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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 謝旭亮 | zh_TW |
| dc.contributor.advisor | Hsu-Liang Hsieh | en |
| dc.contributor.author | 方薇涵 | zh_TW |
| dc.contributor.author | Wei-Han Fang | en |
| dc.date.accessioned | 2024-03-22T16:28:19Z | - |
| dc.date.available | 2024-03-23 | - |
| dc.date.copyright | 2024-03-22 | - |
| dc.date.issued | 2024 | - |
| dc.date.submitted | 2024-01-31 | - |
| dc.identifier.citation | Ahmad, M. and Cashmore, A.R. (1993). HY4 gene of A. thaliana encodes a protein with characteristics of a blue-light photoreceptor. Nature 366: 162–166.
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(2014). Molecular basis for jasmonate and ethylene signal interactions in Arabidopsis. Journal of Experimental Botany 65: 5743–5748. | - |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/92430 | - |
| dc.description.abstract | 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。 | zh_TW |
| dc.description.abstract | 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. | en |
| dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2024-03-22T16:28:19Z No. of bitstreams: 0 | en |
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| dc.description.tableofcontents | 誌謝 II
中文摘要 III Abstract IV Contents VI List of Figures X List of Tables XII Abbreviations XIII Introduction 1 1. Blue light signaling modulates photomorphogenesis 1 2. Plant hormones affect seedling growth and development 2 3. FIN219 integrates blue light and jasmonate signaling 3 4. FIN219 regulatory activity and post-translational modifications 4 5. The blue-light dependent phosphatase candidates of FIN219 5 6. The motivation and objectives of this study. 7 Materials and methods 8 1. Plant materials and growth conditions 8 2. Plasmid construction and generation of transgenic lines 8 3. Light treatment 9 4. Hypocotyl length measurement 9 5. Seedlings treated with MeJA 9 6. RNA isolation and gene expression analysis 9 7. Quantitative real-time PCR (RT-qPCR) 10 8. Anthocyanins content analysis 10 9. Plant total protein extraction 11 10. Western blot analyses 11 11. Co- immunoprecipitation 12 12. Accession Numbers 12 Results 13 1. RCN1 participated in the regulation of hypocotyl growth under blue light. 13 2. FIN219 was dephosphorylated by RCN1 under blue light. 14 3. RCN1 regulated the JA signaling and anthocyanin accumulation in the presence of MeJA under blue light. 15 4. RCN1 did not physically interact with FIN219 in vivo. 17 5. FIN219 is epistatic to RCN1 in seedling development with MeJA under blue light conditions. 18 6. RCN1 and EGR2/PP2CR independently regulated hypocotyl growth and JA responses with MeJA under blue light conditions. 19 7. RCN1 dominantly regulated hypocotyl phenotypes of the seedlings grown in darkness. 20 Discussion 22 1. RCN1 regulated FIN219 dephosphorylation under blue light. 22 2. RCN1 functions in the regulation of seedling development with MeJA treatment in blue light condition. 23 3. RCN1 mediated FIN219-induced JA responses under blue light. 24 4. FIN219 and RCN1 genetically regulated hypocotyl development and JA responses in blue light. 25 5. The phosphatase candidates of FIN219 in blue light. 27 6. RCN1 regulated hypocotyl phenotype in dark-grown seedlings. 28 Figures 30 Figure 1. The rcn1 mutant displayed a shorter hypocotyl phenotype than Col-0 under blue light conditions. 30 Figure 2. RCN1-OE suppressed photomorphogenesis with MeJA in blue light. 31 Figure 3. The protein phosphorylation assay of FIN219 in the rcn1 mutant under blue light and dark conditions. 33 Figure 4. RCN1 regulated FIN219 protein phosphorylation in blue light and MeJA treatment. 34 Figure 5. JAZ1, VSP1, and LOX2 transcript levels were up-regulated in the RCN1-OE lines with MeJA under blue light. 35 Figure 6. RCN1 regulated anthocyanin accumulation in the presence of MeJA under blue light. 36 Figure 7. In vivo co-immunoprecipitation did not show the interaction between FIN219 and RCN1-HA under blue light and darkness. 38 Figure 8. Genetic interaction analysis showed that RCN1 and FIN219 regulated hypocotyl growth and anthocyanin pigments under blue light. 39 Figure 9. Expression levels of JA-responsive genes were downregulated in the rcn1 mutant with MeJA treatment under blue light. 41 Figure 10. The egr2-1 pp2cr-1rcn1 triple mutant had similar hypocotyl lengths to Col-0 under blue light. 42 Figure 11. FIN219 phosphorylation in the egr2-1 pp2cr-1rcn1 triple mutant exposed to blue light and MeJA treatment. 44 Figure 12. The egr2-1 pp2cr-1rcn1 triple mutant had similar expression levels of JA responsive genes to Col-0 in blue light and MeJA treatment. 45 Figure 13. RCN1 regulated hypocotyl growth in response to the dark. 46 Figure 14. The MYC2 expression levels with MeJA in the dark. 48 Figure 15. A proposed model shows how RCN1 dephosphorylated FIN219 to regulate hypocotyl development and JA responses with MeJA under blue light conditions. 49 Table 50 Table 1. Primer sequences used in this study. 50 References 52 | - |
| dc.language.iso | en | - |
| dc.subject | 磷酸化 | zh_TW |
| dc.subject | 茉莉酸訊息傳遞路徑 | zh_TW |
| dc.subject | 藍光型態發生 | zh_TW |
| dc.subject | 去磷酸酶 | zh_TW |
| dc.subject | FIN219 | zh_TW |
| dc.subject | FIN219 | en |
| dc.subject | phosphatases | en |
| dc.subject | blue-light photomorphogenesis | en |
| dc.subject | Jasmonate signal pathway | en |
| dc.subject | phosphorylation | en |
| dc.title | 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 | zh_TW |
| dc.title | Functional study of the regulatory relationships between FIN219/JAR1 and phosphatases upon exposure to blue light and jasmonates in Arabidopsis | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 112-1 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.oralexamcommittee | 涂世隆;蔡皇龍;李金美;鄭梅君 | zh_TW |
| dc.contributor.oralexamcommittee | Shih-Long Tu;Huang-Lung Tsai;Chin-Mei Lee;Mei-Chun Cheng | en |
| dc.subject.keyword | FIN219,去磷酸酶,藍光型態發生,茉莉酸訊息傳遞路徑,磷酸化, | zh_TW |
| dc.subject.keyword | FIN219,phosphatases,blue-light photomorphogenesis,Jasmonate signal pathway,phosphorylation, | en |
| dc.relation.page | 65 | - |
| dc.identifier.doi | 10.6342/NTU202400200 | - |
| dc.rights.note | 同意授權(限校園內公開) | - |
| dc.date.accepted | 2024-02-02 | - |
| dc.contributor.author-college | 生命科學院 | - |
| dc.contributor.author-dept | 植物科學研究所 | - |
| dc.date.embargo-lift | 2029-01-23 | - |
| 顯示於系所單位： | 植物科學研究所 | |
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