Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/21990
標題: 利用ISSR DNA分子標幟分析水稻品種間與品種內之遺傳歧異度
Genetic diversity among and within Rice Varieties Based on ISSR DNA Markers
作者: 柯惠珠
Ke, Hui-Chu
關鍵字: rice;水稻;genetic diversity;ISSR DNA markers;遺傳歧異度;ISSR DNA分子標幟
出版社: 生命科學系
摘要: 
台灣的水稻品種有遺傳基礎狹隘之現象,研究水稻品種間親緣關係有助於日後在育種上選擇親本時的考量。本研究是利用Inter-Simple Sequence Repeats(ISSR) DNA分子標幟來(1)分析13個不同來源的越光品種之遺傳歧異度;(2)觀察兩個世代中八個水稻品種內的變異性,以瞭解ISSR分子標幟的穩定性;以及(3)探討水稻品種間遺傳歧異情形,並在使用的引子中篩選出核心引子。
從80個ISSR引子中,篩選出16個能清楚地產生多型性標幟之引子,以分析品種內(間)的差異。在分析不同來源的越光品種中,發現引子UBC811及UBC848分別在越光品種KHK11及KHK8各增幅出一多型性片段,而此二片段的序列分析結果,並未比對到任何水稻基因序列。將八個稻種經過兩個世代的栽培後做分析,結果每個稻種內均呈現單型性,世代間並無差異。將上述之材料,以11組Simple Sequence Repeats(SSR)引子進行分析。發現有三組引子OSR20、OSR22、RM249,在三個越光品種KHK6、KHK10及KHK11中,共產生五個多型性條帶;以引子OSR22、RM204及RM206分析稻種IR8及台南5號,發現在品種內呈現多型性,但世代間仍無差異。比較這兩種分子標幟,可得SSR的變異性大於ISSR。在兩個世代中,ISSR分子標幟是穩定的。
在探討品種間變異性的分析中,共選取76個原始祖先及栽培品種為材料,包含25個原始祖先與51個栽培品種(25個稉稻品種及26個秈稻品種)。16個ISSR引子共增幅出288個條帶,其中208個條帶具有多型性。經數據處理後得到所有品種的平均相似度為0.29,原始祖先為0.27,稉稻品種為0.66,秈稻品種為0.54。以樹狀圖分析造成多型性的核心引子,除了UBC810,其它15個引子都是核心引子。前人利用相同11組SSR引子,測得76個品種的平均相似度為0.40,原始祖先為0.42,稉稻品種為0.55,秈稻品種為0.63。比較ISSR與SSR所得之結果,對於親源關係較近之台灣水稻品種,ISSR分子標幟具有較佳之區分效果。

Rice (Oryza sativa L.) cultivars of Taiwan were developed on a narrow genetic base. Therefore, the knowledge of relationship among rice varieties is helpful in selecting mates for hybridization breeding program. In this study, we employed Inter-Simple Sequence Repeats (ISSR) DNA markers 1) to analyze the genetic variation of Koshihikri from thirteen seed sources; 2) to detect the stability of ISSR markers from eight rice varieties in two generations of seed propagation; 3) to evaluate the genetic diversity among rice varieties, and to identify the core primers that are informative in estimating diversity.
Sixteen of 80 ISSR primers were sufficient to produce polymorphic banding patterns. We have found specific fragments for KHK11 and KHK8 amplified the by primer UBC811 and UBC848, respectively. The eight rice varieties were propagated for two generations and showed monomorphic banding patterns. Three Simple Sequences Repeats (SSR) primers, OSR20, OSR22, and RM249, amplified five polymorphic banding patterns from eleven primers for SSR analysis. The three SSR primers, OSR22, RM204, and RM206 showed polymorphism within IR8 and Tainan 5, but no variation between two generations. Based on these results, SSR markers revealed more genetic variation than that by ISSR markers, and the ISSR marker is stable in two generations.
The genetic diversity among 76 rice varieties including 25 ancestors and 51 cultivars (25 Japonica and 26 Indica cultivars) were used for ISSR analysis. 16 ISSR primers amplified 288 banding patterns with 208 polymorphic. The averaged similarity estimates were 0.27 among ancestors, 0.66 among Japonica cultivars, 0.54 among Indica cultivars, and 0.29 among all rice varieties. Based on the clustering dendrograph, 16 ISSR primers were the core primer to differentiate varieties tested, except primer UBC810. The previous SSR data from the same material showed that the average similarity estimates among ancestors, Japonica cultivars, Indica cultivars, and all rice varieties were 0.42, 0.55, 0.63, and 0.40, respectively. ISSR showed lower averaged similarity estimates than SSR did. Therefore, ISSR analysis proved to be an efficient method for determining genetic diversity among Taiwanese rice varieties.
URI: http://hdl.handle.net/11455/21990
Appears in Collections:生命科學系所

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