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研究生:周怡彤
研究生(外文):Yi-Tung Chou
論文名稱:利用RAPD研究臺灣紅檜直幹與分叉植株間遺傳上之差異
論文名稱(外文):Using RAPD Makers to Study the Genatic variatoin in straight and folking trees of Chamaecyparis formosensis
指導教授:王亞男王亞男引用關係
指導教授(外文):Ya-Nan Wang
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:森林學研究所
學門:農業科學學門
學類:林業學類
論文種類:學術論文
論文出版年:1999
畢業學年度:88
語文別:中文
論文頁數:56
中文關鍵詞:紅檜直幹分叉遺傳變異逢機擴大多型性核酸
外文關鍵詞:Chamaecyparis formosensisstraightfolkinggenetic variationRandom Amplified Polymorphic DNA (RAPD)
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逢機擴大多形性核酸(Random Amplified Polymorphic DNA, RAPD)分析技術,近年來廣被應用在研究植物的遺傳及分類系統上,本研究即利用RAPD之分析探討紅檜直幹與分叉植株間遺傳上之差異。在棲蘭、溪頭及台東三個紅檜族群中各選取直幹及分叉植株各20株樣本,總共120株樣本進行分析,使用了10個人造序列的引子,共記錄了 131 條DNA片段,其中85 條為多型性(64.9%),其餘的46條為單型性(35.1%)。
結果顯示棲蘭地區直幹紅檜之平均遺傳相似性為0.875474,分叉幹紅檜為0.860263;溪頭地區直幹紅檜之平均遺傳相似性為0.829053,分叉幹紅檜為0.788368;台東地區直幹紅檜之平均遺傳相似性為0.853158,分叉幹紅檜為0.850789。
各地區紅檜直幹與分叉植株單株之間的平均遺傳相似性,棲蘭地區紅檜為0.817615,溪頭地區紅檜為0.786115,而台東地區紅檜為0.813137。再經交叉比對不同地區直幹與分叉幹間的平均遺傳相似性,棲蘭直幹與溪頭分叉幹間為0.799416,棲蘭直幹與台東分叉幹間為0.822449,溪頭直幹與棲蘭分叉幹間為0.824667,溪頭直幹與台東分叉幹間為0.808821,台東直幹與棲蘭分叉幹間之遺傳相似性為0.830416,而台東直幹與溪頭分叉幹間之遺傳相似性為0.805474。
棲蘭、溪頭及台東地區內的平均相似性分別為0.867869,0.808711及0.851974。顯示出台灣紅檜三個地區無族群分化現象。
RAPD (Random Amplified Polymorphic DNA) markers have recently been used to estimate genetic variation and taxonomic relationships in plants. In this study, RAPD analysis was performed to determinate genetic variation between straight and folking trees of Chamaecyparis formosensis. Total 120 samples were used in this study, including 20 straight trees and 20 folking trees from each of the three study area (Chilan, Chitou and Taitung ). Ten arbitary sequenced primers were used and 131 DNA fragments were recorded. Among them, 46 fragments showed monomorphism (35.1%)and 85 showed polymorphism(64.9%).
Average similarity coefficient for straight and folking trees in Chilan were 0.875474 and 0.860263 respectively. Average similarity coefficient for straight and folking trees in Chitou were 0.829053 and 0.788368 respectively .For the Taitung area, average similarity coefficient for straight and folking trees of Chamaecyparis formosensis were 0.853158 and 0.850789 respectively.
The Average similarity coefficient of Chamaecyparis within Chilan, Chitou and Taitung were 0.817615, 0.786115 and 0.813137 respectively. By comparing the average similarity coefficient of straight and folking trees of Chamaecyparis formosensis between different area, the average similarity coefficient between straight trees in Chilan and folking trees in Chitou was 0.799416. The average similarity coefficient between straight trees in Chilan and folking trees in Taitung was 0.822449 and the average similarity coefficient between straight trees in Chitou and folking trees in Chilan was 0.824667. The average similarity coefficient between straight trees in Chitou and folking trees in Taitung was 0.808821. The average similarity coefficient between straight trees in Taitung and folking trees in Chilan was 0.830416 and the average similarity coefficient between straight trees in Taitung and folking trees in Chitou was 0.805747.
Average similarity coefficient for Chilan, Chitou and Taitung area were 0.867869, 0.808711 and 0.851974 respectively. These results suggested that Chamaecyparis formosensis in these three area were not differentiated significantly.
壹、前言………………………………………………………………………1
一、研究動機與目的………………………………………………………2
二、研究流程………………………………………………………………3
貳、前人研究…………………………………………………………………5
一、紅檜之相關研究………………………………………………………5
二、分叉幹之定義、影響枝條型態之因子........................6
三、RAPD技術的發展及原理……………………………………………8
四、RAPD技術在林木上的應用……………………………………11
1.在林木遺傳結構分析方面……………………………………12
2.在林木品種的鑑定方面………………………………………14
參、材料與方法………………………………………………………………17
一、試驗材料之採集………………………………………………………17
二、葉片核酸之萃取………………………………………………………19
三、DNA之定量……………………………………………………………21
四、PCR反應………………………………………………………………22
五、PCR產物之電泳………………………………………………………24
六、電泳結果之判讀與分析……………………………………………25
肆、結果與討論………………………………………………………………27
一、RAPD的分析……………………………………………………………27
二、Bulk segregant 混合總DNA樣本試驗........................46
伍、結論………………………………………………………………………50
陸、引用文獻…………………………………………………………………51
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