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dc.description.abstract植物組織的顏色表現多為花青素累積所致,色素突變體可呈現顯而易見的外表型,為探討遺傳變異與基因調控機制的最佳材料。本研究室利用疊氮化鈉誘變,建立世界廣泛栽培的秈稻品種IR64號的突變庫。本論文以六個植株顏色表現穩定的M6與M7突變品系42231、42231.1、50168、42303、50143、42226為材料,進行突變性狀之遺傳研究與基因定位。六個突變品系與野生型IR64雜交所得F1植株均呈現有色(紫色或紅色)的外表型,說明突變體之花青素生合成基因皆能正常表現,且為顯性遺傳;而F2植株不同部位呈現顏色差異顯示,應有調節基因影響基因之表現。本研究選擇有最多部位表現顏色的突變體42303為主要材料,針對42303/IR64雜交組合F2族群進行遺傳分析,發現葉鞘、葉舌、葉耳、稃尖四個部位,有色:無色植株之分離比均符合3:1,說明這些部位的呈色,均受單一顯性基因座所控制。本試驗篩選簡單序列重複(SSR)分子標誌,進行42303有色突變體色素調控基因之定位。選用84個具多型性之SSR,利用單一標誌策略分析92株42303/IR64 F2子代的基因型,結果顯示第六對染色體上有4個SSR:RM6773、RM5754、RM253、RM2615與顏色性狀呈顯著的連鎖;其中RM253可解釋78% F2的外表型變異,推測控制此四個部位顏色表現的基因可能位在RM253附近。增加分子標誌進一步將此目標區域縮小至RM5754與RM2615間相距約 830kb。利用候選基因策略挑選三個與花青素生合成密切相關的候選基因:bHLH、3GT及Myb,進行基因選殖、定序及序列比對。試驗結果發現,二親本之3GT與Myb基因與胺基酸序列存在顯著變異;對92個F2以基因專一引子組進行基因型分析結果顯示,此二基因均符合單一基因遺傳之卡方檢測,分別解釋68 % 與74 % 的外表型變異,應為共同調控突變體42303外表型變異之主效基因。zh_TW
dc.description.abstractPigmentation in plant is mostly due to accumulation of anthocyanins resulting visible and diverse phenotypes. Therefore, colored mutants provide good materials for studying genetic variation and gene regulation. Six (42231, 42231.1, 50168, 42303, 50143, 42226) stable colored mutants screened from a mutation pool derived from sodium azide mutagenesis on a popular indica rice variety, IR64, were applied to study the inheritance and mapping the gene for colored phenotype. All the F1 plants from mutants crossed with IR64 showed pigmentation (purple or red) suggests that these mutants contain functional anthocyanin pathway and show dominant inheritance. Variations in the F2 tissues imply the involvement of regulatory gene. A mutant, 42303, shows pigmentation in most tissues was crossed to its wild type to obtain the 42303/IR64 F2 population. Genetic analysis on the pigmentations of leaf sheath, ligule, auricle, and apiculus of F2 fits 3 colored : 1 non-colored ratio suggesting that they are controlled by a single dominant locus. Simple sequence repeat (SSR) markers were screened and 84 polymorphic SSRs were applied to map the pigment gene using 92 F2 plants of 42303/IR64 cross. The result indicates thatRM6773, RM5754, RM253, and RM2615 are significantly linked to the colored phenotype. Among them, RM253 explains 78% phenotype variation of F2 suggesting that both are linked. PMore SSRs were further applied to locate the region to 830kb between RM5754 and RM2615. In this region three genes, Myb, 3GT, and bHLH, annotated and related to anthocyanins biosynthesis are cloned, sequenced, and analyzed. Two genes, Myb, and 3GT, show significant variations between wild type and 42303, and genetic analysis on F2 both fit single dominant inheritance model and explain 68% and 74% of phenotype variation, respectively. These results suggest that both 3GT and Myb are together conditioned the phenotype of 42303 mutant.en_US
dc.description.tableofcontents摘要 i 英文摘要 ii 目錄 iii 表目錄 iv 圖目錄 v 第一章 前言 1 第二章 前人研究 4 一、 花青素之生合成途徑 4 二、 花青素生合成調控機制 6 三、 水稻花青素生合成途徑相關基因之研究 8 四、 集群分析與關聯分析 16 五、 分子標誌基因型分析與基因定位 17 六、 花青素性狀基因定位與連鎖分析 20 七、 花青素糖基轉移酶基因之研究 25 第三章 有色突變體性狀調查與純化及遺傳性狀之關聯分析 29 一、 前言 29 二、 材料與方法 30 三、 結果 35 四、 討論 39 第四章 花青素性狀相關基因之定位與選殖基因之探討 60 一、 前言 60 二、 材料與方法 62 四、 結果 71 五、 討論 81 第五章 結論 122 第六章 參考文獻 123 附錄 134zh_TW
dc.subjectgene mapping and cloneen_US
dc.subjectgenetic analysisen_US
dc.titleGenetic analysis and mapping of anthocyanin mutants of IR64 rice varietyen_US
dc.typeThesis and Dissertationzh_TW
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