臺灣博碩士論文加值系統

由Magnaporthe oryzae所引起之水稻稻熱病，為臺灣一期作之主要流行病害，於水稻產區均造成相當危害。使用抗病品種為目前已知有效且對環境友善之防治方法，當前新興的分子標誌輔助選拔技術 (marker-assisted selection, MAS)，可有效節省育種所需時程，因此成為水稻抗性改良之發展趨勢。臺灣良質米雖具有良好農藝特性，但普遍對稻熱病抗性不佳，可藉此方法進行抗性改良。本研究利用帶已知抗性基因且對臺灣稻熱病菌菌株抗幅較廣之13個IRRI-bred blast-resistant lines (IRBLs) 及20個臺灣良質米品種，建立一套可輔助回交育種之分子標誌平台，包括11對R gene分子標誌以及可使用Fluidigm高通量基因型分析之95組背景分子標誌。為了瞭解本土良質米對稻熱病抗性，以兩菌株分別接種13 IRBLs及良質米品種來加以鑑別，結果顯示，台稉4號、台稉9號、台稉10號、台稉14號、台稉15號、台農71號、桃園3號、台南11號、高雄145號、高雄147號等10個品種不帶有Pi9 (IRBL9-W)、Piz5 [IRBLz5-CA、IRBLz5-CA (R)]、Piz (IRBLz-Fu)、Pi1 (IRBL1-CL)、Pi7 (IRBL7-M)、Pik (IRBLk-Ka)、Pik-p (IRBLkp-K60)、Pik-m (IRBLkm-Ts)、Pik-h (IRBLkh-K3)、Pi20 (IRBL20-IR24) 及Pita-2 (IRBLta2-Pi、IRBLta2-Re) 等對臺灣稻熱病菌有抗性之等位基因，適合以13 IRBLs及本研究所開發之多型性分子標誌進行抗性改良。本研究同時運用此分子標誌平台及回交育種策略，實際將10個IRBLs之抗性基因導入高雄145號中，目前IRBLta2-Pi、IRBL9-W、IRBLkp-K60、IRBLkh-K3與高雄145之雜交組合篩選至BC2F2世代，其最高背景回復率分別為80.51%、94.08%、97.27%與95.54%；IRBL7-M、IRBLk-Ka、IRBLz5-CA、IRBLz5-CA(R)、IRBL20-IR24、IRBLta2-Re與高雄145號雜交組合則篩選至BC1F2世代。期望未來選育出高雄145號之抗稻熱病多系品種，並推廣於田間。

Rice blast, caused by Magnaporthe oryzae, is the most important disease commonly occurring in the first crop season in Taiwan. This disease can cause considerable damage in every rice-producing areas. Use of blast resistant varieties is an effective and environmental-friendly method for blast control. The emerging marker-assisted selection (MAS) approach, which can significantly reduce the time needed for the development of new resistant varieties, has become a new trend in rice resistance breeding. In Taiwan, most high-quality rice varieties are with good agronomic traits but lack of blast resistance. The resistance can be improved using this technology. In this study, we used 13 IRRI-bred blast-resistant lines (IRBLs, each carrying a known blast resistance gene effective against more local isolates of M. oryzae) and 20 high-quality rice varieties to develop a marker-assisted backcrossing platform, consisting of 11 polymorphic R gene markers and 95 background selection markers used in Fluidigm high-throughput genotyping. Two M. oryzae isolates were used to determine which Taiwan varieties would be suitable recipients for the 13 R alleles. The result showed that Taikeng 4, Taikeng 9, Taikeng 10, Taikeng 14, Taikeng 15, Tainung 71, Taiyuan 3, Tainan 11, Kaohsiung 145, and Kaohsiung 147 do not carry the R alleles at Pi9 (IRBL9-W), Piz5 [IRBLz5-CA, IRBLz5-CA (R)], Piz (IRBLz-Fu), Pi1 (IRBL1-CL), Pi7 (IRBL7-M), Pik (IRBLk-Ka), Pik-p (IRBLkp-K60), Pik-m (IRBLkm-Ts), Pik-h (IRBLkh-K3), Pi20 (IRBL20-IR24), and Pita-2 (IRBLta2-Pi, IRBLta2-Re). The 13 IRBLs and newly developed markers could be used to improve blast resistance of the 10 high-quality rice varieties. Meanwhile, the marker platform and backcross breeding strategy were utilized to introduce the resistance genes from 10 IRBL lines to Kaohsiung 145. To date, BC2F2 progenies were obtained from the crosses of “IRBLta2-Pi x KH145”, “IRBL9-W x KH145”, “IRBLkp-K60 x KH145”, and “IRBLkh-K3 x KH145”, with the highest background recovery rates of 80.51%, 94.08%, 97.27%, and 95.54%, respectively. For the crosses of “IRBL7-M x KH145”, “IRBLk-Ka x KH145”, “IRBLz5-CA x KH145”, “IRBLz5-CA(R) x KH145”, “IRBL20-IR24 x KH145”, and “IRBLta2-Rex KH145”, BC1F2 progenies were obtained. The work is expected to result in the development and deployment of a Kaohsiung 145 multiline variety carrying various blast resistant genes.

致謝...............................i
中文摘要...........................ii
Abstract..........................iii
前言...............................v
表目錄.............................ix
圖目錄.............................xi
壹、前人研究........................1
一、稻熱病抗性基因...................1
二、IRBL判別品種....................3
三、分子標誌輔助選拔.................4
四、高通量基因型分析.................6
五、抗稻熱病多系品種.................7
貳、材料與方法......................9
一、供試水稻材料與種植...............9
二、水稻雜交........................9
三、GBS定序資料與分析...............10
四、R gene前景分子標誌設計與資料分析.12
五、背景分子標誌設計與資料分析.......14
六、抗性表現檢定....................16
參、結果...........................17
一、GBS資料分析結果.................17
二、IRBLs遺傳背景差異分析............17
三、主成分分析......................17
四、前景基因型分子標誌測試...........18
五、背景分子標誌測試.................18
六、高雄145號與10個IRBLs雜交子代分析..20
七、稻熱病抗性檢定 ...................21
肆、討論............................23
伍、參考文獻........................31
陸、附表............................40
柒、附圖............................85
捌、附錄............................99

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