Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/89182
標題: 扇形文心蘭擬原球體之誘導、增殖及基因轉殖與春石斛蘭品種(系)之親源關係的研究
Studies on protocorm like body induction, multiplication, and gene transformation of Erycina pusilla, and phylogenetic relationships of nobile type Dendrobium orchid
作者: Yu- Sheng Lin
林育聖
關鍵字: 無;NO
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摘要: 
Two subjects were included in this study, studies on protocorm like bodyinduction, multiplication, and gene transformation of Erycina pusilla, andphylogenetic relationships of nobile type Dendrobium orchid. Erycina pusilla is aminiature ornamental flowers, which is suitable as an orchid model plant due toits minimal number of chromosome (2n=12). However, the novel shape,shortening the juvenile phase, and early blooming flowers of Erycina pusilla canbe improved. In this experiment, a suitable production system for protocorm-likebodies (PLBs) induction and propagation of Erycina pusilla was established fromthe seeds and buds. Results showed that the combination of 1 ppm TDZ and 0.2ppm NAA was suitable for PLBs induction, and the 1 ppm BA and 0.2 ppm NAAwas adequate for PLBs maintenance. Subsequently, transformation ofoverexpression vectors of OsMADS14 and OsGA2ox6 genes into the PLBs of theErycina pusilla was conducted via Agrobacterium-mediated gene transformation.Regenerated plantlets were selected by 4 mg/l hygromycin. The regeneratedplantlets were confirmed by PCR and GUS histochemical staining. Resultsindicated that the transformed gene was present in the genome of transformedErycina pusilla.

Nobile dendrobium is a new orchid crop to be promoted in Taiwan due to itspotentials in economic cultivation. Although Taiwan has the potential to promotethis orchid, several problems need to be solved, including limited own varieties,and lacking the genetic information of Taiwan's commercial varieties. RAPD(random amplified polymorphic DNA) technique was used to identify therelationship among twenty commercial varieties (lines) of nobile dendrobium.Twenty-four RAPD primers were selected due to polymorphism, among the 327fragments of all exhibited polymorphism (96.2%). The matrix of pair-wisegenetic similarity coefficient based on Jaccard coefficients, ranged between 0.200to 0.650 among twenty commercial varieties (lines) of nobile dendrobium. Theresults of genetic traits of twenty nobile dendrobium varieties (lines) wereclustered into four groups by UPGMA cluster analysis. The similar clusteringresults were obtained from principal component analysis (PCA).

本研究分為扇形文心蘭擬原球體之誘導、增殖及基因轉殖與春石斛蘭品種(系)之親源關係的研究兩部分。扇形文心蘭屬迷你型蘭花,因其為現已知蘭花染色體數最少者(2n=12),具成為蘭科模式植物的潛能。但育成縮短幼年性、提早開花時間及創新花型與植株型態,藉以提升商品價值,仍有改進的空間 大量表現 OsMADS14 基因可提早水稻抽穗 開花 大量表現 OsGA2ox。、。基因會促進水稻分蘗提早、分蘗數增加及根數目增生之現象、導致植株矮小。本試驗嘗試建立扇形文心蘭擬原球體誘導、增殖系統,並運用農桿菌基因轉殖法,將水稻 OsMADS14 與 OsGA2ox6 基因導入扇形文心蘭,期盼開發出有觀賞價值之創新株型、花型、花期的植株。實驗結果顯示,誘導扇形文心蘭擬原球體宜使用 TDZ 1 ppm  NAA 0.2 ppm 組合,進入擬原球體維持階段則使用 BA 1 ppm  NAA 0.2 ppm 組合為佳。基因轉殖部分,已利用農桿菌法轉殖 OsMADS14 及 OsGA2ox6 基因到扇形文心蘭擬原球體,經過 4mg/l hygromycin 篩選,已獲得擬轉殖再生植株。PCR 分析擬轉殖株可偵測到轉殖之 OsMADS14 及 OsGA2ox6 基因。

春石斛為蘭花產業的明日之星,然商業品種的親源關係研究尚屬不足。本試驗運用逢機增幅片段多型性 DNA (Random Amplified Polymorphic DNA,RAPD)技術對 20 個春石斛品種(系)親源關係進行鑑定。本研究篩選出 24 組具多型性的 RAPD 引子 可產生 327 個具多型性的 DNA 片段(Polymorphism:,96.2%)。經 Jaccard 係數計算分析成對品種(系)間的遺傳相似係數,顯示 20個春石斛品種(系)介於 0.200 至 0.650 之間。利用 UPGMA 群聚分析(clusteranalysis)可大略將其分為 4 個群組,與主成分分析(Principal ComponentAnalysis, PCA)分群結果相似。
URI: http://hdl.handle.net/11455/89182
其他識別: U0005-2805201511442500
Rights: 同意授權瀏覽/列印電子全文服務,2018-06-30起公開。
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