‘红阳’猕猴桃TCP基因家族鉴定及其在果实中的表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2024-0820

摘要/Abstract

摘要：

目的基于全基因组鉴定‘红阳’猕猴桃TCP基因（AcTCPs）家族成员，分析各成员的分类进化关系，研究其组织、果实发育过程和激素处理的表达特性，为深入揭示猕猴桃果实中TCP基因的功能奠定基础。方法利用生物信息学方法分析‘红阳’猕猴桃TCP转录因子的理化性质、基因结构、保守基序、顺式作用元件及物种间共线性关系等。利用转录组数据和RT-qPCR分析‘红阳’猕猴桃中AcTCPs在果实不同发育时期、不同组织中和激素处理下的表达模式。结果在‘红阳’猕猴桃中鉴定出分布于19条染色体上的40个含有完整TCP结构域的AcTCPs基因，分别命名为AcTCP1-AcTCP40。系统发育分析、保守基序和基因结构分析将它们分为PCF（21）、CIN（12）和CYC/TB1（7），且全基因复制或片段复制在‘红阳’猕猴桃TCP基因家族的扩展中发挥了重要作用。组织特异性分析表明，6个候选基因在雄花和果实中高表达，果实不同发育时期RT-qRCR结果显示，AcTCP1在果实发育后期高表达，AcTCP11、AcTCP1、AcTCP35、AcTCP10和AcTCP32在果实发育前期高表达，且表达模式与转录组结果一致。对果实进行外源激素处理结果表明，脱落酸（ABA）和乙烯（ET）处理下调了6个候选AcTCPs的表达；赤霉素（GA₃）处理初期上调了AcTCP1、AcTCP35和AcTCP32的表达，氯吡苯脲（CPPU）处理初期上调了AcTCP35和AcTCP32的表达，其余AcTCPs的表达均被GA₃和CPPU下调。结论从‘红阳’猕猴桃全基因组中系统鉴定出40个TCP基因，6个候选基因在果实和雄花中高表达。在果实中AcTCP35、AcTCP32、AcTCP1、AcTCP10、AcTCP11和AcTCP20的表达与果实发育密切相关且受到外源激素（ABA、ET、GA₃和CPPU）的调控。

关键词: ‘红阳’猕猴桃, TCP转录因子, 全基因组鉴定, 生物信息学, 组织差异, 表达模式分析, 激素处理

Abstract:

Objective The members of TEOSINTE-BRANCHED1/CYCLOIDEA/PCF in kiwifruit ‘Hongyang’ (Actinidia chinensis var. Hongyang) (AcTCPs) gene family were identified based on whole-genome. Taxonomic and evolutionary relationships of the members were analyzed, and the expression profiles of the AcTCP genes in tissue-specific expression, fruit development process, and hormones treatments were explored. These findings provide a basis for further exploring the function of the AcTCP gene in kiwifruit. Method Bioinformatics methods were used to analyze the physicochemical properties, gene structure, conserved motifs, cis-acting elements and inter-species covariance of TCP transcription factors in kiwifruit ‘Hongyang’ were analyzed. RNA-seq and RT-qPCR were used to investigate the expression profiles of AcTCPs in different tissues, fruit development and in response to hormonal stimuli. Result A total of 40 AcTCP genes with complete TCP domains were identified on19 chromosomes in kiwifruit ‘Hongyang’ and named AcTCP1-AcTCP40. Using phylogenetic analyses, conserved motif, and gene structure analyses, the family members were divided into PCF (21), CIN (12) and CYC/TB1 (7). Synteny analysis showed that genome-wide or segment-based replication played an important role in the expansion of the kiwifruit ‘Hongyang’ TCP gene family. Analysis of tissue-specificity showed that the six candidate genes expressed significantly high in male flowers and fruits. The results of RT-qPCR experiments at different stages of fruit development found that AcTCP1 showed elevated expressions at later stages of fruit development, while AcTCP11, AcTCP35, AcTCP10, and AcTCP32 demonstrated high expression during the initial stages of fruit development, and the expression pattern was consistent with the transcriptomic results. In fruit, abscisic acid (ABA) and ethylene (ET) treatments to the fruits down-regulated six candidate AcTCPs, gibberellin (GA₃) treatment up-regulated the expressions of AcTCP1, AcTCP35 and AcTCP32, and the forchlorfenuron (CPPU) treatment up-regulated the expressions of AcTCP1, AcTCP35 and AcTCP32 in initial stages, whereas GA₃ and CPPU decreased the expressions of the others AcTCPs. Conclusion A total of 40 TCP members from kiwifruit ‘Hongyang’ were systematically identified. The expression patterns of six candidate genes highly express in the fruit and male flowers. The expressions of AcTCP35, AcTCP32, AcTCP1, AcTCP10, AcTCP11 and AcTCP20 in the fruit are involved in kiwifruit development and are regulated by various exogenous hormones (ABA, ET, GA₃, and CPPU).

Key words: Actinidia chinensis var. Hongyang, TCP transcription factors, genome-wide identification, bioinformatics, organizational differences, expression pattern analysis, hormone treatment

宋姝熠, 蒋开秀, 刘欢艳, 黄亚成, 刘林娅. ‘红阳’猕猴桃TCP基因家族鉴定及其在果实中的表达分析[J]. 生物技术通报, 2025, 41(3): 190-201.

SONG Shu-yi, JIANG Kai-xiu, LIU Huan-yan, HUANG Ya-cheng, LIU Lin-ya. Identification of the TCP Gene Family in Actinidia chinensis var. Hongyang and Their Expression Analysis in Fruit[J]. Biotechnology Bulletin, 2025, 41(3): 190-201.

图/表 8

图1 猕猴桃（Ac）、拟南芥（At）、草莓（Fv）、菠萝（Aco）和石榴（Pg）TCP转录因子的系统进化分析（A）和AcTCPs 保守结构域的氨基酸序列比对（B）

Fig. 1 Phylogenetic analysis (A) of TCP transcription factors in A. chinensis (Ac), A. thaliana (At), F. vesca (Fv), Ananas comosus (Aco) and P. Granatum L (Pg) and amino acid sequence comparison of conserved structural domains of AcTCPs (B)

图2 AcTCPs的系统发育树（A）、保守基序（B）及基因组结构（C）分析

Fig. 2 Phylogenetic tree (A), conserved motif (B) and genomic structure (C) analysis of AcTCPs

图3 AcTCP基因启动子顺式元件分析

Fig. 3 Cis-element analysis of AcTCP genes promoter region

图4 AcTCPs基因的共线性分析紫色、绿色和黑色的基因名称分别代表PFC基因、CIN基因和CYC/TB1基因。紫色、绿色和黑色线条分别表示PFC基因、CIN基因和CYC/TB1基因的染色体间共线性。蓝线表示CIN基因和CYC/TB1基因之间的染色体间共线性

Fig. 4 Covariance analysis of AcTCPs genesPurple, green and black gene names indicate PFC genes, CIN genes and CYC/TB1 genes, respectively. Purple, green and black lines indicate intrachromosomal covariance of PFC genes, CIN genes and CYC/TB1 genes, respectively. The blue line indicates the intrachromosomal covariance between the CIN gene and the CYC/TB1 gene

图5 果实不同发育时期‘红阳’猕猴桃果实中的TCP基因表达分析

Fig. 5 Expression analysis of TCP genes in fruits at different fruit developmental stages of A. chinensis var. Hongyang

图6 AcTCP基因在‘红阳’猕猴桃中的组织表达模式分析不同的字母表示差异显著（P<0.05），下同

Fig. 6 Analysis of tissue expression pattern of AcTCP genes in A. chinensis var. HongyangDifferent letters indicate significant difference at P<0.05 level. The same below

图7 不同果实发育时期AcTCPs基因家族成员表达模式分析

Fig. 7 Analysis of expression patterns of AcTCP genes in different stages of fruit development

图8 AcTCPs基因在不同激素处理中的表达分析A：ABA；B：ET；C：GA3；D：CPPU；横坐标代表不同处理时间

Fig. 8 Expressions of AcTCP genes in different hormone treatmentsA: ABA treatment; B: ET treatment; C: GA3 treatment; D: CPPU treatment. Horizontal coordinates indicate different treatment times

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