转基因小麦的纯系鉴定及其面团粘弹性的分析研究

发布时间：2018-03-11 20:22

本文选题：转基因小麦　切入点：高分子量麦谷蛋白亚基　出处：《华中科技大学》2016年硕士论文　论文类型：学位论文

【摘要】：高分子量麦谷蛋白亚基对小麦面筋粘弹性起决定性作用。小麦具有三个染色体组1A,1B,1D,分别编码1Ax-型,1Bx-和1By-型,1Dx-和1Dy-型亚基。每种亚基类型对面团粘弹性的影响存在优质和劣质的差异,其中公认的优质亚基或亚基组合为1Ax1,1Bx17+1By18和1Dx5+1Dy10。本人所在实验室通过转基因技术结合传统杂交技术的方式分别获得了外源1Ax1和外源1Dx5超表达的重组自交系和回交导入系。本文主要通过聚丙烯酰胺凝胶电泳(SDS-PAGE)技术,对这批转基因小麦进行全面完整的筛选鉴定,然后进行揉混实验,主要结果如下:筛选出15种转基因型和8种非转基因型,总共23种不同亚基类型是通过自交和回交形成。转基因型分别为将1Ax1和1Dx5通过基因枪法导入受体小麦,使1Ax1和1Dx5亚基超表达。初步获得的种子类型中,各个品种的总数差异显著。本人选取数量足够的7种转基因类型和5种非转基因类型,进行揉混实验,获得揉混曲线和揉混参数,观察各种材料的面粉揉混特性,相互比较,验证不同高分子量麦谷蛋白亚基之间对小麦面粉揉成面团时的影响。通过筛选,获取最优质的高分子量麦谷蛋白亚基组合为1Dx5+1Dy10,1Bx17+1By18,1Ax1超表达;同时,通过比较发现,x-和y-型亚基的比例也会影响揉混特性,影响面筋的形成。小麦种子常年放置后会影响小麦种子的品质,为了保种,同时为了增加几类材料的种子总数,对筛选后所有类型的小麦种子进行扩繁,包括温室扩繁和田间扩繁二步,初期种子较少,在温室中可以不受季节限制,获取足够进行田间播种的种子数量;然后再进行田间扩繁,获得足够的种子,进行保种和以后进行更多不同的实验。
[Abstract]:High molecular weight glutenin subunits play a decisive role in the viscoelasticity of wheat gluten. Wheat has three chromosomes 1A1BX 1D, which encode 1Ax-type 1Bx- and 1By-type 1Dx- and 1Dy- subunits, respectively. The effects of each subunit type on dough viscoelasticity are observed. The difference between good quality and inferior quality, The recognized high quality subunits or combinations were 1Ax1X 1Bx17 1By18 and 1Dx51 Dy10.The recombinant inbred lines of exogenous 1Ax1 and exogenous 1Dx5 overexpression and backcross were obtained by transgenic technology and traditional hybridization in our laboratory, respectively. In this paper, the polyacrylamide gel electrophoresis (SDS-PAGE) technique, After a complete screening and identification of the transgenic wheat, the main results are as follows: 15 transgenic and 8 non-transgenic types were screened. A total of 23 different subunit types were formed by self-crossing and backcrossing. The transgene types of 1Ax1 and 1Dx5 were introduced into recipient wheat by gene gun method, and 1Ax1 and 1Dx5 subunits were overexpressed. There are significant differences in the total number of varieties. I selected 7 transgenic types and 5 non-transgenic types to get blending curves and blending parameters, observe the flour blending characteristics of various materials, and compare with each other. The effects of different high molecular weight glutenin subunits on the dough formation of wheat flour were tested. The best combination of high molecular weight glutenin subunits was obtained as 1Dx51 Dy10Bx17 1By181Ax1 overexpression. It was found by comparison that the proportion of subunits of x- and y- also affected the mixing characteristics and the formation of gluten. The quality of wheat seeds was affected after the wheat seeds were placed all the year round, in order to preserve the seeds and increase the total number of seeds of several kinds of materials. After screening all types of wheat seeds were expanded and propagated, including greenhouse propagation and field propagation. The initial seeds were less, and enough seeds could be obtained in greenhouse without season restriction. Then the field propagation was carried out, enough seeds were obtained, and more different experiments were carried out.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：TS210.1;S512.1

【参考文献】