臺灣博碩士論文加值系統

南瓜 (Cucurbita spp.) 屬於葫蘆科作物，果肉及種子極具營養價值。矮南瓜黃化嵌紋病毒 (Zucchini yellow mosaic virus, ZYMV) 是目前臺灣葫蘆科作物的主要病毒病害之一，感染ZYMV的南瓜，其病徵包括植株生長遲緩，葉片黃化、斑駁、嵌紋及果實畸形，生長初期若遭感染，則會嚴重影響產量。本試驗選擇同為Cucurbita moschata的抗病種原Nigerian Local (NL) 與臺灣市面上常見的感病種原小果木瓜型南瓜 (Local Papaya, PY) 做為親本。將F2和BC1子代接種ZYMV-TN3病毒株後，觀察子代族群病徵的分離比，推測ZYMV之抗性可能由兩個顯性互補基因調控。
此外，為尋找南瓜中與ZYMV抗性基因座連鎖的分子標誌，利用逢機增幅多型性DNA (Randomly Aamplified Polymorphic DNA, RAPD) 配合混合分離族群分析 (bulk segregation analysis, BSA) 進行篩選，在631個RAPD引子中，共找到7個可增幅出多型性片段的引子。將這些多型性片段，分別轉換成序列特徵增幅區域 (Sequence Characterized Amplified Region, SCAR) 或切割增幅多型性序列 (Cleaved Amplified polymorphic sequence, CAPS) 分子標誌，於F2子代進行分析後，發現CA83、CA215及SC522945與ZYMV抗性基因座連鎖。未來期望能再找到與另一個互補基因座連鎖的分子標誌，與本試驗所得到的分子標誌共同進行檢測，將可以提高篩選效率及縮短育成抗病品種的時程。

Squash (Cucurbita spp.) has been grown by humans for fruits and seeds over 10,000 years. There are several viruses infecting squashes in Taiwan, and Zucchini yellow mosaic virus (ZYMV) is one of the most destructive virus. Breeding for ZYMV resistant squash is the best way to control it. The purpose of this study is to tag molecular markers which show the linkage to ZYMV resistance.
A ZYMV resistant line, “Nigerian Local, (NL)”, was crossed to susceptible line, “Local Papaya, (PY)”and a F2, BC1 and BC4 populations were generated for genetic analysis and marker tagging. The result suggested that the inheritance of ZYMV resistance in the near isogenic lines derived from PY x (PY x NL) is controlled by two dominant complemantary genes.
A total of 631 random amplified polymorphic DNA (RAPD) and bulk segregant analysis (BSA) were performed to search useful markers. There were seven polymorphic bands which were converted to sequence characterized amplified region (SCAR) or cleaved amplified polymorphic sequence (CPAS) markers. After linkage analysis, only three markers were shown linkage to ZYMV resistance. In the future, these markers would be useful in marker-assisted selection (MAS) and help us to select the resistant plants more quickly.

中文摘要..................................................................................................i
英文摘要...........................................................................................................................ii
目錄..................................................................................................................................iii
圖目錄...............................................................................................................................v
表目錄..............................................................................................................................vi
第一章 序言 1
第二章 前人研究 2
一、 南瓜之研究 2
（一） 植株形態與經濟價值 2
（二） 常見病毒之危害及影響 3
二、 ZYMV特性 3
（一） ZYMV基因組簡介 4
（二） 病徵及傳播媒介 4
（三） 降低不同病毒危害之防治方式 5
三、 ZYMV抗性南瓜之研究 6
（一） 南瓜抗ZYMV基因之遺傳研究 7
（二） 南瓜分子標誌之研究 8
四、 分子標誌配合BSA法應用於抗病基因之研究 9
第三章 材料與方法 12
一、 試驗材料與族群建構 12
二、 南瓜DNA之萃取 12
三、 ZYMV接種及ELISA病毒檢測 12
四、 RAPD之分析 13
五、 混合族群之建立 14
六、 聚丙醯胺膠體電泳分析 14
七、 多型性之片段回收 14
八、 多型性片段之選殖、篩選與定序 15
九、 SCAR引子之設計 16
十、 CAPS分子標誌之設計 16
十一、 構築遺傳連鎖群與區間定位法 16
第四章 結果 17
一、 抗病親本NL之抗性遺傳分析 17
二、 利用RAPD配合BSA方法篩選與抗性連鎖的分子標誌 17
三、 RAPD多型性片段轉換為SCAR / CAPS標誌 18
四、 SCAR / CAPS標誌於F2族群中之分析 18
五、 SCAR / CAPS標誌於BC4族群 (PYx(PYxNL)) 中之檢測 18
第五章 討論 20
第六章 結語 23
第七章 參考文獻 24
第八章 附錄 35

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