|
|
|
| Biological Characteristics and Thermotolerance Analysis of Heat Shock Transcription Factor TaHsfA1 Subfamily Genes in Wheat (Triticum aestivum) |
| LIU Ran1,2,*, MENG Xiang-Zhao1,*, YUAN Sai-Nan1,2, LI Guo-Liang1, YANG Yang1, DUAN Shuo-Nan1, ZHANG Hua-Ning1,**, GUO Xiu-Lin1,** |
1 Institute of Biotechnology and Food Sciences, Hebei Academy of Agriculture and Forestry Sciences/Plant Genetic Engineering Center of Hebei Province, Shijiazhuang 050051, China;
2 College of Life Sciences, Hebei Normal University, Shijiazhuang 050024, China |
|
|
|
|
Abstract Heat shock transcription factor (Hsf) exists in most plants and plays an important role in the response to heat tolerance and other stresses. Family genes showed diverse characteristics and functions. In this study, 3 homoeologous genes TaHsfA1-1 (GenBank No. MW756130), TaHsfA1-2 (GenBank No. MW756131) and TaHsfA1-3 (GenBank No. MW756132) were obtained by homologous cloning technology from wheat (Triticum aestivum), their cDNA lengths were 1 590, 1 566 and 1 569 bp, respectively. Three protein sequences all contained the typical DNA binding domain (DBD), the same nuclear localization signal (NLS) sequence RRKP/KKRR and transcription activation domain sequence (aromaromatic, large hydrophobic and acidic amino acid residues, AHA) DSFWEQFLCA. The similarity of TaHsfA1-1, TaHsfA1-2 and TaHsfA1-3 to Aegilops tauschii AetHsfA1 was as high as 96%. Chromosome location analysis confirmed that TaHsfA1-1, TaHsfA1-2 and TaHsfA1-3 were located on chromosomes 4A, 5B and 5D, respectively. Through transient reporter assay with tobacco (Nicotiana tabacum) epidermal cells, it was found that the proteins was localized in the nucleus under the normal conditions. The qPCR analysis showed that the expression levels of the 3 genes were up-regulated by heat stress (HS) and salicylic acid (SA) treatment, respectively and down-regulated by abscisic acid (ABA) treatment, and TaHsfA1-3 expression was up-regulated by H2O2. Through assay in Saccharomyces cerevisiae AH109, TaHsfA1-1, TaHsfA1-2 and TaHsfA1-3 all had transcriptional activation activity. According to the expression level responsive to heat stress, TaHsfA1-1 was selected and transformed into Arabidopsis thaliana. Through observation the phenotype, it was found that TaHsfA1-1 could improve the basic thermotolerance and acquired thermotolerance of Arabidopsis thaliana seedlings and the chlorophyll contents of different transgenic lines under high temperature stress. The chlorophyll contents were consistent with the phenotype and survival rate. Hsp genes were induced to expression under the normal conditions and after heat stress in TaHsfA1-1 transgenic A. thaliana plants. The results indicated that TaHsfA1 could regulate thermotolerance, providing a theoretical basis and technical support for further exploring the characteristics and function of heat tolerance of wheat Hsf family.
|
|
Received: 01 April 2021
|
|
|
|
Corresponding Authors:
**myhf2002@163.com;zhn.8888-@163.com
|
|
|
|
[1] 焦淑珍, 姚文孔, 张宁波, 等. 2020. 园艺植物热激转录因子研究进展[J]. 果树学报, 37(3): 419-430.
(Jiao S Z, Yao W K, Zhang N B, et al.2020. Research progress of heat stress transcription factors (Hsfs) in horticultural plants[J]. Journal of Fruit Science, 37(3): 419-430.)
[2] 李慧聪, 李国良, 郭秀林. 2015. 玉米热激转录因子基因(ZmHsf06)的克隆、表达和定位分析[J].农业生物技术学报, 23(1): 41-51.
(Li H C, Li G L, Guo X L.2015. Cloning, expression characteristics and subcellular-location of heat shock transcription factor ZmHsf06 in Zea mays[J]. Journal of Agricultural Biotechnology, 23(1):41-51.)
[3] 张玉杰, 张园园, 张华宁, 等. 2018. 小麦热激转录因子基因TaHsfA2e特性及耐热性功能初探[J]. 作物学报, 44(12): 1818-1828.
(Zhang Y J, Zhang Y Y, Zhang H N, et al.2018. Characterization and regulatory roles in thermotolerance of wheat heat shock transcription factor gene TaHsfA2e[J]. Acta Agronomica Sinica, 44(12): 1818-1828.)
[4] 张园园, 赵慧, 张玉杰,等. 2019. 小麦热激转录因子基因TaHsfA2f生物学特性及耐热性调控作用[J]. 农业生物技术学报, 27(05): 69-79.
(Zhang Y Y, Zhao H, Zhang Y J, et al.2019. Biological characteristics and thermotolerance-regulating roles of wheat (Triticum aestivum) heat shock transcription factor gene TaHsfA2f[J]. Journal of Agricultural Biotechnology, 27(5): 825-835.)
[5] 赵立娜, 刘子会, 段硕楠, 等. 2018. 小麦热激转录因子基因TaHsfB2d的克隆和特性及其对耐热性调控[J]. 作物学报, 44(1): 53-62.
(Zhao L N, Liu Z H, Duan S N, et al.2018. Cloning and characterization of heat shock transcription factor gene TaHsfB2d and its regulating role in thermotolerance[J]. Acta Agronomica Sinica, 44(1): 53-62.)
[6] Almoguera C, Rojas A, Díaz-Martín J, et al.2002. A seed-specific heat-shock transcription factor involved in developmental regulation during embryogenesis in sunflower[J]. Journal of Biological Chemistry, 277(46): 43866-43872.
[7] Aranda M A, Escaler M, Thomas C L, et al.1999. A heat shock transcription factor in pea is differentially controlled by heat and virus replication[J]. The Plant Journal, 20(2): 153-161.
[8] Chan-Schaminet K Y, Baniwal S K, Bublak D, et al.2009. Specific interaction between tomato HsfA1 and HsfA2 creates hetero-oligomeric superactivator complexes for synergistic activation of heat stress gene expression[J]. Journal of Biological Chemistry, 284(31): 20848-20857.
[9] Czarnecka-Verner E, Yuan C X, Fox P C, et al.1995. Isolation and characterization of six heat shock transcription factor cDNA clones from soybean[J]. Plant Molecular Biology, 29(1): 37-51.
[10] Duan S N, Liu B H, Zhang Y Y, et al.2019. Genome-wide identification and abiotic stress-responsive pattern of heat shock transcription factor family in Triticum aestivum L.[J]. BMC Genomics, 20(1): 257-276.
[11] El-Shershaby A, Ullrich S, Simm S, et al.2019. Functional diversification of tomato HsfA1 factors is based on DNA binding domain properties[J]. Gene, 714: 143985.
[12] Fragkostefanakis S, Simm S, Paul P, et al.2015. Chaperone network composition in Solanum lycopersicum explored by transcriptome profiling and microarray meta-analysis[J]. Plant Cell Environment, 38(4): 693-709.
[13] Gagliardi D, Breton C, Chaboud A, et al.1995. Expression of heat shock factor and heat shock protein 70 genes during maize pollen development[J]. Plant Molecular Biology, 29(4): 841-856.
[14] Guo J K, Wu J, Ji Q, et al.2008. Genome-wide analysis of heat shock transcription factor families in rice and Arabidopsis[J]. Genet Genomics, 35(2): 105-118.
[15] Guo M, Liu J H, Ma X, et al.2016. The plant heat stress transcription factors (HSFs): Structure, regulation, and function in response to abiotic stresses[J]. Frontiers in Plant Science, 7: 114.
[16] Guo X L, Yuan S N, Zhang H N, et al.2020. Heat-response patterns of the heat shock transcription factor family in advanced development stages of wheat (Triticum aestivum L.) and thermotolerance-regulation by TaHsfA2-10[J]. BMC Plant Biology, 20(1): 364.
[17] Hahn A, Bublak D, Schleiff E, et al.2011. Crosstalk between Hsp90 and Hsp70 chaperones and heat stress transcription factors in tomato[J]. Plant Cell, 23(2): 741-755.
[18] Hübel A, Schöffl F, 1994. Arabidopsis heat shock factor: isolation and characterization of the gene and the recombinant protein[J]. Plant Molecular Biology, 26(1): 353-362.
[19] Kotak S, Port M, Ganguli A, et al.2004. Characterization of C-terminaldomains 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[J]. The Plant Journal, 39(1): 98-112.
[20] Li G L, Zhang YY, Zhang H N, et al.2019. Characteristics and regulating role in thermotolerance of the heat shock transcription factor ZmHsf12 from Zea mays L.[J]. Journal of Plant Biology, 62(5): 329-341.
[21] Li P S, Yu T F, He G H, et al.2014a. Genome-wide analysis of the Hsf family in soybean and functional identification of GmHsf-34 involvement in drought and heat stresses[J]. BMC Genomics, 15(1): 1009.
[22] Li H C, Zhang H N, Li G L, et al.2015. Expression of maize heat shock transcription factor gene ZmHsf06 enhances the thermotolerance and drought-stress tolerance of transgenic Arabidopsis[J]. Functional Plant Biology, 42(11): 1080-1090.
[23] Li H C, Li G L,Liu Z H, et al., 2014b. Cloning, localization and expression of ZmHsf-like in Zea mays[J]. Journal of Integrative Agriculture, 13(6): 1230-1238.
[24] Lipiec J, Doussan C, Nosalewicz A, et al.2013. Effect of drought and heat stresses on plant growth and yield: A review[J]. International Agrophysics, 27: 463-477.
[25] Liu H C, Liao H T, Charng Y Y, 2011. The role of class A1 heat shock factors (HSFA1s) in response to heat and other stresses in Arabidopsis[J]. Plant Cell Environment, 34(5): 738-751.
[26] Mishra S K, Tripp J, Winkelhaus S, et al.2002. In the complex family of heat stress transcription factors, HsfA1 has a unique role as master regulator of thermotolerance in tomato[J]. Genes & Development, 16(12): 1555-1567.
[27] Pelham H R, 1982. A regulatory upstream promoter element in the drosophila Hsp 70 heat-shock gene[J]. Cell, 30(2): 517-528.
[28] Peteranderl R, Rabenstein M, Shin Y K, et al.1999. Biochemical and biophysical characterization of the trimerization domain from the heat shock transcription factor[J]. Biochemistry, 38(12): 3559-3569.
[29] Scharf K D, Heider H, Höhfeld I, et al.1998. The tomato Hsf system: HsfA2 needs interaction with HsfAl for efficient nuclear import and may be localized in cytoplasmic heat stress granules[J]. Molecular and Cellular Biology, 18(4): 2240-2251.
[30] Shim D, Hwang J U, Lee J, et al.2009. Orthologs of the class A4 heat shock transcription factor HSFA4a confer cadmium tolerance in wheat and rice[J]. The Plant Journal, 21(12): 4031-4043.
[31] Snyman M, Cronjé M J, 2008. Modulation of heat shock factors accompanies salicylic acid-mediated potentiation of Hsp70 in tomato seedlings[J]. Journal of Experimental Botany, 59(8): 2125-2132.
[32] Xue G P, Sadat S, Drenth J, et al.2014. The heat shock factor family from Triticum aestium in response to heat and other major abiotic stresses and their role in regulation of heat shock protein genes[J]. Journal of Experimental Botany, 65(2): 539-557.
[33] Yamanouchi U, Yano M, Lin H X, et al.2002. Rice spotted leaf gene, Spl7, encodes a heat stress transcription factor protein[J]. Proceedings of the National Academy of Sciences of the USA, 99(11): 7530-7535.
[34] Yoshida T, Ohama N, Nakajima J, et al.2011. Arabidopsis HsfA1 transcription factors function as the main positive regulators in heat-shock responsive gene expression[J]. Molecular Genetics and Genomics December, 286(5): 321-332. |
|
|
|
