花苜蓿TCP基因家族的鉴定及其在干旱胁迫下的表达模式分析

doi:10.13560/j.cnki.biotech.bull.1985.2024-1205

摘要/Abstract

摘要：

目的 TCP（Teosinte branched 1/Cincinnata/Proliferating cell factor）是调控植物生长发育和胁迫响应的一类转录因子。解析花苜蓿TCP基因家族成员的序列结构和理化性质，了解花苜蓿MruTCP基因在干旱胁迫下的作用，为牧草分子育种提供基因资源。方法基于全基因组水平鉴定花苜蓿TCP基因，对其进行生物信息学分析和干旱胁迫下的表达模式分析。结果在花苜蓿中共鉴定到20个TCP基因，分布在7条染色体上。系统进化分析结果表明，MruTCP蛋白分为3个亚家族：PCF、CIN和CYC/TB1，各类蛋白的Motif种类和排列顺序有所差异，均含有一个共同的保守基序Motif 1，家族成员间的结构较为简单。共线性分析结果表明，花苜蓿TCP家族内存在2个片段重复事件，且与大豆之间的亲缘关系最近。启动子顺式作用元件分析的结果表明，MruTCP家族成员功能较为复杂，在信号传递、响应非生物胁迫、光信号响应、激素调节等方面均发挥作用。干旱胁迫下的转录组数据和RT-qPCR的结果表明，花苜蓿TCP基因家族成员在不同时间点和不同浓度干旱处理下的表达模式不同。其中，MruTCP05和MruTCP09包含了丰富的激素响应元件和胁迫响应元件，并且受干旱胁迫的强烈诱导。结论结合TCP基因家族的生物信息学分析和表达模式分析结果，推测MruTCP05和MruTCP09是调控花苜蓿耐旱性的候选基因。

关键词: 花苜蓿, TCP基因, 干旱胁迫, 转录表达分析

Abstract:

Objective TCP (Teosinte-branched 1/Cincinnata/Proliferating) is a class of transcription factors that regulate plant growth and development and response to stress. Dissecting the sequence structure and physicochemical properties of the TCP gene family members (MruTCP) in Medicago ruthenica will help to understand the role of MruTCP genes under drought stress and provide genetic resources for molecular breeding of forage. Method MruTCP genes were identified based on the whole genome level and analyzed for bioinformatics and expression patterns under drought stress. Result A total of 20 TCP genes were identified in M. ruthenica, and distributed on seven chromosomes. The results of phylogenetic analysis showed that the MruTCP proteins were categorized into three subfamilies: PCF, CIN and CYC/TB1, with differences in the motif types and order of the proteins, but all of them contained a common conserved motif: Motif 1, and the structure of the family members was relatively simple. The results of the covariance analysis indicated that there were two fragment duplication events within the alfalfa TCP family and the closest affinity with soybean. The results of promoter cis-acting element analysis indicated that the MruTCP family members were more complex in function and played roles in signaling, response to abiotic stress, light signaling response, and hormone regulation. Transcriptomic data and qRT-PCR results under drought stress indicated that the expression patterns of MruTCP gene family members differed at different time points and under different concentrations of drought treatments. Among them, MruTCP05 and MruTCP09 contained abundant hormone-responsive elements and stress-responsive elements and were strongly induced by drought stress. Conclusion Combined with the bioinformatics analysis and expression pattern analysis of the TCP gene family, this study hypothesized that MruTCP05 and MruTCP09 are candidate genes for regulating drought tolerance in M. ruthenica.

Key words: Medicago ruthenica, TCP gene, drought stress, transcriptional expression analysis

王苗苗, 赵相龙, 王召明, 刘志鹏, 闫龙凤. 花苜蓿TCP基因家族的鉴定及其在干旱胁迫下的表达模式分析[J]. 生物技术通报, 2025, 41(6): 179-190.

WANG Miao-miao, ZHAO Xiang-long, WANG Zhao-ming, LIU Zhi-peng, YAN Long-feng. Identification of TCP Gene Family in Medicago ruthenica and Their Expression Pattern Analysis under Drought Stress[J]. Biotechnology Bulletin, 2025, 41(6): 179-190.

图/表 11

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基因名称 Gene name	正向引物Forward primer sequence （5'-3'）	反向引物Reverse primer sequence （5'-3'）
MruActin	ATCCAGGCTGTCCTCTCCCT	ACGAAGGATGGCATGTGGGA
MruTCP01	CCACTCCTGCTTCTTTCTCT	GCCGGAAGGTTTTTCTGTTT
MruTCP02	GGAAACGGAAAACAACAGCC	GGAGCTAGGTCACAAATCCG
MruTCP03	GGAAACGGAAAACAACAGCC	GGAGCTAGGTCACAAATCCG
MruTCP05	CACTTCACTCGACCACAAAC	CGGCCTGATTTTGGACAAAT
MruTCP06	CAGATCTGAACCACCTCCTC	TGTGCAGCTAGAGTTTTCCA
MruTCP09	TGTTGGTGGTGGTAATGGAA	TTAGGTTCAGGAATCGTCGG
MruTCP16	AGGAACAAATTTGGGAGGGA	GGTTCATCAGCAGAATCAGC
MruTCP17	ACTCAAAATCCCAACCACCT	CGTTTCTGTTTCAACCGGTT
MruTCP18	ACCGATCCAACCAACTAGTC	CTTTGCGCTGCTAGAGTTTT

基因名称 Gene name	正向引物Forward primer sequence （5'-3'）	反向引物Reverse primer sequence （5'-3'）
MruActin	ATCCAGGCTGTCCTCTCCCT	ACGAAGGATGGCATGTGGGA
MruTCP01	CCACTCCTGCTTCTTTCTCT	GCCGGAAGGTTTTTCTGTTT
MruTCP02	GGAAACGGAAAACAACAGCC	GGAGCTAGGTCACAAATCCG
MruTCP03	GGAAACGGAAAACAACAGCC	GGAGCTAGGTCACAAATCCG
MruTCP05	CACTTCACTCGACCACAAAC	CGGCCTGATTTTGGACAAAT
MruTCP06	CAGATCTGAACCACCTCCTC	TGTGCAGCTAGAGTTTTCCA
MruTCP09	TGTTGGTGGTGGTAATGGAA	TTAGGTTCAGGAATCGTCGG
MruTCP16	AGGAACAAATTTGGGAGGGA	GGTTCATCAGCAGAATCAGC
MruTCP17	ACTCAAAATCCCAACCACCT	CGTTTCTGTTTCAACCGGTT
MruTCP18	ACCGATCCAACCAACTAGTC	CTTTGCGCTGCTAGAGTTTT

基因名称Gene name	基因ID Gene ID	氨基酸数目 Number of amino acids （aa）	分子量 Molecular weight （kD）	理论等电点 pI	不稳定系数Instability index	脂肪系数Aliphatic index	疏水性 Hydrophobicity	亚细胞定位 Subcellular localization
MruTCP01	MruT001875	225	24.52	8.01	56.29	62.98	-0.76	细胞核
MruTCP02	MruT002689	206	22.20	7.02	81.12	65.29	-0.47	细胞核
MruTCP03	MruT005553	342	35.89	4.97	56.45	62.84	-0.364	细胞核
MruTCP04	MruT005664	390	44.27	6.32	55.17	53.31	-0.766	细胞核
MruTCP05	MruT006387	255	27.23	9.51	48.03	69.76	-0.543	细胞核
MruTCP06	MruT006697	509	54.44	6.43	58.12	51.1	-0.863	细胞核
MruTCP07	MruT011216	328	36.21	6.03	45.67	61.89	-0.74	细胞核
MruTCP08	MruT022062	337	37.82	6.1	43.68	56.05	-0.904	细胞核
MruTCP09	MruT028007	418	44.08	6.26	57.08	59.35	-0.609	细胞核
MruTCP10	MruT031885	329	36.89	6.36	48.26	76.38	-0.604	细胞核
MruTCP11	MruT032111	412	46.55	9.35	54.68	57.31	-0.966	细胞核
MruTCP12	MruT033252	233	25.94	10.08	70.54	64.08	-0.682	细胞核
MruTCP13	MruT034692	351	39.72	7.23	48.98	57.75	-0.948	细胞核
MruTCP14	MruT034836	383	43.73	6.71	53.45	52.53	-1.155	细胞核
MruTCP15	MruT034841	383	43.74	6.71	53.45	52.27	-1.154	细胞核
MruTCP16	MruT035144	286	30.94	8.68	51.03	60.42	-0.851	细胞核
MruTCP17	MruT041006	325	35.38	9.27	62.8	75.94	-0.424	细胞核
MruTCP18	MruT043711	439	47.62	7.01	63.2	53.62	-0.893	细胞核
MruTCP19	MruT043927	390	44.87	8.9	50.63	55.23	-1.029	细胞核
MruTCP20	MruT045760	439	47.62	7.01	63.2	53.62	-0.893	细胞核

基因名称Gene name	基因ID Gene ID	氨基酸数目 Number of amino acids （aa）	分子量 Molecular weight （kD）	理论等电点 pI	不稳定系数Instability index	脂肪系数Aliphatic index	疏水性 Hydrophobicity	亚细胞定位 Subcellular localization
MruTCP01	MruT001875	225	24.52	8.01	56.29	62.98	-0.76	细胞核
MruTCP02	MruT002689	206	22.20	7.02	81.12	65.29	-0.47	细胞核
MruTCP03	MruT005553	342	35.89	4.97	56.45	62.84	-0.364	细胞核
MruTCP04	MruT005664	390	44.27	6.32	55.17	53.31	-0.766	细胞核
MruTCP05	MruT006387	255	27.23	9.51	48.03	69.76	-0.543	细胞核
MruTCP06	MruT006697	509	54.44	6.43	58.12	51.1	-0.863	细胞核
MruTCP07	MruT011216	328	36.21	6.03	45.67	61.89	-0.74	细胞核
MruTCP08	MruT022062	337	37.82	6.1	43.68	56.05	-0.904	细胞核
MruTCP09	MruT028007	418	44.08	6.26	57.08	59.35	-0.609	细胞核
MruTCP10	MruT031885	329	36.89	6.36	48.26	76.38	-0.604	细胞核
MruTCP11	MruT032111	412	46.55	9.35	54.68	57.31	-0.966	细胞核
MruTCP12	MruT033252	233	25.94	10.08	70.54	64.08	-0.682	细胞核
MruTCP13	MruT034692	351	39.72	7.23	48.98	57.75	-0.948	细胞核
MruTCP14	MruT034836	383	43.73	6.71	53.45	52.53	-1.155	细胞核
MruTCP15	MruT034841	383	43.74	6.71	53.45	52.27	-1.154	细胞核
MruTCP16	MruT035144	286	30.94	8.68	51.03	60.42	-0.851	细胞核
MruTCP17	MruT041006	325	35.38	9.27	62.8	75.94	-0.424	细胞核
MruTCP18	MruT043711	439	47.62	7.01	63.2	53.62	-0.893	细胞核
MruTCP19	MruT043927	390	44.87	8.9	50.63	55.23	-1.029	细胞核
MruTCP20	MruT045760	439	47.62	7.01	63.2	53.62	-0.893	细胞核

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