【摘要】：小麥穗發(fā)芽是世界性災(zāi)害,嚴(yán)重影響小麥的產(chǎn)量和品質(zhì)。小麥穗發(fā)芽受數(shù)量性狀控制,目前已鑒定穗發(fā)芽位點(diǎn)遍布于小麥全部21條染色體上,其中位于第三同源群和第四同源群上的較多。影響小麥穗發(fā)芽的因素很多,包括種子休眠性、ABA敏感性、遲熟淀粉酶及籽粒顏色等,盡管已有相關(guān)基因的報(bào)道,但是目前還缺乏關(guān)于這些基因的調(diào)控位點(diǎn)與抗穗發(fā)芽QTL之間關(guān)系的報(bào)道。因而挖掘更多的抗穗發(fā)芽QTL并鑒定其與這些功能基因的團(tuán)控位點(diǎn)之間的關(guān)系對(duì)于有效進(jìn)行分子輔助育種有更大的價(jià)值。本研究利用小麥660K SNP芯片對(duì)抗穗發(fā)芽人工合成小麥SHW-L1(強(qiáng)休眠節(jié)節(jié)麥AS60與四川地方四倍體小麥AS2255合成)與易發(fā)芽小麥川麥32產(chǎn)生的RILs群體(F10,171個(gè)株系)構(gòu)建高密度遺傳圖譜。利用該圖譜進(jìn)行抗穗發(fā)芽QTL定位并在SHW-L1育種后代中對(duì)QTL效應(yīng)進(jìn)行評(píng)價(jià)。同時(shí),對(duì)高等電點(diǎn)淀粉酶編碼基因TaAmyl及ABA信號(hào)相關(guān)基因TaVpl、TaSdr4, TaAIP2和TaSnrk2.6進(jìn)行了表達(dá)模式分析及eQTL定位,分析與抗穗發(fā)芽QTL重疊的eQTL并挖掘重要eQTL位點(diǎn)的候選基因,獲得的主要研究結(jié)果如下：1)利用AxiomTM wheat 660k Arrays基因芯片對(duì)SHW-L1和川麥32重組自交系構(gòu)建高密度遺傳圖譜,平均密度為每個(gè)位點(diǎn)0.148cM和143kb(小麥基因組大小參照IWGSC數(shù)據(jù))。利用該遺傳圖譜在2010年-2015年共10個(gè)獨(dú)立環(huán)境下進(jìn)行抗穗發(fā)芽QTL定位,定位到9個(gè)可在至少兩個(gè)環(huán)境中穩(wěn)定的QTLs,其中qPHS.sicau-1B和qPHS.sicau-3D.2可在8個(gè)環(huán)境中重復(fù)檢測(cè)到,同時(shí)對(duì)該群體籽粒顏色QTL的定位,發(fā)現(xiàn)qPHS.sicau-3D.2與種皮顏色QTL qGC.sica-3D重疊,與調(diào)控顏色的轉(zhuǎn)錄因子Tamyb10-D相近,而另外一個(gè)新主效QTL qPHS.sicau-1B與籽粒顏色無(wú)關(guān)。2)利用SHW-L1與四川主栽品種雜交,創(chuàng)制了一批抗穗發(fā)芽新材料,其中小麥新材料蜀麥580(劉登才、張連全等選育,2016年通過(guò)云南省區(qū)試)的穗發(fā)芽抗性水平與SHW-L1相當(dāng),7天發(fā)芽率比普通小麥降低30%。對(duì)蜀麥580的遺傳背景進(jìn)行分析,其中源自節(jié)節(jié)麥的兩個(gè)抗穗發(fā)芽QTL qPHS.sicau-3D.1和qPHS.sicau-3D.2成功導(dǎo)入蜀麥580中,而另一個(gè)主效位點(diǎn)qPHS.sicau-1B可能也導(dǎo)入(正在精細(xì)定位驗(yàn)證)。3)控制萌發(fā)速率的關(guān)鍵基因TaAmy1屬于大麥AMY2-1亞家族,其在SHW-L1和川麥32間存在6個(gè)SNPs。TaAmyl基因在川麥32籽粒發(fā)育后期的表達(dá)顯著高于抗穗發(fā)芽親本SHW-L1,通過(guò)基因表達(dá)差異在1BS上定位到與主效抗穗發(fā)芽位點(diǎn)qPHS.sicau-1B位置重疊eQTL位點(diǎn)eqamyl. sicau-DPA30,說(shuō)明qPHS.sicau-1B在籽粒發(fā)育后期可能通過(guò)降低TaAny1基因表達(dá)水平從而提升穗發(fā)芽抗性。4) TaVpl基因在SHW-L1中的表達(dá)水平持續(xù)高于川麥32,同時(shí)TaVpl基因eQTL eqABA.sicau-1B.1與穗發(fā)芽QTL qPHS.sicau-1B、α-淀粉酶eQTL eqamy1-DPA30重疊,說(shuō)明TaVp1基因表達(dá)的差異參與了該位點(diǎn)對(duì)穗發(fā)芽抗性的調(diào)控。5)對(duì)調(diào)控TaVp1基因在開(kāi)花后15、20天表達(dá)的eQTL eqABA.sicau-1D進(jìn)行了進(jìn)一步的分析,該區(qū)間共線(xiàn)與水稻5號(hào)染色體上2.8Mb區(qū)間內(nèi),根據(jù)水稻基因的功能注釋,搜索到4個(gè)相關(guān)基因TaABI1、TaGAox20, TaAkinlO和TaRav1。其中TaRav1基因的表達(dá)模式與TaVp1相同,說(shuō)明TaRav1反式調(diào)控TaVp1(與擬南芥中類(lèi)似)。在開(kāi)花后15天和20天控制TaRav1表達(dá)的QTL定位,與TaVp1、 TaSdr4、TaAIP2、TaSnrk2.6及TaAmy1已定位的eQTL位點(diǎn)重疊,說(shuō)明TaRav1基因與其它基因可能同時(shí)受到上游基因的調(diào)控。6)川麥32與SHW-L1中TaRav1-D基因共存在5個(gè)SNPs,分別位于-234bp,-91bp,+18bp,+379bp,+584bp處。其中+379bp,+584bp處的SNPs可導(dǎo)致氨基酸的變異。TaRavl基因序列在52份六倍體普通小麥中非常保守,均與川麥32序列完全一致；節(jié)節(jié)麥AetRavl基因變異類(lèi)型豐富,單倍型與節(jié)節(jié)麥的亞群分類(lèi)相關(guān),其中Ae.strangulata中的一類(lèi)與小麥相同。
[Abstract]:Ear sprouting is a worldwide disaster, which seriously affects the yield and quality of wheat. Ear sprouting is controlled by quantitative traits. It has been identified that ear sprouting sites are located on all 21 chromosomes of wheat, among which there are many factors affecting ear germination, including seed dormancy and ABA. Sensitivity, late-ripening amylase and grain color, although related genes have been reported, there is no report on the relationship between the regulatory sites of these genes and QTLs for panicle germination resistance. In this study, a high-density genetic map of RILs population (F10,171 lines) produced by strong dormant nodal wheat AS60 and Sichuan endemic tetraploid wheat AS2255 and easy-germinating wheat Chuanmai 32 was constructed by wheat 660K SNP chip. At the same time, expression pattern analysis and eQTL mapping of high-point amylase coding gene TaAmyl and ABA signal-related genes TaVpl, TaSdr4, TaAIP2 and TaSnrk2.6 were carried out, and eQTL overlapping resistance to spike germination QTL was analyzed and candidate genes of important eQTL loci were excavated. The results are as follows: 1) High-density genetic maps of SHW-L 1 and Chuanmai 32 inbred lines were constructed using AxiomTM wheat 660k Arrays gene chip. The average densities were 0.148cM and 143KB per locus (the wheat genome size was compared with IWGSC data). QTL mapping of resistance to spike germination was carried out in 10 independent environments from 2010 to 2015. Nine stable QTLs were identified in at least two environments, of which qPHS. sicau-1B and qPHS. sicau-3D.2 could be detected repeatedly in eight environments. At the same time, qPHS. sicau-3D.2 was found to overlap with seed coat color QTL qGC. sica-3D, similar to transcription factor Tamyb10-D and another new major QT. L qPHS.sicau-1B has nothing to do with grain color.2) A number of new materials for ear germination resistance were developed by crossing SHW-L1 with Sichuan main cultivars. Among them, Shumai 580 (Liu Dengcai, Zhang Lianquan, etc.) was selected and bred, and its ear germination resistance level was similar to that of SHW-L1 in 2016. The 7-day germination rate was 30% lower than that of common wheat. Genetic background analysis showed that two QTLs qPHS.sicau-3D.1 and qPHS.sicau-3D.2 derived from Arthropus japonicus were successfully introduced into Shumai 580, while another major locus qPHS.sicau-1B might also be introduced into (being fine-mapping verified). 3) TaAmy1, a key gene controlling the germination rate, belonged to the barley AMY2-1 subfamily, and it was found in SHW-L 1 and Chuanmai 3. Six SNPs. TaAmyl genes were significantly higher than SHW-L1 in the late grain development stage of Chuanmai 32, and the overlapping eQTL site eqamyl. sicau-DPA30 with the main resistant sprouting site qPHS. sicau-1B was located on 1BS by gene expression difference, suggesting that qPHS. sicau-1B might reduce the Any1 base in the late grain development stage. The expression level of TaVpl gene in SHW-L 1 was consistently higher than that in Chuanmai 32, and the expression level of TaVpl gene eQTL eqABA.sicau-1B.1 overlapped with QTL qPHS.sicau-1B and alpha-amylase eQTL eqamy1-DPA30 in spike germination, indicating that the difference of TaVp1 gene expression was involved in the regulation of spike germination resistance. The eQTL eqABA.sicau-1D expressed by aVp1 gene at 15 and 20 days after anthesis was further analyzed. According to the functional annotations of rice genes, four related genes, TaABI1, TaGAox20, TaAkinlO and TaRav1, were found. The expression pattern of TaRav1 gene was the same as that of TaVp1, indicating that TaRav1 was antigenic. TaVp1 (similar to that in Arabidopsis) was regulated by the expression of TaRav1. The QTL mapping of TaRav1 was controlled at 15 and 20 days after anthesis and overlapped with the eQTL loci of TaVp1, TaSdr4, TaAIP2, TaSnrk2.6 and TaAmy1, indicating that TaRav1 gene and other genes might be regulated by the upstream genes at the same time. 6) TaRav1-D gene of Chuanmai 32 and SHW-L1 shared five SNPs, respectively. SNPs located at - 234 bp, - 91 bp, + 18 bp, + 379 bp, + 584 BP could induce amino acid mutation. TaRavl gene sequence was very conserved in 52 hexaploid common wheat, which was completely consistent with Chuanmai 32 sequence; AetRavl gene had abundant variation types, and haplotypes were related to the subgroup classification of Arthroid wheat, including Ae. strangula. One of TA is the same as wheat.
【學(xué)位授予單位】：四川農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：S512.1

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