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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 謝旭亮(Hsu-Liang Hsieh) | |
| dc.contributor.author | Yu-An Chen | en |
| dc.contributor.author | 陳語安 | zh_TW |
| dc.date.accessioned | 2021-06-17T00:23:07Z | - |
| dc.date.available | 2014-07-18 | |
| dc.date.copyright | 2012-07-18 | |
| dc.date.issued | 2012 | |
| dc.date.submitted | 2012-06-03 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/66136 | - |
| dc.description.abstract | 阿拉伯芥中 FAR-RED INSENSIVIVE 219 (FIN219)/JASMONATE RESISTANT 1 (JAR1)與 CORONATINE INSENSITIVE 1 (COI1)是茉莉酸訊息傳遞中兩個正 向調控因子。FIN219/JAR1 催化茉莉酸鍵結上異亮氨酸,而形成具有生理活性的 茉莉酸衍生物,此衍生物為SFCCOI1與JASMONATE-ZIM-DOMAIN PROTEINs (JAZs)結合的配合體,進而促成茉莉酸訊息傳遞抑制者 JAZs 降解。近期的文 獻指出 FIN219 與 COI1 都參與遠紅光訊息傳遞,此研究立基於釐清倆者在遠紅 光訊息傳遞以及茉莉酸訊息傳遞中的調控關係。我們發現 FIN219 與 COI1 在調 控不稔性以及對遠紅光的敏感性上,有加成性的效果;但 FIN219 大量表現造成 的短下胚軸性狀會被 COI1 大量表現抑制。以大腸桿菌表現倆者蛋白質的方式與 酵母菌雙雜合實驗的結果都顯示倆者可互相結合;在阿拉伯芥的原生質體中,倆 者共同坐落在細胞核內。藉由篩選極研究各種大量表現 FLAG-COI1 的轉殖株, 因而發現茉莉酸的感知及訊息傳遞會促進 COI1 的累積,但是 FIN219 卻對此有 抑制的效果。在遠紅光下,COI1 大量表現可以抑制 fin219-2 突變株具有較長下 胚軸的性狀;此外,外加茉莉酸後,倆者可以透過獨立於彼此的方式,促成 HY5 的累積。在以上實驗中,我們呈現 FIN219 與 COI1 在遠紅光及茉莉酸訊息傳導 中相互調控的關係,顯示出訊息傳導中調校與交互作用的現象。 | zh_TW |
| dc.description.abstract | In Arabidopsis, FAR-RED INSENSIVIVE 219 (FIN219)/JASMONATE RESISTANT 1 (JAR1) and CORONATINE INSENSITIVE 1 (COI1) are positive regulators in jasmonate (JA) signaling. FIN219/JAR1 catalyzes the synthesis of JA-isoleucine (JA-Ile), which is a bioactive conjugate of JA that promotes the binding of SFCCOI1 and JASMONATE-ZIM-DOMAIN PROTEINs (JAZs). This binding results in the degradation of JA signaling repressors JAZs by 26S proteasome. Recent publications indicate that both FIN219 and COI1 are involved in phytochrome A mediated high irradiance response during seedling de-etiolation. This study aimed to elucidate the relation of FIN219 and COI1 in FR light and JA signaling. We found that FIN219 and COI1 synergistically regulate fertility and FR light sensitivity. However, COI1 overexpression may suppress the effect of FIN219 overexpression on hypocotyl growth by post-transcriptional regulation. The results of pull-down and yeast two-hybrid assays suggest that FIN219 and COI1 may interact. We also showed both proteins can co-localize in the nuclei of Arabidopsis protoplasts. By generating FLAG-COI1 overexpression plants, we showed that JA perception/signaling promotes COI1 accumulation, whereas FIN219 represses this phenomenon. In FR light, COI1 overexpression suppressed fin219-2 long-hypocotyl phenotype. Both FIN219 and COI1 contribute to HY5 accumulation upon JA activation in FR light, which FIN219 and COI1 may exert their function synergistically. Taken together, the regulatory relationship of FIN219 and COI1 is fine-tuningly modulated and has a crosstalk between FR light and JA signalings. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-17T00:23:07Z (GMT). No. of bitstreams: 1 ntu-101-R98b42011-1.pdf: 29333932 bytes, checksum: d597ebf2219881f66d2e68f2af3999b2 (MD5) Previous issue date: 2012 | en |
| dc.description.tableofcontents | INTRODUCTION 1
Light, Photoreceptors, and Phytochromes 1 Seedling De-etiolation and PhyA Signaling 1 HY5 and COP1 2 FIN219/JAR1 and COP1 3 JA Signaling 4 Crosstalk Between Light and JA Signaling 5 MATERIALS AND METHODS 6 Plant Materials 6 Plasmid Constructions and Transgenic Plants 6 Plant Growth Conditions 7 Total Genomic DNA Isolation for Genomic PCR 7 Genotyping of coi1-16 8 RNA Extraction, DNase Treatment, and Reverse Transcription 8 Yeast Two-Hybrid Assays 9 Protoplast Transfection and BiFC Analysis 9 Total Protein Isolation and Western Blotting 10 Chlorophyll Extraction and Quantification 10 Measurement of Hypocotyl and Roots 10 Statistic Analysis 10 RESULTS 11 FIN219 and COI1 synergistically regulate fertility and FR light sensitivity. 11 COI1 may negatively regulate FIN219 functions under FR light 12 Ectopic expression of COI1 results in the suppression of FIN219 overexpressor phenotype in the dark. 13 Yeast two-hybrid assays reveal that C terminus of FIN219 interacts with COI1, and the interactions are not enhanced by JA or MeJA. 15 COI1-GFP was localized in the nucleus when transiently expressed in the protoplast. 16 FIN219 may have a negative role in the regulation of COI1 levels in response to MeJA or coronatine. 17 COI1 overexpression in fin219-2 complements long-hypocotyl phenotype in FR light. 17 35S::FLAG-COI1 complements coi1-16 phenotype in a quantity-dependent manner. 18 There is a G-box related binding site of MYC2 in the promoter region of HY5. 19 Seedlings grown in the medium supplemented with MeJA can accumulate more chlorophyll after transfer from FR light to white light. 20 DISCUSSION 21 FIN219 is regulated by COI1 in pGR::FIN219 transgenic lines. 22 FR light signaling in pGR::FIN219 transgenic plants: a combinatory effect of FIN219 and COI1-mediated JA signaling. 23 FLAG-COI1 level is affected by JA perception/signaling. 24 JA signaling increases HY5 level in FR light. 25 FIGURES 28 Figure 1. coi1-16 and fin219-2 are insensitive to FR light inhibition of hypocotyl elongation. 28 Figure 2. coi1-16 fin219-2 double mutants exhibit more severe defects in fertility and FR light sensitivity. 29 Figure 3. coi1-16 mutation has little effects on DEX treated pGR::FIN219 lines. 30 Figure 4. COI1 suppresses FIN219 accumulation in pGR::FIN219 transgenic line treated with DEX. 31 Figure 5. COI1 overexpression suppresses FIN219 overexpressor phenotype in the dark. 32 Figure 6. MG132 or MG115 treatment dose not restore FIN219 level in transgenic lines. 33 Figure 7. Ectopic expression of COI1 results in the suppression of FIN219 overexpressor phenotype in the dark and FR light. 34 Figure 8. Ectopic expression of COI1 suppresses FIN219 accumulation in FIN219 overexpressor under darkness and FR light. 35 Figure 9. FIN219 transcript levels in FIN219 overexpressors were suppressed by ectopic expression of COI1. 36 Figure 10. FIN219 interact with COI1 in yeast two-hybrid assay. 37 Figure 11. COI1-GFP localized in the nucleus and COI1 may co-localize with FIN219 in the nucleus. 38 Figure 12. COI1 overexpression suppresses fin219-2 long hypocotyl phenotype in FR light. FLAG-COI1 levels increase in response to MeJA and COR treatment in 35S::FLAG-COI1 /fin219-2 transgenic lines. 39 Figure 13. FIN219 suppresses FLAG-COI1 accumulation under the treatments of MeJA and COR. 40 Figure 14. FLAG-COI1 complements coi1-16 phenotypes. 41 Figure 15. FLAG-COI1 complements long hypocotyl phenotype of coi1-16 in FR light. 42 Figure 16. HY5 may be regulated by MYC2. 43 Figure 17. Treatment of MeJA accelerates chlorophyll synthesis in seedlings transferred from or FR light to white light. 44 Figure 18. Proposed model of regulation between FIN219 and COI1, and crosstalk between JA signaling and light signaling. 45 REFERENCES 46 APPENDIX 51 I. One-step yeast transformation for R919 51 II. Clones and constructs 52 III. qPCR primers and informations 53 | |
| dc.language.iso | en | |
| dc.title | 阿拉伯芥中 FIN219 與 COI1 的交互作用在遠紅光與茉莉酸訊息傳遞中的功能性研究 | zh_TW |
| dc.title | Functional Studies of the Effects of FIN219 and COI1 Interaction on Far-Red Light and Jasmonic Acid Signaling in Arabidopsis thaliana | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 100-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 吳素幸(Shu-Hsing Wu),鄭石通(Shih-Tong Jeng),鄭秋萍(Chiu-Ping Cheng),張英?(Ing-Feng Chang) | |
| dc.subject.keyword | 阿拉伯芥,光型態發生,茉莉酸,遠紅光,交互作用, | zh_TW |
| dc.subject.keyword | Arabidopsis,photomorphogenesis,jasmonate,far-red light,interaction, | en |
| dc.relation.page | 53 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2012-06-04 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 植物科學研究所 | zh_TW |
| 顯示於系所單位： | 植物科學研究所 | |
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