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標題: 利用RAPD與ISSR分子標誌評估楊桃品種間之遺傳歧異度
Assessment of Genetic Diversity among Carambola (Averrhoa carambola L.) Cultivars by RAPD and ISSR Markers
作者: 曾湘琇
Tseng, Shung-Sui
關鍵字: Carambola;楊桃;Molecular;分子標誌
出版社: 園藝學系
本研究以RAPD(random amplified and polymorphic DNA)與ISSR(inter simple sequence repeat)分子標誌進行20個楊桃(Averrhoa carambola L.)栽培品種間遺傳相似度之分析與比較其鑑別能力,試驗結果顯示此二種分析在楊桃栽培品種間均表現高度多型性。
本研究用14支RAPD引子共產生196個增殖DNA片段,其中144個增殖DNA的片段具多型性,所產生的片段大小介於1500 bp-190 bp之間。17支ISSR引子共產生266個增殖DNA片段,其中195個增殖DNA的片段具多型性,所產生的片段大小介於1300 bp-200 bp之間。將14支RAPD引子及17支ISSR引子所產生之多型性DNA片段,經Nei/Li方法計算其相似性,並以UPGMA建構相似度樹狀圖分析,結果以RAPD群叢分析能夠將酸味種分群,而ISSR群叢分析酸味種與酸味厚斂雖未能直接分群,但也有一定相似度。受測楊桃中屬長花柱品種有:秤錘、酸味、酸味厚斂、竹葉、五汴頭反、白絲、石塹、花地及Golden star;短花柱品種有:台農一號、台農二號、二林、大冇、馬來、甜味、Jakarta及Arkin。依楊桃花柱長短則RAPD與ISSR皆可大致將其分群集中。不過,兩種分析方法均無法獲得有效針對果實風味(甜、酸味)及花柱種類(長、短)性狀分群之專屬標誌。
利用RAPD與ISSR所產生之多型性片段DNA的比率分別為73.47%與73.31%,其分析能力相近。RAPD與ISSR引子中產生多型性片段DNA的比率分別介於42.86%-100%與28.57%-100%。在品種鑑定方面,RAPD可鑑別台農一號、甜味、CIND-01、五汴頭反、秤錘、酸味厚斂,ISSR可鑑別台農一號、甜味、白絲、Golden star、Jakarta。

Twenty carambola (Averrhoa carambola L.) cultivars were evaluated and compared by using RAPD (random amplified and polymorphic DNA) and ISSR (inter simple sequence repeat) markers. The object was to assess genetic similarity between cultivars and to search for identification markers for individual cultivars. The results indicated that both methods revealed high degree of polymorphism.
A total 196 amplified DNA fragments DNA were generated by fourteen selected RAPD primers and in which 144 fragments were polymorphic. The sizes of DNA fragments were between 1500 bp and 190 bp. A total 266 amplified DNA fragments, including 195 polymorphic fragments, were generated by seventeen selected ISSR primers. The sizes of DNA fragments were between 1300 bp and 200 bp. The similarity between cultivars were estimated using the Nei/Li formula based on the total amplified and polymorphic DNA fragments by using RAPD and ISSR. The similarity dendrograms of twenty carambola cultivars were generated by UPGMA cluster analysis. The results showed that UPGMA cluster analysis based on RAPD similarity matrixs could separate the sweet and sour flavor carambola cultivars. In contrast, ISSR makers could not effectively separate them. Carambola cultivars with long style in this study were Cheng chui, Suan wei, Sua wei hou lian, Zhu ye, Wu bian tou fan, Pai si, Shi qian, Hua di, Golden star, and cultivars with short style were Arkin, Jakarta, Tainung No.1, Tainung No.2, Er lin, Da mou, ML-F-8, Tian wei. The results of RAPD and ISSR anaysis could separate and group up carambola cultivars based on tow style types. However, the results of both methods did not produce any specific markers for fruit flavors(sweet or sour) and style types(long or short).
The ratios of polymorphic DNA fragments produced by RAPD and ISSR anaysis were 73.47% and 73.31%, respectively. The ratios of polymorphic DNA fragments produced by RAPD and ISSR were between 42.86% and 100%, and 28.57% and 100%, respectively. For identification of individual carambola cultivars, Tainung No.1, Tian wei, CIND-01, Wu bian tou fan, Cheng chui and Suan wei hou lian can be identified by RAPD. Tainung No.1, Tian wei, Pai si, Golden star and Jakarta can be identified by ISSR.
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