膜荚黄芪TCP基因家族全基因组分析及其对非生物胁迫的响应

doi:10.3969/j.issn.1004-1524.20241050

浙江农业学报 ›› 2026, Vol. 38 ›› Issue (2): 269-283.DOI: 10.3969/j.issn.1004-1524.20241050

膜荚黄芪TCP基因家族全基因组分析及其对非生物胁迫的响应

马琦洋^a(), 黄雪莲^b^,^*()

a.浙江中医药大学第一临床医学院, 浙江杭州 310053
b.浙江中医药大学基础医学院, 浙江杭州 310053

收稿日期:2024-12-02 出版日期:2026-02-25 发布日期:2026-03-24
作者简介:马琦洋,主要从事中药资源鉴定与草药分子生理研究。E-mail: mqy20242025@163.com
通讯作者: ^*黄雪莲,E-mail: h25872968110xl@126.com
基金资助:
省级创业训练计划(S202310344003X);2023浙江省新苗人才计划(2023R410004)

Genome-wide analysis of the TCP gene family in Astragalus membranaceus (Fisch.) Bunge and its response to abiotic stress

MA Qiyang^a(), HUANG Xuelian^b^,^*()

a. The First Clinical Medical College, Zhejiang Chinese Medical University, Hangzhou 310053, China
b. School of Basic Medical Sciences, Zhejiang Chinese Medical University, Hangzhou 310053, China

Received:2024-12-02 Online:2026-02-25 Published:2026-03-24

摘要/Abstract

摘要：

本研究旨在鉴定和分析膜荚黄芪TCP基因家族,并揭示其对非生物胁迫的响应,为研究膜荚黄芪TCP基因在非生物胁迫中的功能提供理论依据。通过生物信息学方法对膜荚黄芪TCP基因及其编码的氨基酸序列进行系统分析,利用RNA-seq技术研究膜荚黄芪TCP基因在不同组织(根、茎和叶)中的表达模式,并通过实时荧光定量PCR(RT-qPCR)验证了5个代表性AmTCP基因在非生物胁迫(高温、干旱、盐胁迫)条件下的表达谱。结果表明,共鉴定出24个TCP基因且不均匀分布于膜荚黄芪的8条染色体上。24个AmTCP基因可分为Ⅰ类(PCF)和Ⅱ类(CYC/TB1和CIN)。同一亚家族蛋白具有高度相似的基序组成,大多数AmTCP基因缺乏内含子。片段重复在膜荚黄芪TCP基因家族进化中发挥了重要作用。此外,启动子区域分析发现多个光响应元件和激素响应元件。组织表达分析显示,AmTCP基因具有显著的组织表达特异性。在非生物胁迫下,5个代表性AmTCP基因的表达水平存在显著差异,其中AmTCP18和AmTCP19基因在多种胁迫条件下均表现出较高的表达水平。综上所述,AmTCP18和AmTCP19可能在膜荚黄芪对非生物胁迫的响应中发挥关键作用,本研究为膜荚黄芪TCP基因家族的进一步功能和进化研究奠定了基础。

关键词: 膜荚黄芪, TCP基因家族, 全基因组分析, 非生物胁迫

Abstract:

This study aimed to identify and characterize the TCP gene family in Astragalus membranaceus(Fisch.) Bunge and elucidate its role in abiotic stress responses, and provide a theoretical basis for investigating the functional role of TCP genes in Astragalus membranaceus(Fisch.) Bunge under abiotic stress conditions. A systematic analysis of the TCP genes in Astragalus membranaceus(Fisch.) Bunge and their encoded amino acid sequences was conducted using bioinformatics methods. RNA-seq technology was employed to investigate the expression patterns of TCP genes in different tissues (roots, stems, and leaves) of Astragalus membranaceus(Fisch.) Bunge. Additionally, the expression profiles of five representative AmTCP genes under abiotic stress conditions (high temperature, drought, and salt stress) were validated through real-time quantitative PCR (RT-qPCR). The results showed that a total of 24 TCP genes were identified and unevenly distributed on 8 chromosomes of Astragalus membranaceus(Fisch.) Bunge. The 24 AmTCP genes can be classified into Class Ⅰ (PCF) and Class Ⅱ (CYC/TB1 and CIN). Proteins within the same subfamily had highly similar motif compositions, and most AmTCP genes lack introns. Fragment duplication played an important role in the evolution of the TCP gene family in Astragalus membranaceus(Fisch.) Bunge. In addition, analysis of the promoter region revealed multiple light-responsive elements and hormone-responsive elements. Tissue expression analysis showed that the AmTCP genes had significant tissue expression specificity. Under abiotic stress, there were significant differences in the expression levels of five representative AmTCP genes, among which AmTCP18 and AmTCP19 genes showed higher expression levels under various stress conditions. In summary, AmTCP18 and AmTCP19 may play a key role in the response of Astragalus membranaceus(Fisch.) Bunge to abiotic stress. This study lays the foundation for further functional and evolutionary research on the TCP gene family of Astragalus membranaceus(Fisch.) Bunge.

Key words: Astragalus membranaceus (Fisch.) Bunge, TCP gene family, genome-wide analysis, abiotic stress

中图分类号:

S567

马琦洋, 黄雪莲. 膜荚黄芪TCP基因家族全基因组分析及其对非生物胁迫的响应[J]. 浙江农业学报, 2026, 38(2): 269-283.

MA Qiyang, HUANG Xuelian. Genome-wide analysis of the TCP gene family in Astragalus membranaceus (Fisch.) Bunge and its response to abiotic stress[J]. Acta Agriculturae Zhejiangensis, 2026, 38(2): 269-283.

图/表 11

表1 RT-qPCR引物

Table 1 Primers for RT-qPCR

基因 Gene	正向引物(5'→3') Forward primer (5'→3')	反向引物(5'→3') Reverse primer (5'→3')
Actin	CTCTCTCAGCACCTTCGAGCAG	TCCACATACAACCGCTCCACTG
AmTCP8	CCAGGCGGAACCGAGCATTATTG	TCGCTTCCTGCTGCTGATGATGA
AmTCP16	AACGGCGATGATGGCGATGATG	TGCTGCTGCTGCTGCTGATG
AmTCP18	GCGGCAGCGGCAACTATAACAA	GCTCGGAATATGATGATGATGCGGAAT
AmTCP19	AGCAGCAGCAGCAGCAGAAC	CGTTGTTGCCGGTATCCAGGTT
AmTCP23	AGCCGCACCAACTGGAACATTAC	TCTGATGCTGATGCTGGTTCTGATG

表2 24个AmTCP基因及其编码蛋白的理化性质

Table 2 Physicochemical characteristics of 24 AmTCP genes and their encoded proteins

蛋白名 Protein name	基因名 Gene name	氨基酸数量 Amino acid quantity	相对分子质量/ku Relative molecular weight/ku	等电点 Isoelectric point	不稳定系数 Instability index	脂肪指数 Aliphatic index	亲水性 Hydrophilicity
AmTCP1	Amem01G08650.1	334	36.25	9.62	61.72	72.78	-0.355
AmTCP2	Amem01G26990.1	353	37.44	5.99	52.15	67.73	-0.460
AmTCP3	Amem01G29700.1	412	44.15	7.97	67.32	58.33	-0.720
AmTCP4	Amem01G30870.1	349	39.08	8.44	53.65	55.62	-0.756
AmTCP5	Amem01G31480.1	342	39.30	9.16	57.79	55.85	-1.099
AmTCP6	Amem01G39660.1	329	35.68	6.21	43.87	57.63	-0.681
AmTCP7	Amem02G01750.1	261	27.81	9.56	48.82	70.73	-0.475
AmTCP8	Amem02G07350.1	233	25.13	9.10	52.77	70.52	-0.566
AmTCP9	Amem02G13390.1	203	21.44	8.44	61.98	68.82	-0.265
AmTCP10	Amem02G30910.1	344	36.12	5.46	54.31	65.00	-0.358
AmTCP11	Amem02G31020.1	344	36.13	5.46	53.03	65.00	-0.358
AmTCP12	Amem03G33990.1	375	41.03	6.90	66.54	52.88	-0.784
AmTCP13	Amem03G35070.1	430	47.97	7.42	47.37	59.35	-0.770
AmTCP14	Amem04G15690.1	417	44.13	6.90	56.30	57.15	-0.661
AmTCP15	Amem05G34130.3	450	49.85	7.21	52.74	52.27	-0.986
AmTCP16	Amem06G00490.1	494	52.19	6.15	62.09	50.63	-0.856
AmTCP17	Amem06G06270.2	355	39.83	8.56	58.10	55.55	-1.028
AmTCP18	Amem06G19960.1	177	20.64	10.50	49.17	69.44	-0.921
AmTCP19	Amem06G25360.1	348	37.92	5.67	57.14	59.17	-0.719
AmTCP20	Amem07G03630.1	358	40.03	8.28	49.00	71.84	-0.665
AmTCP21	Amem07G04880.1	391	44.30	9.12	51.58	66.14	-0.874
AmTCP22	Amem09G02360.1	358	40.20	8.26	54.21	55.89	-0.885
AmTCP23	Amem09G03560.1	322	35.47	9.62	52.23	58.79	-0.730
AmTCP24	Amem09G20040.1	362	40.59	8.87	44.80	69.53	-0.735

图1 膜荚黄芪和拟南芥TCP基因家族的系统发育分析

Fig.1 Phylogenetic analysis of the TCP gene family in Astragalus membranaceus(Fisch.) Bunge and Arabidopsis thaliana

图2 AmTCP蛋白多序列比对

Fig.2 Multi sequence alignment of AmTCP protein

图3 AmTCP基因家族进化树(A)、保守基序(B)和基因结构(C)分析

Fig.3 Analysis of the AmTCP gene family evolutionary tree (A), conserved motifs (B) and gene structure (C)

图4 AmTCP基因家族的染色体定位

Fig.4 Chromosomal localisation of the AmTCP gene family

图5 AmTCP基因的共线性内环的绿线代表每条染色体的基因密度,共线性基因用浅灰色线表示,AmTCP的种内共线性基因对用红色曲线表示,字体也标为红色。

Fig.5 Colinearity of the AmTCP gene The green line in the inner ring represents the gene density per chromosome, co-lined genes are indicated by light gray lines, and intraspecific co-lined gene pairs of AmTCP are labeled with red curves and the font is also labeled in red.

图6 AmTCP基因与4种代表性植物的共线性

Fig.6 Colinearity between AmTCP gene and four representative plants

图7 AmTCP基因启动子的顺式作用元件

Fig.7 Cis-acting elements of the AmTCP gene promoter

图8 AmTCP基因在不同组织的表达颜色条代表标准化表达水平(log2转换的倍数变化)。红色和蓝色表示上调和下调的基因,白色表示无表达。右边蓝色、绿色和紫色条带分别表示PCF、CYC/TB1和CIN亚家族。

Fig.8 AmTCP gene expression in different tissues Color bars represent normalised expression levels (log2 converted fold change). Red and blue indicate up- and down-regulated genes, and white indicates no expression. The blue, green and purple bands on the right represent the PCF, CYC/TB1 and CIN subfamilies.

图9 AmTCP基因在非生物胁迫下的表达高温(40 ℃),干旱(15% PEG),盐(10% NaCl)。“*”表示不同处理间在0.05水平差异显著,“**”表示不同处理间在0.01水平差异显著,“***”表示不同处理间在0.000 5水平差异显著,“****”表示不同处理间在0.000 1水平差异显著。

Fig.9 Expression of AmTCP genes under abiotic stress High temperature (40 ℃), drought (15% PEG), salt (10% NaCl). “*” indicates a significant difference at the 0.05 level between different treatments, “**” indicates a significant difference at the 0.01 level between different treatments, “***” indicates a significant difference at the 0.000 5 level between different treatments, “****” indicates a significant difference at the 0.000 1 level between different treatments.

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膜荚黄芪TCP基因家族全基因组分析及其对非生物胁迫的响应

Genome-wide analysis of the TCP gene family in Astragalus membranaceus (Fisch.) Bunge and its response to abiotic stress

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