Please use this identifier to cite or link to this item:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/83587
Title: | 臺灣扁柏枝條之揮發性萜類合成酶基因選殖與功能鑑定 Gene Cloning and Functional Characterization of Volatile-terpene Synthase Gene from Branches of Chamaecyparis obtusa var. formosana |
Authors: | 陳廷瑋 Ting-Wei Chen |
Advisor: | 曲芳華 Fang-Hua Chu |
Keyword: | 臺灣扁柏,次世代定序,萜類合成酶,單萜類化合物,倍半萜類化合物, Chamaecyparis obtusa var. formosana,Next Generation Sequence (NGS),Terpene synthase,Monoterpeniods,Sesquiterpenoids, |
Publication Year : | 2022 |
Degree: | 碩士 |
Abstract: | 臺灣扁柏 (Chamaecyparis obtusa var. formosana),是臺灣珍貴的特有種植物,為日本扁柏 (Chamaecyparis obtusa) 的變種。在利用上非常的廣泛,具有良好質地及材色的主幹適合作為建材和家具,並且整株植物皆富含香氣,可製作成精油及芳香劑等一系列產品。在精油的組成中以揮發性萜類化合物為主,其中不乏只有在臺灣扁柏中才能發現的化合物,為了更加清楚這些產物的生合成機制,因此對其合成酶做更進一步的研究。然而,現今並沒有對於臺灣扁柏萜類合成酶之相關研究及文獻,若能成功的將基因選殖出來並鑑定出功能,除了可與其他植物的揮發性萜類合成酶進行比較外,亦能作為未來為重要成分的半化學合成之設計藍圖。本次實驗自臺灣扁柏的枝條進行轉錄體的次世代定序,並從資料庫中獲得 6 條揮發性萜類合成酶的序列,分別為 Covf_121、Covf_339、Covf_13139、Covf_2194、Covf_16187 及 Covf_10716,並透過 in vitro 及 in vivo 實驗之結果與胺基酸中保守功能序列的分析,可將 6 條揮發性萜類合成酶更進一步區分成單萜及倍半萜合成酶,並且依主要產物對其命名,單萜合成酶分別為 CovfCar (3-Carene synthase)、CovfLin (Linalool synthase) 及 CovfTer (Terpinolene synthase),以及倍半萜類合成酶為 CovfGerA (Germacrene A synthase)、CovfAco (Acoradiene synthase) 與 CovfLon (Longifolene synthase)。 Chamaecyparis obtusa var. formosana, variant of Chamaecyparis obtusa, is a precious and endemic plant in Taiwan. It is very widely used. The trunk with good texture and color is suitable for building materials and furniture, and the whole plant is rich in aroma, which can be made into essential oils, fragrance and a series of related products. Volatile terpenoids are the major compounds in the composition of essential oils, many of which can only be found in C. obtusa var. formosana. Further research on their synthases was carried out to better understand the biosynthesis mechanism of these products. However, there were no reports on terpenoid synthases in C. obtusa var. formosana until now. Thus, we cloned and identified their functions to obtain the information for understanding the relationship between other plant volatile-terpenoid synthases and serving as a blueprint for the semi-chemical synthesis of important components in the future. In this experiment, the sequences of 6 volatile-terpenoid synthases were obtained from the Next Generation Sequence (NGS) database of branch transcripts of C. obtusa var. formosana, including Covf_121, Covf_339, Covf_13139, Covf_2194, Covf_16187 and Covf_10716. Through the results of in vitro and in vivo experiments and the analysis of conserved motif sequences in amino acids, the six volatile terpenoid synthases can be further distinguished into monoterpenoid synthases and sesquiterpenoid synthases, named according to their main products. There are CovfCar (3-Carene synthase), CovfLin (Linalool synthase) and CovfTer (Terpinolene synthase) of monoterpenoid synthases and CovfGerA (Germacrene A synthase), CovfAco (Acoradiene synthase) and CovfLon (Longifolene synthase) of sesquiterpenoid synthases. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/83587 |
DOI: | 10.6342/NTU202202813 |
Fulltext Rights: | 未授權 |
Appears in Collections: | 森林環境暨資源學系 |
Files in This Item:
File | Size | Format | |
---|---|---|---|
ntu-110-2.pdf Restricted Access | 14.58 MB | Adobe PDF |
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.