發(fā)布時(shí)間：2018-03-03 17:49

  本文選題：谷子　切入點(diǎn)：稀碼　出處：《甘肅農(nóng)業(yè)大學(xué)》2017年博士論文　論文類(lèi)型：學(xué)位論文

【摘要】：谷子(Setaria italica(L.)P.Beauv.)具有基因組小、重復(fù)序列少、生育期短、繁殖系數(shù)高等特點(diǎn),并已完成全基因組測(cè)序,正發(fā)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(chǎn)量的主要因素。目前,關(guān)于調(diào)控谷子穗發(fā)育基因的研究鮮有報(bào)道。本研究以EMS誘變谷子全基因組測(cè)序品種Yugu1所獲得的穗發(fā)育異常突變體siaux1和lp1為對(duì)象,圖位克隆目的基因并分析其功能,主要獲得如下結(jié)果:1.與Yugu1相比,siaux1的種子發(fā)芽勢(shì)和發(fā)芽率降低,植株莖節(jié)數(shù)減少、主莖變粗變矮、葉片變長(zhǎng)變寬,主穗變長(zhǎng)、穗碼數(shù)減少、小花數(shù)減少、結(jié)實(shí)率降低;最明顯的特征是穗碼變稀、單穗產(chǎn)量極顯著降低。siaux1的突變性狀由位于5號(hào)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(gè)外顯子區(qū)發(fā)生G→A的突變,導(dǎo)致三聯(lián)體密碼子TGG(UGG)替換為終止密碼子TGA(UGA),造成翻譯蛋白時(shí)在214個(gè)氨基酸處提前終止,缺失了C-端的277個(gè)氨基酸殘基。2.敲除水稻Os AUX1(LOC_Os01g63770)基因突變株的穗密度、側(cè)根密度顯著減小;異源超表達(dá)Si AUX1基因水稻突變株的穗密度、側(cè)根密度顯著增大。50個(gè)代表性谷子品種中,7個(gè)品種的Si AUX1基因3’UTR區(qū)存在38bp的大片段缺失,這些品種的穗長(zhǎng)、第一級(jí)分枝數(shù)、小花數(shù)、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(yīng)2,4-D的誘導(dǎo),表達(dá)量上調(diào)的幅度與2,4-D的處理濃度成正比;Si AUX1基因在生長(zhǎng)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(dá),與生長(zhǎng)素的分布與運(yùn)輸模式吻合。Si AUX1基因突變會(huì)顯著下調(diào)IAA信號(hào)轉(zhuǎn)導(dǎo)途徑多個(gè)基因在穗部的表達(dá)量;siaux1灌漿期生長(zhǎng)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(gè)基因,只有Si AUX1在穗中高表達(dá),具有調(diào)控穗發(fā)育的功能;其它4個(gè)基因Si AUX2(Seita.3G223500)、Si AUX3(Seita.9G471700)、Si AUX4(Seita.9G288200)、Si AUX5(Seita.8G047400)在不同發(fā)育時(shí)期的根中高表達(dá)。4.與Yugu1相比,lp1的植株主莖變矮,主穗變長(zhǎng)、第一級(jí)分枝數(shù)和第二級(jí)分枝數(shù)均減少、小花數(shù)減少、籽粒數(shù)減少;最明顯的特征是穗碼變稀、單穗產(chǎn)量顯著降低,但籽粒變長(zhǎng)變寬,千粒重極顯著增大。lp1的突變性狀由位于2號(hào)染色體屬于WRKY轉(zhuǎn)錄因子家族Ⅰ亞族的基因LP1(Seita.2G369500)控制;lp1的LP1基因第五個(gè)內(nèi)含子末端的堿基發(fā)生G→A的突變,形成3個(gè)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(gè)WRKY結(jié)構(gòu)域的C2H2鋅指結(jié)構(gòu)遭到破壞。5.LP1基因分別在拔節(jié)期的莖節(jié)、孕穗期的穗、灌漿期的籽粒中高表達(dá),這與其調(diào)控株高、穗型和種子大小的功能一致;亞細(xì)胞定位顯示LP1基因在細(xì)胞核中表達(dá),符合轉(zhuǎn)錄因子基因的表達(dá)特征。谷子的穗發(fā)育是受多個(gè)基因控制的復(fù)雜過(guò)程。本研究發(fā)現(xiàn)生長(zhǎng)素輸入載體編碼基因Si AUX1和WRKY家族轉(zhuǎn)錄因子LP1突變均會(huì)導(dǎo)致谷子穗發(fā)育異常,表現(xiàn)出明顯的稀碼特征,且單穗產(chǎn)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(xué)位授予單位】：甘肅農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：S515;Q943.2

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