【作者】 李学军；

【导师】 王辉；

【作者基本信息】 西北农林科技大学 ， 作物遗传育种， 2004， 博士

【摘要】 小麦品质的遗传改良目前多集中于高分子量麦谷蛋白亚基（HMW-GS）的遗传转化。本研究以遗传基础不同、综合性状优良、丰产潜力大的西农2208、西农1718、西农1330三个高产品种（品系）为受体轮回亲本，以16个含有不同HMW-GS组成的国内外材料为供体非轮回亲本，采用生化标记选择回交转育法，培育三个受体亲本的三套不同HMW-GS及其组合的近等基因系，研究高分子量谷蛋白亚基和亚基组合的遗传规律；以遗传基础不同的三套近等基因系为材料，在同一遗传背景条件下研究不同类型HMW-GS及其组合的品质效应，及在不同遗传背景条件下研究同一HMW-GS及其组合品质效应与遗传基础的关系，确定可供小麦品质改良利用的优质HMW-GS及其亚基组合，探讨利用优质亚基的遗传转化进行小麦品质改良的理想的遗传基础，为小麦品质改良提供理论和实践依据。研究结果如下：⑴ 通过对加代条件下半粒无胚籽粒电泳进行研究，创制出了一套适合于小麦半粒分析的高分子量（HMW）麦谷蛋白亚基的SDS-PAGE方法。和前人方法相比有如下改进：①改变了凝胶贮藏液的体积浓度（30%T），在分离胶缓冲液和浓缩胶缓冲液中不加SDS，这些都直接影响凝胶分子筛的有效孔径。通过这种调整，得到了较高分辨率的HMW麦谷蛋白亚基谱带。②改变了电流强度及染色液、脱色液中水、甲醇、冰醋酸的比例，大大缩减了电泳时间。③提出了完善的样品制备程序，摸索出适于半粒小麦的提取液用量及振荡时间、加热时间等，大大提高了电泳成功率。 ⑵ 采用SDS-PAGE方法,对16个含不同HMW-GS的中、法（中国1个，法国15个）材料与陕西关中3个遗传基础不同的农艺亲本及其杂交、正反交、回交籽粒进行小麦高分子量谷蛋白亚基遗传分析表明:①普通小麦品种高分子量谷蛋白亚基受遗传控制,不受环境影响,具有品种稳定的遗传特性；②小麦高分子量谷蛋白亚基在F1代中呈共显性和倾母遗传现象；③控制小麦高分子量谷蛋白亚基的基因遗传行为遵循孟德尔的独立分配和自由组合规律。 ⑶ 本研究采用生化标记选择回交转育法，以高产材料西农1718、2208、1330为受体轮回亲本，以分别含有不同目标HMW-GS的16个材料为供体非轮回亲本，通过回交并<WP=6>结合半籽粒SDS-PAGE电泳分析、筛选含有目标亚基的半籽粒在无菌条件下胚培养、加代并用高产受体材料选择回交，如此循环回交5次，高产受体亲本遗传比重达到98.44%以上，自交分离选择一个世代获得分别含有不同目标亚基的纯合体，单系繁殖，育成分别含有21个不同目标亚基组合的西农1718、2208、1330三套共63个（每套21个）近等基因系（near-isogenic line）。为研究HMW-GS的品质效应、HMW-GS效应与醇溶蛋白、淀粉等有机、生化物质的关系及进一步从分子水平的研究创造了必要的基础材料。 ⑷ 本研究将谷蛋白溶涨指数法(Swelling lndex of Glutenin，简称SIG)应用于小麦不溶性谷蛋白含量的评价，小麦谷蛋白在一定的溶涨时间下所得SIG值反映了谷蛋白的质量或不溶性谷蛋白的含量。 本实验结果表明，谷蛋白溶涨指数（SIG）与微量SDS沉淀值呈极显著正相关，r=0.9620**。而且谷蛋白溶涨指数法具有样品用量小(单粒种子即可满足需要)，实验时间短(一天可以测试100多个样品)，在早代材料中分辨力高(与不溶性谷蛋白的关系大于SDS、Zeleny沉淀值)，仪器要求简单(一台低速离心机即可)，与食品加工品质关系密切，可以代替微量SDS沉淀值（种子用量较大）进行早代品质性状选择，来指导小麦品质育种。 ⑸ 同一遗传背景条件下，各等位亚基对品质的效应为：在Glu-A1位点：2*>1>null；在Glu-B1位点， 7+8与14+15差异不显著（p>0.05），17+18与6+8、6+8与7+9差异不显著（p>0.05），17+18与7+9、7+8与7+9、6+8差异显著（p<0.05），7+8、14+15与17+18、6+8、7+9差异显著；在Glu-D1位点， 5+10与2+12差异极显著， 2+12与3+12差异不显著。即5+10>2+12≈3+12。在A1、B1、D1三个位点的HMW-GS组合中，相对品质效应高的亚基组合有：1 7+8 5+10；2* 7+8 5+10；2* 6+8 5+10；1 7+8 2+12；2* 7+8 2+12；1 14+15 2+12。 ⑹ 本试验应用的三个受体亲本品质性状为1718优于2208，2208优于1330。通过对三个受体品种近等基因系品质指标谷蛋白溶涨指数（SIG）与微量SDS沉淀值的分析表明，2*、1、5+10亚基对品质指标都有增效作用，但在不同品种中对同一品质指标的影响，因品种（遗传背景）不同而存在量的差异。本实验中，以西农1718为遗传背景时，其SIG值和微量SDS沉淀值明显高于西农2208、1330。以西农2208为遗传背景时，其SIG值和微量SDS沉淀值在一定程度上略高于西农1330。表明在优质亚基的遗传转化中，应选择品质基础好的材料做受体，容易得到理想的品质类型。更多还原

【Abstract】 Genetic improvement of wheat quality is centralized in genetic transference of High molecular weight glutentin subunit (HMW-GS) at present. The research takes XiNong 2208,1718,1330 which are highly-product materials of different genetic base, good complex character and great potentialities of gracefully-product as receptor recurrent parents, takes 16 sorts of materials in and abroad which contain different HMW-GS as non- recurrent parents. And it takes the way of biochemical marker 、consecutive backcrossing and transbreeding to nurture 3 suits of near-isogenic line(NIL)of different HMW-GS and its groups of 3 receptor parents and research the laws of HMW-GS and subunits groups. Taking 3 suits of NIL of different genetic base as materials, research the quality efficacy of different HMW-GS and its groups under the same inherited background and research the relation of quality efficacy to inherited background of the same HMW-GS and its groups under different inherited background. Then definite good quality HMW-GS and its subunits groups for the improvement of wheat quality and seek the ideal genetic base for the improvement of wheat quality by genetic transference of good quality subunits. It provided theory and practical base for the improving of wheat quality. The result of the research are as follows：(1)Researching on electrophoresis of the half-seed without embryo of accelerate generation, groped a suit of HMW-GS SDS-PAGE method which adapts to half-seed analysis of wheat. The improvement compared with the method of predecessor are as follows: ①Alter the volume density(30%T)of gel hoarding liquid, and don’t add SDS to separation glue buffer or condensation glue buffer, which effect the valid pore of gel molecular sieve directly. Then obtain the HMW-GS spectrum of higher resolution ratio by the alteration. ②Alter the electric current intension and the proportion of H2O,CH3OH and CH3COOH in dyeing liquid and decolorant, which shorten the time of electrophoresis greatly. ③Raise the consummate procedure of sample establishment, and grope the quality of extract liquid, time of vibration and calefaction which adapt to half-seed of wheat, which elevate the successful ratio of electrophoresis. <WP=8>(2)16 France materials which contain different HMW-GS were used to cross with 3 cultivars of GuanZhong,ShanXi. Different crosses and generation populations were obtained. The grains of 30 reciprocal F1, 15 F2, 30 backcrosses BC1F1,BC1F11 generations and their parents were analysed by SDS-PAGE method for HMW-GS genetic analysis. The results are as follows: ①The band patterns of HMW-GS were genetically controlled and they were not affected by environmental factors. There were large variations among cultivars; ②All HMW-GS can be transmitted to the hybrid F1 as dominance from both parents and a significant effect of female parents was observed in reciprocal F1 hybrid; ③The genes which control HMW-GS were inherited based on Mendel’s laws of gene independent assortment and free combination.(3)The research takes the way of biochemical marker and consecutive backcrossing and transbreeding, takes XiNong 2208,1718,1330 which are highly-product materials as receptor recurrent parents, and takes 16 sorts of materials which contain different objective HMW-GS respectively as non-recurrent parents. In the way of backcrossing and half-seed SDS-PAGE electrophoresis analysis, select the half-seeds which contain the objective subunits and cultivate them under the aseptic condition. Then accelerate generation and backcross with highly-product receptor materials. Such cycle backcross 5 times. The heritability of highly-product receptor parent is up to 98.44%.Select one generation by selfing and segregation and then obtain the homozygote which contain different objective subunits. After apomixis, nurture respectively XiNong1718,2208,1330 3 suits of near-isogenic line which contain 21sorts of different objective subunits. Create good material for the research of HMW-GS quality efficacy, Gliadin and the further research of molecule level.更多还原