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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/17846

Title: 以質譜技術開發找尋植物內生性胜肽荷爾蒙訊息傳遞分析平台
Development of the Plant Signaling Peptides Discovery Platform Using Mass Spectrometry
Authors: Ying-Lan Chen
陳盈嵐
Contributors: NTOU:Institute of Bioscience and Biotechnology
國立臺灣海洋大學:生物科技研究所
Keywords: 番茄;胜肽荷爾蒙;質譜技術;定量;受傷反應
tomato;peptide hormones;mass spectrometry;quantitation;wound response;systemin
Date: 2009
Issue Date: 2011-07-04
Abstract: 有鑑於目前缺乏胜肽荷爾蒙的分析平台,所以已知的植物胜肽荷爾蒙只有十幾個,但植物的胜肽傳遞 (Peptide transporter) 基因卻遠遠多於其他生物,所以此研究首先著重在開發更廣泛的分析平台,結合質譜分析技術來鑑定胜肽質體 (Peptidomics) ,藉此尋找有活性的胜肽片段。以最早被發現的植物胜肽荷爾蒙”systemin”來作為開發分析平台的研究對象。 Systemin 為一個反應機械性傷害或蟲咬傷害的胜肽訊號,最早是從番茄葉中純化出來,目前已被發現在大多數茄科植物上。相較於以往的鑑定胜肽訊號的方法,在此研究中我們能使用較少量的番茄葉樣品以及更簡短的純化步驟,將所得到的胜肽混合物用串聯式液相層析質譜儀去鑑定其序列,再利用自製的胜肽資料庫 (Hypothetical peptide database) 比對,可更快速的檢視植物體內的胜肽質體 (Peptideomics) 表現。加上質譜定量方法,比較受傷刺激前後的胜肽表現,有顯著增加的胜肽即可認為其為受傷反應相關的訊號。在此研究中已發現番茄葉在受傷之後,除了systemin之外,尚有某些胜肽片段會被大量產生,且其前驅蛋白經比對後發現大多為光反應系統 (Photosystem) 或病源體相關 (Phathogenesis-related;PR) 蛋白。此研究平台對於植物的胜肽質體 (Peptideomics) 鑑定及找尋有活性的胜肽訊號能提供更廣泛,更快速的分析平台。
Increasing number of the studies revealed that signaling peptides were involved in plant growth and development. Compared with other organism, plant was predicted to have more signaling peptides and corresponding perceptive receptors. However, the small size of peptide molecules and their cDNA lead to the analysis of signaling peptides more difficult. To date, a limited number of peptide hormones in plant were observed. This is due to the lack of analytical platforms for the study of signaling peptides. In this study, we have developed a comprehensive analytic strategy for discovering the signaling peptides by the combination of liquid chromatography-tandem mass spectrometry (LC-MS/MS), hypothetical peptide databases, and optimized peptide purification protocol. In previous study, the insect attack or mechanical wound will induce systemin in Solanaceae, which is the first discovered peptide hormone in plant. To exam the feasibility of the signaling peptide discovery platform, systemin was used as the model that isolated from tomato wounded leaves. In addition to the previous discovered wound peptide signal (systemin); several new signaling peptides were identified and their precursors mostly are photosystem and phathogensis-related (PR) protein. Therefore, this peptide discovery platform could facilitate the screening of novel signaling peptides and their precursors.
URI: http://ethesys.lib.ntou.edu.tw/cdrfb3/record/#G0M96360038
http://ntour.ntou.edu.tw/ir/handle/987654321/17846
Appears in Collections:[生命科學暨生物科技學系] 博碩士論文

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