臺灣博碩士論文加值系統

本論文是應用RAPD(random amplified polymorphic DNA)標記建立一甜
瓜(Cucumis melo)的遺傳連鎖圖譜(linkage map)。使用單株母本(東方甜
瓜SLK-V-052)與單株父本(洋香瓜PI-414723)雜交後得F1植株,再由單株F1
自交後得F2代植株群。在受篩選的300支含10個鹽基序列的逢機引子(GC含
量百分比從50%-90%)中,共有45個引子在親代與F1代中呈現有多型性的
RAPD標記,而每個引子所衍生出的RAPD標記則通常不只一個,甚至可多達四
個。RAPD標記的長度在200bp至2000bp之間; 75%的標記長度短於1kb。由
初次引子篩選所得之逢機引子,經第二次確認其中14個引子會表現20個多
型性RAPD標誌,觀察這些標記於98株F2代植株群中的分離情形,並將以卡方
檢定(Chi-square)判定是否合乎孟德爾遺傳定律的3:1分離比例,經由軟
體"Linkage 1"的分析計算出這些RAPD標記間的重組機率(recombination
frequency);經由Kosambi''s mapping function公式修正後,計算出各個
RAPD標記間的距離(map distance)建立出一包括七個連鎖群的部份甜瓜連
鎖圖譜,七個連鎖群涵蓋421.66cM。

This research project is intended to construct a RAPD (random
amplified polymorphic DNAs) linkage map of Cucumis melo L.
Conditions such as the amount of DNA template,the concentration
of Mg+2 ions available,the annealing temperature of the primer
used and its duration were optimized to generate reproducible
RAPDpatterns.Furthermore,a F2 population was established from a
F1 plant obtained from a cross of a Eastern melon ♀ parent (cv.
SLK-V-052) and a musk melon ♂ parent (cv. PI-414723) .Fourty
five prescreened from 300 random 10-mer oligonucleotides primers
(with GC% between 50% and 90%) were able to produce DNA
polymorphisms among parental lines and F1 in a preliminary
survey. Their segregation patterns in RAPDs were subsequently
examined on a F2 mapping population comprising of 98
individuals. So far, 20 polymorphic RAPD markers were analyzed.
The mapping distances of 19 RAPD markers were computed using the
software "Linkage1" and their plausible map locations were
determined.In summary, 7 linkage groups spanning 422.61 cM was
recognized.