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標題: 水稻台農六十七號及其感鹽突變體SA0604、耐鹽突變體SM75於初期鹽害逆境下蛋白質體分析
Comparative Proteomic Analysis of Initial Salt-stressed Rice Seedlings TNG67 and its Salt-sensitive Mutant SA0604, Salt-tolerant Mutant SM75
作者: 徐裕凱
Hsu, Yu-Kai
關鍵字: salt stress;鹽害逆境;proteomics;蛋白質體
出版社: 分子生物學研究所
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即使過去在植物耐鹽機制上已有豐碩轉錄體相關研究成果,但有鑑於植物在感應鹽分逆境時複雜的分子調節機制,此一課題仍有很大的空間需持續地被研究著。蛋白質體分析技術於近年來蓬勃發展,此項技術可以窺視基因的終產物-蛋白質的質和量,甚至是蛋白質轉譯後修飾。本實驗利用蛋白質體學與質譜技術,欲探討水稻幼苗於初期鹽害逆境下生理以及分子反應機制。選用的水稻材料為TNG67 以及經由疊氮化鈉誘變而成的耐鹽水稻 SM75 與感鹽水稻 SA0604,而具有不同耐鹽程度的植物往往是拿來研究鹽逆境反應機制的最佳材料。生長19天的三種水稻幼苗(三葉齡)以250 mM 之氯化鈉處理0.5小時,再將萃取的葉部及根部蛋白質以二維膠體電泳分析,並以比對軟體分析於初期鹽逆境下是否具有表現量變化之蛋白質點,經由胰蛋白酶水解後再以 LC-MS/MS 分析,並搭配蛋白質資料庫比對鑑定具表現量差異之蛋白質身份。
實驗結果發現,TNG67 的葉部蛋白體於初期鹽害逆境下共有31個蛋白質表現差異點,而 SM75 及 SA0604 分別有33及16個蛋白質表現差異點。其鑑定的結果中,有若干蛋白質在三種不同耐鹽程度的水稻植株中,有不同的消長現象,此些蛋白質可能決定了水稻生理上不同耐鹽程度,包含了 Cysteine synthase、Fructokinase 2、3-O-methyltransferase 1、ATP synthase 等等。此外,有鑑於轉譯後修飾如泛素修飾及磷酸化修飾蛋白被多篇研究證實會參與植物耐鹽分子機制,我們以質譜儀技術找出數個有被修飾的蛋白質。希望此研究成果有利於未來植物生理之研究,甚至可以提供基因改造策略以增進水稻於鹽害逆境下耐受性。

Rice (Oryza sativa L.) serves as a staple food source for more than one-third of the world’s population. Rice growth and productivity are limited by salt stress which is a major abiotic stress in agriculture worldwide. Increased salinization of arable land is predicted with 50% land loss by the year 2050; hence it is important to understand plant tolerance to salinity that is fundamental to develop stress tolerant crops.
Although this topic has long been studied by transcriptome-based studies, it is still required for dissecting due to the complicated regulatory mechanism of salt adaptation in plant. Recently the proteomics have been developed more widespread, the techniques can offer insights into the quantity and quality of the final gene products even the post-translaitonal modifications. In this study, we aim to elucidate the physiological and molecular mechanism response to initial salt stress in rice by using proteomics method. The chosen rice strains TNG67, salt-tolerant mutant SM75 and salt-sensitive strain SA0604 derived from TNG67 by sodium azide (NaN3) mutagenesis were valuable for this research of plant salt-responsive mechanism. 19 days-old rice seedlings of three strains were treatrd with 250 mM NaCl for 0.5 h. Total proteins of leaves and roots were extracted for comparative proteomics analysis via 2D electrophoresis and LC-MS/MS analysis. The result showed 31 differentially expressed proteins spots of leaf proteome in TNG67, 33 in SM75 and 16 in SA0604. Several identified proteins like Cysteine synthase、Fructokinase 2、3-O-methyltransferase 1、ATP synthase may be potential for gene improved strategy of salt tolerance. Further, the post-translational modification such as phosphorylation and ubiquitination in proteins which have been discussed involved in plant salt adaptation were also analyzed in our research. These identified proteins may be helpful in elucidating the molecular basis of salt tolerance in plant.
其他識別: U0005-1808201017233500
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