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標題: | 探討阿拉伯芥在藍光與茉莉酸處理下去磷酸酶與FIN219/JAR1調控關係的功能性研究 Functional study of the regulatory relationships between FIN219/JAR1 and phosphatases upon exposure to blue light and jasmonates in Arabidopsis |
作者: | 方薇涵 Wei-Han Fang |
指導教授: | 謝旭亮 Hsu-Liang Hsieh |
關鍵字: | FIN219,去磷酸酶,藍光型態發生,茉莉酸訊息傳遞路徑,磷酸化, FIN219,phosphatases,blue-light photomorphogenesis,Jasmonate signal pathway,phosphorylation, |
出版年 : | 2024 |
學位: | 碩士 |
摘要: | 阿拉伯芥可以利用感知藍光的光受體Cryptochromes (CRYs)與植物荷爾蒙茉莉酸(JAs)訊息傳遞,影響植物幼苗的生長發育。活化態茉莉酸(JA-Ile)關鍵的合成酵素FAR-RED INSENSITIVE 219/JASMONATE RESISTANT1 (FIN219/JAR1)扮演著整合藍光與茉莉酸調控幼苗發育的生理功能;先前研究發現黑暗生長並轉移到藍光照射的阿拉伯芥幼苗中的FIN219蛋白質存在磷酸化狀態的改變,但是轉譯後修飾是如何調節FIN219的功能與酵素活性,進而調控藍光及茉莉酸訊息的分子機制目前尚未清楚。先前分析出可能參與在藍光下調控FIN219的去磷酸酶包括屬於蛋白質去磷酸酶2A骨架次單元A的RCN1,以及屬於蛋白質去磷酸酶2C基因家族的E支的EGR2與H支的PP2CR。本研究結果中,將rcn1突變株以及RCN1大量表現的轉殖株幼苗從黑暗轉移到藍光時,發現造成FIN219蛋白質的磷酸化狀態的改變。分析下胚軸外表型,顯示rcn1突變株幼苗對藍光照射以及外源性甲基化茉莉酸(MeJA)抑制下胚軸延長有較敏感的反應。相反地,在相同的處理下大量表達RCN1比起野生型呈現長下胚軸的外表型。此外,在藍光照射以及MeJA處理下rcn1突變株體內花青素含量以及茉莉酸反應的相關基因表現量相似於野生型。進一步檢測fin219-2 rcn1雙突變株的外表型,顯示RCN1與FIN219參與藍光與茉莉酸調控下胚軸生長與茉莉酸相關的反應。綜合目前實驗結果推測,在阿拉伯芥的幼苗生長與發育中,去磷酸酶RCN1透過交互作用改變FIN219磷酸化狀態進而影響FIN219在藍光以及茉莉酸訊息傳遞的調控機制。 Both blue light photoreceptors cryptochromes and the phytohormone jasmonates (JAs) can regulate the growth and development of Arabidopsis seedlings. FAR-RED INSENSITIVE 219/JASMONATE RESISTANT1 (FIN219/JAR1) is a JA-conjugating enzyme and plays a crucial role in the crosstalk between blue light and JA signaling pathways. Previous studies revealed that the phosphorylated FIN219 protein exists in the seedlings transferred from dark to blue light. However, it is unclear that how the posttranslational modifications of FIN219 affect its function and enzymatic activity to control JA signaling upon exposure to blue light. We previously identified phosphatase candidates of FIN219, including ROOTS CURL IN NPA1 (RCN1), a scaffold subunit of Type 2A Protein Phosphatase (PP2A) holoenzymes, the clade H Type 2C protein phosphatase PP2CR, and Clade E Growth-Regulating Type 2C protein phosphatase-2 (EGR2). Here, we primarily conducted immunoblot analysis, indicating that the phosphorylation levels of FIN219 were affected in the rcn1 mutant and RCN1-overexpression line 35S: RCN1-YFP-HA / Col-0 (RCN1-OE) under blue light conditions. The phenotypic analyses of the rcn1 mutant seedlings revealed a hypersensitive response to blue light and methyl-JA (MeJA)-mediated inhibition of hypocotyl growth. In contrast, the RCN1-OE lines had a longer hypocotyl than Col-0 under the same conditions. Furthermore, the rcn1 mutant contained similar anthocyanin content and expression levels of JA-responsive genes compared to Col-0 with MeJA treatment under blue light conditions. Moreover, the fin219-2 rcn1 double mutant showed that RCN1 and FIN219 participated in the control of hypocotyl growth and JA responses with MeJA under blue light conditions. Taken together, these findings reveal that RCN1 phosphatase modulates FIN219 functions in the integration of blue light and JA signaling pathways through direct interaction in Arabidopsis. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/92430 |
DOI: | 10.6342/NTU202400200 |
全文授權: | 同意授權(限校園內公開) |
顯示於系所單位: | 植物科學研究所 |
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