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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 林長平(Chan-Pin Lin) | |
| dc.contributor.author | Ying-Chi Huang | en |
| dc.contributor.author | 黃盈綺 | zh_TW |
| dc.date.accessioned | 2022-11-25T05:33:45Z | - |
| dc.date.available | 2026-08-18 | |
| dc.date.copyright | 2021-11-11 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-08-26 | |
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Frontiers in Plant Science, 8(1598). doi:10.3389/fpls.2017.01598 | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/81994 | - |
| dc.description.abstract | Phyllody symptoms1 (PHYL1) 是花生簇葉菌質體中主要造成花器葉化的效應子。在先前研究顯示，PHYL1 效應子能夠抑制 microRNA396 (miR396) 的表現，因 miR396 的表現量減少，進而使其目標基因 SHORT VEGETATIVE PHASE (SVP) 調控不當。SVP 為負調控開花之重要調節基因，因為 SVP 受到miR396 表現量下降的影響，無法受到妥善的調控，造成 SVP 表現過量，導致葉狀花之形成。但 PHYL1 是如何調控 MIR396 基因表現？PHYL1 效應子如何與寄主因子之相互作用導致之病徵發展？這些機制尚不明確。其中，SVP 受 miR396 調控的機制在降解組的定序資料中顯示，miR396 並非經由 mRNA 剪切方式來調控 SVP。是否是經由轉譯抑制或其他方式來調控 SVP的表現，並無文獻報導。在阿拉伯芥中，miR396 具有兩種同功型 (miR396a及miR396b)，目前兩種同功型調控 SVP 的貢獻度尚不明確，也沒有證據直接證明 miR396 會影響 SVP 之蛋白表現。PHYL1、SVP 和 miR396 之間的關係仍然有很多尚未解惑的問題，因此，我們利用 CRISPR/Cas9 系統在阿拉伯芥中建構了 miR396a 及 miR396b 之單突變株及雙突變株。本研究也生產出 SVP Keratin-like domain (SVP_K) 之特異性抗體，以確認在不同突變株中之 SVP 蛋白表現量。目前我們已經獲得 miR396 純合子之單突變株及雙突變株，且 α-SVP 特異性抗體可以測得 SVP 蛋白。後續研究中，我們也在菸草中暫時性表現野生型及突變之 SVP，並以 real-time RT-PCR 及西方墨點法比較有無外加 miR396 處理之 SVP 表現量。結果顯示，突變型的 SVP mRNA 加入 miR396 後，有較低的 SVP 蛋白表現，但其 RNA 表現量卻無明顯下降。顯示，miR396 對於調控 SVP 之蛋白是具有影響力並與先前推測藉由轉譯抑制調控有相符之處。期望本研究可以幫助未來對於 miR396、SVP 及 PHYL1 之關係能有更深入之了解。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-25T05:33:45Z (GMT). No. of bitstreams: 1 U0001-2508202109534400.pdf: 2517290 bytes, checksum: 8bd2e033f129cfbb9683cdb8117f5155 (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | 口試委員審定書 I 謝辭 II 中文摘要 IV Abstract VI Contents VIII List of Tables X List of Figures XI Introduction 1 Materials and Methods 8 Plant materials and growth conditions 8 MIR396a and MIR396b genes editing with CRISPR/Cas9 8 Genotyping 9 Prediction of pre-miR396 structure 9 Small RNA Detection 10 SVP antibody production 10 Western blotting 12 Protein degradation inhibiting assay 12 Real-time RT-PCR 13 Transient expression 13 Results 14 MIR396 genes knock out by CRISPR/Cas9 14 The miR396 precursor structures of mutants 15 Mutate MIR396a and MIR396b causing reduction of miR396 expression level 16 The mir396 mutants showed bigger leaf and long petiole phenotypes 17 Detection of endogenous SVP protein 17 Evaluation SVP and GRF1 gene expression in miR396 mutants 18 Demonstration of miR396-mediated SVP translation-inhibition 19 Discussion 22 CRISPR/Cas9 system can successfully generate miR396 mutants 22 SVP is an essential element in PHYL1-mediated leafy flower development 22 More mismatches on the target site can trigger miRNA-mediated translation inhibition 24 Working Hypothesis 26 Conclusions 27 References 28 | |
| dc.language.iso | en | |
| dc.subject | SHORT VEGETATIVE PHASE | zh_TW |
| dc.subject | CRISPR/Cas9 系統 | zh_TW |
| dc.subject | microRNA396 | zh_TW |
| dc.subject | 葉狀花 | zh_TW |
| dc.subject | 轉譯抑制 | zh_TW |
| dc.subject | translation inhibition | en |
| dc.subject | microRNA396 | en |
| dc.subject | SHORT VEGETATIVE PHASE | en |
| dc.subject | CRISPR/Cas9 | en |
| dc.subject | leafy flower | en |
| dc.title | 探討轉譯抑制對miRNA396a/b介導 SHORT VEGETATIVE PHASE (SVP)調控機制之影響 | zh_TW |
| dc.title | Investigation of the translation inhibition effects in miRNA396a/b-mediated SHORT VEGETATIVE PHASE (SVP) regulation | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.coadvisor | 林詩舜(Shih-Shun Lin) | |
| dc.contributor.oralexamcommittee | 詹富智(Hsin-Tsai Liu),楊俊逸(Chih-Yang Tseng),郭志鴻 | |
| dc.subject.keyword | microRNA396,SHORT VEGETATIVE PHASE,CRISPR/Cas9 系統,葉狀花,轉譯抑制, | zh_TW |
| dc.subject.keyword | microRNA396,SHORT VEGETATIVE PHASE,CRISPR/Cas9,leafy flower,translation inhibition, | en |
| dc.relation.page | 46 | |
| dc.identifier.doi | 10.6342/NTU202102710 | |
| dc.rights.note | 同意授權(限校園內公開) | |
| dc.date.accepted | 2021-08-27 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 植物病理與微生物學研究所 | zh_TW |
| dc.date.embargo-lift | 2026-08-18 | - |
| 顯示於系所單位： | 植物病理與微生物學系 | |
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