中国樱桃 WRKY 基因家族鉴定及其表达分析

Identification and expression analysis of the WRKY gene family in Chinese cherry

摘要

【目的】WRKY 转录因子在园艺植物果实发育过程中发挥关键调控作用。本研究旨在探究中国樱桃 （Prunus pseudocerasus (Lindl.) G. Don）WRKY 转录因子家族的全基因组特征及其在果实发育过程中的表达模式， 并筛选调控果实发育与成熟的关键候选基因。【方法】从中国樱桃全基因组中鉴定 WRKY 基因家族成员，利用生物 信息学方法系统分析其理化性质和进化关系等，并结合 3 个中国樱桃品种果实不同发育时期的转录组数据，分析其 表达模式。【结果】共鉴定到 58 个中国樱桃 WRKY 基因家族成员，命名为 PpseWRKY1～PpseWRKY58。这些基因 不均匀分布在 8 条染色体上，编码蛋白长度为 117～644 aa，等电点范围为 4.94～9.92，且均为亲水性蛋白。系统 进化分析结果显示，58 个 PpseWRKY 基因可分为Ⅰ、Ⅱ和Ⅲ族，分别包含 12、39 和 7 个成员。保守基序和基因结构 分析表明，所有中国樱桃 WRKY 基因均含有保守的 W RKYGQK 基序，且同一亚族内基因结构和保守基序相似性 较高。PpseWRKY 基因家族启动子区富集了大量与激素调控、光响应及胁迫响应相关的顺式作用元件。共线性分析 发现，中国樱桃种内存在 15 对 PpseWRKY 基因片段复制事件。转录组数据与 RT-qPCR 验证结果表明，中国樱桃 WRKY 基因的表达在不同品种及果实不同发育阶段存在显著差异。多数基因（如 PpseWRKY27、PpseWRKY37、 PpseWRKY49 和 PpseWRKY53）在绿果期的表达量最高，随果实成熟进程显著下调，推测其参与果实前期生长发育 调控；而 PpseWRKY5 和 PpseWRKY23 表达量随果实发育逐渐升高，推测其正向调控果实成熟与品质形成过程。【结 论】本研究首次系统解析了中国樱桃 WRKY 转录因子家族的全基因组特征及其在果实发育中的表达模式，为深入 解析其在果实发育与成熟中的功能提供了理论基础。

Abstract

[Objective] WRKY transcription factors play crucial regulatory roles in fruit development in horti- cultural plants. This study aimed to systematically characterize the genome-wide features of the WRKY transcription factor family in Chinese cherry (Prunus pseudocerasus (Lindl.) G. Don), analyze its expression patterns during fruit development, and screen for key candidate genes involved in regulating fruit development and ripening. [Method] WRKY gene family members were identified from the Chinese cherry genome, and comprehensive bioinformatics analyses were conducted to explore their physicochemical properties and phylogenetic relationships. Expression pat- terns were investigated using transcriptome data from different fruit developmental stages of three Chinese cherry cul- tivars and validatedvia RT-qPCR. [Result] Fifty-eight WRKYgene family members were identified in the P. pseu- docerasus genome and designated as PpseWRKY1–PpseWRKY58. These genes were unevenly distributed across eight chromosomes and encoded proteins ranging from 117 to 644 amino acids with isoelectric points of 4.94 – 9.92; all predicted proteins were hydrophilic. Phylogenetic analysis classified the 58 PpseWRKY genes into three groups: Group I (12), Group II (39), and Group III (7). Conserved motif and gene structure analyses revealed that all PpseWRKY genes contained the conserved WRKYGQK motif, with high similarity in gene structure and conserved motifs within the same group. Numerous cis-acting elements related to hormone regulation, light response, and stress response were enriched in the promoters of PpseWRKY genes. Collinearity analysis identified 15 segmental duplica- tion pairs among PpseWRKY genes. Transcriptome and RT-qPCR analyses showed distinct expression patterns of PpseWRKY genes among different cultivars and fruit developmental stages. Most genes, such as PpseWRKY27, PpseWRKY37, PpseWRKY49, and PpseWRKY53, were highly expressed at the green fruit stage and significantly down-regulated during ripening, suggesting roles in early fruit development. In contrast, the expression levels of PpseWRKY5 and PpseWRKY23 gradually increased during fruit development, implying positive regulatory functions in fruit ripening and quality formation. [Conclusion] This study provides the first systematic characterization of the genome-wide features of the WRKY transcription factor family and their expression profiles during fruit development in Chinese cherry, establishing a theoretical foundation for further functional analysis of fruit development and ripening.

关键词

中国樱桃 / 果实发育 / WRKY 转录因子 / 基因家族 / 表达分析

Key words

Chinese cherry / fruit development / WRKY transcription factor / gene family / expression analysis

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基金资助

2025 年甘薯、草莓等品种资源保存与利用项目(1021302504/004)

2025 年南京市地方果树资源收集、保存与评价(1041302505)