目的 克隆山茱萸MYB62基因（Cornus officinalis MYB62，CoMYB62）的全长cDNA序列，对其编码蛋白的结构特征、系统进化关系、亚细胞定位以及组织表达特性进行分析，为阐明MYB转录因子在山茱萸生长发育及环境适应过程中的潜在功能提供分子基础。方法 基于山茱萸转录组数据筛选CoMYB62基因序列，采用RT-PCR技术克隆其全长cDNA序列；利用生物信息学方法对CoMYB62编码蛋白的理化性质、保守结构域、二级和三级结构及系统进化关系进行预测分析；构建pCAMBIA1300-CoMYB62-GFP融合表达载体并侵染烟草叶片，激光共聚焦倒置显微镜观察CoMYB62蛋白的亚细胞定位；采用实时荧光定量PCR（quantitative real-time PCR，qRT-PCR）技术检测CoMYB62在根、茎、叶和果实4种不同组织中的表达特征。结果 成功克隆获得CoMYB62基因全长cDNA，序列分析表明其编码蛋白属于典型的R2R3-MYB转录因子，N端含有高度保守的DNA结合结构域，预测主要定位于细胞核。系统进化分析显示，CoMYB62与多种植物MYB同源蛋白具有较高进化保守性。激光共聚焦显微镜下观察到CoMYB62蛋白定位于细胞核中。qRT-PCR结果表明，CoMYB62在山茱萸不同组织中均有表达，但表达水平存在明显差异。结论 CoMYB62是山茱萸中一类结构保守、表达具有组织差异性的MYB转录因子，可能参与其生长发育及逆境响应相关调控过程，为后续深入解析CoMYB62的生物学功能奠定了基础。

Objective To clone the full-length cDNA sequence of the CoMYB62 gene from Cornus officinalis, and analyze the structural characteristics, phylogenetic relationships, subcellular localization, and tissue expression patterns of its encoded protein, thereby providing a molecular basis for elucidating the potential roles of MYB transcription factors in the growth, development, and environmental adaptation of C. officinalis. Methods Based on transcriptome data of C. officinalis, the CoMYB62 gene sequence was screened and its full-length cDNA sequence was cloned using RT-PCR. Bioinformatics analyses were conducted to predict the physicochemical properties, conserved domains, secondary and tertiary structures, and phylogenetic relationships of the CoMYB62-encoded protein. A pCAMBIA1300-CoMYB62-GFP fusion expression vector was constructed and transiently expressed in tobacco leaves, and the subcellular localization of the CoMYB62 protein was observed using a laser confocal inverted microscope. In addition, quantitative real-time PCR (qRT-PCR) was performed to analyze the expression patterns of CoMYB62 in four tissues, including root, stem, leaf, and fruit. Results The full-length cDNA of the CoMYB62 gene was successfully cloned. Sequence analysis indicated that the encoded protein belonged to a typical R2R3-MYB transcription factor and contained a highly conserved DNA-binding domain at the N-terminus, which was predicted to be mainly localized in the nucleus. Phylogenetic analysis revealed that CoMYB62 shared high evolutionary conservation with MYB homologous proteins from various plant species. Laser confocal microscopy further confirmed that the CoMYB62 protein was localized in the nucleus. The qRT-PCR results showed that CoMYB62 was expressed in all examined tissues of C. officinalis, although the expression levels varied significantly among different tissues. Conclusion CoMYB62 is a conserved MYB transcription factor in C. officinalis with tissue-specific expression characteristics, which may participate in the regulation of plant growth, development, and stress responses to provide a foundation for further biological functional analysis of CoMYB62 in C. officinalis.

R282.12

河南省中央引导地方科技发展资金项目（Z20241471030）；中央本级重大增减支项目“名贵中药资源可持续利用能力建设项目”（2060302）；河南省科技攻关项目（242102110325,262102110326）；河南省中药材产业科技特派员服务团项目