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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 羅筱鳳 | |
dc.contributor.author | Yu-Heng Lin | en |
dc.contributor.author | 林煜恆 | zh_TW |
dc.date.accessioned | 2021-06-15T00:16:32Z | - |
dc.date.available | 2013-08-16 | |
dc.date.copyright | 2011-08-16 | |
dc.date.issued | 2011 | |
dc.date.submitted | 2011-08-15 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/41354 | - |
dc.description.abstract | 番茄黃化捲葉病毒(Tomato yellow leaf curl virus, TYLCV)所誘發之番茄黃化捲葉病(Tomato yellow leaf curl disease, TYLCD)為近年來影響世界番茄生產的嚴重病害,常造成番茄生產上100%的經濟損失。防治TYLCD目前除了抗病育種,尚無有效之防治方法。由於番茄植株感染TYLCD後之生理尚不明瞭,故本研究針對植株染病後之抗氧化生理及蛋白質體兩方面深入探討,期有助於瞭解番茄TYLCD之抗病機制。植物材料使用由亞洲蔬菜研究發展中心選育出之番茄抗病品系CLN2777G-1-5及感病品種Tomatoll,病毒材料為番茄黃化捲葉泰國病毒種(Tomato yellow leaf curl Thailand virus, TYLCTHV)。試驗結果顯示TYLCTHV不能藉由機械法接種至番茄植株,以粉蝨為媒介接種TYLCTHV至感病品種植株後,葉片數、株高、SPAD值及葉綠素螢光Fv/Fm值皆顯著下降,於第3天所有植株皆檢測出病毒,第7天植株表現出病徵;抗病品系則於接種後第28天方有83%植株檢測出病毒,且仍無病徵表現。雖然抗病品系於接種TYLCTHV後 1及3天超氧歧化酶(superoxide dismutase, SOD)活性顯著低於感病品種,但是抗病品系之過氧化氫酶(catalase, CAT)、抗壞血酸過氧化酶(ascorbate peroxidase, APX)、穀胱甘肽還原酶(glutathione reductase, GR)、過氧化物酶(peroxidase)活性皆可持續升高,而有助於植株抗病毒侵害之逆境。兩番茄品種(系)於接種TYLCTHV後葉片蛋白質總量皆增加。利用全自動二維蛋白快速層析分離系統(ProteomeLabTM PF-2D)及基質輔助雷射脫附游離飛行時間質譜儀(Matrix-assisted laser desorption ionization time-of-flight, MALDI-TOF)分析,比對出番茄兩品種(系)感染TYLCTHV後具差異表現之30個蛋白質,這些蛋白質參與植物光合作用、逆境防禦系統、蛋白質運輸、蛋白質轉譯、蛋白質裂解及氧化還原相關反應。本研究結果或可作為番茄抗TYLCD育種策略及抗病篩選指標之參考。 | zh_TW |
dc.description.abstract | Tomato yellow leaf curl disease (TYLCD) induced by Tomato yellow leaf curl virus is one of the most severe diseases of tomato. TYLCD defeats tomato production worldwide and often causes 100% economic loss. There is no efficient method to control TYLCD, except for breeding resistant cultivars. However, the plant physiology concerning TYLCD infection remains totally unknown. This research studied the antioxidation and proteomics of tomato plants after infection of TYLCD to understand the resistance mechanism. The resistant tomato line CLN2777G-1-5 and susceptible cultivar Tomatoll which were selected and bred by Asian Vegetable Research and Development Center, and Tomato yellow leaf curl Thailand virus (TYLCTHV) were used as experimental materials. The results indicated that TYLCTHV is transmitted by whitefly that not mechanical. The leaf number, plant height, relative chlorophyll content (SPAD value) and chlorophyll fluorescence Fv/Fm of the infected ‘Tomatoll’ plant, were significantly decreased. All plants of susceptible cultivar were detected the occurrence of TYLCTHV DNA and first symptom appeared on 3 and 7 days after inoculation. While eighty three percent plants of resistant line were not detected the occurrence of virus DNA until 28 days and did not show symptom. Activity of superoxide dismutase (SOD) in leaves of resistant line was significantly lower than susceptible cultivar on 1 and 7 days after inoculation. However, the activities of catalase (CAT)、ascorbate peroxidase (APX)、glutathione reductase (GR) and peroxidase of resistant line raised successively after inoculation. The raised activities of antioxidant enzymes may induce the resistance of CLN2777G-1-5 plants to TYLCTHV. Proteomics of leaves analyzed by ProteomeLabTM PF-2D and matrix-assisted laser desorption ionization time-of-flight, thirty proteins with differential expression were identified from CLN2777G-1-5 and‘Tomatoll’after inoculation with TYLCTHV. These proteins involve photosynthesis, stress defense, protein transport, translation, protein lysis, and oxidation-reduction reaction. The results may be helpful to plan strategy and select index for breeding TYLCTHV resistant tomato. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T00:16:32Z (GMT). No. of bitstreams: 1 ntu-100-R98628116-1.pdf: 1200681 bytes, checksum: 2f5141d94928ca157b6c8625613a780d (MD5) Previous issue date: 2011 | en |
dc.description.tableofcontents | 目 錄
論文口試委員審定書………………………………………………………………… 致謝……………………………………………………………………………………I 中文摘文……………………………………………………………………………II 英文摘要……………………………………………………………………………III 目錄…………………………………………………………………………………IV 表目錄……………………………………………………………………………….VI 圖目錄………………………………………………………………………………VII 附表目錄……………………………………………………………………………X 前言……………………………………………………………………………………1 第壹章 前人研究……………………………………………………………………3 一、番茄黃化捲葉病毒病……………………………………………………………3 二、植物感染病毒後之抗病生理研究………………………………………………6 三、蛋白質體分析應用於植物逆境生理之研究……………………………………8 四、番茄黃化捲葉病於臺灣發生情形………………………………………………9 第貳章 材料方法……………………………………………………………………11 一、試驗材料………………………………………………………………………11 二、病毒接種方式…………………………………………………………………11 三、生理調查分析項目……………………………………………………………12 四、病毒DNA偵測………………………………………………………………12 (一) 病毒DNA萃取……………………………………………………………12 (二) 聚合酶連鎖反應…………………………………………………………13 (三) 電泳………………………………………………………………………13 五、抗氧化酵素活性分析…………………………………………………………13 (一) 超氧歧化酶…………………………………………………………………13 (二) 過氧化氫酶…………………………………………………………………14 (三) 抗壞血酸過氧化酶…………………………………………………………14 (四) 穀胱甘肽還原酶……………………………………………………………15 (五) 過氧化物酶…………………………………………………………………15 六、蛋白質體分析……………………………………………………………………15 (一) 蛋白質萃取…………………………………………………………………15 (二) 蛋白質回溶置換……………………………………………………………16 (三) 蛋白質定量…………………………………………………………………16 (四) 一維電泳……………………………………………………………………16 (五) 二維電泳……………………………………………………………………17 (六) 胰蛋白脢分解………………………………………………………………18 (七) 蛋白質去鹽類………………………………………………………………18 (八) 蛋白質質譜分析……………………………………………………………19 七、統計分析………………………………………………………………………19 第參章 結果…………………………………………………………………………20 一、接種試驗………………………………………………………………………20 二、抗氧化酵素活性分析…………………………………………………………22 三、蛋白質體分析…………………………………………………………………23 第肆章 討論…………………………………………………………………………25 一、接種試驗………………………………………………………………………25 二、抗氧化酵素活性分析…………………………………………………………27 三、蛋白質體分析…………………………………………………………………29 第伍章 結論…………………………………………………………………………33 參考文獻……………………………………………………………………………35 表目錄 表1. 番茄黃化捲葉泰國種病毒以機械接種至兩番茄品種(系)後各時期之發病情形……………………………………………………………………………………44 表2. 番茄黃化捲葉泰國病毒種以粉蝨接種至兩番茄品種(系)後各時期之發病情形……………………………………………………………………………………45 表3. 兩番茄品種(系)接種番茄黃化捲葉泰國病毒種1天後蛋白質體之差異表現……………………………………………………………………………………46 表4. 兩番茄品種(系)接種番茄黃化捲葉泰國病毒種3天後蛋白質體之差異表現……………………………………………………………………………………47 表5. 兩番茄品種(系)接種番茄黃化捲葉泰國病毒種7天後蛋白質體之差異表現……………………………………………………………………………………48 表6. 兩番茄品種(系)接種番茄黃化捲葉泰國病毒種28天後蛋白質之差異表現……………………………………………………………………………………50 圖目錄 圖1. 兩番茄品種(系)以粉蝨接種番茄黃化捲葉泰國病毒種前及接種1天後之植株外觀………………………………………………………………………………51 圖2. 兩番茄品種(系)以粉蝨接種番茄黃化捲葉泰國病毒種後3及7天之植株外 觀……………………………………………………………………………………52 圖3. 兩番茄品種(系)以粉蝨接種番茄黃化捲葉泰國病毒種後14及28天之植株外觀…………………………………………………………………………………53 圖4. 番茄兩品種(系)以粉蝨接種番茄黃化捲葉泰國病毒種前與接種1天後葉片中之病毒DNA………………………………………………………………………54 圖5. 番茄兩品種(系)以粉蝨接種番茄黃化捲葉泰國病毒種3與7天後葉片中之病毒DNA……………………………………………………………………………55 圖6. 番茄兩品種(系)以粉蝨接種番茄黃化捲葉泰國病毒種14與28天後葉片中之病毒DNA…………………………………………………………………………56 圖7. 番茄抗病品系CLN2777G-1-5機械接種番茄黃化捲葉泰國病毒種後各時期之葉片數……………………………………………………………………………57 圖8. 番茄抗病品系CLN2777G-1-5機械接種番茄黃化捲葉泰國病毒種後各時期之植株高度…………………………………………………………………………57 圖9. 番茄抗病品系CLN2777G-1-5機械接種番茄黃化捲葉泰國病毒種後各時期之植株莖徑…………………………………………………………………………58 圖10. 番茄抗病品系CLN2777G-1-5機械接種番茄黃化捲葉泰國病毒種後各時期之葉片葉綠素相對含量(SPAD值)…………………………………………………58 圖11. 番茄抗病品系CLN2777G-1-5機械接種番茄黃化捲葉泰國病毒種後各時期之葉綠素螢光Fv/Fm………………………………………………………………59 圖12. 番茄感病品種Tomatoll機械接種番茄黃化捲葉泰國病毒種後各時期之葉片數…………………………………………………………………………………59 圖13. 番茄感病品種Tomatoll機械接種番茄黃化捲葉泰國病毒種後各時期之植株高度變化…………………………………………………………………………60 圖14. 番茄感病品種Tomatoll機械接種番茄黃化捲葉泰國病毒種後各時期之植株莖徑………………………………………………………………………………60 圖15. 番茄感病品種Tomatoll機械接種番茄黃化捲葉泰國病毒種後各時期之葉片葉綠素相對含量(SPAD值)………………………………………………………61 圖16. 番茄感病品種Tomatoll機械接種番茄黃化捲葉泰國病毒種後各時期之葉片葉綠素螢光Fv/Fm………………………………………………………………61 圖17. 番茄抗病品系CLN2777G-1-5以粉蝨接種番茄黃化捲葉泰國病毒種後各時期之葉片數…………………………………………………………………………62 圖18. 番茄抗病品系CLN2777G-1-5以粉蝨接種番茄黃化捲葉泰國病毒種後各時期之植株高度………………………………………………………………………62 圖19. 番茄抗病品系CLN2777G-1-5以粉蝨接種番茄黃化捲葉泰國病毒種後各時期之莖徑……………………………………………………………………………63 圖20. 抗病番茄品系CLN2777G-1-5以粉蝨接種番茄黃化捲葉泰國病毒種後各時期之葉綠素相對含量(SPAD)………………………………………………………63 圖21. 番茄抗病品系CLN2777G-1-5以粉蝨接種番茄黃化捲葉泰國病毒種後各時期之葉綠素螢光Fv/Fm………………………………………………………………………………64 圖22. 番茄感病品種Tomatoll以粉蝨接種番茄黃化捲葉泰國病毒種後各時期之葉片數………………………………………………………………………………64 圖23. 番茄感病品種Tomatoll以粉蝨接種番茄黃化捲葉泰國病毒種後各時期之植株高度……………………………………………………………………………65 圖24. 番茄感病品種Tomatoll以粉蝨接種番茄黃化捲葉泰國病毒種後各時期之莖徑…………………………………………………………………………………65 圖25. 番茄感病品種Tomatoll以粉蝨接種番茄黃化捲葉泰國病毒種後各時期之葉綠素相對含量(SPAD)……………………………………………………………66 圖26. 番茄感病品種Tomatoll以粉蝨接種番茄黃化捲葉泰國病毒種後各時期之葉綠素螢光Fv/Fm…………………………………………………………………66 圖27. 番茄感病品種Tomatoll以粉蝨接種番茄黃化捲葉泰國病毒種後各時期之超氧歧化酶活性……………………………………………………………………67 圖28. 番茄抗病品系CLN2777G-1-5以粉蝨接種番茄黃化捲葉泰國病毒種後各時期之超氧歧化酶活性………………………………………………………………67 圖29. 番茄兩品種(系)以粉蝨接種番茄黃化捲葉泰國病毒種後各時期超氧歧化酶活性之比較…………………………………………………………………………68 圖30. 番茄感病品種Tomatoll以粉蝨接種番茄黃化捲葉泰國病毒種後各時期之過氧化氫酶活性……………………………………………………………………………………68 圖31. 番茄抗病品系CLN2777G-1-5以粉蝨接種番茄黃化捲葉泰國病毒種後各時期之過氧化氫酶活性……………………………………………………………………………………69 圖32. 番茄兩品種(系)以粉蝨接種番茄黃化捲葉泰國病毒種後各時期過氧化氫酶活性之比較……………………………………………………………………………………69 圖33. 番茄感病品種Tomatoll以粉蝨接種番茄黃化捲葉泰國病毒種後各時期之抗壞血酸過氧化酶活性……………………………………………………………………………………70 圖34. 番茄抗病品系CLN2777G-1-5以粉蝨接種番茄黃化捲葉泰國病毒種後各時期之抗壞血酸過氧化酶活性……………………………………………………………………………………70 圖35. 番茄兩品種(系)以粉蝨接種番茄黃化捲葉泰國病毒種後各時期抗壞血酸過氧化酶活性之比較……………………………………………………………………………………71 圖36. 番茄感病品種Tomatoll以粉蝨接種番茄黃化捲葉泰國病毒種後各時期之穀胱甘肽還原酶活性……………………………………………………………………………………71 圖37. 番茄抗病品系CLN2777G-1-5以粉蝨接種番茄黃化捲葉泰國病毒種後各時期之穀胱甘肽還原酶活性……………………………………………………………………………………72 圖38. 番茄兩品種(系)以粉蝨接種番茄黃化捲葉泰國病毒種後各時期穀胱甘肽還原酶活性之比較……………………………………………………………………………………72 圖39. 番茄感病品種Tomatoll以粉蝨接種番茄黃化捲葉泰國病毒種後各時期之過氧化物酶活性……………………………………………………………………………………73 圖40. 番茄抗病品系CLN2777G-1-5以粉蝨接種番茄黃化捲葉泰國病毒種後各時期之過氧化物酶活性……………………………………………………………………………………73 圖41. 番茄兩品種(系)以粉蝨接種番茄黃化捲葉泰國病毒種後各時期過氧化物酶活性之比較……………………………………………………………………………………74 圖42. 番茄感病品種Tomatoll以粉蝨接種番茄黃化捲葉泰國病毒種後各時期之蛋白質總量……………………………………………………………………………………74 圖43. 番茄抗病品系CLN2777G-1-5以粉蝨接種番茄黃化捲葉泰國病毒種後各時期之蛋白質總量……………………………………………………………………………………75 圖44. 比較番茄兩品種(系)以粉蝨接種番茄黃化捲葉泰國病毒種後各時期蛋白質總量…………………………………………………………………………………75 附表目錄 附表1. 胰蛋白酶分解實驗參照表……………………………………………………………………………………76 | |
dc.language.iso | zh-TW | |
dc.title | 番茄植株感染番茄黃化捲葉泰國病毒種後之抗氧化生理及蛋白質體研究 | zh_TW |
dc.title | The study on antioxidation and proteomics of tomato plants infected by Tomato yellow leaf curl Thailand virus | en |
dc.type | Thesis | |
dc.date.schoolyear | 99-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 張英?,葉開溫,楊雯如,蔡文錫 | |
dc.subject.keyword | 番茄黃化捲葉泰國病毒種,超氧歧化酶,過氧化氫酶,抗壞血酸過氧化酶,穀胱甘肽,還原酶,過氧化物酶,蛋白質體, | zh_TW |
dc.subject.keyword | Tomato yellow leaf curl Thailand virus,superoxide dismutase,catalase,ascorbate peroxidase,glutathione reductase,peroxidase,proteomics, | en |
dc.relation.page | 76 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2011-08-15 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 園藝學研究所 | zh_TW |
顯示於系所單位: | 園藝暨景觀學系 |
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ntu-100-1.pdf 目前未授權公開取用 | 1.17 MB | Adobe PDF |
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