本文選題：谷子　切入點：稀碼　出處：《甘肅農業大學》2017年博士論文　論文類型：學位論文

【摘要】：谷子(Setaria italica(L.)P.Beauv.)具有基因組小、重復序列少、生育期短、繁殖系數高等特點,并已完成全基因組測序,正發展成為禾本科新的模式植物。谷子屬于稀播作物,穗部性狀是決定產量的主要因素。目前,關于調控谷子穗發育基因的研究鮮有報道。本研究以EMS誘變谷子全基因組測序品種Yugu1所獲得的穗發育異常突變體siaux1和lp1為對象,圖位克隆目的基因并分析其功能,主要獲得如下結果:1.與Yugu1相比,siaux1的種子發芽勢和發芽率降低,植株莖節數減少、主莖變粗變矮、葉片變長變寬,主穗變長、穗碼數減少、小花數減少、結實率降低;最明顯的特征是穗碼變稀、單穗產量極顯著降低。siaux1的突變性狀由位于5號染色體編碼IAA輸入載體(AUX1)的基因Si AUX1(Seita.5G387200)控制;siaux1的Si AUX1基因第四個外顯子區發生G→A的突變,導致三聯體密碼子TGG(UGG)替換為終止密碼子TGA(UGA),造成翻譯蛋白時在214個氨基酸處提前終止,缺失了C-端的277個氨基酸殘基。2.敲除水稻Os AUX1(LOC_Os01g63770)基因突變株的穗密度、側根密度顯著減小;異源超表達Si AUX1基因水稻突變株的穗密度、側根密度顯著增大。50個代表性谷子品種中,7個品種的Si AUX1基因3’UTR區存在38bp的大片段缺失,這些品種的穗長、第一級分枝數、小花數、主穗粒重顯著小于其它品種,而穗碼密度顯著大于其它品種。3.Si AUX1基因能夠響應2,4-D的誘導,表達量上調的幅度與2,4-D的處理濃度成正比;Si AUX1基因在生長旺盛的胚芽、心葉、幼穗等幼嫩組織和主根的髓、穎殼的脊等維管組織中高表達,與生長素的分布與運輸模式吻合。Si AUX1基因突變會顯著下調IAA信號轉導途徑多個基因在穗部的表達量;siaux1灌漿期生長旺盛的穗部IAA的含量(46.17 mg/g)極顯著高于野生型(27.61 mg/g)。谷子AUX/LAX家族有5個基因,只有Si AUX1在穗中高表達,具有調控穗發育的功能;其它4個基因Si AUX2(Seita.3G223500)、Si AUX3(Seita.9G471700)、Si AUX4(Seita.9G288200)、Si AUX5(Seita.8G047400)在不同發育時期的根中高表達。4.與Yugu1相比,lp1的植株主莖變矮,主穗變長、第一級分枝數和第二級分枝數均減少、小花數減少、籽粒數減少;最明顯的特征是穗碼變稀、單穗產量顯著降低,但籽粒變長變寬,千粒重極顯著增大。lp1的突變性狀由位于2號染色體屬于WRKY轉錄因子家族Ⅰ亞族的基因LP1(Seita.2G369500)控制;lp1的LP1基因第五個內含子末端的堿基發生G→A的突變,形成3個不同于野生型的蛋白翻譯提前終止型突變的可變剪切,造成LP1蛋白C-末端第二個WRKY結構域的C2H2鋅指結構遭到破壞。5.LP1基因分別在拔節期的莖節、孕穗期的穗、灌漿期的籽粒中高表達,這與其調控株高、穗型和種子大小的功能一致;亞細胞定位顯示LP1基因在細胞核中表達,符合轉錄因子基因的表達特征。谷子的穗發育是受多個基因控制的復雜過程。本研究發現生長素輸入載體編碼基因Si AUX1和WRKY家族轉錄因子LP1突變均會導致谷子穗發育異常,表現出明顯的稀碼特征,且單穗產量顯著下降。
[Abstract]:Millet (Setaria Italica (L.) P.Beauv.) has a small genome, repeat, short growth period, high propagation coefficient, and whole genome sequencing has been completed, is developing into a new plant. The model of gramineous crops of millet is rare, panicle traits are the main factors determining yield. At present, there are few reports on on gene regulation millet ear development. In this study, whole genome sequencing EMS mutagenesis millet varieties Yugu1 acquired ear development mutant siaux1 and Lp1 as the object, map based gene cloning and function analysis, the main results are as follows: 1. compared with Yugu1 siaux1, the seed germination potential and germination rate decreased, the stem section the number of plants decreased, stem thick and short blade of variable length and variable width, variable length of main spike, spike number decreased, the number of florets decreased, the seed setting rate decreased; the most obvious feature is the spike code thinning, yield per spike decreased significantly in.Siaux1 process Shaped by degeneration of chromosome 5 IAA encoding the input vector (AUX1) gene Si AUX1 (Seita.5G387200) siaux1 Si control; AUX1 gene mutation of fourth exons of G to A promoter, resulting in three CIS codon TGG (UGG) to replace the termination codon TGA (UGA), caused by protein translation in 214 amino acids at the early termination, the deletion of 277 amino acid residues of.2. C- end of the knockout AUX1 rice Os (LOC_Os01g63770) mutant panicle density, lateral root density decreased significantly; heterologous over expression of Si gene AUX1 in rice mutant panicle density, root density increased significantly.50 representative millet varieties there, a large fragment deletion of 38bp Si AUX1 gene of 7 varieties of 3 UTR region, these varieties of ear length, the first branch number, floret number, main spike grain weight was significantly less than other varieties, and spike code density was significantly greater than the other varieties of.3.Si AUX1 gene in response to 2,4-D By the concentration of 2,4-D was increased and the expression magnitude is proportional to the leaf; Si AUX1 gene in the germ, vigorous growth, such as young spike tissues and root pulp, high expression of tissue glume ridge dimension, consistent with the auxin distribution and transport patterns of.Si mutations in the AUX1 gene can significantly reduce IAA the signal transduction pathways of multiple gene expression in panicle weight; content of strong IAA growth siaux1 ear filling stage (46.17 mg/g) was significantly higher than that of the wild type (27.61 mg/g). Millet AUX/LAX family has 5 genes, only Si high expression of AUX1 in Guangzhou, with regulation of ear development of other functions; 4 genes Si AUX2 (Seita.3G223500), Si AUX3 (Seita.9G471700), Si AUX4 (Seita.9G288200), Si AUX5 (Seita.8G047400) at different developmental stages in the root of the high expression of.4. compared with Yugu1, Lp1 of the plant stem short, main spike becomes longer, the first branch number and level second The number of branches were reduced, the number of florets decreased, grain number decreased; the most obvious feature is the spike code thinning, significant yield per spike decreased, but the grain grow wider, the mutation of LP1 gene traits significantly increased by.Lp1 were located on chromosome 2 belongs to WRKY transcription factor family 1 subfamily (Seita.2G369500) control LP1 gene mutation; fifth introns end Lp1 base G and A, the formation of 3 different from the wild type protein translation termination variable shear mutation, C2H2 zinc caused LP1 protein C- terminal second WRKY domain refers to the destruction of the.5.LP1 gene structure respectively in stem elongation stage. At the booting stage of panicle, high expression in grain filling period, which related to the regulation of plant height, panicle and seed size consistent function; subcellular localization showed that LP1 gene expression in the nucleus, consistent with the expression pattern of transcription factor gene of millet ear hair. Fertility is a complex process controlled by multiple genes. We found that auxin input vector coding gene Si AUX1 and WRKY family transcription factor LP1 mutation will lead to abnormal development of millet panicle, showing obvious sparse code characteristics, and the yield of single spike decreased significantly.

【學位授予單位】：甘肅農業大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：S515;Q943.2

【參考文獻】

相關期刊論文 前10條

1 杜彥修;季新;陳會杰;彭廷;張靜;李俊周;孫紅正;趙全志;;基于CRISPR/Cas9系統的OsbHLH116基因編輯及其脫靶效應分析[J];中國水稻科學;2016年06期

2 陶洋;李金城;廖奇;;CRISPR相關的生物信息學基礎[J];中國生物化學與分子生物學報;2016年10期

3 王芳權;范方軍;李文奇;朱金燕;王軍;仲維功;楊杰;;利用CRISPR/Cas9技術敲除水稻Pi21基因的效率分析[J];中國水稻科學;2016年05期

4 李雯;智慧;張碩;房雪嬌;王海龍;賈冠清;韓淵懷;刁現民;;谷子Si-SP1小穗突變基因的遺傳分析和定位[J];植物遺傳資源學報;2015年03期

5 謝圣男;王宏光;楊振;劉春燕;蔣洪蔚;辛大偉;胡國華;陳慶山;;大豆綏農14突變體庫構建及株高性狀分析[J];核農學報;2013年03期

6 李偉;智慧;王永芳;李海權;刁現民;;谷子EMS誘變的處理條件分析[J];河北農業科學;2010年11期

7 賈小平;譚賢杰;李永祥;王天宇;黎裕;;用SSR標記研究谷子品種的遺傳多樣性[J];江西農業大學學報;2009年04期

8 張娟;;生長素信號轉導途徑及參與的生物學功能研究進展[J];生命科學研究;2009年03期

9 劉進平;;生長素受體與信號轉導機制研究進展[J];生物技術通報;2007年03期

10 張超;張暉;李冀新;;小米的營養以及應用研究進展[J];中國糧油學報;2007年01期

相關碩士學位論文 前1條

1 李靜;毛白楊PtAUX1基因的分離及超表達植株的表型分析[D];山東農業大學;2007年

，

本文編號：1562064