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研究生:張育維
研究生(外文):Yu-Wei Chang
論文名稱:土肉桂葉部揮發性成分及根部微生物多樣性之研究
論文名稱(外文):Study on Volatile Compounds from Leaves and Microbial Diversity of Roots of Cinnamomum osmophloeum Kanehira
指導教授:林瑞進林瑞進引用關係
指導教授(外文):Lei-Chen Lin
學位類別:碩士
校院名稱:國立嘉義大學
系所名稱:森林暨自然資源學系研究所
學門:農業科學學門
學類:林業學類
論文種類:學術論文
畢業學年度:102
語文別:中文
論文頁數:80
中文關鍵詞:土肉桂揮發性成分土壤微生物暗色隔膜內生菌變性梯度凝膠電泳分析
外文關鍵詞:Cinnamomum osmophloeum Kanehiravolatile compoundssoil microorganismsdark septate fungal endophytesdenaturing gradient gel electrophoresis
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土肉桂(Cinnamomum osmophloeum Kanehira)為台灣原生樹種,其精油(Essential oil)含量豐富,常散發出令人愉悅的芳香氣味,而芳香氣味為植物重要特徵之一,因此本試驗即利用固相微萃取技術(Solid-phase microextraction,SPME)配合氣相層析質譜儀(Gas chromatography-mass spectrometry,GC/ MS)分析不同甜度土肉桂與印尼肉桂(C. burmannii Blume)葉部揮發性成分的組成及差異,以簡便法來快速區分與鑑定不同甜味土肉桂及印尼肉桂;並利用分子生物技術探討土肉桂不同品系間之土壤微生物多樣性。

首先是土肉桂精油的部分,經發現越甜者肉桂醛(Cinnamaldehyde)含量越高,以最甜土肉桂(Co1)之Cinnamaldehyde (81.08%)為最多,而越不甜者則呈現相反趨勢,並發現越不甜者則Linalool含量越高,以不甜土肉桂(Co5)之Linalool (94.36%)為最多,印尼肉桂則兩者成分含量都很低,其成分中以Borneol (63.32%)及1,8-Cineole (17.68%)兩成分含量為最高,接著是揮發性成分的部分,與精油有相同的趨勢,以最甜土肉桂(Co1)之Cinnamaldehyde含量74.87%為最多,而不甜土肉桂(Co5)之Linalool 含量64.14%為最多。此外,在日照時間試驗上亦發現各試驗樹種主要成分於夜照時間所萃取的含量皆高於日照時間所測得的含量。

其次在根菌結合體方面,於顯微構造觀察中無發現外生菌根的特徵,但於染根觀察中發現具有暗色隔膜內生菌之構造。另於微生物相部分,經變性梯度凝膠電泳分析(Denaturing gradient gel electrophoresis, DGGE)分析得知,在這2種土肉桂品系中,其土壤微生物之固氮菌及細菌的菌相皆相當豐富,因此目前初步以DGGE分析得知,土肉桂植株中Cinnamaldehyde含量之多寡無法與固氮菌相及細菌相找到相關性。
In Taiwan, Cinnamomum osmophloeum Kanehira is an endemic species that is rich in essential oil. It often emits a fragrant odor that makes people feel pleasant, and this fragrant odor is one of the important characteristics of the plant.

In this study, we used solid-phase micro-extraction and gas chromatography-mass spectrometry to analyze the composition of volatiles from leaves of C. osmophloeum and C. burmannii Blume that taste sweet and non-sweet. Also, we compared the differences in leaf volatiles between C. osmophloeum and C. burmannii, which is a simple method to distinguish between them due to their different sweetnesses. We also used molecular biotechnology to explore the microbial diversity between different strains of C. osmophloeum.

First, based on pretesting, the content of cinnamaldehyde is higher when C. osmophloeum leaves taste sweet, whereas non-sweet leaves show an opposite trend. We also found that non-sweet leaves of C. osmophloeum have a higher content of linalool. In addition, based on the results obtained from the volatiles analysis in daytime and at night, we found that the content of major volatile constituents at night were higher in both species.

The second part of our study focused on the fungus-root association of C. osmophloeum. This research demenstrated no ectomycorrhizal structure under SEM, but had the DSE structure under light microscopy. A molecular analysis of soil microorganisms using DGGE indicated that nitrogen-fixing bacteria and bacterial microflora were abundant. Therefore, the result of this study indicated the nitrogen-fixing bacteria and the bacterial microflora from the DGGE analysis did not clearly show relationship to the amount of cinnamaldehyde in C. osmophloeum.
摘要......................................................I
Abstract................................................III
目次.....................................................V
表目次...................................................VII
圖目次..................................................VIII
I.前言....................................................1
II.前人研究...............................................3
(I)樹種介紹...............................................3
(II)土肉桂與印尼肉桂形態特徵的比較............................6
(III)芬多精(Phytocide)及精油(Essential oil)................8
(IV)天然物有效成分之萃取方法.................................10
(V)天然物成分之分析........................................17
(VI)土肉桂天然有效成分之應用.................................18
(VII)森林土壤.............................................25
(VIII)土壤有益微生物.......................................26
(IX)微生物菌相結構分析技術...................................27
(X)菌根...................................................29
(XI)暗色隔膜內生菌(Dark septate fungal endophytes, DSE )........30
(XII)微生物與化學成分的關係..................................33
III.研究目的..............................................35
IV.材料與方法.............................................36
( I )實驗材料及儀器........................................36
(II)試驗方法..............................................38
V.結果與討論..............................................49
(I)水蒸餾法分析............................................49
(II)葉片破碎萃取分析.......................................52
(III)葉片非破壞萃取分析.....................................57
(IV)土壤性質測定...........................................60
(V)土壤微生物分析..........................................62
(VI)根菌結合體形態觀察:....................................65
VI.結論..................................................68
VII.參考文獻..............................................70
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