| 研究生: |
吳紀瑄 Wu, Chi-Hsuan |
|---|---|
| 論文名稱: |
台灣原生種白花蝴蝶蘭粒線體基因表現之分析 The analysis of mitochondrial gene expression in Phalaenopsis aphrodite subsp. formosana |
| 指導教授: |
張清俊
Chang, Ching-Chun |
| 學位類別: |
碩士 Master |
| 系所名稱: |
生物科學與科技學院 - 生物科技研究所 Institute of Biotechnology |
| 論文出版年: | 2014 |
| 畢業學年度: | 102 |
| 語文別: | 中文 |
| 論文頁數: | 139 |
| 中文關鍵詞: | 台灣白花蝴蝶蘭 、粒線體RNA 、轉錄體 |
| 外文關鍵詞: | P. aphrodite subsp. formosana, mitochondrial RNA, transcriptomes |
| 相關次數: | 點閱:116 下載:6 |
| 分享至: |
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台灣原生種白花蝴蝶蘭(Phalaenopsis aphrodite subsp. formosana),為野外瀕臨絕種的植物與商業育種上的重要親本。本論文主要著重於探討白花蝴蝶蘭粒線體的轉錄體、RNA編輯與基因表現。藉由純化蝴蝶蘭花部組織的粒線體RNA,再利用Illumina次世代定序系統將蝴蝶蘭粒線體的轉錄子進行解序。註解出具有粒線體特異性的3個rRNA基因,6個tRNA基因與25個蛋白質編碼基因。進一步將轉錄體與基因組做比較後,在25個編碼蛋白質的轉錄子中發現有393個RNA編輯位點。另外,3個內插子發現了8個RNA編輯位置。此外,不同轉錄子的RNA編輯位點在不同組織的編輯效率上有些差異。另外,利用即時定量PCR系統分析在不同生長發育時期的根、葉與花部組織,發現部分粒線體基因的表現量與細胞核ubiquitin 10基因的表現量相當,有些卻少於ubiquitin 10的表現量,但是葉綠體基因的表現量皆高於粒線體基因。
The moth orchid, P. aphrodite subsp. formosana is the endemic species in Taiwan, but now it is endangered in the original habitat. This moth orchid has been commonly used as parental strains in commercial breeding program. In this study, the mitochondrial RNA (mtRNA) was isolated from the floral tissues. Subsequently, the mitochondrial transcriptomes was determined by next generation sequencing through Illumina methods. By BLAST mtRNA sequence to NCBI database, 3 rRNA, 6 tRNA and 25 protein-coding transcripts specifically to mitochondria were annotated. In addition, the mitochondrial RNA editing sites were determined by comparing the sequence of genomic DNA versus transcriptomes. In total, 393 editing sites from 25 transcripts were identified. Additionally, 8 RNA editing sites from 3 introns were found. Differential frequency of RNA editing pattern in transcripts were observed at different tissues. When the mitochondrial gene expression were examined from roots, leafs or floral tissues at different developmental stages in mitochondrial and chloroplast gene by real time RT-PCR, some of the mitochondrial transcripts show as abundant as nuclear ubiquitin 10 mRNA, but some transcripts are less than that of ubiquitin 10 mRNA. In general, mitochondrial transcripts are less abundant than that of chloroplast transcripts in all tested tissues.
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