臺灣博碩士論文加值系統

利用植株形態、RAPD (Random Amplified Polymorphic DNA)和SCAR (Sequence Characterized Amplified Regions)三種方法進行萵苣31個crisphead形態品系及26個butterhead形態品系之遺傳歧異度研究。以19項外表性狀指數所得資料進行UPGMA群叢分析，可將57個品系區分成二個群組，遺傳相似度為0.38。第一群組中21個品系皆為crisphead品系，不僅包含全部15個台南結球萵苣品系還包含3個美國品系，第一群組可細分5個亞群，遺傳相似度在0.61以上；第二群組中包含了10個crisphead品系與26個butterhead品系，第二群組可分為6個亞群，遺傳相似度在0.70以上。
RAPD分析中，自130個逢機引子中挑選出29個可產生多型性之引子，共產生271個條帶，平均每引子可產生9.3個條帶，多型性比率為53.9%。57個品系依形態分成三個群組，第一群組有13個品系，全為選自國家作物種原中心13個crisphead品系，相似度為0.86；第二群組有21個品系，包含15個台南區農業改良場crisphead品系、3個美國crisphead品系及3個butterhead品系，相似度為0.85；第三群組包含23個品系全為butterhead品系，相似度為0.82。第二群組包含3個butterhead品系，分布於第三、五亞群，crisphead品系分布於第一、二、四亞群。
以59組SCAR引子分析57個萵苣品系，選出21個具多型性之引子，共產生100個條帶，平均每組引子可產生3.5個多型性條帶，多型性比率為73%。57個品系可被分為三大群組，第一群組由13個國家作物種原中心crisphead品系組成，相似度為0.76；第二群組由15個crisphead品系組成，相似度為0.88；第三群組由26個butterhead品系與3個美國crisphead品系組成，相似度為0.88。比較二分子標誌技術所得結果，RAPD標誌分析樹狀圖各群組間相似度0.78~0.86，而SCAR標誌分析者則為0.68~0.78，顯示RAPD相似度較高。各亞群內品系間分群，RAPD標誌分析樹狀圖之相似度0.84~0.96，SCAR標誌分析者則為相似度0.88~0.97，顯示SCAR相似度較高。
本實驗結果可知，萵苣種原鑑別方法可由園藝性狀如葉色、葉緣齒狀、結球性等進行初步分群，再利用SCAR分子標誌進行鑑別出形態上不同，利用RAPD分析相同形態之品系，使crisphead及butterhead萵苣可有效被鑑別。

The genetic diversity of 31 butterhead (BUT) and 26 crisphead (CRP) accessions were evaluated by morphological traits, RAPD (Random Amplified Polymorphic DNA) and SCAR (Sequence Characterized Amplified Regions). Data for 19 morphological traits were subjected to a genetic diversity analysis, after which a UPGMA cluster analysis was performed. The 57 accessions were clustered into 2 groups according to type with genetic similarity of 0.38. The first group included 15 accessions form Tainan and 3 accessions form the National Germplasm of the USA, which were clustered into 5 subgroups with the similarity beyond 0.61. The second group included 10 crisphead and 26 butterhead accessions, which were clustered into 6 subgroups with the similarity beyond 0.70.
A total of 271 RAPD bands, with a mean band of 9.3 for each primer, were generated using 29 out of 130 primers in the RAPD analysis. The polymorphism was 53.9%. All accessions were clustered into 3 groups in the RAPD analysis. The first group included 13 crisphead accessions form the Taiwan agricultural research institute(TARI) with the similarity 0.86. The second group including 15 accessions form the Tainan district agricultural research and extention station(TDARES)、3 accessions form USA and 3 butterhead accessions with the similarity 0.88. The first, second and fourth subgroups included crisphead accessions. The third and fifth subgroups included 3 butterhead accessions. The third group was composed of 23 butterhead accessions with a the similarity of 0.82.
In analyzing results from SCAR of butterhead and crisphead accessions, a total of 100 bands were generated using 21 out of 51 primer pairs. The means of bands generated by a primer pairs was 4.8 and the polymorphism was 73%. The 57 accessions were clustered into 3 groups. The first group included 13 crisphead accessions form the TARI with the similarity 0.76. The second group including 15 accessions form TDARES with a similarity of 0.88. The third group was composed of 26 butterhead and 3 accessions the USA with a similarity of 0.88. The analysis of RAPD and SCAR result in the main group of the dendrogram showed a similarity of 0.78~0.86 based on RAPD markers and which the SCAR markers displayed a similarity 0.68~0.78. The analysis of RAPD and SCAR result in the subgroup of the dendrogram showed a similarity of 0.84~0.96 based on RAPD markers and which the SCAR markers displayed a similarity 0.88~0.97.
Lettuce germplasm could be identified by morphological characteristics such as leaf color, leaf texture and head type. The molecular markers could be used to cluster accessions into groups by SCAR and subgroups by RAPD successfully.

中文摘要.............................................................................................................i
英文摘要.............................................................................................................ii
壹、 前言......................................................................................................................1
貳、 前人研究
一、 萵苣之起源、分類與植株性狀....................................................................2
二、 品種鑑定技術之發展..................................................................................5
三、 RAPD標誌技術之原理與特色..................................................................6
四、 SCAR標誌技術之原理與特色...................................................................8
五、 RAPD及SCAR分子標誌在農業上之應用...............................................8
參、 材料方法
一、材料與植株形態性狀調查..........................................................................13
二、逢機增殖多型性DNA標誌(DNA markers)分析. .....................................15
三、SCAR多型性標誌分析...............................................................................16
肆、 結果
一、 以形態性狀鑑別萵苣種原…....................................................................18
二、 RAPD標誌分析…………........................................................................48
三、 SCAR標誌分析.......................................................................................119
伍、 討論
一、 以形態性狀鑑別萵苣種原......................................................................170
二、 RAPD標誌分析......................................................................................172
三、 SCAR標誌分析.......................................................................................175
四、 結論..........................................................................................................177
陸、 參考文獻..........................................................................................................178
柒、 附錄..................................................................................................................186

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