摘要
【目的】比较蜂王浆高产蜜蜂(浆蜂,Apis mellifera liguatica)和意大利蜜蜂(意蜂,Apis mellifera liguatica)哺育蜂头唾腺、胸唾腺的蛋白质组差异,揭示唾液腺调控蜂王浆生产的分子基础,为解析浆蜂蜂王浆高产机理提供依据。【方法】解剖浆蜂、意蜂哺育蜂头唾腺和胸唾腺,提取蛋白质、液内酶切并进行液相色谱与串联质谱蛋白质组分析。采用MaxQuant软件对质谱数据定量和定性分析,利用Perseus软件对结果进行生物信息学分析。使用SignalP预测分泌性蛋白。利用ClueGO软件对唾液腺蛋白质组进行生物学进程和代谢通路富集分析。【结果】浆蜂和意蜂哺育蜂唾液腺共鉴定到2335个蛋白,其中头唾腺1823个,胸唾腺1922个。浆蜂、意蜂头唾腺和胸唾腺核心蛋白表达谱相似,主要参与RNA代谢、核酸代谢、ATP代谢、蛋白质的翻译、翻译调控和分解代谢,为腺体行使生物学功能提供了必要的代谢能和核酸、蛋白质等原料。浆蜂、意蜂哺育蜂头唾腺和胸唾腺主成分分析(PCA)显示二者唾液腺分子基础在选育过程中出现了一定程度的分化。意蜂、浆蜂头唾腺分别上调表达254和333个蛋白,意蜂小分子、碳水化合物代谢等通路上调,浆蜂有机氮化合物合成、细胞氧化还原稳态、氨基酸代谢、氧化还原等通路上调,证明浆蜂头唾腺细胞蛋白质合成、氨基酸代谢旺盛、供能加强。意蜂、浆蜂胸唾腺分别上调表达412和162个蛋白,意蜂氧化磷酸化、翻译调控等通路上调,浆蜂氧化磷酸化、对有毒物质的反应等通路上调,说明浆蜂胸唾腺细胞抗逆水平提高。浆蜂、意蜂唾液腺共鉴定到43个分泌性蛋白,其中有15个也在蜂王浆中得到鉴定,蜂王浆主蛋白1、2、3、4、5、7同时在头唾腺和胸唾腺中检出,表明头唾腺和胸唾腺均参与了蜂王浆主蛋白的合成;参与花蜜转化的α-葡萄糖苷酶和参与化学信息素合成与释放的气味结合蛋白3、13、17、21同时在头唾腺和胸唾腺中检出,为花蜜转化和信息素合成奠定了基础。蜂王浆主蛋白1、2、3、7,昆虫储存蛋白70a、110,气味结合蛋白3、13、17、21以及与幼虫先天免疫紧密相关的转铁蛋白、载脂蛋白Ⅲ样蛋白均在浆蜂唾液腺中高表达,说明浆蜂唾液腺信息素和蜂王浆蛋白质合成较意蜂旺盛。【结论】浆蜂、意蜂唾液腺具有相似的核心蛋白质组以保证蜂王浆蛋白质、信息素和转化酶的合成与分泌。经过长期选育,浆蜂、意蜂唾液腺分子基础出现差异,浆蜂哺育蜂唾液腺较意蜂蛋白质合成、氨基酸代谢旺盛,细胞供能、抗逆性加强且分泌性蛋白普遍在浆蜂唾液腺上调表达,为浆蜂提供了更加持久和高效的蛋白质合成系统,促成了蜂王浆的高产。
【Objective】The objective of this study is to investigate the proteome profile of the postcereberal gland(PGld)and thoracic gland(ThGld)between high royal jelly producing bees(Apis mellifera liguatica,RJBs)and Italian bees(Apis mellifera liguatica,ITBs)with the aim of revealing the molecular basis of salivary gland regulating royal jelly production,and to provide a basis for analyzing the high-yield mechanism of royal jelly.【Method】PGld and ThGld were dissected from nurse bees of RJBs and ITBs.After protein extraction and enzyme digestion,the peptide samples were analyzed by liquid chromatography coupled with tandem mass spectrometry.Furthermore,the mass spectral data were qualified and quantified by MaxQuant software,and the following bioinformatic analysis was conducted using Perseus software,prediction of secretory protein was achieved by SignalP database,biological process and KEGG pathway were enriched by Cluego software.【Result】Totally 2335 proteins were identified in salivary glands of RJBs and ITBs nurse bees,including 1823 proteins in PGld and 1922 proteins in ThGld.The expression profiles of the core proteins in the PGld and ThGld of RJBs and ITBs were similar,mainly involved in RNA metabolism,nucleic acid metabolism,ATP metabolism,protein translation,translation regulation and catabolism.The principal component analysis(PCA)showed that the molecular basis of salivary gland of RJBs and ITBs had exerted varying extent of differentiation during the selective breeding.Quantitatively,the PGld of ITBs and RJBs expressed 254 and 333 up-regulated proteins,respectively,corresponding to the small molecule and carbohydrate metabolism pathway in ITBs,and the organic nitrogen compound synthesis,cell redox homeostasis,amino acid metabolism in RJBs.Those proved that the protein synthesis,amino acid metabolism and energy supply of salivary gland cells in RJBs were more active than ITBs.In the same way,the up-regulated expressions of 412 and 162 proteins were detected in the ThGld of ITBs and RJBs,respectively,which involved in the pathways of oxidative phosphorylation,translation regulation in ITBs,and oxidative phosphorylation,response to toxic substances in RJBs,indicating that the level of resistance of RJBs ThGld cells was increased.A total of 43 secretory proteins were identified in the salivary glands of RJBs and ITBs,in which 15 were detected in royal jelly.Major royal jelly proteins 1,2,3,4,5 and 7 were detected in PGld and ThGld,indicating that both of the glands were involved in the synthesis of royal jelly main protein.The identification ofα-glucosidase related to nectar transformation and odorant binding proteins 3,13,17 and 21 involved in the synthesis and release of chemical pheromones in both PGld and ThGld indicated their basic function of nectar transformation and pheromone synthesis.The enhanced expression of major royal jelly proteins 1,2,3 and 7,hexamerin 70a and 110,odorant binding proteins 3,13,17 and 21,transferrin and apolipophorin-Ⅲ-like protein in the salivary gland of RJBs demonstrated that the synthesis of pheromone and royal jelly protein was stronger than ITBs.【Conclusion】The core proteome with similar pattern in salivary glands of RJBs and ITBs ensures the synthesis and secretion of royal jelly protein,pheromone and invertase.Molecular variation between the salivary glands of RJBs and ITBs was developed after long-term selective breeding.Relative to ITBs,the salivary glands of RJBs enhance the ability of protein synthesis,amino acid metabolism,cell energy supply and stress resistance,and upregulate the expression of most secretory proteins,which benefits to a better protein synthesis system with promising efficiency and lasting,and contributes to the high producing of royal jelly.
作者
王荣华
孟丽峰
冯毛
房宇
魏俏红
马贝贝
钟未来
李建科
WANG RongHua;MENG LiFeng;FENG Mao;FANG Yu;WEI QiaoHong;MA BeiBei;ZHONG WeiLai;LI JianKe(Institute of Apicultural Research,Chinese Academy of Agricultural Science,Beijing 100091)
出处
《中国农业科学》
CAS
CSCD
北大核心
2022年第13期2667-2684,共18页
Scientia Agricultura Sinica
基金
国家现代农业产业技术体系(蜜蜂)(CARS-44)
中国农业科学院科技创新工程(CAAS-ASTIP-2015-IAR)。