【作者】 刘春；

【导师】 麻浩；

【作者基本信息】 南京农业大学 ， 作物遗传育种， 2006， 硕士

【摘要】 本研究以1400份大豆品种为材料，在筛选到蛋白亚基特异种质的基础上，进行了亚基含量年份间稳定性分析、亚基缺失类型的进一步鉴定和遗传分析以及种质创新、亚基特异种质的蛋白功能性评价等方面的研究。以期为进一步发掘我国丰富的大豆基因资源、筛选出亚基组成优良的大豆材料，明确亚基变异类型遗传规律，阐明亚基变异对大豆蛋白功能性的影响奠定基础。主要结果如下： 1、1400份参试品种的11S／7S比值范围为0.44～3.64，平均值为1.68±0.37，鉴定出34份亚基含量特异材料，其中包括α＇和α亚基含量低、β亚基含量低、A₃B₄亚基缺失、A₅A₄B₃亚基缺失和A_1aB_1b亚基缺失种质。理化诱变结果表明：化学诱变率为2.92％，物理诱变率为5.71％，对大豆蛋白的诱变效果0.4％EMS处理优于0.2％EMS处理，而仰⁶⁰Co γ射线慢照射处理优于化学诱变处理。并从M₃代种子中筛选出了211粒亚基变异种质。 2、分析了2003年、2004年和2005年种植的280份大豆种质的7S和11S组分及其亚基含量间的差异，结果表明：品种对大豆贮藏蛋白7S、11S组分及其亚基含量、7S+11s和11S／7S值的影响均达极显著水平，年份除对α亚基和A₄亚基含量的影响达极显著水平外，对其它亚基含量及7S、11S、7S+11S和11S／7S值的影响均不显著；品种×年份对7S、11S、11S／7S值、α亚基、A₄亚基和A₅亚基含量的影响达极显著水平，对其它组分和亚基含量的影响均不显著。 3、在SDS-PAGE筛选到蛋白亚基特异种质的基础上，运用双向聚丙烯酰胺凝胶电泳（2-DE）分析大豆贮藏亚基正常品种南农大黄豆和亚基变异品种桂阳紫金豆种子总蛋白的蛋白质组。差异蛋白质组学显示，等电点和分子量分别约为5.40和37.85kDa、5.24和37.2kDa、5.15和37.05kDa的蛋白质点在正常品种种子中表达而在缺失品种中未表达。对这3个蛋白质点用基质辅助激光解析电离飞行时间质谱（MALDI-TOF-MS）测定其胶内酶解后的肽质量指纹谱（PMF），对获得的PMF用Mascot软件在NCBI数据库中查询比对，鉴定出这3个蛋白质点均为大豆球蛋白A_1aB_1b亚基同源三聚体，表明桂阳紫金豆种子贮藏蛋白缺少A_1ab_1b亚基。 4、研究了4个不同蛋白亚基类型(A_1ab_1b亚基正常、A_1ab_1b亚基缺失；A₃B₄亚基正常、A₃B₄亚基缺失)的品种所配成的6个杂交组合的F₁、F₂、F₃种子亚基表现及其遗更多还原

【Abstract】 The contents of 7S and 11S globulins and their respective subunits, and the 11S/7S ratios in seeds of 1400 soybean varieties were analyzed using SDS-PAGE, the affection of varieties and years on the major fractions and their subunits, further identification the kind of subunits deficiency, the genetic analysis of A_1aB_1b-subunit and A₃B₄-subunit in soybean seeds, integration null mutations for A_1aB_1b-subunit, A₃B₄-subunit and A₅A₄B₃-subunit of the storage protein were by crossbreeding, induced variation germplasm of storage proteins by physical and chemical method, and the relationships between subunits variation and functional properties of soybean protein were analyzed. The main results are as follows:1. The average and variation range of the 11S/7S ratios of 1400 soybean varieties were 1.68±0.37 and 0.44-3.64, and 34 variation accessions of subunits content were identified, including low level of α’-,α-subunit, low level of β-subunit, absence of A₃B₄-subunit, absence of A₅A₄B₃-subunit and A_1aB_1b-subunit. The results of induced variation germplasm of storage proteins by physical and chemical method indicated: the chemical induced rate is 2. 92%, while the physical induced rate is 5.71%, ⁶⁰Co γ-ray irradiation treatment > 0.4%EMS treatment > 0.2% EMS treatment, and 211 seeds of subunits variation were detected in the M₃ seed.2. The content of 11S and 7S fractions and their major subunits in 280 soybean varieties storage protein during 2003, 2004 and 2005 years were compared with SDS-PAGE. The results showed that there were significant differences on the content of α’, α, β, A₃, A₄, A₂A_1aA_1b, B₃, B_1aB_1bB₂, B₄, A₅ subunits, 11S and 7S, 11S +7S and 11S/7S between difference varieties, while except A₄-subunit there no significant differences among 2003, 2004 and 2005 years.3. Basis on successfully selected a mutant line which characterized by the lack of the A_1aB_1b or A_1bB₂ or A₂B_1a -subunits of glycinin from cultivar variety "Guiyangzijindou", the proteome of the seed protein of soybean varieties "Nannongdahuangdou" and更多还原