西方蜜蜂Cytochrome c基因的分子特征、系统进化及时空表达谱

doi:10.19586/j.2095-2341.2025.0141

生物技术进展 ›› 2026, Vol. 16 ›› Issue (1): 115-122.DOI: 10.19586/j.2095-2341.2025.0141

西方蜜蜂Cytochrome c基因的分子特征、系统进化及时空表达谱

李婧娴¹(), 宓诗雨¹(), 左真祯¹, 王伟¹, 李紫诺¹, 龚海宁¹, 陈大福¹^,²^,³, 郭睿¹^,²^,³(), 邱剑丰¹^,²^,³()

^1.福建农林大学蜂学与生物医药学院，福州 350002
^2.福建农林大学天然生物毒素国家地方联合工程实验室，福州 350002
^3.福建农林大学蜂疗研究所，福州 350002

收稿日期:2025-10-16 接受日期:2025-11-13 出版日期:2026-01-25 发布日期:2026-02-12
通讯作者: 郭睿,邱剑丰
作者简介:李婧娴E-mail: 18236228430@163.com
宓诗雨E-mail: Missy7728@163.com第一联系人：（并列第一作者）
基金资助:
国家自然科学基金面上项目(32172792);国家自然科学基金面上项目(32372943);国家现代农业产业技术体系专项资金(CARS-44-KXJ7);福建省自然科学基金面上项目(2025J01616);福建省大学生创新创业训练计划项目(S2025103890105);福建省大学生创新创业训练计划项目(S2025103890101);福建省大学生创新创业训练计划项目(S202510389089)

Molecular Characteristics, Phylogeny and Spatiotemporal Expression Profile of the Cytochrome c Gene in Apis mellifera

Jingxian LI¹(), Shiyu MI¹(), Zhenzhen ZUO¹, Wei WANG¹, Zinuo LI¹, Haining GONG¹, Dafu CHEN¹^,²^,³, Rui GUO¹^,²^,³(), Jianfeng QIU¹^,²^,³()

^1.College of Bee Science and Biomedicine，Fujian Agriculture and Forestry University，Fuzhou 350002，China
^2.National & Local United Engineering Laboratory of Natural Biotoxin，Fujian Agriculture and Forestry University，Fuzhou 350002，China
^3.Apitherapy Research Institute of Fujian Agriculture and Forestry University，Fuzhou 350002，China

Received:2025-10-16 Accepted:2025-11-13 Online:2026-01-25 Published:2026-02-12
Contact: Rui GUO,Jianfeng QIU

摘要/Abstract

摘要：

研究聚焦西方蜜蜂(Apis mellifera)的细胞色素c基因(cytochrome c，CytC)，解析其分子特征、系统进化及时空表达模式。通过PCR扩增CytC的编码序列(coding sequence，CDS)并进行Sanger测序比对；利用ProtParam、Porter5、SWISS-MODEL、STRING数据库等预测CytC蛋白的理化性质、结构域及磷酸化修饰位点并构建蛋白互作网络；采用MEGA软件对10个物种的CytC蛋白进行多序列比对与系统进化分析；通过RT-qPCR检测CytC在工蜂成虫7个组织、幼虫发育阶段和成蜂不同日龄的表达谱。结果表明，CytC的CDS全长327 bp，共编码108个氨基酸，分子量约为11 kD，等电点为9.50。CytC蛋白无信号肽与跨膜结构，含13个磷酸化位点；二级结构以无规则卷曲(48.15%)和α-螺旋(25.93%)为主。系统进化显示，西方蜜蜂与小蜜蜂和中华蜜蜂的CytC蛋白聚为一支，与熊蜂科物种的分化显著。时空表达谱表明，CytC基因在脑组织中表达最高，咽下腺中最低；CytC在卵期的表达量最高，且显著高于其他时期(P<0.05)，在工蜂幼虫体内的表达水平总体表现为持续下降趋势；成蜂中，CytC表达量随成虫期日龄增加逐渐上升，在18日龄最高，在1日龄最低。综上，CytC是亲水性的胞内蛋白，具有保守的分子特征与功能，其可能在西方蜜蜂发育过程中发挥着重要作用。

关键词: 西方蜜蜂, 时空表达谱, 细胞色素c, 基因表达

Abstract:

This study focused on the cytochrome c gene （CytC） in Apis mellifera， with the aim of characterizing its molecular features， phylogenetic evolution， and spatiotemporal expression patterns. The coding sequence （CDS） of CytC was amplified by PCR and verified through Sanger sequencing alignment. Bioinformatics tools including ProtParam， Porter5， SWISS-MODEL and STRING were employed to predict the physicochemical properties， structural domains， phosphorylation sites， and protein-protein interaction networks of CytC. Phylogenetic analysis of CytC proteins from 10 species was conducted using MEGA software. RT-qPCR was used to examine the expression profiles of CytC across seven tissues of worker bees， larval developmental stages， and different ages of adult bees. The results indicated that a 327 bp CytC CDS encoding 108 residues， with predicted molecular mass of 11 kD and isoelectric point at 9.50. The CytC polypeptide lacks signal peptides and transmembrane domains but harbors 13 putative phosphorylation sites. And its secondary structure is primarily composed of random coils （48.15%） and α-helices （25.93%）. Phylogenetic analysis revealed that the CytC protein of A. mellifera， is clustered with that of A. florea and A. cerana， exhibiting marked divergence from Bombus species. Spatiotemporal expression profiling indicated that CytC expression was highest in the brain and lowest in the hypopharyngeal glands. During development， CytC expression was highest at the egg stage， significantly exceeding that at all other stages （P<0.05）， and gradually decreased throughout larval development. In adult bees，CytC expression increased with age， peaking at 18 days after eclosion and reaching the lowest level at 1 day. In summary， CytC is a hydrophilic intracellular protein with conserved molecular characteristics and functions， potentially playing a significant role in the development of western honey bees.

Key words: Apis mellifera, spatiotemporal expression pattern, cytochrome c, gene expression

中图分类号:

Q965.9

李婧娴, 宓诗雨, 左真祯, 王伟, 李紫诺, 龚海宁, 陈大福, 郭睿, 邱剑丰. 西方蜜蜂Cytochrome c基因的分子特征、系统进化及时空表达谱[J]. 生物技术进展, 2026, 16(1): 115-122.

Jingxian LI, Shiyu MI, Zhenzhen ZUO, Wei WANG, Zinuo LI, Haining GONG, Dafu CHEN, Rui GUO, Jianfeng QIU. Molecular Characteristics, Phylogeny and Spatiotemporal Expression Profile of the Cytochrome c Gene in Apis mellifera[J]. Current Biotechnology, 2026, 16(1): 115-122.

图/表 6

图1 CytC基因编码序列扩增产物的琼脂糖凝胶电泳和Sanger测序结果A:CytC基因扩增；B：CytC基因的Sanger测序

Fig. 1 Results of agarose gel electrophoresis and sanger sequencing of the CytC gene coding sequence amplicon

图2 西方蜜蜂CytC蛋白结构特征A：亲水性；B：信号肽；C：跨膜结构域；D：磷酸化位点分布

Fig. 2 Structural characterization of Apis mellifera CytC protein

图3 CytC蛋白的结构和互作网络A:CytC蛋白的二级结构；B:CytC蛋白的三级结构；C:CytC蛋白的互作网络

Fig. 3 The structure and interaction network of CytC protein

图4 西方蜜蜂与其他9个物种CytC蛋白系统进化树

Fig. 4 Evolutionary relationships of CytC proteins across Apis mellifera and nine representative species

图5 西方蜜蜂工蜂7个组织中CytC基因的相对表达量注：图中数据为平均值±标准误；柱上不同小写字母表示在P<0.05水平上具有统计学意义（Tukey检验)。

Fig. 5 Relative expression level of CytC in seven tissues of Apis mellifera worker

图6 西方蜜蜂工蜂不同发育阶段CytC的相对表达量A:幼虫发育阶段；B：成虫阶段。柱上不同小写字母表示在P<0.05水平上具有统计学意义（Tukey检验)。

Fig. 6 Relative expression level of CytC in various developmental stages of Apis mellifera worker

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西方蜜蜂Cytochrome c基因的分子特征、系统进化及时空表达谱

Molecular Characteristics, Phylogeny and Spatiotemporal Expression Profile of the Cytochrome c Gene in Apis mellifera

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