研究论文 RESEARCH PAPER

基于相对和绝对定量同位素标记技术分析2种成熟度桑叶对家蚕脂肪体蛋白质组的影响

  • 谢岩 ,
  • 王晖 ,
  • 李娜 ,
  • 贾漫丽 ,
  • 黄露 ,
  • 孙志超 ,
  • 李季生
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  • 承德医学院蚕业研究所, 河北省高校特产蚕桑应用技术研发中心, 承德 067000
谢岩(1985—),女,河北承德人,助理研究员,博士,从事家蚕分子生物学研究工作。E-mail:308229509@qq.com

收稿日期: 2021-10-10

  网络出版日期: 2022-05-14

基金资助

河北省自然科学基金青年项目(C201806033);河北省高等学校科学技术研究项目(BJ2018032);河北省科技厅"技术创新引导专项-科技工作会商"项目;承德医学院高层次人才科研启动基金项目(201604)

Analysis on Effects of Mulberry Leaves at Two Maturities on Fat Body Proteome in Silkworm Based on Isobaric Tags for Relative and Absolute Quantitation Technology

  • XIE Yan ,
  • WANG Hui ,
  • LI Na ,
  • JIA Manli ,
  • HUANG Lu ,
  • SUN Zhichao ,
  • LI Jisheng
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  • Applied Technology R&D Center of Sericulture and Specialty of Hebei Universities, Institute of Sericulture of Chengde Medical University, Chengde 067000, China

Received date: 2021-10-10

  Online published: 2022-05-14

摘要

本试验旨在探索家蚕体适应桑叶营养物质变动的微观机制。首先检测老、嫩2种成熟度桑叶的水分、粗蛋白质、粗脂肪、可溶性糖、还原性糖、多糖、总黄酮、总多酚和1-脱氧野尻霉素(1-DNJ)含量,然后采用相对和绝对定量同位素标记(iTRAQ)技术分析由这2种桑叶饲喂3 d的5龄家蚕脂肪体蛋白质组表达谱,并对差异表达蛋白进行生物信息学分析。结果显示:嫩桑叶的粗蛋白质、粗脂肪、还原性糖、总黄酮、总多酚、1-DNJ含量极显著高于老桑叶(P<0.01)。iTRAQ定量分析共鉴定到家蚕脂肪体蛋白2 172个,2组间差异表达蛋白173个。相对于摄食老桑叶的家蚕,摄食嫩桑叶的家蚕脂肪体中上调表达蛋白为85个,下调表达蛋白为88个。对其中的111个差异表达蛋白[差异表达倍数(FC)>1.25或<0.80]的功能富集分析发现,GO和KEGG通路注释到这些蛋白主要参与物质代谢、运输和贮存,应激和免疫反应,机体发育等生物学过程。差异表达蛋白互作网络分析筛选到核糖体蛋白和神经前体细胞表达发育下调蛋白8(NEDD8)的网络中心性高。综上可知,桑叶成熟度显著影响其营养物质含量,引起摄食老、嫩桑叶家蚕脂肪体蛋白质组灵敏、广泛的差异响应。

本文引用格式

谢岩 , 王晖 , 李娜 , 贾漫丽 , 黄露 , 孙志超 , 李季生 . 基于相对和绝对定量同位素标记技术分析2种成熟度桑叶对家蚕脂肪体蛋白质组的影响[J]. 动物营养学报, 2022 , 34(5) : 3271 -3285 . DOI: 10.3969/j.issn.1006-267x.2022.05.053

Abstract

This research was to explore the micromechanism of silkworm body adapting to the changes of nutrients in mulberry leaves. The contents of nutrients and active components of tender and aged mulberry leaves, these components including moisture, crude protein, crude fat, soluble sugar, reduced sugar, polysaccharide, total flavonoid, total polyphone and 1-deoxynojirimycin (1-DNJ), were detected and compared. The proteomic profile of fat body from the 5th instar silkworm fed for 3 days with tender and aged mulberry leaves were obtained using isobaric tags for relative and absolute quantitation (iTRAQ) technology, the differentially expressed proteins were also screened and analyzed by bioinformatics method. The results showed that the contents of crude protein, crude fat, reduced sugar, total flavonoid, total polyphone and 1-DNJ tender mulberry leaves were extremely significantly higher than those in aged mulberry leaves (P<0.01). The total of 2 172 fat body proteins were detected by iTRAQ technology, and there were 173 differentially expressed proteins between two groups. Compared with the aged mulberry leaves group, the tender mulberry leaves group had 88 down-regulated expressed proteins and 85 up-regulated expressed proteins. For 111 differentially expressed proteins[fold change (FC)>1.25 or <0.80] of them were used for functional enrichment analysis. GO and KEGG pathway annotation showed these proteins were mainly involved in the biological processes such as substance metabolism, transportation and storage, stress and immune response, body development. Differentially expressed protein interaction analysis found that the ribosomal proteins and neural precursor cell-expressed developmentally down-regulated ptotein 8 (NEDD8) protein had high centrality in interaction network. In conclusion, the maturity of mulberry leaves significantly affects its nutrient contents, resulting the sensitive and extensive differential responses of fat body proteome of silkworm fed tender or aged mulberry leaves.

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