作為重要的糧食作物,水稻從傳統(tǒng)的經(jīng)驗(yàn)育種向現(xiàn)代精準(zhǔn)育種轉(zhuǎn)變的過程中,株型是影響水稻產(chǎn)量最為重要因素之一,它取決于植株高度、分蘗數(shù)目、分蘗角度、穗形態(tài)等因素。對(duì)于水稻株型的機(jī)制研究,是短期內(nèi)難以突破的科學(xué)問題和技術(shù)難題,其中矮化突變體是非常理想的研究對(duì)象,但是調(diào)控機(jī)制知之甚少。本文在前期研究的工作基礎(chǔ)上,對(duì)水稻矮化、少分蘗D26突變體進(jìn)行系統(tǒng)研究,揭示它參與水稻油菜素內(nèi)酯激素調(diào)控的信號(hào)通路機(jī)制。本文從水稻突變體庫中組織培養(yǎng)變異獲得了2個(gè)農(nóng)藝性狀表現(xiàn)矮化、少分蘗的突變材料D26-1和D26-2,通過圖位克隆實(shí)現(xiàn)精細(xì)定位和進(jìn)一步的功能分析,揭示了D26突變體矮化表型的遺傳機(jī)理,并從互作蛋白篩選和轉(zhuǎn)錄組測(cè)序分析角度,進(jìn)一步挖掘水稻矮化機(jī)制。主要結(jié)果如下:D26突變體一個(gè)隱性單基因控制表型。利用D26突變體做母本和93-11親本做父本,構(gòu)建雜交F₂群體,通過425個(gè)群體和531個(gè)群體將其分別定位在3號(hào)染色體短臂上63 kb區(qū)間,該區(qū)間內(nèi)包含12個(gè)候選基因;分析預(yù)測(cè)和克隆、測(cè)序表明D26基因是一個(gè)編碼GRAS家族蛋白的轉(zhuǎn)錄因子,兩個(gè)突變體在D26基因540 bp和560 bp... 

【文章來源】：華僑大學(xué)福建省

【文章頁數(shù)】：143 頁

【學(xué)位級(jí)別】：博士

【部分圖文】：

水稻油菜素內(nèi)酯相關(guān)基因研究網(wǎng)絡(luò)(Tong,2018)

D26基因分別從水稻秈稻明恢86和D26突變體的c DNA中擴(kuò)增獲得全長(zhǎng),片段回收與pMD18-T連接、測(cè)序比對(duì)分析。互補(bǔ)實(shí)驗(yàn)載體構(gòu)建:一個(gè)含有D26基因啟動(dòng)子、c DNA全長(zhǎng)和3"UTR非編碼區(qū)3563 bp全長(zhǎng)片段從中間載體p UPGWPR上由HindIII/Eco RV酶切,與Bam HI酶切p CAMBIA1300載體分別回收,兩片段經(jīng)平端連接,最終構(gòu)建互補(bǔ)載體p CAMBIAGWP26(見圖1)。2.2.8 植物葉片總RNA提取-TRIzol法

轉(zhuǎn)錄組測(cè)序流程

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