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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 張皓巽 | zh_TW |
| dc.contributor.advisor | Hao-Xun Chang | en |
| dc.contributor.author | 陳政諺 | zh_TW |
| dc.contributor.author | Jheng-Yan Chen | en |
| dc.date.accessioned | 2023-09-22T16:59:04Z | - |
| dc.date.available | 2023-11-09 | - |
| dc.date.copyright | 2023-09-22 | - |
| dc.date.issued | 2023 | - |
| dc.date.submitted | 2023-08-10 | - |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/89995 | - |
| dc.description.abstract | 植物可辨識植物促生根際細菌 (plant growth-promoting rhizobacteria, PGPR) 釋出的激發子 (elicitor) 以預啟 (prime) 植物防禦反應相關基因，進而提升植物的抗病能力。在預啟反應所表現的蛋白中，病程相關蛋白 (pathogenesis-related protein, PR-protein) 之表現扮演了重要的角色。其中隸屬病原性相關蛋白PR-3、PR-4、PR-8及PR-11的幾丁質酶 (chitinase) 為一種廣泛存在於多種生物的糖苷水解酶 (glycoside hydrolase)，可透過水解幾丁質之糖苷鍵，進而影響真菌細胞壁等由幾丁質所構成的組織。由於幾丁質酶提供良好抗病潛力，加上近年高通量定序技術的發展，目前已有數種植物以全基因體角度切入探討幾丁質酶相關基因。此外，結合植株受病原菌感染之轉錄體分析，亦可進一步研究受到土壤微生物預啟的幾丁質酶基因是否受到病原菌調控，並以基因轉殖技術驗證其功能性。大豆 (Glycine max) 作為重要經濟作物之一，目前鮮有研究對其基因體中的幾丁質酶基因數量進行預測，故本研究透過大豆全基因體分析找到37個幾丁質酶基因，並經由大豆接種Burkholderia. ambifaria及Fusarium oxysporum之轉錄體分析，發現了5個幾丁質酶基因會受二者影響而上調。結合來自22種植物中已受功能性驗證之幾丁質酶序列進行親緣樹分析後，本研究挑選3個大豆幾丁質酶基因 (GmChi01、GmChi02及GmChi16) 透過農桿菌介導的阿拉伯芥轉殖技術，獲得了三個大豆基因的過表達子代。相較於阿拉伯芥Col-0，過表達大豆幾丁質酶基因之阿拉伯芥在整體生長勢上並無顯著差異，然而以離葉接種及土壤接種F. oxysporum f.sp. rapae後，計算葉片黃化程度換算病害發生進程下之面積 (AUDPC) 及幼苗存活率，發現過表達GmChi02及GmChi16之阿拉伯芥葉片可展現較強之的抗病能力。雖然GmChi02與GmChi16皆具提升抗病能力，但僅有GmChi02受到B. ambifaria預啟，說明幾丁質酶基因可能受到不同PGPR影響。透過接種6種不同PGPR之轉錄體分析，發現9個幾丁質酶基因不受到所測試的6種PGPR影響，另有10個幾丁質酶基因受到至少一種PGPR影響而顯著上調。綜合上述，本研究以全基因體分析大豆幾丁質酶基因，並搭配轉錄體學及阿拉伯芥轉殖技術驗證大豆幾丁質酶之抗真菌能力，同時紀錄6種PGPR預啟大豆幾丁質酶基因表現能力具有差異性。 | zh_TW |
| dc.description.abstract | Plants recognize elicitors released by plant growth-promoting rhizobacteria (PGPR) to initiate pathogenic-related proteins (PR-proteins) responses in disease priming. PR-proteins such as PR-3, PR-4, PR8, and PR-11 are chitinases that hydrolyze the glycosidic bond of chitin, which is the main component of fungal cell wall. In advantage of high-throughput sequencing technologies, it becomes powerful to perform genome-wide characterization of chitinases in plant genomes. Together with transcriptomic analyses to find PR-protein genes under biotic stresses such as fungal infection, validating gene functions in resistance through heterogeneous transformation has been robust and approachable. Soybean (Glycine max) is one of the most important economic crops, but limited researches have focused on soybean chitinase genes in the genome or validated their functions in defense responses. In this study, we identified 37 chitinase genes in the soybean genome, and we integrated the transcriptomic data in response to Burkholderia ambifaria and Fusarium oxysporum infection to prioritize 5 candidate chitinase genes. According to the phylogenetic analysis with previously functionally characterized chitinase genes from 22 plant species, we chose 3 genes (GmChi01, GmChi02, and GmChi16) to obtain the over-expression Arabidopsis lines by Agrobacterium‐mediated transformation. Compare to the Col-0, transgenic Arabidopsis lines did not show significant differences in phenotypes of agronomic traits. Furthermore, the results of detached leaf assay and soil inoculation showed that over-expression of GmChi02 and GmChi16 in Arabidopsis reduced area under disease progress curve (AUDPC) and enhanced the survival rate of seedlings upon F. oxsporum f.sp. rapae infection. Although GmChi02 and GmChi16 significantly enhanced defense to F. oxsporum f.sp. rapae, only GmChi02 was significantly primed by B. ambifaria. This observation indicated plant chitinases may be induced by different PGPR in defense responses. According to the transcriptomic analysis of soybean in response to 6 PGPR, 9 chitinases were not induced by any of PGPR and 10 chitinases were significantly primed by at least one of PGPR. In conclusion, this study verified the functions of GmChi01, GmChi02 or GmChi16 in disease responses and provided a comprehensive analysis of 37 soybean chitinase genes in PGPR-induced defense priming. | en |
| dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2023-09-22T16:59:04Z No. of bitstreams: 0 | en |
| dc.description.provenance | Made available in DSpace on 2023-09-22T16:59:04Z (GMT). No. of bitstreams: 0 | en |
| dc.description.tableofcontents | 目錄
摘要 I Abstract II 目錄 III 表目錄 V 圖目錄 VI 第壹章 前人研究 1 1.1 導論 1 1.2 植物促生根際細菌與防禦預啟及誘導系統性抗病 4 1.3 病程相關蛋白 6 1.4 幾丁質酶 10 1.5 幾丁質酶與植物抗病反應 11 1.6 高通量定序於幾丁質酶參與抗病反應之研究 12 1.7 幾丁質酶於防禦預啟反應之研究探索 13 1.8 研究動機 14 第貳章 材料方法 15 2.1 大豆幾丁質酶基因之辨識 15 2.2 大豆幾丁質酶胺基酸序列之親緣樹分析 16 2.3 大豆轉錄體分析 16 2.4 大豆幾丁質酶基因擴增與純化 17 2.5 大豆幾丁質酶基因之表現載體構築 17 2.6 農桿菌勝任細胞備製與轉型 19 2.7 阿拉伯芥之栽培與花序浸漬轉殖法 20 2.8 阿拉伯芥葉片之RNA萃取及cDNA生合成 21 2.9 阿拉伯芥之反轉錄酶連鎖反應 22 2.10 阿拉伯芥之農藝性狀 22 2.11 阿拉伯芥接種Fusarium oxysporum f.sp. rapae之抗病反應 22 2.12 植物促生根際細菌之培養、定量與接種 23 2.13 大豆主根之RNA萃取與轉錄體分析 24 2.14 實驗數據統計與分析 25 第參章 結果 26 3.1 大豆幾丁質酶基因之辨識 26 3.2 大豆轉錄體中幾丁質酶之基因表現 27 3.3 大豆幾丁質酶與功能性驗證幾丁質酶之親緣樹分析 28 3.4 阿拉伯之芥農藝性狀 29 3.5 阿拉伯芥對Fusarium oxysporum f.sp. rapae之抗病反應 30 3.6 六種植物促生根際細菌預啟大豆幾丁質酶之基因表現 31 第肆章 討論 33 4.1 應用基因體分析探討大豆幾丁質酶基因 33 4.2 參與抗病反應與B. ambifaria預啟之大豆幾丁質酶基因 34 4.3 GmChi02及GmChi16增強阿拉伯芥對F. oxysporum之抗病能力 35 4.4 六種植物促生根際細菌預啟大豆幾丁質酶基因具差異性 36 4.5 研究總結與未來展望 37 參考文獻 38 表 63 圖 73 附錄 88 | - |
| dc.language.iso | zh_TW | - |
| dc.subject | 大豆 | zh_TW |
| dc.subject | 轉錄體學 | zh_TW |
| dc.subject | 阿拉伯芥轉殖技術 | zh_TW |
| dc.subject | 基因體學 | zh_TW |
| dc.subject | 幾丁質酶 | zh_TW |
| dc.subject | Defense priming | en |
| dc.subject | Genome-wide characterization analysis | en |
| dc.subject | Plant growth-promoting rhizobacteria (PGPR) | en |
| dc.subject | Chitinase | en |
| dc.subject | Arabidopsis transformation | en |
| dc.subject | Soybean | en |
| dc.title | 探討大豆幾丁質酶於植物促生根際細菌預啟防禦反應之基因表現與功能 | zh_TW |
| dc.title | Genome-wide Characterization of Soybean Chitinase Expression and Function in Plant Growth-Promoting Rhizobacteria-induced Defense Priming | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 111-2 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.oralexamcommittee | 吳志航;林柏安 | zh_TW |
| dc.contributor.oralexamcommittee | Chih-Hang Wu;Po-An Lin | en |
| dc.subject.keyword | 大豆,阿拉伯芥轉殖技術,基因體學,轉錄體學,幾丁質酶, | zh_TW |
| dc.subject.keyword | Arabidopsis transformation,Chitinase,Defense priming,Genome-wide characterization analysis,Plant growth-promoting rhizobacteria (PGPR),Soybean, | en |
| dc.relation.page | 93 | - |
| dc.identifier.doi | 10.6342/NTU202303890 | - |
| dc.rights.note | 同意授權(限校園內公開) | - |
| dc.date.accepted | 2023-08-12 | - |
| dc.contributor.author-college | 生物資源暨農學院 | - |
| dc.contributor.author-dept | 植物病理與微生物學系 | - |
| 顯示於系所單位： | 植物病理與微生物學系 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| ntu-111-2.pdf 授權僅限NTU校內IP使用（校園外請利用VPN校外連線服務） | 15.51 MB | Adobe PDF |
系統中的文件，除了特別指名其著作權條款之外，均受到著作權保護，並且保留所有的權利。