摘要
【目的】探讨移虫龄期对西方蜜蜂Apis mellifera蜂王卵巢发育的分子调控机制。【方法】运用相对和绝对定量同位素标记(iTRAQ)技术对不同龄期移虫培育的蜂王卵巢组织蛋白质进行定量分析,筛选差异表达蛋白。利用蛋白质免疫印迹法对结果进行验证。【结果】iTRAQ定量分析共鉴定到蜂王卵巢组织蛋白质二级图谱452 966个,最终获得3 642个蛋白。GO(Gene Ontology)富集结果表明,不同龄期移虫发育的蜂王卵巢组织差异表达蛋白主要富集在细胞代谢、分裂以及蛋白质合成。Pathway富集分析表明,1龄幼虫期移虫和2龄幼虫期移虫发育的蜂王卵巢组织差异表达蛋白主要富集在碳水化合物代谢、脂代谢和外源降解类群;1龄幼虫期移虫和3龄幼虫期移虫发育蜂王卵巢组织差异蛋白主要富集在有机体发育通路、核糖体通路和溶酶体代谢。与此同时,两个差异表达蛋白即储存蛋白Hexamerin110和Hexamerin70b的蛋白免疫印迹检测结果表明,随着移虫龄期的增加,Hexamerin110和Hexamerin70b的表达量均呈现降低的趋势。【结论】对不同龄期移虫的蜂王之间差异表达蛋白进行鉴定,为进一步研究蜂王生殖发育及级型分化的调控机制提供理论依据。
【Aim】To explore the molecular regulation mechanism of grafted larvae on ovarian development of queens in the Western honey bee,Apis mellifera.【Methods】The isobaric tags for relative and absolute quantitation（i TRAQ）method was used to quantify the proteins in the ovaries of queens reared from the larvae grafted at different instars and screen the differentially expressed proteins（DEPs）.The results were validated by Western blotting.【Results】A total of 452 966 spectra were obtained and 3 642 proteins were identified from ovaries of A.mellifera queens.Gene Ontology（GO）analysis revealed that the DEPs in the ovaries of queens reared from the larvae grafted at different instars are associated with cell metabolism,cell division,and protein biosynthesis.The DEPs in the ovaries of queens reared from the larvae grafted between the 1st and 2nd instars are enriched in such pathways as carbohydrate metabolism,lipid metabolism,and exogenous degradation,while those reared from the larvae grafted between the 1st and 3rd instars mainly enriched in developmental pathways,ribosome pathways,and lysosome metabolism.The Western blotting results of the two differentially expressed storage proteins hexamerin 110 and hexamerin 70b showed that their expression levels decreased with the grafted larval instar.【Conclusion】The identification of differentially expressed proteins in the ovaries of A.mellifera queens reared from the larvae grafted at different instars provides a foundation for the regulatory mechanism of queen reproductive development and caste differentiation.
作者
庞倩
沈芳
王康
张文文
殷玲
吉挺
PANG Qian SHEN Fang WANG Kang ZHANG Wen-Wen YIN Ling JI Ting(College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu 225009, China Fuyang Animal Husbandry and Veterinary Burean, Fuyang, Anhui 236000, China Jiangsu Agri-Animal Husbandry Vocational College, Taizhou, Jiangsu 225300, China)
出处
《昆虫学报》
CAS
CSCD
北大核心
2017年第7期760-771,共12页
Acta Entomologica Sinica
基金
国家自然科学基金项目(31502020,31372382)
关键词
西方蜜蜂
移虫
龄期
卵巢
蜂王
蛋白质组学
差异表达蛋白
Apis mellifera
grafted larva
instar
ovary
queen
proteomics
differentially expressed proteins
