tolerant Vitis davidii and Vitis quinquangularis. Genetic evidence demonstrates VdHSFA2 and VvHSFA2 are positive regulators in grape thermotolerance, and the former can confer higher thermotolerance than the latter. Moreover, VdHSFA2 can regulate more target genes than VvHSFA2. As a target gene of both VdHSFA2 and VvHSFA2, overexpression of MBF1c enhanced the grape thermotolerance whereas dysfunction of MBF1c resulted in thermosensitive phenotype. Together, our results revealed that VdHSFA2 confers higher thermotolerance than VvHSFA2, and MBF1c acts as their target gene to induce thermotolerance. The VdHSFA2 may be adopted for molecular breeding in grape thermotolerance.

Acknowledgement

This work was supported by the National Key Research and Development Program of China (Grant No. 2018YFD1000300), the National Natural Science Foundation of China (Grant No. 31872065 and U21A20227), and the Strategic Priority Research Program of the Chinese Academy of Sciences (Grant No. XDA23080602). We would like to thank Professors Xianjun Song and Jingbo Jin from the Institute of Botany, Chinese Academy of Sciences. This research was conducted as part of the LIA INNOGRAPE International Associated Laboratory.

Author contributions

X.L. performed the work and initiated the draft. H.C. measured heat tolerance of grapevine accessions. S.L. helped to clone promoters. D.L. critically reviewed the manuscript. W.D., P.F., and Z.L. helped to design the experiment. L.W. obtained funding, designed the experiment and revised the final version.

Data availability

The data used to support the findings of this study are available from the corresponding authors upon reasonable request.

Conflict of interests

The authors declare no conflict of interest.

Supplementary data

Supplementary data is available at Horticulture Research online.

References

[1] Gong Z, Xiong L, Shi H et al. Plant abiotic stress response and nutrient use efficiency. Sci China Life Sci. 2020; 63: 635-74.

[2] Ohama N, Sato H, Shinozaki K et al. Transcriptional regulatory network of plant heat stress response. Trends Plant Sci. 2017; 22: 53-65.

[3] Katano K, Honda K, Suzuki N . Integration between ROS regulatory systems and other signals in the regulation of various types of heat responses in plants. Int J Mol Sci. 2018; 19: 3370.

[4] 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.

[5] 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.

[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] Liu HC, Liao HT, Charng YY . 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.

[8] Wu Z, Liang J, Wang C et al. Alternative splicing provides a mechanism to regulate LlHSFA3 function in response to heat stress in lily. Plant Physiol. 2019; 181: 1651-67.

[9] Hu Y, Mesihovic A, Jiménez-Gómez JM et al. Natural variation in HsfA2 pre-mRNA splicing is associated with changes in thermotolerance during tomato domestication. New Phytol. 2020; 225: 1297-310.

[10] Liu GT, 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.

[11] Xue GP, Sadat S, Drenth J et al. The heat shock factor family from Triticum aestivum in response to heat and other major abiotic stresses and their role in regulation of heat shock protein genes. J Exp Bot. 2014; 65: 539-57.

[12] Wang C, Zhang Q, Shou HX . Identification and expression analysis of OsHsfs in rice. J Zhejiang Univ Sci B. 2009; 10: 291-300.

[13] Zang D, Wang J, Zhang X et al. Arabidopsis heat shock transcription factor HSFA7b positively mediates salt stress tolerance by binding to an E-box-like motif to regulate gene expression. J Exp Bot. 2019; 70: 5355-74.

[14] Nover L, Bharti K, Doring 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.

[15] Fragkostefanakis S, Mesihovic A, Simm S et al. HsfA2 controls the activity of developmentally and stress-regulated heat stress protection mechanisms in tomato male reproductive tissues. Plant Physiol. 2016; 170: 2461-77.

[16] Charng YY, Liu HC, Liu NY et al. A heat-inducible transcription factor, HsfA2, is required for extension of acquired thermotolerance in Arabidopsis. Plant Physiol. 2007; 143: 251-62.

[17] Ogawa D, Yamaguchi K, Nishiuchi T . High-level overexpression of the Arabidopsis HsfA2 gene confers not only increased themotolerance but also salt/osmotic stress tolerance and enhanced callus growth. J Exp Bot. 2007; 58: 3373-83.

[18] Gu L, Jiang T, Zhang C et al. Maize HSFA2 and HSBP2 antagonistically modulate raffinose biosynthesis and heat tolerance in Arabidopsis. Plant J. 2019; 100: 128-42.

[19] Pillet J, Egert A, Pieri P et al. VvGOLS1 and VvHsfA2 are involved in the heat stress responses in grapevine berries. Plant Cell Physiol. 2012; 53: 1776-92.

[20] Jaimes-Miranda F, Chávez Montes RA . The plant MBF1 protein family: a bridge between stress and transcription. J Exp Bot. 2020; 71: 1782-91.

[21] Li FQ, Ueda H, Hirose S . Mediators of activation of fushi tarazu gene transcription by BmFTZ-F1. Mol Cell Biol. 1994; 14: 3013-21.

[22] Takemaru K, Harashima S, Ueda H et al. Yeast coactivator MBF1 mediates GCN4-dependent transcriptional activation. Mol Cell Biol. 1998; 18: 4971-6.

[23] Tsuda K, Tsuji T, Hirose S et al. Three Arabidopsis MBF1 homologs with distinct expression profiles play roles as transcriptional co-activators. Plant Cell Physiol. 2004; 45: 225-31.

[24] Suzuki N, Rizhsky L, Liang H et al. Enhanced tolerance to environmental stress in transgenic plants expressing the transcriptional coactivator multiprotein bridging factor 1c. Plant Physiol. 2005; 139: 1313-22.

[25] Suzuki N, Bajad S, Shuman J et al. The transcriptional co-activator MBF1c is a key regulator of thermotolerance in Arabidopsis thaliana. J Biol Chem. 2008; 283: 9269-75.

[26] Arce DP, Godoy AV, Tsuda K et al. The analysis of an Arabidopsis triple knock-down mutant reveals functions for MBF1 genes under oxidative stress conditions. J Plant Physiol. 2010; 167: 194-200.

[27] Zandalinas SI, Balfagón D, Arbona V et al. ABA is required for the accumulation of APX1 and MBF1c during a combination of water deficit and heat stress. J Exp Bot. 2016; 67: 5381-90.

[28] Zegzouti H, Jones B, Frasse P et al. Ethylene-regulated gene expression in tomato fruit: characterization of novel ethylene-responsive and ripening-related genes isolated by differential display. Plant J. 1999; 18: 589-600.

[29] Qin D, Wang F, Geng X et al. Overexpression of heat stress-responsive TaMBF1c, a wheat (Triticum aestivum L.) multiprotein bridging factor, confers heat tolerance in both yeast and rice. Plant Mol Biol. 2015; 87: 31-45.

[30] Zou L, Yu B, Ma XL et al. Cloning and expression analysis of the BocMBF1c gene involved in heat tolerance in Chinese kale. Int J Mol Sci. 2019; 20: 5637.

[31] Ding L, Wu Z, Teng R et al. LlWRKY39 is involved in thermotolerance by activating LlMBF1c and interacting with LlCaM3 in lily (Lilium longiflorum). Hortic Res. 2021; 8: 36.

[32] Yan Q, Hou H, Singer SD et al. The grape VvMBF1 gene improves drought stress tolerance in transgenic Arabidopsis thaliana. Plant Cell Tiss Org. 2014; 118: 571-82.

[33] Rienth M, Torregrosa L, Luchaire N et al. Day and night heat stress trigger different transcriptomic responses in green and ripening grapevine (vitis vinifera) fruit . BMC Plant Biol. 2014; 14: 108.

[34] Rienth M, Torregrosa L, Sarah G et al. Temperature desynchronizes sugar and organic acid metabolism in ripening grapevine fruits and remodels their transcriptome. BMC Plant Biol 2016; 16: 164.

[35] Lecourieux F, Lecourieux D, Vignault C et al. A sugar-inducible protein kinase, VvSK1, regulates hexose transport and sugar accumulation in grapevine cells. Plant Physiol. 2010; 152: 1096-106.

[36] 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.

[37] Tu M, Wang X, Zhu Y et al. VlbZIP30 of grapevine functions in dehydration tolerance via the abscisic acid core signaling pathway. Hortic Res. 2018; 5: 49.

[38] Jiang J, Xi H, Dai Z et al. VvWRKY8 represses stilbene synthase genes through direct interaction with VvMYB14 to control resveratrol biosynthesis in grapevine. J Exp Bot. 2019; 70: 715-29.

[39] Sun X, Zhang L, Wong DCJ et al. The ethylene response factor VaERF092 from Amur grape regulates the transcription factor VaWRKY33, improving cold tolerance. Plant J. 2019; 99: 988-1002.

[40] 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.

[41] 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.

[42] Liu Y, Jiang J, Fan X et al. Identification of heat tolerance in Chinese wildgrape germplasm resources. Horticulturae. 2020; 6: 68.

[43] George IS, Pascovici D, Mirzaei M et al. Quantitative proteomic analysis of cabernet sauvignon grape cells exposed to thermal stresses reveals alterations in sugar and phenylpropanoid metabolism. Proteomics. 2015; 15: 3048-60.

[44] Nishizawa A, Yabuta Y, Yoshida E et al. Arabidopsis heat shock transcription factor A2 as a key regulator in response to several types of environmental stress. Plant J. 2006; 48: 535-47.

[45] Li H, Durbin R . Fast and accurate short read alignment with burrows-wheeler transform. Bioinformatics. 2009; 25: 1754-60.

[46] Kim D, Langmead B, Salzberg SL . HISAT: a fast spliced aligner with low memory requirements. Nat Methods. 2015; 12: 357-60.

[47] Yokotani N, Ichikawa T, Kondou Y et al. Expression of rice heat stress transcription factor OsHsfA2e enhances tolerance to environmental stresses in transgenic Arabidopsis. Planta. 2008; 227: 957-67.

[48] Jiang Y, Zheng Q, Chen L et al. Ectopic overexpression of maize heat shock transcription factor gene ZmHsf04 confers increased thermo and salt-stress tolerance in transgenic Arabidopsis . Acta Physiol Plant. 2018; 40: 9.

[49] Ding X, Guo Q, Li Q et al. Comparative transcriptomics analysis and functional study reveal important role of high-temperature stress response gene GmHSFA2 during flower bud development of CMS-based F₁ in soybean . Front Plant Sci. 2020; 11: 600217.

[50] Liu Z, Li G, Zhang H et al. TaHsfA2-1, a new gene for thermotolerance in wheat seedlings: characterization and functional roles. J Plant Physiol. 2020; 246-247: 153135.

[51] Friedrich T, Oberkofler V, Trindade I et al. Heteromeric HSFA2/HSFA3 complexes drive transcriptional memory after heat stress in Arabidopsis. Nat Commun. 2021; 12: 3426.

[52] Venios X, Korkas E, Nisiotou A et al. Grapevine responses to heat stress and global warming. Plants (Basel). 2020; 9: 1754.

[53] Gomes E, Maillot P, Duchene E . Molecular tools for adapting viticulture to climate change. Front Plant Sci. 2021; 12: 633846.

[54] Evrard A, Kumar M, Lecourieux D et al. Regulation of the heat stress response in Arabidopsis by MPK6-targeted phosphorylation of the heat stress factor HsfA2. PeerJ. 2013; 1: e59.

[55] Li XM, Chao DY, Wu Y et al. Natural alleles of a proteasome alpha2 subunit gene contribute to thermotolerance and adaptation of African rice. Nat Genet. 2015; 47: 827-33.

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

[57] Kidokoro S, Watanabe K, Ohori T et al. Soybean DREB1/CBF-type transcription factors function in heat and drought as well as cold stress-responsive gene expression. Plant J. 2015; 81: 505-18.

[58] Yun SD, Kim MH, Oh SA et al. Overexpression of C-repeat binding factor1 (CBF1) gene enhances heat stress tolerance in Arabidopsis. J Plant Biol. 2022; 65: 253-60.

[59] Li C, Qiao Z, Qi W et al. Genome-wide characterization of cis-acting DNA targets reveals the transcriptional regulatory framework of opaque2 in maize. Plant Cell. 2015; 27: 532-45.

[60] Zhang H, Zhang Q, Zhai H et al. IbBBX24 promotes the jasmonic acid pathway and enhances fusarium wilt resistance in sweet potato. Plant Cell. 2020; 32: 1102-23.

[61] Matsushita Y, Miyakawa O, Deguchi M et al. Cloning of a tobacco cDNA coding for a putative transcriptional coactivator MBF1 that interacts with the tomato mosaic virus movement protein. J Exp Bot. 2002; 53: 1531-2.

[62] Arce DP, Tonón C, Zanetti ME et al. The potato transcriptional co-activator StMBF1 is up-regulated in response to oxidative stress and interacts with the TATA-box binding protein. J Biochem Mol Biol. 2006; 39: 355-60.

[63] Guo WL, Chen RG, Du XH et al. Reduced tolerance to abiotic stress in transgenic Arabidopsis overexpressing a Capsicum annuum multiprotein bridging factor 1 . BMC Plant Biol. 2014; 14: 138.

[64] 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.

[65] Ma X, Liu YG . CRISPR/Cas9-based multiplex genome editing in monocot and dicot plants. Curr Protoc Mol Biol. 2016; 115: 31.6.1-21.

[66] Xu M, Tong Q, Wang Y et al. Transcriptomic analysis of the grapevine LEA gene family in response to osmotic and cold stress reveals a key role for VamDHN3. Plant Cell Physiol. 2020; 61: 775-86.

[67] Landt SG, Marinov GK, Kundaje A et al. ChIP-seq guidelines and practices of the ENCODE and modENCODE consortia. Genome Res. 2012; 22: 1813-31.

[68] Lin R, Ding L, Casola C et al. Transposase-derived transcription factors regulate light signaling in Arabidopsis. Science 2007; 318: 1302-5.