臺灣博碩士論文加值系統

物種的鑑定有助於種原資料的收集，亦可保護育種者的智慧財產權。本研究以25個台灣現有落花生品系的乾燥成熟種子為試驗材料，以SDS-PAGE蛋白質電泳圖譜與逢機增殖多型性DNA (random amplified polymorphic DNA, 簡稱RAPD) 標誌進行落花生品系的鑑定。蛋白質圖譜分析的結果，產生的多型性不多，但分子量19.0 KDa為Virginia bunch型的特有分子標誌，可將立枝仔與其他品系鑑別；分子量20.0 KDa為Virginia runner型的特有分子標誌，可將澎湖二號、澎湖三號以及大冇與其他品系鑑別，然而同一植物型態之間各品系則無法區別。
以10個逢機引子對25個落花生品系進行RAPD分析，直接偵測DNA序列的差異，共產生184條DNA條帶，其中有128個分子標誌在品系間具有多型性，其中有19個分子標誌為特定品系的特有標誌。RAPD分子標誌技術操作簡易、快速、安全，以乾燥成熟種子作為試驗材料，可節省大量時間，唯宜降低萃取DNA時高含油量的干擾，提升萃取DNA的純度，提高RAPD反應的再現性，以達快速鑑定落花生品系之目的。

Species identification has contribution to the collection of germplasm information and can also protect intellectual property of plant breeders. The research is to identify 25 peanut lines available in Taiwan by SDS-PAGE protein electrophoresis and random amplified polymorphic DNA (RAPD) markers using dry mature seeds. There is not much polymorphism of the outcome of protein profile analysis. However, 19.0 KDa protein molecular marker is specific to peanut of Virginia bunch type, and can identify line Lichitsai from the rest of 25 peanut lines tested. And 20.0 KDa protein molecular marker is specific to peanut of Virginia runner type, and can identify line Ponghu 1, Ponghu 2 and Taimo from the rest of 25 peanut lines tested. Nevertheless, peanut lines within the same morphology type cannot be identified by protein profiles.
Ten random primers were used to detect differences of DNA sequence of 25 peanut lines by RAPD reactions. Hundred and eighty four DNA fragments were produced, and 128 molecular markers among them were polymorphic between different lines. And 19 molecular markers were unique to specific lines. RAPD technique is quick, safe and easy to operate. It will save much time using dry mature seeds as experiment material as long as reducing oil content of DNA extraction from seeds in order to raise DNA purity and enhance reduplication of RAPD reaction.

壹、前言．．．．．．．．．．．．．．．．．．．．．．． 1
貳、前人研究．．．．．．．．．．．．．．．．．．．．． 3
參、材料與方法．．．．．．．．．．．．．．．．．．．． 6
一、材料．．．．．．．．．．．．．．．．．．．．． 10
二、方法．．．．．．．．．．．．．．．．．．．．． 10
三、資料分析．．．．．．．．．．．．．．．．．．． 18
肆、結果．．．．．．．．．．．．．．．．．．．．．．． 20
一、蛋白質圖譜分析．．．．．．．．．．．．．．．． 20
二、RAPD分子標誌分析．．．．．．．．．．．．．．． 20
伍、討論．．．．．．．．．．．．．．．．．．．．．．． 60
陸、參考文獻．．．．．．．．．．．．．．．．．．．．． 64

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