油茶MYB转录因子家族鉴定及CoMYB107基因在雄蕊发育中的功能分析

doi:10.13560/j.cnki.biotech.bull.1985.2026-0115

摘要/Abstract

摘要：

目的鉴定油茶（Camellia oleifera）MYB基因家族成员，解析其花药发育的分子调控网络，为阐明油茶生殖发育机制提供理论依据。方法基于油茶基因组数据，通过生物信息学方法鉴定MYB基因家族成员，解析其序列特征、理化性质、进化关系，并预测亚细胞定位；利用转录组与RT-qPCR技术，分析候选MYB基因在不同组织和不同发育阶段的表达模式，筛选关键基因，以烟草叶盘法异源过表达基因，验证候选基因功能。结果从油茶基因组鉴定到137个MYB基因，其氨基酸长度为102-1 752 aa，分布于45条染色体中，大多数成员的亚细胞定位预测定位在细胞核中。存在55次片段复制事件，共有44对有效基因，38对基因受纯化选择影响，5对基因受正选择影响。与拟南芥存在95对共线性基因对。系统发育分析显示，油茶与拟南芥的MYB家族可分为26个亚家族。启动子顺式作用元件主要与光响应、激素响应和胁迫应答相关，与激素相关的顺式作用元件主要有赤霉素、脱落酸和茉莉酸元件。转录组数据与RT-qPCR表达模式分析发现，8个基因在花药中呈现特异性高表达特征。此外，蛋白互作网络预测提示，CoMYB107、CoMYB55和CoMYB136基因可能参与特定的蛋白互作调控网络；CoMYB107过表达烟草的花粉活力显著下降、花丝长度伸长和花药延迟开裂。结论 CoMYB107可能通过GA-JA信号通路调控雄蕊发育，表现为促进花丝伸长、延迟花药开裂以及降低花粉活力。

关键词: 油茶, CoMYB107, 转录组, 花药发育, 表达分析, MYB基因家族, 赤霉素, 全基因组鉴定

Abstract:

Objective To identify members of the MYB gene family in Camellia oleifera and analyze the molecular regulatory network of its anther development, providing theoretical support for elucidating the reproductive development mechanism of C. oleifera. Method Based on the C. oleifera genome data, members of the MYB gene family were identified using bioinformatics methods, and their sequence characteristics, physicochemical properties, and evolutionary relationships were analyzed, and subcellular localization was predicted. Using transcriptome data and RT-qPCR technology, the expression patterns of candidate MYB genes in different tissues and at different developmental stages were investigated, key genes were screened, and heterologous overexpression of genes was performed using the tobacco leaf disc method to verify the functions of candidate genes. Result A total of 137 MYB genes were identified from the C. oleifera genome, with amino acid lengths ranging from 102 to 1 752 aa, distributed on 45 chromosomes, and the majority of members were predicted to localize in the nucleus. There were 55 segmental duplication events, involving 44 valid gene pairs. Among these, 38 gene pairs were under purifying selection, and 5 gene pairs were under positive selection. There were 95 collinear gene pairs with Arabidopsis. Phylogenetic analysis indicated that the MYB family in C. oleifera and Arabidopsis could be classified into 26 subfamilies. Promoter cis-acting elements were mainly associated with light response, hormone response, and stress response, with hormone-related cis-elements mainly including gibberellin, abscisic acid, and jasmonic acid elements. Transcriptome data and RT-qPCR expression pattern analysis showed that 8 genes exhibited specifically high expression in anthers. In addition, protein interaction network prediction suggested that CoMYB107, CoMYB55, and CoMYB136 may be involved in a specific protein interaction regulatory network; overexpression of CoMYB107 in tobacco significantly reduced pollen viability, elongated filament length, and delayed anther dehiscence. Conclusion CoMYB107 may regulate stamen development through the GA-JA signaling pathway, which manifests as promoting filament elongation, delaying anther dehiscence, and decreasing pollen viability.

Key words: Camellia oleifera, CoMYB107, transcriptome, anther development, expression analysis, MYB gene family, gibberellin, whole-genome identification

谭澍, 沈红建, 高晓磊, 尹倩, 邹锋. 油茶MYB转录因子家族鉴定及CoMYB107基因在雄蕊发育中的功能分析[J]. 生物技术通报, 2026, 42(9): 196-209.

TAN Shu, SHEN Hong-jian, GAO Xiao-lei, YIN Qian, ZOU Feng. Genome-wide Identification of the MYB Transcription Factor Family in Camellia oleifera and Functional Analysis of CoMYB107 in Stamen Development[J]. Biotechnology Bulletin, 2026, 42(9): 196-209.

图/表 13

图1 不同时期油茶花芽外观形态T1-T5表示花芽采集日期（2024年）；T1：9月5日；T2：9月20日；T3：10月5日；T4：10月20日；T5：11月4日。下同

Fig. 1 The morphology of flower buds in different periods of C. oleiferaT1-T5 represent the dates of flower bud collection (2024). T1: September 5. T2: September 20. T3: October 5. T4: October 20. T5: November 4. The same below

图2 油茶MYB基因结构分析A：进化树；B：motif结构；C：保守结构域；D：UTR和CDS

Fig. 2 Structural analysis of C. oleiferaMYB genesA: Phylogenetic tree. B: Motif structure. C: Conserved domain. D: UTR and CDS

图3 油茶MYB基因染色体定位

Fig. 3 Chromosomal localization of C. oleiferaMYB genes

图4 MYB基因共线性分析A：油茶MYB基因共线性分析；B：拟南芥MYB与油茶MYB基因共线性分析

Fig. 4 Collinearity analysis of MYB genesA: Collinearity analysis of C. Oleifera MYB genes. B: Collinearity analysis of A. thaliana MYB and C. oleifera MYB genes

图5 油茶MYB基因启动子分析

Fig. 5 Analysis of C. oleiferaMYB gene promoters

图6 拟南芥和油茶MYB蛋白的系统发育树

Fig. 6 Phylogenetic tree of MYB proteins from A. thaliana and C. oleifera

图7 CoMYB基因不同时期表达量热图

Fig. 7 Heatmap of CoMYB gene expression levels in different periods

图8 油茶MYB基因在不同组织中的相对表达量R：根；S：茎；L：叶；C：花萼；PE：花瓣；A：花药；PI：雌蕊；数据为3个生物学重复（n=3）的均值±标准误（SE）。不同小写字母表示差异显著（P<0.05）。下同

Fig. 8 Relative expression analysis of C. oleifera MYB genes in different tissueR: Root; S: stem; L: leaf; C: calyx; PE: petal; A: anther; PI: pistil. Data are presented as the mean ± standard error (SE) of three biological replicates (n=3). Different lowercase letters indicate significant differences (P<0.05). The same below

图9 油茶MYB基因在不同发育时期表达分析

Fig. 9 Expression analysis of C. oleiferaMYB genes at different developmental stages

图10 蛋白质互作网络预测

Fig. 10 Protein interaction network prediction

图11 CoMYB107蛋白的亚细胞定位

Fig. 11 Subcellular localization of CoMYB107 protein

图12 过表达CoMYB107转基因烟草表型A：野生型烟草花粉活力；B：CoMYB107转基因烟草花粉活力；C：烟草雄蕊表型（C1：野生型烟草；C2：CoMYB107转基因烟草）；D：野生型与CoMYB107转基因烟草表型统计；** P<0.01，下同

Fig. 12 Phenotype of OE-CoMYB107Nicotiana tabacumA: Pollen viability of wild-type N. tabacum. B: Pollen viability of CoMYB107 transgenic N. tabacum. C: N. tabacum stamen phenotype (C1: Wild-type N. tabacum. C2: CoMYB107 transgenic N. tabacum). D: Statistics of pollen viability in wild-type and overexpression N. tabacum. ** P<0.01. The same below

图13 8个花发育相关基因在烟草花中的表达分析

Fig. 13 Expression analysis of 8 flower development-related genes in tobacco flowers

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