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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 李勇毅 | zh_TW |
| dc.contributor.advisor | Yung-I Lee | en |
| dc.contributor.author | 徐菡憶 | zh_TW |
| dc.contributor.author | Han-I Hsu | en |
| dc.date.accessioned | 2025-08-18T01:16:29Z | - |
| dc.date.available | 2025-08-18 | - |
| dc.date.copyright | 2025-08-15 | - |
| dc.date.issued | 2025 | - |
| dc.date.submitted | 2025-08-06 | - |
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Ovary and gametophyte development are coordinately regulated by auxin and ethylene following pollination. Plant Cell 5:403-418. | - |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/98665 | - |
| dc.description.abstract | 蝴蝶蘭(Phalaenopsis)的種子在成熟時缺乏頂端分生組織，於種子萌芽後才逐步發育出頂端分生組織以及葉片與根系。此發育模式有別於大多數被子植物於胚胎早期即完成分生組織定位。為探討此特殊時序下頂端分生組織建立的分子調控機制，本研究分析WUSCHEL-related homeobox（WOX）家族與生長素訊號在蝴蝶蘭種子及原球莖發育階段中的表現與分布。首先，根據胚珠與原球莖的形態特徵與組織學觀察來定義不同發育階段，並透過quantitative real-time PCR與轉基因報導植株（DR5::GUS）進行WOX基因與生長素的不同發育階段表現分析。結果顯示，PaWUS以及PaWOX3a、PaWOX4、PaWOX5、PaWOX9等基因於胚珠發育期間表現量顯著上升，顯示其可能參與組織分化與分生組織形成之調控。在原球莖發育期間，PaWOX3a、PaWOX4 與 PaWOX5 於頂端區域有較高的表現，推測其可能參與頂端分生細胞群的建立。DR5::GUS染色亦顯示生長素在原球莖內呈現極性分佈，並於葉原基與根原基高度累積，顯示生長素在器官發育中所扮演的角色。外施 IAA 與 NAA實驗顯示生長素可促進原球莖生長與不定根形成，反之TIBA處理則導致莖頂、葉片與根部發育異常。此外本研究亦成功建立 WOX 啟動子::GUS 之轉基因構築並初步應用於蝴蝶蘭轉殖。整體而言，本論文研究探討WOX家族與生長素在蝴蝶蘭發育過程中的表現，為進一步研究蘭科植物特殊的發育模式提供重要的基礎資料。 | zh_TW |
| dc.description.abstract | Unlike most angiosperms, which complete meristem specification during early embryogenesis, Phalaenopsis lacks an apical meristem at seed maturity and gradually establishes meristems and organs only after germination. To explore the molecular regulation of apical meristem formation in orchids, this study examined the expression patterns of WUSCHEL-related homeobox (WOX) genes and auxin accumulation during seed and protocorm development in Phalaenopsis. Different developmental stages of ovules and protocorms were defined based on their morphological and histological characteristics. Expression patterns of WOX genes and auxin accumulation were analyzed using quantitative real-time PCR and transgenic reporter plants (DR5::GUS). The expression of PaWUS, PaWOX3a, PaWOX4, PaWOX5, and PaWOX9 significantly increased during ovule development, suggesting roles in tissue differentiation and meristem formation. During protocorm development, PaWOX3a, PaWOX4, and PaWOX5 were highly expressed in the apical region, implying involvement in maintaining undifferentiated apical cells. DR5::GUS staining revealed polarized auxin distribution with accumulation in leaf and root primordia, supporting its role in organ differentiation. IAA and NAA promoted growth and adventitious root formation, while TIBA caused developmental abnormalities. WOX promoter::GUS constructs were also successfully established and preliminarily applied in Phalaenopsis transformation. Together, these results advance our understanding of the molecular regulation underlying apical meristem formation in orchids. | en |
| dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2025-08-18T01:16:29Z No. of bitstreams: 0 | en |
| dc.description.provenance | Made available in DSpace on 2025-08-18T01:16:29Z (GMT). No. of bitstreams: 0 | en |
| dc.description.tableofcontents | 口試委員會審定書 i
誌謝 ii 中文摘要 iii ABSTRACT iv CONTENTS v LIST OF TABLES ix LIST OF FIGURES x LIST OF SUPPLEMENTAL FIGURES xii Chapter 1 Introduction 1 1.1 Apical meristems and their functions in plant growth 1 1.2 WUSCHEL-RELATED HOMEOBOX (WOX) family 1 1.3 Regulation of WOX gene expression by auxin 2 1.4 A brief introduction to Orchidaceae 3 1.5 Development of embryos and protocorms in orchids 4 1.6 Aims of this study 5 Chapter 2 Materials and Methods 7 2.1 Plant materials 7 2.2 Seed germination and seedling cultures 7 2.3 Morphological and histological studies 8 2.4 Identification of WOX genes and phylogenetic analysis 9 2.5 Quantitative real-time PCR (qRT-PCR) experiments 9 2.5.1 Plant sample preparation for gene expression analysis 9 2.5.2 RNA extraction and reverse transcription 10 2.5.3 Primer design and cDNA concentration determination 10 2.5.4 Primer efficiency validation 11 2.5.5 qRT-PCR procedure 12 2.6 Cloning of promoters from Phalaenopsis WOX genes 12 2.6.1 Genomic DNA extraction 12 2.6.2 Promoter isolation and plasmid construction 13 2.7 Preparation of Agrobacterium competent cells and transformation by freeze-thaw method 13 2.8 DR5::GUS construct and transformation 14 2.9 Antibiotic sensitivity test 15 2.10 Agrobacterium-mediated transformation 15 2.11 Evaluation of DR5::GUS activity during Phalaenopsis ontogeny 16 2.12 Histochemical staining for GUS activity 16 2.13 Vibratome Sectioning 17 2.14 Effect of auxin on protocorm development 17 2.15 Effect of auxin on adventitious root formation in stem cuttings 18 2.16 Graphical visualization and statistical analysis 18 Chapter 3 Results 20 3.1 Ovule and embryo development in Phalaenopsis. 20 3.2 Morphological changes during protocorm development 20 3.3 Histological changes during protocorm development. 21 3.4 Phylogenetic analysis of WOX family proteins in Phalaenopsis 22 3.5 Relative expression levels of WOX genes during ovule and embryo development 22 3.6 Relative expression levels of WOX genes during protocorm development 23 3.7 Transformation experiments 24 3.8 Antibiotic sensitivity test 25 3.9 Optimization of post-transformation culture conditions 25 3.10 Confirmation of transgenic orchids via PCR and GUS staining. 26 3.11 Evaluation of DR5::GUS activity during Phalaenopsis ontogeny 27 3.11.1 Establishment of DR5::GUS transgenic lines 27 3.11.2 Analysis of DR5::GUS expression during Phalaenopsis development 28 3.11.3 GUS staining of vibratome sections reveals SAM and RAM formation 29 3.12 Effects of auxins on the growth and development of Phalaenopsis protocorms 31 3.12.1 Growth responses of 18 DAS protocorms to auxin and TIBA 32 3.12.2 Growth responses of 55 DAS protocorms to auxin and TIBA 33 3.12.3 TIBA disrupts auxin distribution and induces abnormal leaf morphologies in Phalaenopsis protocorms 34 3.13 Adventitious rooting of Phalaenopsis stem cuttings under NAA and TIBA treatments 35 3.14 Auxin accumulation patterns during adventitious root formation in Phalaenopsis stem cuttings 36 Chapter 4 Discussion 37 4.1 Expression profiles of WOX genes during embryogenesis and protocorm development in Phalaenopsis 37 4.2 Dynamics of auxin accumulation during protocorm development 39 4.3 Effect of auxin on protocorm development and adventitious root formation 41 4.4 Challenges of genetic transformation in orchids 43 Chapter 5 Conclusion 44 Reference 45 Table 51 Figure 53 Supplemental figure 83 | - |
| dc.language.iso | en | - |
| dc.subject | 蝴蝶蘭 | zh_TW |
| dc.subject | WOX 基因 | zh_TW |
| dc.subject | 生長素 | zh_TW |
| dc.subject | 分生組織 | zh_TW |
| dc.subject | 原球莖 | zh_TW |
| dc.subject | 轉基因系統 | zh_TW |
| dc.subject | auxin | en |
| dc.subject | Phalaenopsis | en |
| dc.subject | WOX genes | en |
| dc.subject | transgenic system | en |
| dc.subject | protocorm | en |
| dc.subject | meristem | en |
| dc.title | WOX基因家族與生長素在蝴蝶蘭種子及原球莖發育時的表現 | zh_TW |
| dc.title | Expression of WOX Gene Family and Auxin During Seed and Protocorm Development in Phalaenopsis | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 113-2 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.oralexamcommittee | 呂維茗;傅士峯 | zh_TW |
| dc.contributor.oralexamcommittee | Wei- Ming Leu;Shih-Feng Fu | en |
| dc.subject.keyword | 蝴蝶蘭,WOX 基因,生長素,分生組織,原球莖,轉基因系統, | zh_TW |
| dc.subject.keyword | Phalaenopsis,WOX genes,auxin,meristem,protocorm,transgenic system, | en |
| dc.relation.page | 86 | - |
| dc.identifier.doi | 10.6342/NTU202504132 | - |
| dc.rights.note | 未授權 | - |
| dc.date.accepted | 2025-08-12 | - |
| dc.contributor.author-college | 生命科學院 | - |
| dc.contributor.author-dept | 生命科學系 | - |
| dc.date.embargo-lift | N/A | - |
| 顯示於系所單位： | 生命科學系 | |
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