‘红阳’猕猴桃BGAL基因家族的全基因组鉴定与表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2025-0816

摘要/Abstract

摘要：

目的对‘红阳’猕猴桃β-半乳糖苷酶基因（AcBGALs）家族进行全基因组鉴定和表达模式分析，为深入解析该基因家族功能提供理论依据。方法基于‘红阳’猕猴桃基因组数据对AcBGALs进行鉴定，利用生物信息学方法对其理化性质、染色体定位、基因结构、系统进化关系及基因间的共线性进行分析，结合转录组数据和实时荧光定量PCR技术分析基因在猕猴桃不同组织、果实不同发育时期和后熟软化及激素响应过程中的表达模式。结果在‘红阳’猕猴桃全基因组中共鉴定出21个BGAL家族成员，分属6个亚家族，不均匀分布在14条染色体上。顺式作用元件分析发现，该基因家族启动子区含有丰富的生长发育及激素响应相关元件。表达分析结果显示，除AcBGAL3外，其余5个候选基因在果实中均呈现较高表达水平；随着果实的发育，AcBGAL2与AcBGAL8呈明显上调表达，而AcBGAL13表达下调，该表达趋势与转录组分析结果一致。在果实后熟软化过程中，除AcBGAL12外，其他5个AcBGAL基因均表现出上调表达的趋势。此外，在果实中，乙烯（ET）和脱落酸（ABA）处理下调AcBGAL2、AcBGAL8和AcBGAL12的表达；氯吡脲（CPPU）和赤霉素（GA₃）处理下调AcBGAL2和AcBGAL12的表达，但在处理前期上调AcBGAL3、AcBGAL10和AcBGAL13的表达。结论在‘红阳’猕猴桃基因组中共鉴定出21个AcBGAL基因，6个候选基因的表达与果实发育和后熟软化密切相关，并受外源激素（ET、ABA、CPPU和GA₃）调控，为进一步挖掘AcBGAL基因功能奠定基础。

关键词: ‘红阳’猕猴桃, β-半乳糖苷酶（BGAL）, 全基因组鉴定, 生物信息学, 表达分析, 果实生长发育, 果实后熟软化, 激素处理

Abstract:

Objective This study is aimed to conduct a genome-wide identification and expression pattern analysis of the β-galactosidase gene family (AcBGALs) in ‘Hongyang’ kiwifruit (Actinidia chinensis var. Hongyang), providing a theoretical basis for further elucidating the functions of this gene family. Method The AcBGAL genes wereidentified based on the whole-genome sequence of ‘Hongyang’ kiwifruit. Their physicochemical properties, chromosomal localizations, gene structures, phylogenetic relationships, and gene collinearity were analyzed using bioinformatics methods. Transcriptomic data and quantitative real-time PCR (RT-qPCR) were combined to investigate the expression patterns of these genes in different tissues, various fruit developmental stages, postharvest softening, and hormone response processes. Result A total of 21 BGAL family members were identified in the ‘Hongyang’ kiwifruit genome, which were belonged to six subfamilies and unevenly distributed across 14 chromosomes. Cis-acting element analysis revealed that the promoter regions of this gene family were enriched with elements related to growth/development and hormone response. Expression analysis showed that five of the six candidate genes (excluding AcBGAL3) were highly expressed in the fruit. Throughout fruit development, the expressions AcBGAL2 and AcBGAL8 were significantly upregulated, whereas that of AcBGAL13 was downregulated, a trend consistent with transcriptomic analysis. During ripening and softening, five AcBGAL genes (excluding AcBGAL12) showed an upward trend in expression. Additionally, ethylene (ET) and abscisic acid (ABA) treatments downregulated the expressions of AcBGAL2, AcBGAL8 and AcBGAL12 in fruit. Treatment with forchlorfenuron (CPPU) and gibberellin (GA₃) downregulated the expressions of AcBGAL2 and AcBGAL12 but upregulated the expressions of AcBGAL3, AcBGAL10 and AcBGAL13 in the early stages of treatment. Conclusion A total of 21 AcBGAL genes were identified in the ‘Hongyang’ kiwifruit genome. Among them, six candidate genes showed expression patterns closely associated with fruit development and post-ripening softening, and were regulated by exogenous hormones (ET, ABA, CPPU, and GA₃). These findings provide a foundation for further functional characterization of AcBGAL genes.

Key words: Actinidia chinensis var. Hongyang, β-galactosidase (BGAL), genome-wide identification, bioinformatics, expression analysis, fruit growth and development, fruit ripening and softening, hormone treatment

刘林娅, 刘欢艳, 梁鑫钰, 宋姝熠, 何斌, 王绪英, 黄亚成. ‘红阳’猕猴桃BGAL基因家族的全基因组鉴定与表达分析[J]. 生物技术通报, doi: 10.13560/j.cnki.biotech.bull.1985.2025-0816.

LIU Lin-ya, LIU Huan-yan, LIANG Xin-yu, SONG Shu-yi, HE Bin, WANG Xu-ying, HUANG Ya-cheng. Genome-wide Identification and Expression Analysis of BGAL Family in Actinidiachinensis var. Hongyang[J]. Biotechnology Bulletin, doi: 10.13560/j.cnki.biotech.bull.1985.2025-0816.

图/表 9

表1 实时荧光定量PCR引物

Table 1 Primers for RT-qPCR

引物名称 Primer name	引物序列 Primer sequence （5′‒3′）
AcBGAL2-F/R	AGCTTGGCTGTGGTCTGC/GGTCCCCTCCGAGTTCCT
AcBGAL3-F/R	AGCACTGAGTCTTCATTCTCT/CTCCCCCTGGAGCATTGAAAA
AcBGAL8-F/R	TGGTCATGAGCCTTCTCCTGGA/GGGGCCAATTCGGAGATGAA
AcBGAL10-F/R	CGGAGGCTTAGATGTGATCG/TAAACCAGCTTCAGCGACCA
AcBGAL12-F/R	TACCTCCCTCGGAAGCAAC/CCATTGAACTTGACCGTAGC
AcBGAL13-F/R	ACGAAGCACTCCCGAAATGT/AGGGCTCATGCACATTCCAA
18S-F/R	GAATCATCAGAGCAACGGGC/TTTGATGGTATTTGCTACTCGGA
GAPDH-F/R	CGTCCTTCCCTCCCTCAAA/TCTTCTTGGTGACCTGGACGA

图1 AcBGALs基因的染色体定位（A）及共线性（B）分析

Fig. 1 Chromosomal localization (A) and covariance analysis (B) of AcBGALs genes

图2 猕猴桃（Ac）、桃（Pp）、苹果（Md）和拟南芥（At）BGAL同源蛋白的系统发育树

Fig. 2 Phylogenetic tree of BGAL homologous protein in A. chinensi (Ac), P. persica (Pp), M. domestica (Md)and A. thaliana (At)

图3 AcBGALs的保守基序和基因结构分析A：AcBGALs系统发育树；B：AcBGAL保守基序；C：AcBGAL基因结构

Fig. 3 Analysis of conserved motifs and gene structure of AcBGALsA: Phylogenetic tree of AcBGALs. B: Conserved motifs of AcBGAL. C: Structure of AcBGAL gene

图4 AcBGAL基因启动子顺式作用元件分析A：顺式作用元件预测统计；B：各类顺式元件的数量分布

Fig. 4 Cis-acting element analysis of AcBGAL gene promotersA: Prediction statistics of cis-acting elements. B: The number distribution of various cis-acting elements

图5 果实不同发育时期‘红阳’猕猴桃果实中的BGAL基因表达分析DPA：授粉后天数

Fig. 5 Expression analysis of BGAL genes in fruits at different fruit developmental stages of A. chinensis var. HongyangDPA: Days after pollination

图6 AcBGAL基因在不同组织中的表达不同的字母表示差异显著（P<0.05），下同

Fig. 6 Expressions of AcBGAL genes in different tissuesDifferent letters indicate significant difference at P<0.05 level. The same below

图7 AcBGAL基因在果实不同发育时期（A）和不同贮藏天数（B）的表达

Fig. 7 Expressions of AcBGAL genes in fruit at different developmental stages (A) and storage days (B)

图8 AcBGAL基因在不同激素胁迫下的表达

Fig. 8 Expressions of AcBGAL genes in different hormone treatments

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