白栎TCP基因家族的鉴定及功能分析

doi:10.13560/j.cnki.biotech.bull.1985.2025-0818

摘要/Abstract

摘要：

目的 TCP（teosinte branched 1/cincinnata/proliferating cell factor）是植物特有的转录因子，在植物整个生长发育过程中都发挥着重要作用。通过鉴定白栎TCP基因家族，研究QfTCP基因在白栎分枝发育中的表达模式，验证关键基因QfTCP22的功能，为白栎株型改良提供分子基础。方法基于白栎基因组数据，通过生物信息学方法对白栎TCP基因家族进行鉴定，并分析该家族成员的理化性质、染色体分布、系统进化、基因结构、顺式作用元件和基因共线性。结合转录组数据和RT-qPCR对QfTCP基因家族成员在腋芽发育中的表达模式进行分析，并对QfTCP22进行异源过表达拟南芥验证基因功能。结果在白栎中共鉴定23个TCP基因，分为PCE、CIN和CYC/TB1 3个亚家族，不均匀地分布在10条染色体上，其中，第10染色体含有的成员数量最多。所有QfTCP成员都含有一个共同的保守基序Motif 1，基因结构较为简单。共线性分析发现10对QfTCP基因存在共线性关系。顺式作用元件分析发现QfTCP基因的功能较为复杂，在光信号响应、激素调控和逆境胁迫等方面均发挥作用。腋芽发育转录组数据和RT-qPCR结果表明，在腋芽发育中，CYC/TB1亚族的QfTCP1和QfTCP22均呈显著下调趋势，在拟南芥brc1突变体中过表达QfTCP22，能显著减少其分枝数量。结论鉴定白栎TCP家族成员，揭示QfTCP22是植物分枝发育中的抑制因子。

关键词: 白栎, TCP基因, 分枝发育, 生物信息学, 表达模式分析

Abstract:

Objective TCP (teosinte branched 1/cincinnata/proliferating cell factor) is a plant-specific transcription factor that plays a significant role in the growth and development of plants. By identifying the TCP gene family in Quercus fabri, this study aims to investigate the expression patterns of QfTCP genes in the branching development, validate the function of the key gene QfTCP22, and provide a molecular basis for the improvement of plant architecture in Q. fabri. Method Bioinformatics methods were used to identify the TCP gene family based on the genomic data of Q. fabri, and to analyze the physicochemical properties, chromosomal distribution, phylogenetic relationships, gene structures, cis-acting elements, and gene collinearity of the family members. RT-qPCR combined with transcriptome data was to examine the expression patterns of QfTCP gene family members during axillary bud development, while the function of QfTCP22 was verified through heterologous overexpression in Arabidopsis. Result A total of 23 TCP genes were identified in Q. fabri, classified into three subfamilies: PCE, CIN, and CYC/TB1. These genes were unevenly distributed across 10 chromosomes, with chromosome 10 containing the highest number of members. All QfTCP members possessed a common conserved motif Motif 1, and presented relatively simple gene structures. Collinearity analysis revealed that 10 pairs of QfTCP genes had collinear relationships. Analysis of cis-acting elements indicated that the functions of QfTCP genes were relatively complex, involving light signal response, hormone regulation, and stress adversity. Transcriptomic data of axillary bud development and RT-qPCR results showed that QfTCP1 and QfTCP22 of the CYC/TB1 subfamily showed a significant downregulation trend during axillary bud development. Furthermore, the overexpression of the QfTCP22 gene in the Arabidopsis brc1 mutant significantly reduced the number of branches. Conclusion Members of the Q. fabriTCP gene family was identified, revealing that QfTCP22 acts as a suppressor in plant branch development.

Key words: Quercus fabri, TCP gene, branch development, bioinformatics, analysis of expression pattern

熊仕发, 陈益存, 吴立文, 施翔, 张盛剿, 彭方有, 陈涛梅, 汪阳东. 白栎TCP基因家族的鉴定及功能分析[J]. 生物技术通报, doi: 10.13560/j.cnki.biotech.bull.1985.2025-0818.

XIONG Shi-fa, CHEN Yi-cun, WU Li-wen, SHI Xiang, ZHANG Sheng-jiao, PENG Fang-you, CHEN Tao-mei, WANG Yang-dong. Identification and Functional Analysis of the TCP Gene Family in Quercus fabri[J]. Biotechnology Bulletin, doi: 10.13560/j.cnki.biotech.bull.1985.2025-0818.

图/表 8

表1 引物列表

Table 1 Primer list

引物名称 Primer name	序列 Sequence （5′‒3′）	用途 Usage
QfTCP1-F	TCACACTGGGCTCCTCTTTT	RT-qPCR
QfTCP1-R	TTTGGACCACCATTGCTTTT	RT-qPCR
QfTCP22-F	TTCCAACAAAAATCACATCA	RT-qPCR
QfTCP22-R	TTCAAGAGTAACCATTCAACG	RT-qPCR
QfTCP22-OE-F	TCAGCAGTCGAAGAGC	过表达
QfTCP22-OE-R	TTAGCGTGTGAAGAGC	过表达
QfActin-F	GATTCTGGTGATGGTGTGAGC	内参
QfActin-R	ATGAGAGATGGCTGGAAGAGT	内参

图1 白栎TCP家族成员在染色体上的定位

Fig. 1 Chromosomal distribution of TCP genes in Q. fabri

图2 白栎、蒙古栎、栓皮栎、拟南芥TCP蛋白的系统发育进化树

Fig. 2 Phylogenetic tree of TCP proteins from Q. fabri, Q. mongolica, Q. variabilis and A. thaliana

图3 白栎TCP蛋白的系统进化和基因结构

Fig. 3 Phylogenetic relationships and gene structure of TCP protein in Q. fabri

图4 白栎TCP基因家族物种内共线性分析

Fig. 4 Synteny analysis of TCP gene family in Q. fabri

图5 白栎TCP基因家族启动子顺式作用元件分析

Fig. 5 Analysis of cis-acting elements in the promoter region of the TCP gene family in Q. fabri

图6 白栎TCP基因家族在腋芽发育中的表达热图（A）及部分成员荧光定量PCR验证（B）* P<0.05， ** P<0.01. The same below

Fig. 6 Heatmap of TCP gene family expression in axillary bud development of Q. fabri (A) and RT-qPCR validation of some members (B)

图7 过表达QfTCP22拟南芥植株的表型及RT-qPCRA：brc1突变体和QfTCP22回补brc1突变体的植株表型；B：PCR鉴定转基因阳性植株（M：DL2000 marker；WT：阴性对照；OE1‒OE3：转基因植株）；C：brc1突变体和QfTCP22回补brc1突变体植株的分枝数及RT-qPCR

Fig. 7 Plant phenotype and RT-qPCR of Arabidopsis plants overexpressing QfTCP22A: Phenotypes of the brc1 mutant and QfTCP22-complemented brc1 mutant plants. B: PCR identification of transgenic positive plants (M: DL2000 marker; WT: negative control; OE1-OE3: transgenic lines). C: Branch number of the brc1 mutant and QfTCP22-complemented brc1 mutant plants and RT-qPCR

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