Identification and Expression Analysis of Trehalose-6- Phosphate Synthase Family in Casuarina equisetifolia
ZHANG Gui-Hua1,*, YUE Chun-Lei1, LI He-Peng1, ZHANG Xiao-Mian1, YU Zhen-Ming2
1 Zhejiang Academy of Forestry, Hangzhou 310023, China; 2 School of Pharmaceutical Sciences, Zhejiang Chinese Medical University, Hangzhou 311402, China
Abstract：Trehalose-6-phosphate synthase (TPS) is the crucial rate-limiting enzyme for the trehalose biosynthesis, and plays a significant role in the stress responsiveness. To investigate the function of TPS regarding growth and development, stress response in Casuarina equisetifolia, physicochemical features, chromosomal localization, phylogenetic tree, conserved motifs, gene structure, cis-regulatory elements and collinearity were analyzed in this study. Their expression patterns in different tissues, and exposed to drought, salt stress and abscisic acid (ABA) treatment were determined. Results indicated that 7 CeTPS genes were randomly distributed in 4 chromosomes, encoding 826~998 amino acids that were hydrophilic. The CeTPS proteins were divided into 2 subfamilies through phylogenetic analysis, showing relatively close to dicotyledonous plants, and harbored gene duplications. The promoter regions of CeTPS genes were rich in cis-acting elements involved in hormonal responsiveness, growth and development, and stress responsiveness. CeTPS genes showed tissue-specific expression, and were differentially expressed under drought, salt stress, and ABA treatment. Especially, CeTPS4 might play an essential role in stress responsiveness. The results lay a foundation for in-depth study of the CeTPS genes associated with abiotic stress responsiveness.
[1] 杜姣林, 蔺新兰, 马豫皖, 等. 2023. 植物海藻糖-6-磷酸合成酶基因研究进展[J]. 植物科学学报, 41(3): 411-420. (Du J L, Lin X L, Ma Y W, et al.2023. Research progress in plant trehalose-6-phosphate synthase genes[J]. Plant Science Journal, 41(3): 411-420.) [2] 孙梦利, 徐子健, 王世豪, 等. 2018. 西瓜TPS家族基因的鉴定、分类与分析[J]. 分子植物育种, 16(9): 2778-2785. (Sun M L, Xu Z J, Wang S H, et al.2018. Identification, classification and analysis of TPS family genes in Citrullus lanatus[J]. Molecular Plant Breeding. 16(9): 2778-2785.) [3] 杨永超, 张海斐, 杨小振, 等. 2017. 甜瓜海藻糖-6-磷酸合成酶基因鉴定及表达[J]. 西北植物学报, 37(6): 1066-1072. (Yang Y C, Zhang H F, Yang X Z, et al.2017. Expression analysis and genome-wide identification of trehalose-6-phosphate synthase gene family in melon (Cucumis melo L.)[J]. Acta Botanica Boreali Occidentalia Sinica. 37(6): 1066-1072.) [4] 郑飞雄, 陈俊宇, 江林琪, 等. 2024. 掌叶覆盆子4CL基因家族鉴定及表达分析[J]. 农业生物技术学报, 32(2): 311-321. (Zheng F X, Chen J Y, Jiang L Q, et al.2024. Identification and expression analysis of 4CL gene family in Rubus chingii[J]. Journal of Agricultural Biotechnology, 32(2): 311-321.) [5] Ai D, Wang Y J, Wei Y C, et al.2022. Comprehensive identification and expression analyses of the SnRK gene family in Casuarina equisetifolia in response to salt stress[J]. BMC Plant Biology, 22(1): 572. [6] Chen C, Chen H, Zhang Y, et al.2020. TBtools: An integrative toolkit developed for interactive analyses of big biological data[J]. Molecular Plant, 13(8): 1194-1202. [7] Chen J H, Fan J P, Liu W D, et al.2022. Trehalose-6-phosphate synthase influences polysaccharide synthesis and cell wall components in Ganoderma lucidum[J]. Journal of Basic Microbiology, 62(11): 1337-1345. [8] Fichtner F, Olas J J, Feil R, et al.2020. Functional features of trehalose-6-phosphate synthase1, an essential enzyme in Arabidopsis[J]. Plant Cell, 32(6): 1949-1972. [9] Gao Y H, Yang X Y, Yang X, et al.2021. Characterization and expression pattern of the trehalose-6-phosphate synthase and trehalose-6-phosphate phosphatase gene families in Populus[J]. International Journal of Biological Macromolecules, 187: 9-23. [10] Göbel M, Fichtner F.2023. Functions of sucrose and trehalose 6-phosphate in controlling plant development[J]. Journal of Plant Physiology, 291: 154140. [11] Lin M F, Jia R H, Li J C, et al.2018. Evolution and expression patterns of the trehalose-6-phosphate synthase gene family in drumstick tree (Moringa oleifera Lam.)[J]. Planta, 248(4): 999-1015. [12] Livak K J, Schmittgen T D.2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-∆∆CTmethod[J]. Methods, 25(4): 402-408. [13] Lu S, Wang J, Chitsaz F, et al.2020. CDD/SPARCLE: The conserved domain database in 2020[J]. Nucleic Acids Research, 48(1): 265-268. [14] Morabito C, Secchi F, Schubert A.2021. Grapevine TPS (trehalose-6-phosphate synthase) family genes are differentially regulated during development, upon sugar treatment and drought stress[J]. Plant Physiology and Biochemistry, 164: 54-62. [15] Potter S C, Luciani A, Eddy S R, et al.2018. HMMER web server: 2018 update[J]. Nucleic Acids Research, 46(1): 200-204. [16] Raza A, Bhardwaj S, Rahman M A, et al.2024. Trehalose: A sugar molecule involved in temperature stress management in plants[J]. The Crop Journal, 12(1): 1-16. [17] Reichelt N, Korte A, Krischke M, et al.2023. Natural variation of warm temperature-induced raffinose accumulation identifies trehalose-6-phosphate synthase 1 as a modulator of thermotolerance[J]. Plant Cell and Environment, 46(11): 3392-3404. [18] Tamura K, Stecher G, Kumar S.2021. MEGA11: Molecular evolutionary genetics analysis version 11[J]. Molecular Biology and Evolution, 38(7): 3022-3027. [19] Thumuluri V, Almagro Armenteros J J, Johansen A R, et al.2022. DeepLoc 2.0: Multi-label subcellular localization prediction using protein language models[J]. Nucleic Acids Research, 50(1): 228-234. [20] Vandesteene L, López-Galvis L, Vanneste K, et al.2012. Expansive evolution of the trehalose-6-phosphate phosphatase gene family in Arabidopsis[J]. Plant Physiology, 160(2): 884-896. [21] Vishal B, Krishnamurthy P, Kumar P P.2024. Arabidopsis class II TPS controls root development and confers salt stress tolerance through enhanced hydrophobic barrier deposition[J]. Plant Cell Reports, 43(5): 115. [22] Wang Y P, Tang H B, Debarry J D, et al.2012. MCScanX: A toolkit for detection and evolutionary analysis of gene synteny and collinearity[J]. Nucleic Acids Research, 40(7): e49. [23] Wei X R, Ling W, Ma Y W, et al.2022. Genome-wide analysis of the trehalose-6-phosphate synthase gene family in rose (Rosa chinensis) and differential expression under heat stress[J]. Horticulturae, 8(5): 429. [24] Xu X Y, Chen L R, Luo Y C, et al.2022. Discovery of cyclic diarylheptanoids as inhibitors against influenza a virus from the roots of Casuarina equisetifolia[J]. Journal of Natural Products, 85(9): 2142-2148. [25] Xu Z T, Zhang G H, Chen J Y, et al.2024. Role of Rubus chingii BBX gene family in anthocyanin accumulation during fruit ripening[J]. Frontiers in Plant Science, 15, 1427359. [26] Yang Y J, Ma K F, Zhang T X, et al.2020. Characteristics and expression analyses of trehalose-6-phosphate synthase family in Prunus mume reveal genes involved in trehalose biosynthesis and drought response[J]. Biomolecules, 10(10): 1358. [27] Ye G F, Zhang H X, Chen B H, et al.2019. De novo genome assembly of the stress tolerant forest species Casuarina equisetifolia provides insight into secondary growth[J]. Plant Journal, 97(4): 779-794. [28] Yuan G P, Liu J P, An G L, et al.2021. Genome-wide identification and characterization of the trehalose-6-phosphate synthetase (TPS) gene family in watermelon (Citrullus lanatus) and their transcriptional responses to salt stress[J]. International Journal of Molecular Sciences, 23(1): 276. [29] Yuan P, Zhou G L, Yu M Z, et al.2024. Trehalose-6-phosphate synthase 8 increases photosynthesis and seed yield in Brassica napus[J]. Plant Journal, 118(2): 437-456. [30] Zang B S, Li H W, Li W J, et al.2011. Analysis of trehalose-6-phosphate synthase (TPS) gene family suggests the formation of TPS complexes in rice[J]. Plant Molecular Biology, 76(6): 507-522. [31] Zhang S K, He C M, Wei L, et al.2023a. Transcriptome and metabolome analysis reveals key genes and secondary metabolites of Casuarina equisetifolia ssp. incana in response to drought stress[J]. BMC Plant Biology, 23(1): 200. [32] Zhang S K, Wang G B, Yu W W, et al.2024. Multi-omics analyses reveal the mechanisms underlying the responses of Casuarina equisetifolia ssp. incana to seawater atomization and encroachment stress[J]. BMC Plant Biology, 24(1): 854. [33] Zhang Y, Wei Y C, Meng J X, et al.2023b. Chromosome-scale de novo genome assembly and annotation of three representative Casuarina species: C. equisetifolia, C. glauca, and C. cunninghamiana[J]. Plant Journal, 114(6): 1490-1505. [34] Zhao X H, Qi G N, Liu J H, et al.2024. Genome-wide identification of WRKY transcription factors in Casuarina equisetifolia and the function analysis of CeqWRKY11 in response to NaCl/NaHCO3 stresses[J]. BMC Plant Biology, 24(1): 376. [35] Zhong C, He Z H, Liu Y, et al.2024. Genome-wide identification of TPS and TPP genes in cultivated peanut (Arachis hypogaea) and functional characterization of AhTPS9 in response to cold stress[J]. Frontiers in Plant Science, 14: 1343402. [36] Zhou W Y, Sheng C, Dossou S S K, et al.2023. Genome-wide identification of TPS genes in sesame and analysis of their expression in response to abiotic stresses[J]. Oil Crop Science, 8(2): 81-88.
