水稻小分子量熱休克蛋白質—OsHSP16.9A關鍵胺基酸分析;Analysis for key amino acid of OsHSP16.9A, a rice small heat shock protein

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Title:	水稻小分子量熱休克蛋白質—OsHSP16.9A關鍵胺基酸分析;Analysis for key amino acid of OsHSP16.9A, a rice small heat shock protein
Authors:	陳韻竹;Chen, Yun-Zhu
Contributors:	生命科學系
Keywords:	熱休克蛋白質;熱逆境;熱休克蛋白質101;HSPs;sHSPs;HSP101
Date:	2022-09-15
Issue Date:	2022-10-04 11:03:39 (UTC+8)
Publisher:	國立中央大學
Abstract:	近年來全球暖化對於作物產量造成嚴重威脅，水稻是世界上重要的糧食作物，如何幫助水稻抵抗惡劣環境以提升作物產量實為現在重要的議題。小分子量熱休克蛋白質（Small heat shock proteins; sHSPs）是植物體內最豐富的一群熱休克蛋白質，其可在熱逆境下降低不正常的蛋白質聚集。實驗室先前發現過量表現OsHSP16.9A水稻轉殖株種子（OsHSP16.9A–OE）較野生型水稻（WT）具更高耐熱性，亦發現當OsHSP16.9A第74個胺基酸從Glutamate （Glu）突變成Aspartate（Asp）時，其與OsHSP101結合的能力就會大幅下降。據此推論，OsHSP16.A的第74個胺基酸對於與OsHSP101形成交互作用十分重要。為了進一步了解它們的特性，我們建構OsHSP16.9A第74個胺基酸Glutamate （Glu；E）為Aspartate （Asp；D）之水稻轉殖株（OsHSP16.9AE74D）。從種子、幼苗耐熱性試驗結果發現， OsHSP16.9A-E74D水稻轉植株在種子及幼苗階段，其耐熱性皆低於OsHSP16.9A–OE，大致與WT相近。此外，我們觀察熱逆境（46℃, 110 min；48℃, 110 min）與低溫逆境（4℃, 3– 14 days）處理後幼苗的電解質滲漏度（Ion leakage），結果發現OsHSP16.9A-E74D轉殖株對高溫及低溫的細胞膜耐受性均較OsHSP16.9A–OE 轉殖株低，綜上所述，我們的結果顯示OsHSP16.9A– OsHSP101間的交互作用在OsHSP16.9A之熱保護機制中扮演重要角色。;In recent years, global warming has posed a serious threat to crop yields. Rice is an important food crop in the world. How to help rice resist harsh environments to improve crop yields is an important issue now. Small heat shock proteins (sHSPs) are the most abundant group of heat shock proteins in plants, which can reduce abnormal protein aggregation under thermal stress. We previously found that OsHSP16.9A overexpressing rice transgenic seeds (OsHSP16.9A–OE) had higher heat tolerance than wild-type rice (WT), and also found that when the 74th amino acid of OsHSP16.9A was mutated from Glutamate (Glu) into Aspartate (Asp), its ability to bind to OsHSP101 was greatly reduced. According to this inference, the 74th amino acid of OsHSP16.A is very important for the interaction with OsHSP101. To further understand their properties, we constructed a rice transgenic line (OsHSP16.9AE74D–OE) in which the 74th amino acid Glutamate (Glu; E) of OsHSP16.9A was Aspartate (Asp; D). Seed and seedling heat tolerance results showed that the heat tolerance of OsHSP16.9AE74D–OE rice transgenic plants was lower than OsHSP16.9A-OE at the seed and seedling stage, and was roughly similar to WT. In addition, we observed the electrolyte leakage (Ion leakage) of seedlings treated under thermal stress (46°C, 110 min; 48°C, 110 min) and low-temperature stress (4°C, 3– 14 days), and found that OsHSP16.9AE74D-OE cell membrane tolerance of transgenic lines to high temperature and low temperature is lower than that of OsHSP16.9A–OE transgenic lines. In conclusion, our results show that the interaction between OsHSP16.9A–OsHSP101 is involved in the thermal protection mechanism of OsHSP16.9A and plays an important role in it.
Appears in Collections:	[Graduate Institute of Life Science] Electronic Thesis & Dissertation

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