Abstract：The dehydration responsive element binding (DREB) gene family plays an important role in plant response to abiotic stresses such as low temperature, drought, and high salinity, and currently, there are few reports on the DREB gene family in Phalaenopsis spp. In this study, Phalaenopsis aphrodite was used as the reference genome for the identification. In addition, the subcellular localization, bioinformatics, and gene expression pattern of DREB family members under 3 abiotic stress conditions: low temperature stress, drought treatment, and salt stress, and abscisic acid treatment were studied and analysed. Using conserved structural domain analysis and BLAST comparison, 31 DREB family gene members were found from the P. aphrodite genome database. PaDREB transcription factors were divided into 6 subgroups, numbered A1 to A6, according to phylogenetic research. Conserved motif analysis revealed that all of PaDREB transcription factors had a AP2 conserved structural domain. The transcriptome expression pattern study of P. aphrodite revealed that different PaDREBs showed differential tissue expression and most of the genes were highly expressed in the roots. The 4 members of subgroup A1 were isolated and named PaDREB1A, PaDREB1B, PaDREB1C, and PaDREB1D, respectively, and subcellular localization was carried out on them, and the results showed that all of them were localized in the nucleus. Meanwhile, the results of expression analysis of the above 4 members showed that PaDREB1A mainly responded to drought and high salt and ABA treatments; PaDREB1B mainly responded to low temperature and high salt treatments; PaDREB1C mainly responded to drought and ABA treatments; and PaDREB1D mainly responded to low temperature, drought and ABA treatments. The above results suggested that the DREB family of Phalaenopsis had an important role in the response to abiotic stress pathway, and this study provides a theoretical basis for further research on the mechanism of the DREB family's response to abiotic stress in Phalaenopsis.
胡子昂, 罗平, 辛静静, 崔永一. 蝴蝶兰DREB基因家族鉴定及非生物胁迫表达分析[J]. 农业生物技术学报, 2024, 32(8): 1715-1728.
HU Zi-Ang, LUO Ping, XIN Jing-Jing, CUI Yong-Yi. Identification of DREB Gene Family in Phalaenopsis aphrodite and its Expression Analysis Under Abiotic Stresses. 农业生物技术学报, 2024, 32(8): 1715-1728.
[1] 王嘉文. 2022. 蝴蝶兰MADS和AP2基因家族的全基因组鉴定和分析[D]. 硕士学位论文, 南昌大学, 导师: 罗火林, pp. 24-28. (Wang J W.2022. Genome-wide identification and analysis of MADS and AP2 gene families in Phalaenopsis aphrodite[D]. Thesis for M.S., Nanchang University, Suppervisor: Luo H L, pp. 24-28.) [2] Cao S L, Wang Y, Li X T, et al.2020. Characterization of the AP2/ERF transcription factor family and expression profiling of DREB subfamily under cold and osmotic stresses in Ammopiptanthus nanus[J]. Plants (Basel), 9(4): 455-475. [3] Chao Y T, Chen W C, Chen C Y, et al.2018. Chromosome-level assembly, genetic and physical mapping of Phalaenopsis aphrodite genome provides new insights into species adaptation and resources for orchid breeding[J]. Plant Biotechnology Journal, 16(12): 2027-2041. [4] Chao Y T, Yen S H, Yeh J H, et al.2017. Orchidstra 2.0-A transcriptomics resource for the orchid family[J]. Plant & Cell Physiology, 58(1): e9(1-13). [5] Chen C J, Chen H, Zhang Yet al.2020. TBtools: An integrative toolkit developed for interactive analyses of big biological data[J]. Molecular Plant, 13(8): 1194-1202. [6] Donde R, Gupta M K, Gouda G, et al.2019. Computational characterization of structural and functional roles of DREB1A, DREB1B and DREB1C in enhancing cold tolerance in rice plant[J]. Amino Acids, 51(5): 839-853. [7] Dong C, Xi Y, Chen X L, et al.2021. Genome-wide identification of AP2/EREBP in Fragaria vesca and expression pattern analysis of the FvDREB subfamily under drought stress[J]. BMC Plant Biology, 21: 295. [8] Feng K, Hou X L, Xing G M, et al.2020. Advances in AP2/ERF super-family transcription factors in plant[J]. Critical Reviews in Biotechnology, 40(6): 750-776. [9] Gilmour S J, Sebolt A M, Salazar M P, et al.2000. Overexpression of the Arabidopsis CBF3 transcriptional activator mimics multiple biochemical changes associated with cold acclimation[J]. Plant Physiology, 124(4): 1854-1865. [10] Goldberg T, Hecht M, Hamp T, et al.2014. LocTree3 prediction of localization[J]. Nucleic Acids Research, 42(Web Server issue): W350-355. [11] Han Y L, Cai M H, Zhang S Q, et al.2022. Genome-wide identification of AP2/ERF transcription factor family and functional analysis of DcAP2/ERF#96 associated with abiotic stress in Dendrobium catenatum[J]. International Journal of Molecular Sciences, 23(21): 13603. [12] Hu Y R, Jiang L Q, Wang F, et al.2013. Jasmonate regulates the inducer of cbf expression-C-repeat binding factor/DRE binding factor1 cascade and freezing tolerance in Arabidopsis[J]. The Plant Cell, 25(8): 2907-2924. [13] Jaglo-Ottosen K R, Gilmour S J, Zarrka D G, et al.1998. Arabidopsis CBF1 overexpression induces COR genes and enhances freezing tolerance[J]. Science, 280(5360): 104-106. [14] Jin R, Kim B H, Ji C Y, et al.2017. Overexpressing IbCBF3 increases low temperature and drought stress tolerance in transgenic sweet potato[J]. Plant Physiology and Biochemistry, 118: 45-54. [15] Kidokoro S, Watanabe K, Ohori T, et al.2015. Soybean DREB1/CBF-type transcription factors function in heat and drought as well as cold stress-responsive gene expression[J]. The Plant Journal, 81(3): 505-518. [16] Kolachevskaya O O, Omin S N, Arkhipov D V, et al.2019. Auxins in potato: Molecular aspects and emerging roles in tuber formation and stress resistance[J]. Plant Cell Reports, 38(6): 681-698. [17] Li J, Wang Y Q, Yu B, et al.2018. Ectopic expression of StCBF1 and ScCBF1 have different functions in response to freezing and drought stresses in Arabidopsis[J]. Plant Science, 270: 221-233. [18] Liu Q, Kasuga M, Sakuma Y, et al.1998. Two transcription factors, DREB1 and DREB2, with an EREBP/AP2 DNA binding domain separate two cellular signal transduction pathways in drought- and low-temperature-responsive gene expression, respectively, in Arabidopsis[J]. The Plant Cell, 10(8): 1391-1406. [19] Maqsood H, Munir F, Amir R, et al.2022. Genome-wide identification, comprehensive characterization of transcription factors, cis-regulatory elements, protein homology, and protein interaction network of DREB gene family in Solanum lycopersicum[J]. Frontiers in Plant Science, 13: 1031679. [20] Nakano T, Suzuki K, Fujimura T, et al.2006. Genome-wide analysis of the ERF gene family in Arabidopsis and rice[J]. Plant Physiology, 140(2): 411-432. [21] Nicole F, Daniele G, Kerstin N, et al.2007. DRE-1: An evolutionarily conserved F box protein that regulates C. elegans developmental age[J]. Developmental Cell, 12(3):443-455. [22] Niu X, Luo T, Zhao H, et al.2020. Identification of wheat DREB genes and functional characterization of TaDREB3 in response to abiotic stresses[J]. Gene, 740: 144514. [23] Novillo F, Medina J, Salinas J.2007. Arabidopsis CBF1 and CBF3 have a different function than CBF2 in cold acclimation and define different gene classes in the CBF regulon[J]. Proceedings of the National Academy of Sciences of the USA, 104(52): 21002-21007. [24] Reiser L, Subramaniam S, Li Det al.2017. Using the Arabidopsis information resource (TAIR) to find information about Arabidopsis genes[J]. Current Protocols in Bioinformatics, 60(11): 1-45. [25] Sakuma Y, Liu Q, Dubouzet J G, et al.2002. DNA-binding specificity of the ERF/AP2 domain of Arabidopsis DREBs, transcription factors involved in dehydration- and cold-inducible gene expression[J]. Biochemical and Biophysical Research Communications, 290(3): 998-1009. [26] Stockinger E J, Gilmour S J, Thomashow M F.1997. 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[J]. Proceedings of the National Academy of Sciences of the USA, 94(3): 1035-1040. [27] Verma V, Ravindran P, Kumar P P.2016. Plant hormone-mediated regulation of stress responses[J]. BMC Plant Biology, 16: 86. [28] Wilkins M R, Gasteiger E, Bairoch A, et al.1999. Protein identification and analysis tools in the ExPASy server[J]. Methods in Molecular Biology, 112(2): 531-552. [29] Xie J, Chen Y, Cai G, et al.2023. Tree visualization by one table (tvBOT): A web application for visualizing, modifying and annotating phylogenetic trees[J]. Nucleic Acids Research, 51(W1): W587-W592. [30] Yamaguchi K, Shinozaki K.1994. A novel cis-acting element in an Arabidopsis gene is involved in responsiveness to drought, low-temperature, or high-salt stress[J]. The Plant Cell, 6(2): 251-264. [31] Yamaguchi K, Shinozaki K.2005. Organization of cis-acting regulatory elements in osmotic- and cold-stress-responsive promoters[J]. Trends in Plant Science, 10(2): 88-94. [32] Yoshihiro N, Kazuzo N, Zabta K S, et al.2003. Interaction between two cis-acting elements, ABRE and DRE, in ABA-dependent expression of Arabidopsis rd29A gene in response to dehydration and high-salinity stresses[J]. The Plant Journal, 34(2):134-148. [33] Yu C S, Chen Y C, Lu C H, et al.2006. Prediction of protein subcellular localization[J]. Proteins: Structure, Function and Bioinformatics, 64(3): 643-651. [34] Zhu J K.2016. Abiotic stress signaling and responses in plants[J]. Cell, 167(2): 313-324.
