本文選題：高分子量谷蛋白亞基 + SDS-聚丙烯酰胺凝膠電泳��； 參考：《南京農(nóng)業(yè)大學(xué)》2016年碩士論文

【摘要】：小麥貯藏蛋白主要由麥谷蛋白和醇溶蛋白組成,谷蛋白由高分子量谷蛋白亞基(HMW-GS)和低分子量谷蛋白亞基組成(LMW-GS)。雖然HMW-GS只占小麥貯藏蛋白總量的10%,但對小麥品質(zhì)或面筋質(zhì)量具有決定性的作用。利用特異麥谷蛋白亞基改良小麥品質(zhì)是小麥育種的重要手段之一。本研究選用攜帶不同外緣染色體的小麥材料進行研究,鑒定其HMW-GS組成,分析小麥品質(zhì),探討外緣染色體的HMW-GS導(dǎo)入對于小麥加工品質(zhì)的影響,同時對外緣染色體的HMW-GS基因編碼區(qū)進行分子克隆,以明確其影響品質(zhì)的分子基礎(chǔ),從而為小麥品質(zhì)改良提供參考依據(jù)。主要研究內(nèi)容及研究結(jié)果如下:(1)長穗偃麥草代換系和添加系與普通小麥之間籽粒蛋白含量、濕面筋含量、GMP含量和乳酸SRC,以及面團流變學(xué)特性均有顯著差異。DS1E/1A的面包總分81.5±0.71,顯著大于中國春的52±2.83。除DS1E/1D以外,其他長穗偃麥草代換系和添加系,顯著增加了面團的形成時間,峰高峰寬,8分鐘帶寬和乳酸SRC,從而顯著增加了面包體積。不同品質(zhì)指標(biāo)相關(guān)性研究表明,面包體積與GMP含量、乳酸SRC、峰高、峰寬以及八分鐘帶寬存在顯著性相關(guān),與GMP相關(guān)系數(shù)最高,為0.938。研究還發(fā)現(xiàn)碳酸鈉SRC與和面時間、峰高、峰寬和八分鐘帶寬存在顯著負(fù)相關(guān)。因此在早代育種中可以根據(jù)GMP含量、乳酸SRC以及峰高等參數(shù)的快速測定準(zhǔn)確篩選出適合面包加工品質(zhì)的材料。(2)克隆了長穗偃麥草1E亞基基因序列,該基因CDS序列全長1512個堿基對,推導(dǎo)的氨基酸序列全長為502個氨基酸殘基。基因結(jié)構(gòu)與已發(fā)表的HMW-GS亞基基因的結(jié)構(gòu)一致,不含內(nèi)含子,以信號肽、N-端保守區(qū)、中央重復(fù)區(qū)、C-末端構(gòu)成。與已克隆的HMW-GC基因序列比對,進化樹分析將該基因與其他已登錄的長穗偃麥草y型基因和中間偃麥草基因聚在一起,與普通小麥的HMW-GS亞基x或y型基因相似度較低,實驗材料中的1E基因為y型高分子量麥谷蛋白亞基。(3)對來源于野生粗山草D組染色體的50個人工合成小麥品系進行品質(zhì)篩選,發(fā)現(xiàn)品質(zhì)性狀間存在廣泛的遺傳變異。以HPLC微量測定谷蛋白大聚體和麥谷蛋白與醇溶蛋白比值為指標(biāo),取最高和最低的各3-4份品系進行進一步品質(zhì)分析,結(jié)果表明SE43、SE63和SE76等3個品系在濕面筋含量、籽粒硬度、乳酸SRC及揉混參數(shù)等方面與SE32、SE66、SE75和SE77等品系存在顯著差異,前3個品系為屬強筋硬質(zhì),后4個品系屬于弱筋軟質(zhì)。(4)克隆了人工合成小麥Glu-D1位點上HMW-GS的x型和y型亞基基因,分析HMW-GS的基因序列和推導(dǎo)的氨基酸序列,結(jié)果表明SE43、SE63和SE76等3個品系的Dy亞基基因序列一致,與SE32、SE66、SE75和SE77等品系的Dy亞基基因序列存在較大差異,基因進化樹分析,SE63的Dy亞基與擬斯卑爾脫山羊草(A.speltoides)克隆的Dy亞基基因最近,而與粗山羊草中克隆的Dy亞基基因較遠(yuǎn)。研究HMW-GS的分子結(jié)構(gòu)與小麥面粉品質(zhì)之間的聯(lián)系時發(fā)現(xiàn)小麥加工品質(zhì)產(chǎn)生的差異與HMW-GS基因的分子結(jié)構(gòu)有關(guān)系,可能是發(fā)生β-轉(zhuǎn)角的重復(fù)序列和谷氨酰胺殘基數(shù)量的差異引起的。本研究比較了人工合成小麥2個亞基(Dx和Dy)的中部重復(fù)區(qū)域中發(fā)生β-轉(zhuǎn)角的序列,結(jié)果表明,x-亞基含有的四種重復(fù)序列多于y-亞基含有的四種重復(fù)序列,這表明HMW-GS的x-型亞基的中央重復(fù)區(qū)含有更多的β-轉(zhuǎn)角結(jié)構(gòu),對面團加工品質(zhì)的影響比Y-型亞基要大,預(yù)示它們能使面團具有較強的彈性。比較了 2個亞基含有的谷氨酰胺(Q)的數(shù)量及其百分含量,結(jié)果顯示,Dx亞基含有的谷氨酰胺(Q)的數(shù)量和摩爾百分含量均比較高,表明在Dx亞基中部重復(fù)區(qū)域彼此之間以氫鍵相結(jié)合形成長鏈的能力較強,Dy亞基含有的酪氨酸(Y)的數(shù)量較少,但其百分含量較高,達到6.42%。實驗中發(fā)現(xiàn)品質(zhì)性狀較好的SE63材料在Dx的中部重復(fù)序列發(fā)生β-轉(zhuǎn)角的序列數(shù)量高于品質(zhì)較差的小麥材料,同樣的結(jié)論也出現(xiàn)在谷氨酰胺百分比和酪氨酸百分之上,其中篩選出來的SE63材料的谷氨酰胺和酪氨酸百分比分別高達39.91%和6.29%。
[Abstract]:Wheat storage protein is composed mainly of glutenin and gliadin, and glutenin is composed of high molecular weight glutenin subunit (HMW-GS) and low molecular weight glutenin subunit (LMW-GS). Although HMW-GS only accounts for 10% of the total amount of wheat storage protein, it plays a decisive role in wheat quality or gluten quality. Wheat quality is one of the important means of wheat breeding. In this study, wheat materials with different external chromosomes were selected to study the HMW-GS composition, analyze the quality of wheat, and explore the effect of the HMW-GS introduction of the outer chromosome on the processing quality of wheat, and the molecular cloning of the HMW-GS gene coding region of the external chromophore. The molecular basis of its influence quality was clarifying, thus providing reference for the improvement of wheat quality. The main research contents and results are as follows: (1) the grain protein content, the wet gluten content, the GMP content and the lactic acid SRC, and the rheological properties of the dough have significant differences in the.DS1E/1A surface. The total score of the package was 81.5 + 0.71, which was significantly greater than that of 52 + 2.83. in China's spring, except for DS1E/1D. Other long panicle intermedium and adding lines significantly increased the formation time of the dough, the peak peak width, the 8 minute bandwidth and the lactic acid SRC, thus significantly increasing the volume of bread. The correlation of different quality indexes showed that the volume and GMP content of bread, SRC, There is a significant correlation between peak height, peak width and eight minute bandwidth. The correlation coefficient with GMP is the highest. The 0.938. study also found that there is a significant negative correlation between SRC and surface time, peak height, peak width and eight minute bandwidth. Therefore, in the early generation breeding, it can be accurately selected according to the rapid determination of GMP content, lactic acid SRC and peak parameters. Materials for processing quality of bread. (2) cloned the sequence of 1E subunit gene of Agropyron long panicle, the whole length of the gene CDS sequence was 1512 base pairs. The total length of the deduced amino acid sequence was 502 amino acid residues. The structure of the deduced amino acid was consistent with the published HMW-GS subunit gene, containing no intron, the signal peptide, the N- terminal conservative region, the central repeat area, and the end of C-. Compared with the sequence of the cloned HMW-GC gene, the phylogenetic tree analyses the gene with other logged y and Agropyron intermedium gene and the HMW-GS subunit X or Y gene of common wheat, and the 1E based in the experimental material is derived from the Y type high molecular weight glutenin subunit (3). 50 synthetic wheat lines from the D group of wild wild grasses were selected for quality screening. There were extensive genetic variations among the quality traits. The ratio of glutenin and glutenin to gliadin was measured by HPLC as the index, and the highest and lowest 3-4 strains were taken into one step quality analysis. The results showed that SE43, SE63 and The 3 strains of SE76, such as the content of wet gluten, grain hardness, lactic acid SRC and mixing parameters, were significantly different from those of SE32, SE66, SE75 and SE77. The first 3 strains were strong tendons and the latter 4 belonged to weak tendons. (4) the X and Y subunits of HMW-GS in the Glu-D1 site of the synthetic wheat were cloned, and the HMW-GS gene was analyzed. The sequence and deduced amino acid sequence showed that the sequence of the Dy subunits of the 3 strains of SE43, SE63 and SE76 was the same, and there were great differences with the Dy subunit sequences of SE32, SE66, SE75, SE77 and other strains. The Dy subunit cloned in Aegilops was more distant. The study of the relationship between the molecular structure of HMW-GS and the quality of wheat flour found that the difference in processing quality of wheat was related to the molecular structure of the HMW-GS gene, which may be caused by the difference of the repeated sequence of beta angle and the difference of the number of glutamine residues. The sequence of beta rotation in the middle repeat region of 2 subunits of wheat (Dx and Dy) shows that the four repeat sequences of x- subunits contain more than four repeat sequences containing the y- subunit, which indicates that the central repeat region of the HMW-GS x- subunit contains more beta rotation structure, and the effect of the processing quality of the dough is larger than that of the Y- subunit. The number of glutamine (Q) and its percentage content in the 2 subunits were compared. The results showed that the number of glutamine (Q) and the content of mole percentage of Dx subunits were high, indicating that the ability to form a long chain with hydrogen bonds in the repeating region of the central Dx subunit was stronger, Dy The number of subunits containing tyrosine (Y) is relatively small, but its percentage is higher. The number of SE63 materials with better quality traits in the 6.42%. experiment is higher than that of poor quality wheat materials in the middle repeat sequence of Dx. The same conclusion also appears on the percentage of glutamine and the percentage of tyrosine. The percentages of glutamine and tyrosine of the selected SE63 materials were as high as 39.91% and 6.29%. respectively.
【學(xué)位授予單位】：南京農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S512.1

【參考文獻】

相關(guān)期刊論文 前10條

1 宋杰;李小軍;郎艷民;任翠翠;茹振鋼;;普通小麥-長穗偃麥草異附加系的分子標(biāo)記鑒定[J];西北農(nóng)業(yè)學(xué)報;2014年10期

2 程西永;吳少輝;李海霞;董中東;任妍;詹克慧;許海霞;;小麥高、低分子量麥谷蛋白亞基對品質(zhì)性狀的影響[J];麥類作物學(xué)報;2014年04期

3 劉麗;楊金華;胡銀星;程耿;;麥谷蛋白亞基與小麥品質(zhì)的關(guān)系研究進展[J];中國農(nóng)業(yè)科技導(dǎo)報;2012年01期

4 張樹華;楊學(xué)舉;張彩英;;小麥高分子量麥谷蛋白亞基重組對面團流變學(xué)特性的影響[J];華北農(nóng)學(xué)報;2011年02期

5 劉春雷;劉澤平;張麗琴;王世杰;;小麥高分子量谷蛋白亞基研究進展[J];河南教育學(xué)院學(xué)報(自然科學(xué)版);2010年04期

6 劉琦;張懷剛;劉寶龍;王欣;劉登才;;青海省審定的小麥品種高分子量麥谷蛋白亞基遺傳多樣性研究[J];新疆農(nóng)業(yè)科學(xué);2010年11期

7 姚金保;Edward Souza;馬鴻翔;張平平;姚國才;楊學(xué)明;任麗娟;張鵬;;小麥品種(系)間溶劑保持力差異及軟質(zhì)小麥資源篩選[J];麥類作物學(xué)報;2010年01期

8 楊玉雙;龐斌雙;王蘭芬;張學(xué)勇;于元杰;;小麥高分子量谷蛋白亞基間的互補效應(yīng)對面包加工品質(zhì)的影響[J];作物學(xué)報;2009年08期

9 夏云祥;馬傳喜;司紅起;喬玉強;;溶劑保持力與小麥部分品質(zhì)性狀的關(guān)系[J];中國糧油學(xué)報;2009年05期

10 夏云祥;喬玉強;司紅起;常成;馬傳喜;;小麥粉溶劑保持力與籽粒硬度及蛋白質(zhì)含量的關(guān)系[J];麥類作物學(xué)報;2009年03期

相關(guān)博士學(xué)位論文 前6條

1 何方;小麥—長穗偃麥草雜種后代的分子細(xì)胞遺傳學(xué)分析及種質(zhì)材料的篩選鑒定[D];山東農(nóng)業(yè)大學(xué);2014年

2 房敬業(yè);粗山羊草高分子量麥谷蛋白新亞基基因的克隆、分子進化及其遺傳轉(zhuǎn)化研究[D];華中科技大學(xué);2008年

3 安學(xué)麗;小麥近緣種谷蛋白新亞基鑒定與編碼基因克隆及其分子進化分析[D];首都師范大學(xué);2006年

4 劉勇;普通小麥和粗山羊草Glu-1位點遺傳多樣性研究[D];華中科技大學(xué);2006年

5 鄧志英;小麥高分子量谷蛋白亞基表達量及其對加工品質(zhì)的影響[D];山東農(nóng)業(yè)大學(xué);2005年

6 楊學(xué)舉;小麥蛋白成分和淀粉特性對面包品質(zhì)的影響及品質(zhì)改良應(yīng)用[D];中國農(nóng)業(yè)大學(xué);2004年

相關(guān)碩士學(xué)位論文 前6條

1 黃澤峰;長穗偃麥草分子標(biāo)記開發(fā)及硬粒小麥—長穗偃麥草附加系創(chuàng)建[D];揚州大學(xué);2013年

2 何坤;小麥麥谷蛋白亞基基因克隆及其功能研究[D];河北師范大學(xué);2012年

3 陳平;國內(nèi)外小麥種質(zhì)資源麥谷蛋白亞基組成及其對品質(zhì)的影響[D];河南農(nóng)業(yè)大學(xué);2009年

4 張超;長穗偃麥草E組染色體特異PCR標(biāo)記開發(fā)[D];揚州大學(xué);2009年

5 劉艷玲;小麥面筋指數(shù)及其它品質(zhì)指標(biāo)對面包烘焙的影響[D];山東農(nóng)業(yè)大學(xué);2004年

6 王建軍;小麥高分子量麥谷蛋白7+8與17+18亞基的遺傳特性和對品質(zhì)的影響[D];甘肅農(nóng)業(yè)大學(xué);2003年

，

本文編號：1935400