巴戟天萜烯合成酶基因家族鉴定、进化及表达模式分析

doi:10.13560/j.cnki.biotech.bull.1985.2025-0465

摘要/Abstract

摘要：

目的探究巴戟天萜烯合成酶（terpene synthases, TPS）基因家族的基本特征、进化及其潜在的功能。方法基于巴戟天全基因组和转录组数据，通过成员鉴定、结构特征、进化关系、基因表达和蛋白互作（protein-protein interaction, PPI）分析，系统解析巴戟天TPS基因家族的结构、进化与表达模式。结果在巴戟天中共鉴定到33个MoTPS基因，不均匀分布在7条染色体上，均为定位在叶绿体的亲水性蛋白。系统发育分析表明MoTPS基因家族成员分为6个亚族，其中TPS-b和TPS-a亚家族中的基因最多。在MoTPS基因家族中存在6对串联复制基因和2对片段复制基因。PPI预测表明MoTPS1、MoTPS32和MoTPS33可能通过与ent-KOD蛋白互作参与赤霉素前体合成，而MoTPS18、MoTPS19、MoTPS20和MotPS21可能与萜类修饰合成的CYP450蛋白互作。顺式作用元件分析表明MoTPS基因家族成员广泛参与光响应、激素响应以及非生物胁迫响应。基因表达相关性分析表明19个MoTPS家族基因与萜类合成途径上游基因呈现显著正相关性。其中，参与环烯醚萜成分合成的关键基因香叶醇合酶MoTPS31与MEP途径的DXS1的表达量呈正相关。结论串联复制和片段复制是MoTPS家族基因进化和扩张的主要动力。MoTPS基因通过调控萜类物质的生物合成广泛参与光暗反应、激素信号转导和非生物胁迫响应。

关键词: 巴戟天, 萜烯合成酶, 基因家族, 生物信息学分析, 表达模式

Abstract:

Objective To explore the basic characteristics, evolution and potential functions of the terpene synthases (TPS) gene family in Morinda officinalis. Method Based on the whole genome and transcriptome data of M. officinalis, the gene structure, evolution and expression pattern of MoTPS were systematically analyzed through screening of gene family members, examination of structural features, construction of evolutionary relationships, gene expression analysis, and protein-protein interaction (PPI) prediction. Result A total of 33 MoTPS genes were identified in M. officinalis, unevenly distributed on 7 chromosomes, all of which were hydrophilic proteins located in chloroplasts. Phylogenetic analysis showed that the MoTPS gene family was divided into six subfamilies, among which the TPS-b and TPS-a subfamilies contained the most genes. There were six pairs of tandem replication genes and two pairs of segmental replication genes in MoTPS gene family. PPI prediction indicated that MoTPS1, MoTPS32, and MoTPS33 participated in the synthesis of gibberellins precursors by interacting with ent-KOD proteins, while MoTPS18, MoTPS19, MoTPS20, and MotPS21 interacted with CYP450 proteins synthesized by terpenoid modification. Cis-acting element analysis showed that the MoTPS gene family was widely involved in light, hormone and abiotic stress responses. The correlation analysis of gene expression showed that 19 MoTPS family genes were significantly positively correlated with upstream genes in terpenoid biosynthesis pathway. Among them, the expression of MoTPS31, which encoded geraniol synthase, a key gene involved in the biosynthesis of iridoid components, was positively correlated with DXS1 in the MEP pathway. Conclusion Tandem duplication and segmental duplication events are the main driving forces for the evolution and expansion of MoTPS family genes. The MoTPS family genes are widely involved in light, hormone signal transduction and abiotic stress responses by regulating the biosynthesis of terpenoids.

Key words: Morinda officinalis, terpene synthase, gene family, bioinformatics analysis, expression pattern

李旭龙, 徐世强, 张龙, 李静宇, 何伟贤, 陈立凯, 王继华. 巴戟天萜烯合成酶基因家族鉴定、进化及表达模式分析[J]. 生物技术通报, doi: 10.13560/j.cnki.biotech.bull.1985.2025-0465.

LI Xu-long, XU Shi-qiang, ZHANG Long, LI Jing-yu, HE Wei-xian, CHEN Li-kai, WANG Ji-hua. Genome-wide Identification， Evolution and Expression Pattern Analysis of Terpene Synthase Gene Family in Morinda officinalis[J]. Biotechnology Bulletin, doi: 10.13560/j.cnki.biotech.bull.1985.2025-0465.

图/表 7

图1 巴戟天、拟南芥和长叶薄荷TPS基因家族系统进化树

Fig. 1 Phylogenetic tree of the TPS gene family in M. officinalis, A. thaliana, and M. longifolia

图2 巴戟天TPS基因家族保守基序（A）、基因结构（B）染色体分布（C）及与拟南芥、咖啡和喜树的共线性分析（D）

Fig. 2 Conserved motifs (A), gene structure (B), chromosomal distribution (C) of TPS gene family in M. officinalis, and collinearity analysis of TPS gene in M. officinalis, A. thaliana, C. canephora and C. acuminata (D)

图3 巴戟天TPS基因家族启动子顺式作用元件预测分布（A）以及激素、光和胁迫响应元件中每种顺式作用元件所占比重（B）

Fig. 3 Prediction of cis-acting element distribution in the promoter of TPS gene family in M. officinalis (A), and the proportion of each cis-acting element in hormone, light response, and stress response elements (B)

图4 巴戟天TPS蛋白互作网络预测分析

Fig. 4 Prediction and analysis of the protein-protein interaction network analysis of TPS proteins in M. officinalis

图5 非生物胁迫（A）和MeJA（B）诱导下巴戟天TPS基因家族表达模式分析

Fig. 5 Analysis of expression pattern of TPS gene family in M. officinalis under abiotic stress (A) and MeJA induction (B)

图6 胁迫和MeJA处理下巴戟天TPS基因RT-qPCR验证*P<0.05， **P<0.01 （compared with CK， Student’s t-test）

Fig. 6 Expression verification of TPS gene by RT-qPCR in M. officinalis under abiotic stress and MeJA treatment

图7 巴戟天TPS家族基因与萜类合成通路结构基因表达量相关性热图（A）以及部分显著相关基因间的相关性网络图（B）

Fig. 7 Heatmap of the expression correlation between TPS family genes in M. officinalis and structure genes of terpenoid synthesis pathway (A), and correlation network diagram among some significantly correlated genes (B)

参考文献 31

[1]	Zhang JH, Xin HL, Xu YM, et al. Morinda officinalis How.-A comprehensive review of traditional uses, phytochemistry and pharmacology [J]. J Ethnopharmacol, 2018, 213: 230-255.
[2]	沈燚, 张奇, 刘梦琴, 等. 巴戟天属植物环烯醚萜类化学成分及生物活性研究进展 [J]. 药学实践杂志, 2020, 38(2): 110-114, 119.
	Shen Y, Zhang Q, Liu MQ, et al. Research on chemical components and biological activities of the iridoids in Morinda genus [J]. J Pharm Pract, 2020, 38(2): 110-114, 119.
[3]	Chen F, Tholl D, Bohlmann J, et al. The family of terpene synthases in plants: a mid-size family of genes for specialized metabolism that is highly diversified throughout the Kingdom [J]. Plant J, 2011, 66(1): 212-229.
[4]	Aubourg S, Lecharny A, Bohlmann J. Genomic analysis of the terpenoid synthase (AtTPS) gene family of Arabidopsis thaliana [J]. Mol Genet Genom, 2002, 267(6): 730-745.
[5]	Zhou F, Pichersky E. The complete functional characterisation of the terpene synthase family in tomato [J]. New Phytol, 2020, 226(5): 1341-1360.
[6]	Martin DM, Aubourg S, Schouwey MB, et al. Functional annotation, genome organization and phylogeny of the grapevine (Vitis vinifera) terpene synthase gene family based on genome assembly, FLcDNA cloning, and enzyme assays [J]. BMC Plant Biol, 2010, 10(1): 226.
[7]	零唯, 覃艳红, 黄鼎, 等. 绞股蓝萜类合成酶(TPS)基因家族鉴定及其在非生物胁迫下的表达分析 [J]. 中国中药杂志, 2023, 48(4): 930-938.
	Ling W, Qin YH, Huang D, et al. Identification of terpene synthase gene family in Gynostemma pentaphyllum and expression pattern analysis under abiotic stresses [J]. China J Chin Mater Med, 2023, 48(4): 930-938.
[8]	Bohlmann J, Meyer-Gauen G, Croteau R. Plant terpenoid synthases: molecular biology and phylogenetic analysis [J]. Proc Natl Acad Sci U S A, 1998, 95(8): 4126-4133.
[9]	Jia QD, Brown R, Köllner TG, et al. Origin and early evolution of the plant terpene synthase family [J]. Proc Natl Acad Sci U S A, 2022, 119(15): 1-9.
[10]	Jiang SY, Jin JJ, Sarojam R, et al. A comprehensive survey on the terpene synthase gene family provides new insight into its evolutionary patterns [J]. Genome Biol Evol, 2019, 11(8): 2078-2098.
[11]	许文杰, 娄千, 陈启桢, 等. 基于全基因组的栀子花香成分合成TPS基因系统分析 [J]. 植物科学学报, 2024, 42(1): 85-95.
	Xu WJ, Lou Q, Chen QZ, et al. Systematic analysis of TPS genes for the synthesis of floral aroma components of Gardenia jasminoides J.Ellis based on the whole genome [J]. Plant Sci J, 2024, 42(1): 85-95.
[12]	贾聪玲, 舒娟, 党静洁, 等. 荆芥萜类合成酶(TPS)基因家族鉴定与分析 [J]. 中国中药杂志, 2023, 48(22): 6039-6050.
	Jia CL, Shu J, Dang JJ, et al. Identification and analysis of terpene synthase (TPS) gene family in Schizonepeta tenuifolia [J]. China J Chin Mater Med, 2023, 48(22): 6039-6050.
[13]	王刚, 崔媛, 李启东, 等. 忍冬TPS基因家族鉴定及其在蚜虫危害下的表达分析 [J]. 中国中药杂志, 2025, 50(8): 2116-2129.
	Wang G, Cui Y, Li QD, et al. Identification of Lonicera japonica TPS gene family and expression analysis under aphid damage [J]. China J Chin Mater Med, 2025, 50(8): 2116-2129.
[14]	周成城, 谢德金, 叶友杰, 等. 巴戟天MoTPS基因及其启动子克隆与分析 [J]. 分子植物育种, 2021, 19(9): 2889-2898.
	Zhou CC, Xie DJ, Ye YJ, et al. Cloning and analysis of MoTPS and its promoter in Morinda officinalis how [J]. Mol Plant Breed, 2021, 19(9): 2889-2898.
[15]	Wang JH, Xu SQ, Mei Y, et al. A high-quality genome assembly of Morinda officinalis, a famous native southern herb in the Lingnan region of Southern China [J]. Hortic Res, 2021, 810.1038: s41438-21-00551-w.
[16]	Chen CJ, Chen H, Zhang Y, et al. TBtools: an integrative toolkit developed for interactive analyses of big biological data [J]. Mol Plant, 2020, 13(8): 1194-1202.
[17]	Cannon SB, Mitra A, Baumgarten A, et al. The roles of segmental and tandem gene duplication in the evolution of large gene families in Arabidopsis thaliana [J]. BMC Plant Biol, 2004, 4(1): 10.
[18]	Chen ZQ, Vining KJ, Qi XW, et al. Genome-wide analysis of terpene synthase gene family in Mentha longifolia and catalytic activity analysis of a single terpene synthase [J]. Genes, 2021, 12(4): 518.
[19]	Chen CJ, Wu Y, Li JW, et al. TBtools-II: a “one for all, all for one” bioinformatics platform for biological big-data mining [J]. Mol Plant, 2023, 16(11): 1733-1742.
[20]	Singh S, Chhatwal H, Pandey A. Deciphering the complexity of terpenoid biosynthesis and its multi-level regulatory mechanism in plants [J]. J Plant Growth Regul, 2024, 43(10): 3320-3336.
[21]	李金燃, 张麒功, 陈丝雨, 等. 栀子TPS基因家族鉴定及与萜类物质代谢的相关性分析 [J]. 沈阳农业大学学报, 2024, 55(1): 66-78.
	Li JR, Zhang QG, Chen SY, et al. Identification of TPS genes family in Gardenia jasminoides and its relationship with terpenoid metabolism [J]. J Shenyang Agric Univ, 2024, 55(1): 66-78.
[22]	Diao S, Zhang YN, Luan QF, et al. Identification of TPS-d subfamily genes and functional characterization of three monoterpene synthases in slash pine [J]. Ind Crops Prod, 2022, 188: 115609.
[23]	Guo J, Zhou XT, Dai KL, et al. Comprehensive analysis of YABBY gene family in foxtail millet (Setaria italica) and functional characterization of SiDL [J]. J Integr Agric, 2022, 21(10): 2876-2887.
[24]	Jin KM, Zhuo RY, Xu D, et al. Genome-wide identification of the expansin gene family and its potential association with drought stress in moso bamboo [J]. Int J Mol Sci, 2020, 21(24): 9491.
[25]	Qiao X, Yin H, Li LT, et al. Different modes of gene duplication show divergent evolutionary patterns and contribute differently to the expansion of gene families involved in important fruit traits in pear (Pyrus bretschneideri) [J]. Front Plant Sci, 2018, 9: 161.
[26]	Zhang Z, Tian YL, Qiao X, et al. Integrated analysis of terpenoid profiles and full-length transcriptome reveals the central pathways of sesquiterpene biosynthesis in Atractylodes chinensis (DC.) koidz [J]. Int J Mol Sci, 2025, 26(3): 1074.
[27]	Valifard M, Mohsenzadeh S, Kholdebarin B, et al. Effect of salt stress on terpenoid biosynthesis in Salvia mirzayanii: from gene to metabolite [J]. J Hortic Sci Biotechnol, 2019, 94(3): 389-399.
[28]	Bergman ME, Dudareva N. Plant specialized metabolism: Diversity of terpene synthases and their products [J]. Curr Opin Plant Biol, 2024, 81: 102607.
[29]	Wang Q, Jiang J, Liang YW, et al. Expansion and functional divergence of terpene synthase genes in angiosperms: a driving force of terpene diversity [J]. Hortic Res, 2025, 12(1): uhae272.
[30]	Spyropoulou EA, Haring MA, Schuurink RC. RNA sequencing on Solanum lycopersicum trichomes identifies transcription factors that activate terpene synthase promoters [J]. BMC Genom, 2014, 15(1): 402.
[31]	Nieuwenhuizen NJ, Chen XY, Wang MY, et al. Natural variation in monoterpene synthesis in kiwifruit: transcriptional regulation of terpene synthases by NAC and ETHYLENE-INSENSITIVE3-like transcription factors [J]. Plant Physiol, 2015, 167(4): 1243-1258.