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| 研究生： |
許惠嵐 Hsu, Hui-Lan |
|---|---|
| 論文名稱： |
台灣白花蝴蝶蘭第七條染色體上遺傳標誌的染色體圖譜 Chromosome mapping of genetic markers on chromosome 7 Phalaenopsis aphrodite subsp. formosana |
| 指導教授： |
張松彬
Chen, Song-Bin |
| 學位類別： |
碩士 Master |
| 系所名稱： |
生物科學與科技學院 - 生命科學系 Department of Life Sciences |
| 論文出版年： | 2016 |
| 畢業學年度： | 104 |
| 語文別： | 英文 |
| 論文頁數： | 48 |
| 中文關鍵詞： | 蝴蝶蘭屬 、螢光原位雜合 、粗絲期染色體 、染色體圖譜 |
| 外文關鍵詞： | Phalaenopsis orchid, Fluorescent in situ hybridization (FISH), pachytene chromosome, chromosome mapping |
| 相關次數： | 點閱：102 下載：2 |
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蘭科(Orchidaceae)是顯花植物中最大的一科，包含25000個以上的物種。蝴蝶蘭屬(Phalaenopsis)為其中一屬，在全球園藝市場中屬高價經濟作物。研究蝴蝶蘭屬植物的基因體可增進我們對於蘭科植物的了解。然而截至今日，只有一個蝴蝶蘭屬物種的基因體組被大致解序。台灣白花蝴蝶蘭(Phalaenopsis aphrodite subsp. formosana)在蝴蝶蘭育種中是一重要親本，也是蘭科植物的模式物種之一。雖然其基因及基因組資訊已可得，但由於不同版本的遺傳圖譜(genetic map)有所差異，導致台灣白花蝴蝶蘭序列拼裝上的困難。染色體圖譜(chromosome map)可扮演一個檢驗序列拼裝正確性的角色，但台灣白花蝴蝶蘭的中期染色體小，若用於檢驗序列拼裝會受限於解析度偏低而無法區分不同遺傳標誌(genetic markers)的相對位置。本研究建立了一個改良過的滴落法(drop method)，可用於製備台灣白花蝴蝶蘭高解析度粗絲期(pachytene)染色體，並且檢驗粗絲期染色體於螢光原位雜合(fluorescent in situ hybridization, FISH)技術上的適用性。藉由定位屬於D_L10連鎖群的四個鷹架(scaffold) DNA至台灣白花蝴蝶蘭粗絲期染色體上，我們檢測不同版本的遺傳圖譜其正確性，並確認D_L10連鎖群位於台灣白花蝴蝶蘭第七條染色體上。同時，我們觀察到在第七條染色體不同區域，其重組頻率(recombination frequency)會有所差異。另外，台灣白花蝴蝶蘭粗絲期染色體也可應用至雷射捕捉顯微切割(laser capture microdissection, LCM)技術上，未來有潛力建構個別染色體資料庫(chromosome-specific library)。由滴落法所製備的台灣白花蝴蝶蘭粗絲期染色體可以應用在染色體圖譜的建立上，可整合遺傳圖譜及實質圖譜，加速台灣白花蝴蝶蘭的基因體組及細胞遺傳學研究，進而增進我們對於蝴蝶蘭屬植物的了解。
Orchidaceae is the largest flowering family, which consists of at least 25,000 species. Phalaenopsis, an Orchidaceae genus, is a high-value ornamental plant in global horticulture market. Studying Phalaenopsis genomes will benefit our knowledge of orchid biology. Up to now, there is only one species that its draft genome sequence has been determined in Phalaenopsis. Phalaenopsis aphrodite subsp. formosana is an important parental material in breeding and one of model organisms for studying orchid biology. Due to different assembly strategies, they resulted in inconsistent linkage groups. Cytogenetic maps can ensure the accuracy of linkage groups, but small-size metaphase chromosomes of P. aphrodite limit cytogenetic researches and applications. In this study, we developed a modified drop method for pachytene chromosomes preparation in P. aphrodite and examined its applicability for fluorescent in situ hybridization (FISH) technique. By mapping four scaffolds of D_L10 linkage group to high-resolution pachytene chromosomes, we validated the genome assembly of P. aphrodite, and found that there are differences of recombination frequency among different chromosome regions. Also, the chromosome which D_L10 linkage group located on was identified as chromosome 7 of P. aphrodite. Additionally, we examined that pachytene chromosomes of P. aphrodite can apply on chromosome laser capture microdissection (LCM) which could establish chromosome-specific library. The development of modified drop method for pachytene chromosome preparation would accelerate the integration of genetic map and chromosome map, and also benefit the genomic and cytogenetic researches in P. aphrodite.
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校內：2021-01-01公開校外：2021-04-01公開