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研究生: 林斯賢
Lin, Szu-Hsien
論文名稱: 長葉木薑子蟲癭轉錄體之研究
Transcriptomic profiling of insect gall in Litsea acuminata
指導教授: 孫智雯
Sun, Chih-Wen
楊棋明
Yang, Chi-Ming
學位類別: 碩士
Master
系所名稱: 生命科學系
Department of Life Science
論文出版年: 2016
畢業學年度: 104
語文別: 中文
論文頁數: 82
中文關鍵詞: 蟲癭長葉木薑子次世代定序轉錄體光合作用植物激素細胞壁植物防禦系統
英文關鍵詞: insect gall, Litsea acuminata, next generation sequencing, transcriptome, photosynthesis, plant hormone, cell wall, plant defense system
DOI URL: https://doi.org/10.6345/NTNU202203615
論文種類: 學術論文
相關次數: 點閱:119下載:3
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  • 蟲癭(insect gall)是植物經過造癭昆蟲刺激後產生不正常增生組織。至今蟲癭的形成機制仍不明瞭。先前研究著重於分類學、動物學、生態學等,隨著次世代定序發展,使得探討基因表現等轉錄體研究也越快捷。本論文研究長葉木薑子(Litsea acuminata)蟲癭之轉錄體,以了解蟲癭的形成發育機制。研究蟲癭與其宿主葉基因表現差異,一共發現1,452個差異表現基因,其中包含細胞壁、植物激素代謝、植物逆境、光合作用、二次代謝、四吡咯生合成等類群。資料顯示蟲癭組織光合作用相關基因表現量降低,推測蟲癭組織無法提供足夠養分構築蟲癭結構或是提供造癭昆蟲,營養則可能由宿主葉而來,進而顯示植物組織從葉片轉變成蟲癭組織是一種營養流的改變,即從供源(source)轉變成積儲(sink)。本研究還發細胞壁相關基因表現上升,以及植物激素如油菜素類固醇、生長素與吉貝素相關基因表現上升,並推測上述幾種植物激素在蟲癭形成中可能扮演重要角色。造癭昆蟲不但誘導蟲癭構形還啟動植物防禦系統,如致病過程相關蛋白質(pathogenesis-related proteins, PR protein)基因表現上升,此可能誘發植物系統性誘導抗病機制(systemic acquired resistance)以阻礙病原侵入。

    Insect galls are the abnormal growth of plant tissue induced by herbivore, bite or egg laying from insects. To date, the mechanisms of gall formation are still undetermined. Previous researches used taxonomical, zoological and ecological approaches to study insect galls. In this study the insect galls were investigated with the view of transcriptome. We collected cup-shaped galls on host leaves of Litsea acuminata, and characterized the differential gene expression between galls and its host leaves by using next generation sequencing and transcriptomic approaches. A total of 1,452 differentially expressed genes (DEGs) were found between insect gall and it host leaf. These DEGs are classified into functional category of cell wall, hormone metabolism, abiotic or biotic stress, photosynthesis, secondary metabolism, tetrapyrrole synthesis and so on. The data showed that the gall shift from a source (host leaves) to a sink (gall tissues) with decreased gene expression of photosynthetic-related genes. I also analyzed the cell wall synthesis and plant hormone like brassinosteroids, auxin and gibberellin. These hormones may play important role in gall development. Insects not only induce the structure of galls but also stimulate plant defense system. For example that increased expression of genes related to pathogenesis-related proteins (PR protein). They are induced as part of systemic acquired resistance (SAR) to resist disease.

    目錄 中文摘要 i Abstract ii 目錄 iv 圖目錄 vi 表目錄 vii 緒論 1 研究材料與方法 5 一、研究材料 5 二、RNA萃取、純化與轉cDNA 5 三、轉錄組定序 5 四、數據分析 6 五、即時聚合酶鏈式反應(RT-qPCR) 6 六、顯微結構分析 7 結果 8 一、重疊序列組合 8 二、序列功能及細胞位置推測 8 三、序列比對 8 四、移除重複序列 9 五、TAIR資料庫比對 9 六、蟲癭及宿主葉差異表現分析 9 (一)光合作用相關基因 10 (二)粒線體電子電鏈相關基因 11 (三)細胞壁生合成相關基因 11 (四)植物激素相關基因 12 (五)植物逆境相關基因 12 (六)水分傳輸相關基因 13 (七)葉片結構生長發育相關基因 13 七、即時聚合酶鏈式反應結果分析 13 八、顯微結構分析 14 討論 15 一、光合作用 15 二、蟲癭的生長發育 16 三、植物防禦系統 18 四、營養流 18 五、定序品質 19 六、即時聚合酶鏈式反應驗證 19 結論 20 未來展望 21 參考文獻 22 檢索表 82   圖目錄 圖1、長葉木薑子與其蟲癭 35 圖2、Contigs長度與對應到蛋白質數量分布 36 圖3、蟲癭及宿主葉間轉錄體水平變化之關係 37 圖4、Gene Ontology 38 圖5、比對14種植物蛋白質資料庫之各物種比對到基因數比例 39 圖6、MapMan基因表現 40 圖7、PageMan分析 41 圖8、光合作用基因表現 42 圖9、粒線體電子傳遞鏈基因表現 43 圖10、細胞壁相關基因表現 44 圖11、四吡咯(tetrapyrrole)生合成(葉綠素a及b)基因表現 45 圖12、油菜素類固醇生合成基因表現 46 圖13、即時聚合酶鏈式反應與轉錄體資料迴歸關係 47 圖14、光合作用相關基因之即時聚合酶鏈式反應結果 48 圖15、四吡咯相關基因之即時聚合酶鏈式反應結果 49 圖16、色素蛋白複合體用相關基因之即時聚合酶鏈式反應結果 50 圖17、粒線體電子傳遞鏈相關基因之即時聚合酶鏈式反應結果 51 圖18、細胞壁相關基因之即時聚合酶鏈式反應結果 52 圖19、油菜素類固醇相關基因之即時聚合酶鏈式反應結果 53 圖20、長葉木薑子葉片及蟲癭之光學顯微鏡分析 54 圖21、長葉木薑子葉片及蟲癭之穿透式電子顯微鏡分析 55 圖22、長葉木薑子葉片及蟲癭之類囊膜間距差異 56  表目錄 表1、重疊序列組合及Contigs組裝品質 57 表2、各植物功能類群比對基因數與差異表現基因數及所佔比例 58 表3、光合作用相關基因數與DEGs數及所佔比例 60 表4、光合作用相關DEGs及其所屬TAIR ID與倍數差異 61 表5、粒線體電子傳遞鏈相關基因數與DEGs數及所佔比例 62 表6、粒線體電子傳遞鏈相關DEGs及其所屬TAIR ID與倍數差異 63 表7、細胞壁相關基因數與DEGs數及所佔比例 64 表8、細胞壁相關DEGs及其所屬TAIR ID與倍數差異 65 表9、植物激素相關基因數與DEGs數及所佔比例 68 表10、植物激素相關DEGs及其所屬TAIR ID與倍數差異 69 表11、植物逆境相關基因數與DEGs數及所佔比例 71 表12、植物逆境相關DEGs及其所屬TAIR ID與倍數差異 72 表13、水分傳輸相關基因數與DEGs數及所佔比例 75 表14、水分傳輸相關DEGs及其所屬TAIR ID與倍數差異 76 表15、葉片結構生長發育相關基因數與DEGs數及所佔比例 77 表16、葉片結構生長發育相關DEGs及其所屬TAIR ID與倍數差異 78 表17、即時聚合酶鏈式反應之引子設計 79 表18、次世代定序數據與即時聚合酶鏈式反應資料比對 81

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