謝旭亮 潘素美　臺灣大學：植物科學研究所陳英謙Chen, Ing-ChienIng-ChienChen2007-11-272018-07-062007-11-272018-07-062007http://ntur.lib.ntu.edu.tw//handle/246246/57959本論文探討三個參與光線訊息傳遞分子的功能，這三個分子為：綠豆VrGIR1 (gibberellic acid [GA]- and IR [infrared]-induced gene 1)、阿拉伯芥AtGASA4 (gibberellic acid-stimulated transcripts in Arabidopsis) 與 AtFIP1 (FIN219[far-red insensitive 219]-interacting protein 1, AtGSTU20)。 在第一部分研究中，由差異表現法篩選到2-5The functions of three light signaling components, VrGIR1 (gibberellic acid [GA]- and IR [infrared]-induced gene 1) in mungbean (Vigna radiata), AtGASA4 (gibberellic acid-stimulated transcripts in Arabidopsis) and AtFIP1 (FIN219[far-red insensitive 219]-interacting protein 1, AtGSTU20) in Arabidopsis (Arabidopsis thaliana) were characterized in this study. In part I, VrGIR1 was isolated by differential display from mungbean seedlings irradiated with 2 to 5誌謝……………………………………………………………………… i 目錄……………………………………………………………………… ii PART I: Both VrGIR1 and AtGASA4, GA- and IR-Stimulated Genes, Participate in Light Signaling in Mungbean and Arabidopsis 中文摘要…………….…………………………………………………… 2 Abstract…………………………………………………………………… 3 Introduction…………………………………………………………….. 4 Materials and methods………………………………………………… 10 Plant Material and Light Treatment………………………………..... 10 RNA Extraction, RNA Gel Blot and RT-PCR Analyses…………….. 10 mRNA Differential Display System………………………………… 11 Complementation Test of the gasa4 Mutant………………………… 12 Response of the gasa4 T-DNA Inserted Mutant to Exogenous Application of GA………………………………………………… 13 Determination of Flowering Time for the gasa4 T-DNA Inserted Line………………………………………………………………… 13 Results………………………………………………………………….. 14 Isolation of IR-Induced Genes VrGIR1 from Mungbean……………. 14 VrGIR1 Is Induced by GA and Displayed Tissue Specific Expression………………………………………………………… 15 VrGIR1 Is an AtGASA-Related Protein……………………………… 16 Expressions of VrGIR1 and AtGASA4 Are Differentially Regulated by Light…………………………………………………………… 17 AtGASA4 Is Regulated by Photoreceptors and Light Signaling Transducers……………………………………………………… 18 The Null Mutant of gasa4 Exhibits a Hyposensitive Phenotype under R and FR Light……………………………………………. 19 The Null Mutant of gasa4 Can Be Rescued by the Ectopic Expressions of AtGASA4……………………………………………. 19 AtGASA4 Represses Several Key Enzyme-Corresponding Genes Involved in GA Metabolism……………………………………… 20 The gasa4 Mutant Exhibits Increased Response to Exogenous GA... 21 The gasa4 Mutant Shows an Earlier Flowering Phenotype under Long-Day Conditions……………………………………………… 22 Discussion……………………………………………………………….. 24 Table………………………………………………………………………. 33 Figures…………………………………………………………………… 34 References………………………………………………………………… 53 Appendix……………………………………………………………….. 60 PART II: Glutathione S-Transferase Interacting with FIN219 Is Involved in Phytochrome A-Mediated Signaling in Arabidopsis 中文摘要………………………………………………………………… 62 Abstract………………………………………………………………….. 63 Introduction……………………………………………………………… 64 Materials and Methods………………………………………………….. 69 Plant Materials and Growth Conditions…………………………….. 69 Yeast Two-Hybrid Assay…………………………………………….. 70 Pull-Down Assay…………………………………………………….. 71 Protein Co-Immunoprecipitation……………………………………… 73 Recombinant Plasmids for Plant Transformation…………………….. 73 Glutathione S-Transferase Activity Assay……………………………. 74 RNA Gel Blot and RT-PCR Analyses……………………………….. 75 Results…………………………………………………………………… 77 Isolation of FIN219-Interacting Partners by Yeast Two-Hybrid Method…………………………………………………………….. 77 FIP1 Encodes a Plant Glutathione S-Transferase with Affinities on the Substrates Glutathione (GSH) and 1-Chloro-2,4- Dinitrobenzene (CDNB)…………………………………………. 79 Transgenic Seedlings Overexpressing or Showing Reduced FIP1 Expression Exhibit a Hyposensitive Phenotype under cFR……… 81 FIP1 Expression Is Regulated by Light Signaling Transducers…….. 83 Transgenic Plants Overexpressing or Reducing FIP1 Expression Displayed a Delayed Flowering Phenotype under Long-Day conditions………………………………………………………… 84 Discussion………………………………………………………………… 86 Figures……………………………………………………………………. 91 Table…………………………………………………………………….. 101 References……………………………………………………………….. 1033159528 bytesapplication/pdfen-US阿拉伯芥遠紅光FIN219FIP1GASA4GibberellinGIR1Glutathione S-transferase紅外光綠豆ArabidopsisFar-redInfraredMungbeanotherhttp://ntur.lib.ntu.edu.tw/bitstream/246246/57959/1/ntu-96-D87226003-1.pdf