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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 陳昭瑩 | |
| dc.contributor.author | Yu-Ting Shih | en |
| dc.contributor.author | 施侑廷 | zh_TW |
| dc.date.accessioned | 2021-06-17T03:33:59Z | - |
| dc.date.available | 2020-03-01 | |
| dc.date.copyright | 2018-03-01 | |
| dc.date.issued | 2018 | |
| dc.date.submitted | 2018-02-12 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/69917 | - |
| dc.description.abstract | 百合為具有高經濟價值的球根花卉，除了土壤傳播性病蟲害外，影響其生產最鉅者為Botrytis elliptica (Berk.) Cooke所引起的百合灰黴病，如何有效地防治灰黴病，便成為農友最關心的問題。植物防禦相關蛋白LsGRP1 (Lilium ‘Star Gazer’ glycine-rich protein 1) 為水楊酸誘發葵百合Lilium cv. Star Gazer 對灰黴病抗性過程大量表現的蛋白。根據前人的研究，LsGRP1可能透過直接抑制微生物而抑制病原菌的侵染，本研究則探討LsGRP1是否能透過誘導抗病機制幫助植物抵抗灰黴病菌，並探討其應用價值與最適化之施用條件。首先將大腸桿菌生產的LsGRP1重組蛋白注射於百合中位葉，評估此處理是否能誘發系統葉組織對灰黴病的抗性，並同時測試施用時間、施用濃度及施用位置對LsGRP1重組蛋白誘導植物抗病之影響。接著為降低防治成本，將胞內表現LsGRP1重組蛋白的大腸桿菌以超音波破菌後直接處理植物，測試其誘發系統抗病性的作用，並評估其用以防治其他台灣重要作物病害，如草莓及番茄灰黴病的可行性。期本研究能協助發展新穎的植物病害防治資材，用以保障農作物生產，促進環境永續發展。 | zh_TW |
| dc.description.abstract | Lily is a bulbous flower crop with high economic importance. Beside of soil-borne diseases and pests, gray mold disease caused by Botrytis elliptica, affects lily production enormously. How to effectively control gray mold disease is a main concern of lily farmers. Defense-related protein LsGRP1 (Lilium ‘Star Gazer’ glycine-rich protein 1) is a protein with increased expression in lily which exhibits salicylic acid-induced resistance against gray mold disease. According to previous reports, LsGRP1 may enhance plant immunity via its antimicrobial activity. In this study, the potential of LsGRP1 to induce plant defense against gray mold pathogen and optimal application methods were investigated. At first, LsGRP1-derived recombinant protein produced by Escherichia coli was infiltrated into the middle leaves of lily plants to evaluate its effects on the induction of disease resistance in upper and lower systemic leaves. The time points of application, optimal concentrations, and application sites were determined by comparing the efficacy of disease suppression. In order to reduce the cost, the total lysate of the LsGRP1-derived recombinant protein from sonicated E. coli cells was applied directly to lily plants to examine the effects of systemic resistance induction. On the other hand, the protection effect of crude LsGRP1-derived recombinant protein on the gray mold disease of strawberry and tomato were evaluated. Expectedly, this study would facilitate the development of novel disease control agents suitable for crop production and environmental sustainability. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-17T03:33:59Z (GMT). No. of bitstreams: 1 ntu-107-R04645006-1.pdf: 3034287 bytes, checksum: 4201ba6fdbed392a16681d966b2a04e1 (MD5) Previous issue date: 2018 | en |
| dc.description.tableofcontents | 目錄
壹、中文摘要 VII 貳、英文摘要 VIII 參、前言 1 肆、前人研究 3 一、百合 3 二、百合灰黴病 4 三、植物富含甘胺酸蛋白質 (glycine-rich proteins, GRPs) 5 四、葵百合之水楊酸誘導性富含甘胺酸防禦相關蛋白LsGRP1 6 五、植物誘導性抗病及防禦反應 8 六、植物系統性抗病之誘導物質 11 七、胜肽及蛋白質誘導性抗病之植物病害防治應用 12 伍、材料與方法 15 一、供試植物之栽培 15 1.栽培介質調配 15 2.百合 15 3.番茄 15 4.草莓 15 二、供試真菌之培養、保存與接種源製備 16 1.供試真菌之培養及保存 16 2.供試真菌之接種源製備 16 三、大腸桿菌Escherichia coli表現LsGRP1融合蛋白 17 1.大腸桿菌表現蛋白株之建構與保存 17 2.誘導大腸桿菌表現融合蛋白及蛋白質粗萃取 18 3.蛋白質純化及透析 19 4.冷凍乾燥法濃縮蛋白質 19 5.蛋白質定量及保存 20 四、百合蛋白質粗萃取 20 五、十二烷基硫酸鈉聚丙烯醯胺膠體電泳偵測蛋白 21 1.十二烷基硫酸鈉聚丙烯醯胺膠體電泳 21 2.考馬斯蘭染色法 22 3.西方墨點法 22 六、注射SUMO-LsGRP1ΔSS融合蛋白後誘導百合系統葉抑制灰黴病病斑實驗 23 1.注射SUMO-LsGRP1ΔSS融合蛋白於百合葉片及病原接種 23 2.定量百合葉片病斑面積 23 3.定量百合葉片中真菌族群相對生物量 23 七、注射SUMO-LsGRP1ΔSS融合蛋白促進百合系統葉防禦相關物質及反應測試 25 1.植物細胞死亡造成離子滲漏之測量 25 2.觀察植物葉片接種B.elliptica後累積之癒傷葡聚醣 25 3.觀察植物葉片接種B.elliptica後活性氧物質之產生 26 八、NADPH氧化酶抑制劑二苯基氯化碘鹽抑制活性氧物質影響百合灰黴病病斑發展觀察 26 九、SUMO-LsGRP1ΔSS融合蛋白有效誘導抗病活性之耐熱性測試 26 1.蛋白溶液於熱處理之誘導抗病活性耐熱實驗 26 2.蛋白質乾燥粉末熱處理之誘導抗病活性耐熱實驗 27 十一、澆灌水楊酸及含有SUMO-LsGRP1∆SS融合蛋白的大腸桿菌溶解產物施用原液誘導植物抗病測試 27 1.製備澆灌處理所需之含有SUMO-LsGRP1∆SS融合蛋白的大腸桿菌溶解產物之施用原液 27 2.澆灌處理實驗用植物 28 3.百合栽培種澆灌實驗之抑制病率計算 28 4.草莓罹病度分級 29 十二、評估澆灌水楊酸及SUMO-LsGRP1∆SS施用原液誘導百合葉片LsGRP1相對累積量 29 十三、澆灌LsGRP1∆SS施用原液對植物葉綠素含量及光合作用的影響 30 1.葉綠素相對含量測定 30 2.葉綠素螢光測定 30 十四、將LsGRP1∆SS OAF在田間環境中應用於植物之效果 30 1.亞洲型雜交群食用姬百合之施用效果測試 30 2.草莓苗之施用效果測試 31 十五、數據統計及分析 31 陸、結果 32 一、大腸桿菌表現之LsGRP1衍生性融合蛋白 32 二、注射大腸桿菌表現之SUMO-LsGRP1ΔSS融合蛋白誘導百合系統葉抑制灰黴病病徵發展之效果 32 三、注射SUMO-LsGRP1ΔSS融合蛋白減少百合系統葉於灰黴病菌侵染時之植物細胞死亡現象 34 四、注射SUMO-LsGRP1ΔSS融合蛋白提升百合系統葉於灰黴病菌侵染時之癒傷葡聚醣累積量 35 五、注射SUMO-LsGRP1ΔSS融合蛋白促進百合系統葉於灰黴病菌侵染時之活性氧物質累積 35 六、以二苯基氯化碘鹽抑制活性氧物質產生可抑制百合葉片於接種早期之活性氧物質累積 36 七、SUMO-LsGRP1ΔSS融合蛋白誘導百合抗病活性之耐熱性測試 37 八、澆灌含有SUMO-LsGRP1∆SS融合蛋白的大腸桿菌溶解產物之誘導百合抗灰黴病效果 39 九、澆灌水楊酸及含有SUMO-LsGRP1∆SS融合蛋白的大腸桿菌溶解產物誘導百合自體累積LsGRP1 40 十、澆灌含有SUMO-LsGRP1∆SS融合蛋白的大腸桿菌溶解產物對百合葉片之葉綠素相對含量及光合作用效率之影響 41 十一、澆灌含有SUMO-LsGRP1∆SS融合蛋白的大腸桿菌溶解產物誘導不同百合栽培種抑制灰黴病的效果 42 十二、澆灌含有SUMO-LsGRP1∆SS融合蛋白的大腸桿菌溶解產物於不同植物之抑制灰黴病效果 43 十三、含SUMO-LsGRP1∆SS融合蛋白的大腸桿菌溶解產物於田間環境盆栽植物之抑病效果 44 柒、討論 45 捌、參考文獻 52 玖、圖表集 69 表一、澆灌含有SUMO-LsGRP1∆SS融合蛋白之大腸桿菌溶解產物對不同百合栽培種灰黴病之抑制率 70 圖一、以十二烷基硫酸鈉聚丙烯醯胺膠體電泳分析大腸桿菌表現之LsGRP1衍生性融合蛋白 71 圖二、注射SUMO-LsGRP1ΔSS融合蛋白誘導百合系統葉抑制灰黴病效果 72 圖三、注射不同濃度SUMO-LsGRP1ΔSS融合蛋白誘導百合系統葉抑制灰黴病效果 73 圖四、百合前處理注射SUMO-LsGRP1ΔSS融合蛋白可抑制系統葉中Botrytis elliptica之生物量 74 圖五、注射SUMO-LsGRP1ΔSS融合蛋白誘導上下位系統葉抑制灰黴病之效果 75 圖六、SUMO-LsGRP1ΔSS融合蛋白處理後灰黴病菌於系統葉造成的死細胞之導電度 77 圖七、百合中位葉注射SUMO-LsGRP1ΔSS融合蛋白促進系統葉於灰黴病感染時葡聚醣累積狀況 78 圖八、SUMO-LsGRP1ΔSS融合蛋白處理對系統葉活性氧物質累積之影響 80 圖九、活性氧物質抑制劑二苯基氯化碘鹽對灰黴病菌感染百合之影響 81 圖十、熱處理對SUMO-LsGRP1ΔSS融合蛋白誘導抗病活性之影響 84 圖十一、不同時間之55oC熱處理後SUMO-LsGRP1ΔSS融合蛋白溶液之沉澱情況 85 圖十二、100oC熱處理30分鐘對SUMO-LsGRP1ΔSS融合蛋白溶液誘導抗病活性之影響 86 圖十三、不同溫度處理對SUMO-LsGRP1ΔSS融合蛋白粉末誘導抗病活性之影響 88 圖十四、澆灌處理時間表 89 圖十五、不同稀釋倍率之LsGRP1∆SS OAF對百合灰黴病病斑之影響 90 圖十六、單次澆灌處理誘導百合抑制灰黴病病斑之有效持續時間 91 圖十七、澆灌處理後百合葉片之LsGRP1累積量 92 圖十八、澆灌處理後之百合葉片葉綠素相對含量及光合作用效率 95 圖十九、澆灌LsGRP1∆SS OAF誘導不同百合栽培種抑制灰黴病病斑效果 97 圖二十、澆灌LsGRP1∆SS OAF對番茄灰黴病病斑面積的影響 98 圖二十一、澆灌LsGRP1∆SS OAF對草莓灰黴病罹病度之影響 100 圖二十二、單次澆灌LsGRP1∆SS OAF於田間環境之亞洲型雜交群姬百合誘導其抗灰黴病效果 101 圖二十三、單一次澆灌LsGRP1∆SS OAF於田間環境之草莓苗誘導其抗炭疽病效果 103 | |
| dc.language.iso | zh-TW | |
| dc.title | 百合防禦相關蛋白LsGRP1誘導植物抗灰黴病之應用 | zh_TW |
| dc.title | Application of Lilium defense-related protein LsGRP1 to induce plant resistance against gray mold disease | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 106-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 沈偉強,鍾嘉綾,林乃君,黃秀珍 | |
| dc.subject.keyword | Botrytis elliptica,誘導抗病,百合,LsGRP1,灰黴病, | zh_TW |
| dc.subject.keyword | Botrytis elliptica,Induced resistance,Lily,LsGRP1,Gray mold disease, | en |
| dc.relation.page | 103 | |
| dc.identifier.doi | 10.6342/NTU201800537 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2018-02-13 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 植物醫學碩士學位學程 | zh_TW |
| 顯示於系所單位： | 植物醫學碩士學位學程 | |
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