| 李卫锦 1，张静文 2，袁长春 1，张 颖 1,2.基于 iTRAQ 技术筛选红榄李种子响应败育差异表达蛋白[J].广东农业科学,2020,47(7):68-78 |
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基于 iTRAQ 技术筛选红榄李种子响应败育差异表达蛋白 |
| Identification of Differentially Expressed Proteins in Abortive Seeds of Lumnitzera littorea Based on iTRAQ |
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| DOI：10.16768/j.issn.1004-874X.2020.07.009 |
| 中文关键词: 红榄李 濒危红树植物 败育机制 蛋白质组 |
| 英文关键词: Lumnitzera littorea endangered mangrove abortion mechanism proteome |
| 基金项目:海南省自然科学基金（418QN240）；国家自然科学基金（41776148，31760119）；岭南师范学院理科重点项目（LZ1906）；湛江市科技计划项目（2018A03024） |
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| 中文摘要: |
| 【目的】探索濒危红树植物红榄李败育种子的分子机制。【方法】借助 iTRAQ 定量蛋白质组学技术分析红榄李不同育性种子蛋白质组的表达差异。对质检合格的两个蛋白质组进行酶解，iTRAQ 标记后进行高效液相色谱、质谱检测。利用 Mascot 软件进行蛋白质鉴定，通过 iTRAQ 定量寻找差异表达蛋白，并进行 GO 功能注释、KEGG 代谢通路和蛋白互作分析，并通过实时荧光定量 PCR 对 8 个差异候选表达基因进行验证。【结果】红榄李种子蛋白质组共鉴定出 2 380 个蛋白，其中差异表达蛋白 448 个，包括上调表达蛋白 238 个和下调表达蛋白 210个。差异蛋白中的 1.86% 与生殖发育的生物学过程密切相关。GO 分析表明，败育种子的上调蛋白主要参与到内质网蛋白的加工、RNA 转运、RNA 降解、囊泡运输中 SNARE 相互作用及 mRNA 调控等生物学过程。差异蛋白中的 UGGT 蛋白、HSP 蛋白和囊泡相关膜蛋白可能在败育种子的发育过程中发挥重要调控作用。【结论】参与种子萌发和植物生长发育的转录因子出现不同程度的差异表达，可能是导致红榄李种子败育的直接或间接原因，研究结果为红榄李濒危机制和种质资源保护研究提供了科学支撑。 |
| 英文摘要: |
| 【Objective】The study was conducted to explore the molecular mechanism of the abortive seeds of endangered mangrove Lumnitzera littorea.【Method】The expression difference in proteome of different fertility seeds of L. littorea was analyzed by iTRAQ quantitative proteomics technology. Enzymolysis was conducted for two qualified proteomes, which were labelled by iTRAQ and then detected by HPLC and MS. Protein identification was carried out by using Mascot software. Differentially expressed proteins were found by iTRAQ. Then, GO function annotation, KEGG metabolic pathway and protein interaction analysis were performed. Furthermore, eight candidate genes were validated by quantitative realtime PCR.【Results】In the two proteomes, 2 380 proteins were identified, and 448 proteins were differentially expressed in the abortive seeds of L. littorea. Among the differentially expressed proteins, 238 of them were up-regulated proteins and 210 were down-regulated ones. 1.86% of the differentially expressed proteins were closely related to the biological processes of reproduction. GO analysis indicated that most of the up-regulated proteins in the abortive seeds were mainly involved into endoplasmic reticulum protein processing, RNA transport, RNA degradation, SNARE interactions in vesicular transport and mRNA regulation. The UGGT proteins, HSP proteins and the vesicle-associated membrane proteins probably play important regulation roles in the development of abortive seeds. These results reflect the possible biochemical and physiological processes of germination of the abortion L. littorea seeds.【Conclusion】Some transcription factors involved in seed germination and plant
growth expressed differently, which may be the direct or indirect reasons for seed abortion. The study results provided scientific support for the research on the mechanism of endangered L. littorea and the conservation of such germplasm resources. |
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