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標題: | 阿拉伯芥熱休克轉錄因子HsfA7a和HsfA7b參與非生物逆境反應之功能性研究 Functional study of heat shock transcription factors HsfA7a and HsfA7b required for abiotic stress responses in Arabidopsis |
作者: | 許宇豪 Yu-Hao Xu |
指導教授: | 靳宗洛 Tsung-Luo Jinn |
關鍵字: | 熱休克轉錄因子,阿拉伯芥,非生物逆境,耐熱性,核糖核酸定序, Heat shock factor,Arabidopsis,abiotic stress,thermotolerance,RNA-seq, |
出版年 : | 2023 |
學位: | 碩士 |
摘要: | 植物熱休克轉錄因子(HSFs)調節包括高溫、乾旱、低溫、鹽度和氧化壓力逆境在內等非生物逆境反應。HSFA7a 和 HSFA7b 在演化上具有親緣關係,暗示著這兩個基因可能共同參與調控非生物逆境反應。利用CRISPR-Cas9技術建立HSFA7a 和 HSFA7b的雙突變株,其在75 mM氯化鈉或100 mM甘露醇琼脂培養基上生長的根長度較野生型短。此外,HSFA7b缺失株在滲透壓逆境下呈現不敏感的表現型。關於下胚軸延長的實驗結果顯示,雙突變植物對於先天耐熱性(BT)不敏感,但在後天短期耐熱性(SAT)條件下呈現敏感的表型。耐熱性分析的結果顯示,在SAT和梯度熱逆境(GHS)下,HSFA7a和HSFA7b共同負調控植物耐熱性;然而HSFA7b單獨正向調控長期後天耐熱性(LAT)。有鑒於這些表型,植株在GHS和LAT熱處理後進行了RNA-seq轉錄體分析,以尋找由HSFA7a和HSFA7b所調控的下游基因。我們觀察到在兩個數據集中關於光合作用相關的上調差異表達基因(DEGs)重疊,暗示這兩個基因可能參與光合作用過程。總而言之,我們的結果突顯了HSFA7a和HSFA7b在不同非生物逆境下的複雜調控機制,並識別出潛在的下游基因,將進一步進行分析。 Plant heat shock factors (HSFs) are crucial in orchestrating responses to various abiotic stresses, including high and low temperatures, drought, salinity, and oxidative stress. HSFA7a and HSFA7b exhibit a shared evolutionary lineage, suggesting their possible collaboration in the regulation of abiotic stress responses. Leveraging CRISPR-Cas9 technology, we generated double mutant lines of HSFA7a and HSFA7b, which exhibited reduced root lengths when exposed to 75 mM sodium chloride or 100 mM mannitol treatments compared to wild-type plants. Furthermore, HSFA7b-deficient mutants displayed insensitivity to osmotic stress. Our experimental findings on hypocotyl elongation indicated that these double mutant plants were less responsive to basal thermotolerance (BT) yet displayed heightened sensitivity under short-term acquired thermotolerance (SAT) conditions. Heat tolerance analyses unveiled that HSFA7a and HSFA7b jointly exerted a negative regulatory influence on plant thermotolerance during SAT and gradient heat stress (GHS) scenarios, with HSFA7b demonstrating independent positive regulation of long-term acquired thermotolerance (LAT). Given these distinctive phenotypes, we conducted RNA-seq transcriptome analyses on plants subjected to GHS and LAT heat treatments to identify downstream genes under the regulatory influence of HSFA7a and HSFA7b. Remarkably, we observed a convergence of upregulated differentially expressed genes (DEGs) associated with photosynthesis in both datasets, suggesting the potential involvement of these genes in the photosynthesis process. In summary, our findings shed light on the complex regulatory mechanisms governing HSFA7a and HSFA7b responses to diverse abiotic stresses and identify candidate downstream genes warranting further investigation. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/91199 |
DOI: | 10.6342/NTU202304437 |
全文授權: | 同意授權(全球公開) |
顯示於系所單位: | 植物科學研究所 |
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