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研究生:張光毅
研究生(外文):Chang Kuang-Yi
論文名稱:甘藷遺傳連鎖圖譜與數量性狀基因座定位
論文名稱(外文):Genetic Linkage Mapping and Quantitative Trait Loci Determination of Sweetpotato
指導教授:黃士穎
指導教授(外文):Hwang Shih-Ying
學位類別:碩士
校院名稱:中國文化大學
系所名稱:生物科技研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
論文頁數:85
中文關鍵詞:ISSR甘藷數量性狀基因座標誌輔助選拔遺傳圖譜連鎖群
外文關鍵詞:ISSRSweetpotatoQuantitative trait loci (QTLs)Marker-assisted selection (MAS)Genetic maplinkage group
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本研究利用Inter-Simple Sequence Repeat(ISSR)分子標誌,對多倍體物種甘藷進行遺傳圖譜繪製。標誌輔助選拔(Marker-assisted selection, MAS)在作物的研究發展中,扮演Genotyping及各種作物育種價(Breeding value)的建立。台農27號(TN 27)與Nancy Hall親本互交方式,產生兩F1雜交子代(TN 27 x Nancy Hall、Nancy Hall x TN 27),連鎖群分析採用Haldane’s map function、LOD值為3 ~ 10及重組互換率(r)≤ 0.35方式進行。12組ISSR Primers可擴增出48(TN 27)與36(Nancy Hall)個標誌,繪製甘藷遺傳圖譜13與10個連鎖群(Linkage groups),得到最終遺傳連鎖圖譜,此外沒有得到Repulsion phase出現在連鎖群中。標誌間平均距離為12.3 cM,平均3個標誌構築1個連鎖群。另以CIM檢驗數量性狀基因座(QTLs),得到TN 27有4個(塊根產量、皮色及肉色)及Nancy Hall有11個(塊根皮色、肉色及形狀)數量性狀基因座。各有學者提出甘藷為同源多倍體(Autopolyploid)與異源多倍體(Allopolyploid)的說法,在本研究中同源與異源多倍體分析皆為非常顯著差異(p<0.001),若以85.4 % Simplex markers解釋,可推論出甘藷為同源多倍體。另將兩親本13與10個遺傳連鎖圖譜結合,連鎖群上所含有相同的29個Double-simplex markers可推導出三個相關圖譜,因此認為,ISSR可將不同分子標誌技術所得之標誌相互結合,形成準確性更高之遺傳連鎖圖譜。
Genetic linkage mapping of sweetpotato (Ipomoea batata (L.) Lam.) based on Inter-Simple Sequence Repeat (ISSR) markers. MAS in agriculture research was to build the rule of genotyping and to improve breeding value of crops. Two F1 progenies obtained from ‘TN 27’ x ‘Nancy Hall’ reciprocol testcross. Linkage groups analysis at a Haldane’s map function, LOD scores of 3 ~10 and recombination (r) ≤ 0.35. A total of 48 and 36 markers were ordered in 13 and 10 linkage groups and didn’t find repulsion phase in linkage groups. The average distance between markers was 12.3 cM and the average 3 markers were constructed one linkage group. Respectively, QTLs analysis in this study found 4 QTLs (Root weight, Root skin color and Root color ) in ‘TN 27’ and 11 QTLs (Root skin color, Root color and Root shape ) in ‘Nancy Hall’. Type of polyploidy analysis of sweetpopato, our data explained that neither autopolyploidy nor allopolyploidy measured indicated significant at P<0.001, but autopolyploidy was suggested at 85.4% of simplex markers. 13 and 10 linkage groups of ‘TN 27’ and ‘Nancy Hall’ were combined with 29 same markers of Double-simplex markers that joined into three relative maps. ISSR marker was to implicate that combined the other molecular markers into genetic linkage maps and improved precision and accuracy of maps.
圖目錄----------------------------------------------------------------------------------Ⅰ表目錄----------------------------------------------------------------------------------Ⅱ中文摘要-------------------------------------------------------------------------------Ⅳ英文摘要-------------------------------------------------------------------------------Ⅴ壹、前言----------------------------------------------------------------------------------1貳、前人研究----------------------------------------------------------------------------4
一、甘藷染色體(Chromosome)-------------------------------------------------4
二、甘藷配子分離率(Segregation Ratio)--------------------------------------5
三、染色體遺傳圖譜與實體圖譜(Genetic and Physical Maps)------------6
四、連鎖群(Linkage groups)分析----------------------------------------------7
五、分子標誌技術(Marker technique)之發展與應用-----------------------7
六、QTLs分析方法--------------------------------------------------------------12
七、連鎖群分析軟體-------------------------------------------------------------12
八、QTLs分析軟體--------------------------------------------------------------13
參、材料與方法-----------------------------------------------------------------------14
一、試驗材料----------------------------------------------------------------------14
二、DNA萃取---------------------------------------------------------------------15
三、DNA定量與稀釋------------------------------------------------------------16
四、ISSR標誌分析---------------------------------------------------------------16
五、電泳----------------------------------------------------------------------------18
六、資料分析----------------------------------------------------------------------21
肆、結果--------------------------------------------------------------------------------26
一、性狀變異數分析-------------------------------------------------------------26
二、ISSR標誌分析---------------------------------------------------------------37
三、甘藷配子分離率分析-------------------------------------------------------41
四、甘藷多倍體基因型分析----------------------------------------------------42
五、連鎖群分析-------------------------------------------------------------------43
六、數量性狀基因座(QTLs)---------------------------------------------------50
七、總基因長度(Genome size)估算------------------------------------------54
伍、討論--------------------------------------------------------------------------------55
一、雜交族群分析----------------------------------------------------------------55
二、性狀變異數分析-------------------------------------------------------------56
三、ISSR標誌分析---------------------------------------------------------------58
四、連鎖群與QTLs分析--------------------------------------------------------60
五、總基因長度估算-------------------------------------------------------------67
陸、結論--------------------------------------------------------------------------------69
柒、參考文獻--------------------------------------------------------------------------71
附圖-------------------------------------------------------------------------------------80
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