过表达森林草莓FvBBX22促进拟南芥花青素合成和延迟开花时间

doi:10.13560/j.cnki.biotech.bull.1985.2026-0204

摘要/Abstract

摘要：

目的 BBX转录因子在植物生长发育过程中发挥着重要作用，探究森林草莓FvBBX22的生物学功能，为草莓分子育种提供基因资源。方法以森林草莓‘Ruegen’为试验材料，通过RT-PCR技术克隆出FvBBX22基因，并利用生物信息学方法分析其蛋白的理化性质、结构特征和系统进化树；构建FvBBX22基因的过表达载体，并通过花序侵染法获得转基因拟南芥；观察转基因拟南芥的开花表型和测定叶片花青素含量，通过RT-qPCR技术检测花青素合成和开花途径中相关基因的表达水平，以及分析FvBBX22基因在森林草莓中的组织特异性。结果 FvBBX22基因CDS全长为864 bp，编码287个氨基酸，含有2个B-box保守结构域，属于第Ⅳ亚组成员。系统进化树分析发现，BBX22蛋白在森林草莓和蔷薇科的尖齿黑莓之间的亲缘关系较近。组织特异性结果显示，FvBBX22基因在森林草莓各个组织中均有表达，其中在茎中表达量最高，其次是叶片。在过表达FvBBX22的拟南芥中，叶片呈现浅紫色，叶片花青素含量显著高于野生型拟南芥，检测花青素合成通路中相关基因的表达水平发现，AtCHS、AtCHI、AtF3H、AtDFR、AtANS和AtUF3GT基因的表达水平显著升高。然而，转基因拟南芥的开花时间延迟，检测开花途径中相关基因的表达水平发现，AtCO、AtFT、AtFUL和AtLFY基因的表达量显著降低。结论森林草莓FvBBX22能够促进拟南芥花青素的积累，并能延迟开花时间。

关键词: 草莓, BBX22, 花青素, 开花, 功能分析, 基因克隆, 生物信息学分析, 表达分析

Abstract:

Objective BBX transcription factors play important roles in plant growth and development, and exploring the biological functions of FvBBX22 provides genetic resources for strawberry molecular breeding. Method Using Fragaria vesca ‘Ruegen’ as experimental material, the FvBBX22 gene was cloned by RT-PCR. The physicochemical properties and structural characteristics of its protein, as well as its phylogenetic tree, were analyzed using bioinformatics methods. An overexpression vector for the FvBBX22 gene was constructed, and transgenic Arabidopsis plants were obtained via the floral dip method. The flowering phenotype of transgenic Arabidopsis was observed, and anthocyanin content in leaves was measured. The expression levels of genes related to anthocyanin biosynthesis and flowering pathways were detected by RT-qPCR. Additionally, the tissue-specific expression pattern of FvBBX22 in Fragaria vesca was analyzed. Result The full-length CDS of FvBBX22 was 864 bp, encoding 287 amino acids. It contained two conserved B-box domains and belonged to subgroup Ⅳ of the BBX family. Phylogenetic analysis revealed that the BBX22 proteins from F. vesca and Rubus argutus (of the Rosaceae family) are closely related. Tissue-specific expression analysis showed that FvBBX22 is expressed in all tissues of F. vesca, with the highest expression in stems, followed by leaves. In Arabidopsis plants overexpressing FvBBX22, leaves appeared light purple, and anthocyanin content was significantly higher than that in wild-type Arabidopsis. Analysis of the expression of genes involved in the anthocyanin biosynthesis pathway revealed significantly elevated levels of AtCHS, AtCHI, AtF3H, AtDFR, AtANS, and AtUF3GT. However, the flowering time of transgenic Arabidopsis was delayed. Analysis of the expression of genes involved in the flowering pathway revealed significantly reduced levels of AtCO, AtFT, AtFUL, and AtLFY. Conclusion FvBBX22 promotes anthocyanin accumulation and delays flowering time in Arabidopsis.

Key words: strawberry, BBX22, anthocyanin, flowering, functional analysis, gene cloning, bioinformatics analysis, expression analysis

董向向, 侯威威, 缪百灵, 陈娟娟, 李亮杰, 翟敏, 朱庆松, 刘松虎. 过表达森林草莓FvBBX22促进拟南芥花青素合成和延迟开花时间[J]. 生物技术通报, 2026, 42(9): 231-237.

DONG Xiang-xiang, HOU Wei-wei, MIAO Bai-ling, CHEN Juan-juan, LI Liang-jie, ZHAI Min, ZHU Qing-song, LIU Song-hu. Overexpression of FvBBX22 from Fragaria Vesca Promotes Anthocyanin Synthesis and Delays Flowering Time in Arabidopsis thaliana[J]. Biotechnology Bulletin, 2026, 42(9): 231-237.

图/表 4

图1 不同物种BBX22蛋白的系统进化分析

Fig. 1 Phylogenetic analysis of BBX22 proteins from different species

图2 FvBBX22在不同组织的表达分析（A）、过表达FvBBX22拟南芥的DNA水平鉴定（B）和基因表达水平分析（C）不同小写字母表示差异显著（P<0.05）。下同

Fig. 2 Expression analysis of FvBBX22 in different tissues (A), DNA-level identification (B) and gene expression level analysis (C) of A. thaliana overexpressing FvBBX22Different lowercase letters indicate significant differences (P<0.05). The same below

图3 过表达FvBBX22拟南芥对花青素含量的影响及花青素合成相关基因的表达水平分析A：叶片表型；B：叶片花青素含量的测定；C‒H：花青素合成相关基因的表达水平

Fig. 3 Effects of FvBBX22 overexpression in A. thaliana on anthocyanin content and expression levels of anthocyanin-synthesis-related genesA: Leaf phenotype. B: Determination of anthocyanin content in leaves. C‒H: Expression levels of anthocyanin synthesis related genes

图4 过表达FvBBX22拟南芥的开花表型以及开花相关基因的表达水平分析A：开花表型；B：开花时间统计；C‒F：开花相关基因的表达水平

Fig. 4 Flowering phenotype and expression analysis of flowering-related genes in A. thaliana overexpressing FvBBX22A: Flowering phenotype. B: Statistics of flowering time. C‒F: Expression levels of flowering-related genes

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