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請用此 Handle URI 來引用此文件: http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/71357
完整後設資料紀錄
DC 欄位值語言
dc.contributor.advisor張育森(Yu-Sen Chang)
dc.contributor.authorPo-Yen Chenen
dc.contributor.author陳伯彥zh_TW
dc.date.accessioned2021-06-17T05:59:19Z-
dc.date.available2024-02-19
dc.date.copyright2019-02-19
dc.date.issued2019
dc.date.submitted2019-02-13
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dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/71357-
dc.description.abstract民族藥用植物對於傳統醫學及現代醫學皆為重要的資源,現今許多保健食品及臨床用藥來自天然植物及其衍生物,如何有效運用及發展天然資源成為重要的議題。枸杞(Lycium chinense Mill.)葉為傳統的中草藥,其葉片經常受到癭螨叮咬而形成癭,使植物形態、生理及組織內的生化組成產生重大的改變。受癭感染後,葉片扭曲變形導致光合作用能力下降,影響植株生長勢,對此農民主要噴施農藥來減輕癭害的問題。然而,研究顯示葉片受造癭生物感染後,受感染組織的機能性成分含量增加。其中,癭在傳統民俗療法已被作為藥物使用,許多研究也證實其功效性,若葉用枸杞受癭螨感染後其保健功效有所提升,可增加其商業價值並減少農藥用量。本研究利用葉用枸杞進行分析,透過測量正常葉、感染葉、癭組織的植化物質及抗氧化功能性,探討癭組織與寄主葉片保健功效之差異。以及利用葉綠素螢光影像與非破壞性光生理檢測來暸解癭組織與寄主葉片間光合作用及其生理狀態。最後透過RNA-Seq分析癭組織、感染葉及正常葉的差異表現基因,配合生理生化分析試驗,瞭解癭形成對於光合作用及二級代謝物於基因層面的調控。結果顯示,癭的形成隨著不同發育階段的生長改變了葉片形態及組織結構,其葉綠素及類胡蘿蔔素含量較正常葉低,且隨著感染程度越高其下降程度越高。葉綠素螢光影像及非破壞性光生理檢測,顯示葉用枸杞受癭感染後光合效能下降,推測可能為葉片形態改變以及細胞間隙產生的空洞、光合色素含量下降以及PS II相關基因受到限制所影響,透過轉錄體分析,光反應和卡爾文循環功能類群中,癭與葉片相比大多數的基因皆顯著下調。癭的形成誘導植物開啟防禦系統,使植物產生化學信號,開啟內部的代謝過程,透過轉錄體分析結果發現生物逆境、植物激素以及二級代謝物相關基因表現上升。其中,受癭螨感染後水楊酸有升高趨勢,且主要累積在癭組織,而二級代謝物含量也有明顯變動,癭中花青素及類黃酮含量降低,但酚類含量顯著提升且於受感染組織上亦有顯著上升。進一步分析羥基肉桂酸衍生物,發現綠原酸受癭感染後樣本具有提升;而黃酮醇衍生物之芸香苷在癭中具有較高的程度。在保健功效方面,清除DPPH自由基結果顯示感染葉片其抗氧化能力較正常葉佳。綜合以上結果,葉用枸杞受癭感染後,光合作用能力下降,但酚類化合物以及抗氧化能力顯著提升,因此,可將受癭感染的葉用枸杞做為保健食品應用與開發之方向,減少農藥施用並友善農業施作環境。由本研究之轉錄體結果對於植物與昆蟲間的交互作用,可以更深入的探討,其對於應用上可作為後續的發展。zh_TW
dc.description.abstractExploit and develop the usage of natural resources as traditional medicine and health food are becoming more popular in modern society. The leaf of wolfberry (Lycium chinense Mill.) is a traditional Chinese medical herb for their known compounds, such as rutin and chlorogenic acid. However, leaves of wolfberry are often suffered from the gall mite infection, resulting in a declined harvest. Regarding that the gall tissues in other plant species were found to contain higher functional biochemical compounds that benefit for human health, it is worthy to examine the content change of functional compounds in the infected leaves of wolfberry. In this study, therefore, galled leaves of wolfberry were selected to be the target for developing new health food. Anatomy and photosynthesis activities of infected leaves were examined firstly follow by the analysis of the contents of secondary metabolite. Transcriptomes of galled leaves were analyzed to reveal the genetic regulation of the change of infection. Results showed that the photo-physiological status of galled leaves was significantly decreased, as well as the contents of chlorophyll and carotenoids. Transcriptomic analysis also showed the expressional decrease of photosynthesis-related genes. The level of anthocyanin and flavonoid were declined in gall leaves, but the levels of phenol-related metabolites were increased. In the analysis of functional compounds, the contents of rutin and chlorogenic acid were elevated in the infected leaves. The efficiency of eliminating free radical in galled leaves was also found to be higher than that in uninfected leaves. This study provided the data that supports the feasibility of using the galled leaves to develop a health food.en
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dc.description.tableofcontents誌謝…………………………………………………………i
摘要…………………………………………………………iii
Abstract……………………………………………………v
目錄…………………………………………………………vi
圖目錄………………………………………………………ix
表目錄………………………………………………………xi
檢索表………………………………………………………xii
第一章、前言………………………………………………………1
一、緒論……………………………………………………………1
二、葉用枸杞的概況………………………………………………2
三、癭………………………………………………………………3
四、癭的光合作用…………………………………………………5
五、癭的植生物質與抗氧化能力…………………………………6
六、次世代定序……………………………………………………9
七、研究目的………………………………………………………10
第二章、材料與方法………………………………………………11
一、試驗材料………………………………………………………11
二、試驗處理………………………………………………………11
(一) 細胞組織結構分析及非破壞性光生理檢測…………………11
(二) 植化物質含量、抗氧化功能性以及二級代謝物衍生物分析…11
(三) RNA-Seq分析…………………………………………………12
三、分析項目………………………………………………………12
(一) 細胞組織結構分析……………………………………………12
(二) 非破壞性光生理檢測…………………………………………15
(三) 植化物質………………………………………………………17
(四) 抗氧化功能指標………………………………………………18
(五) 超高效能液相層析法…………………………………………20
(六) RNA萃取與純化………………………………………………21
(七) 轉錄組定序……………………………………………………21
(八) RNA-Seq分析…………………………………………………22
四、統計方法………………………………………………………22
第三章、結果………………………………………………………23
一、癭形態變化……………………………………………………23
二、非破壞性光生理檢測…………………………………………23
(一) 非破壞性光生理檢測及葉綠素螢光…………………………23
(二) 葉綠素螢光影像………………………………………………23
三、植化物質含量…………………………………………………24
(一) 葉綠素…………………………………………………………24
(二) 類胡蘿蔔素……………………………………………………24
(三) 花青素…………………………………………………………25
(四) 類黃酮…………………………………………………………25
(五) 多酚……………………………………………………………25
四、抗氧化功能性…………………………………………………25
(一) 螯合亞鐵離子能力……………………………………………25
(二) 清除DPPH自由基能力………………………………………26
五、羥基肉桂酸衍生物及黃酮醇衍生物分析……………………27
六、水楊酸分析……………………………………………………28
七、RNA-Seq分析…………………………………………………29
(一) Contig序列組裝組裝…………………………………………29
(二) 序列功能及細胞位置推測……………………………………29
(三) TAIR資料庫比對………………………………………………29
(四) 差異表現分析…………………………………………………30
(五) 光合作用相關基因……………………………………………30
(六) 二級代謝相關基因……………………………………………32
(七) 生物逆境相關基因……………………………………………33
(八) 植物激素相關基因……………………………………………34
(九) 四吡咯生合成相關基因………………………………………37
第四章、討論………………..……………………………………38
一、葉用枸杞光合作用能力的變化………………………………38
二、葉用枸杞植化物質與植物防禦………………………………40
三、葉用枸杞植化物質與抗氧化能力……………………………43
四、葉用枸杞RNA-Seq分析………………………………………45
第五章、結論與展望………………………………………………49
第六章、參考文獻…………………………………………………50
圖 目 錄
圖1、葉用枸杞葉片及其癭與市售果實。…………………………………………59
圖2、葉用枸杞癭的細胞組織結構。………………………………………………60
圖3、葉用枸杞不同感染程度葉片及不同發育階段癭之葉綠素螢光影像分析。…61
圖4、葉用枸杞葉片、癭與市售果實之光合色素含量。…………………………62
圖5、葉用枸杞葉片、癭與市售果實之花青素含量。……………………………63
圖6、葉用枸杞葉片、癭與市售果實之類黃酮含量。……………………………64
圖7、葉用枸杞葉片、癭與市售果實之多酚類含量。……………………………65
圖8、葉用枸杞不同感染程度葉片、癭與市售果實之螯合亞鐵離子能力。…66
圖9、葉用枸杞不同感染程度葉片、癭與市售果實之清除DPPH自由基能力。…67
圖10、葉用枸杞不同感染程度葉片、癭與市售枸杞果實之綠原酸相對含量。…68
圖11、葉用枸杞不同感染程度葉片、癭與市售枸杞果實之芸香苷相對含量。…69
圖12、葉用枸杞不同感染程度葉片、癭與市售枸杞果實之對香豆酸相對含量。…70
圖13、葉用枸杞不同感染程度葉片、癭與市售枸杞果實之咖啡酸相對含量。…71
圖14、葉用枸杞不同感染程度葉片、癭與市售枸杞果實之阿魏酸相對含量。…72
圖15、葉用枸杞不同感染程度葉片、癭與市售枸杞果實之水楊酸相對含量。…73
圖16、葉用枸杞轉錄組Contigs長度分佈。………………………………………74
圖17、葉用枸杞葉及癭基因本體論。………………………………………………75
圖18、葉用枸杞葉及癭PageMan分析。……………………………………………76
圖19、葉用枸杞癭與正常葉間轉錄差異的MapMan基因表現總圖。……………77
圖20、葉用枸杞癭與感染葉間轉錄差異的MapMan基因表現總圖。……………78
圖21、葉用枸杞感染葉與正常葉間轉錄差異的MapMan基因表現總圖。……79
圖22、葉用枸杞癭、感染葉及正常葉間轉錄差異的光反應基因表現。………80
圖23、葉用枸杞癭、感染葉及正常葉間轉錄差異的卡爾文循環基因表現。…81
圖24、葉用枸杞癭、感染葉及正常葉間轉錄差異的二級代謝物基因表現。…82
圖25、葉用枸杞癭與正常葉間轉錄差異的四吡咯生合成基因表現。…………83
 
表 目 錄
表1、葉用枸杞癭於不同發育階段之形態分析。………………………………84
表2、葉用枸杞不同感染程度葉片、癭之光合作用參數。…………………85
表3、葉用枸杞不同感染程度葉片、癭與市售果實甲醇萃取物抗氧化活性之IC50。…86
表4、不同季節葉用枸杞葉片、癭與市售果實之酚類化合物積分面積比值。…87
表5、葉用枸杞轉錄組重疊序列組合及Contigs組裝品質。………………………88
表6、各植物功能群比對基因數與差異表現基因數(DEGs)以及其所佔比例。…89
表7、葉用枸杞葉及癭光合作用相關基因數與差異表現基因數(DEGs)以及其所佔比例。…91
表8、葉用枸杞葉及癭光合作用相關功能類別及倍數差異。……………………92
表9、葉用枸杞葉及癭二級代謝物相關基因數與差異表現基因數(DEGs)以及其所佔比例。…95
表10、葉用枸杞葉及癭二級代謝物相關相關功能類別及倍數差異。……………96
表11、葉用枸杞葉及癭生物逆境相關基因數與差異表現基因數(DEGs)以及其所佔比例。…98
表12、葉用枸杞葉及癭生物逆境相關相關功能類別及倍數差異。……………99
表13、葉用枸杞葉及癭植物激素相關基因數與差異表現基因數(DEGs)以及其所佔比例。…102
表14、葉用枸杞葉及癭植物激素相關相關功能類別及倍數差異。…………103
表15、葉用枸杞葉及癭四吡咯相關基因數與差異表現基因數(DEGs)以及其所佔比例。…106
表16、葉用枸杞葉及癭四吡咯相關相關功能類別及倍數差異。………………107
dc.language.isozh-TW
dc.subject枸杞zh_TW
dc.subject癭zh_TW
dc.subject光合作用zh_TW
dc.subject植化物質zh_TW
dc.subject抗氧化能力zh_TW
dc.subject次世代定序zh_TW
dc.subjectantioxidant abilityen
dc.subjectgallen
dc.subjectLycium chinenseen
dc.subjectphotosynthesisen
dc.subjectnext generation sequencingen
dc.subjectphytochemicalsen
dc.title蟲癭對枸杞葉光合作用及機能性成分與其轉錄體之影響zh_TW
dc.titleEffects of Gall on Photosynthesis and Functional Compounds and Their Transcriptomes of Wolfberry Leafen
dc.typeThesis
dc.date.schoolyear107-1
dc.description.degree碩士
dc.contributor.coadvisor楊棋明(Chi-Ming Yang)
dc.contributor.oralexamcommittee羅筱鳳(Hsiao-Feng Lo),林冠宏(Kuan-Hung Lin),黃文達(Wen-Dar Huang)
dc.subject.keyword枸杞,癭,光合作用,植化物質,抗氧化能力,次世代定序,zh_TW
dc.subject.keywordLycium chinense,gall,photosynthesis,phytochemicals,antioxidant ability,next generation sequencing,en
dc.relation.page120
dc.identifier.doi10.6342/NTU201900547
dc.rights.note有償授權
dc.date.accepted2019-02-14
dc.contributor.author-college生物資源暨農學院zh_TW
dc.contributor.author-dept園藝暨景觀學系zh_TW
顯示於系所單位:園藝暨景觀學系

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