转基因小麦外源优质亚基基因的杂交转育研究

在获得外源品质基因1Dx5和1Ax1超量表达的转基因小麦的基础上,利用小麦转基因品系‘B72-8-11b’和‘B102-1-2’为父本,主要以湖北省栽培品种‘鄂麦12’为母本,配置杂交组合。杂交后代中采用系谱选择法,结合HMW-GS鉴定,研究了转基因小麦外源品质基因在F1、F2、F3、F4代的传递,并筛选出外源1Dx5或1Ax1基因保持超表达的2个新型转基因株系;同时证明了将外源品质基因向栽培品种转育,是提高小麦优质亚基含量和提高HMW-GS总量的有效方法之一。     

两对高分子量麦谷蛋白亚基等位基因及其相互作用对小麦面筋品质的影响

小麦谷蛋白赋于面筋弹性,其亚基组成类型对加工品质有着重要的决定作用。采用一对杂交组合(烟农19×安农9914)的后代,随机选择至F3,种植871个F4穗系,分别检测了高分子量麦谷蛋白亚基(high molecular weight glutenin subunits,HMW-GS)、低分子量麦谷蛋白亚基(low molecular weight glutenin subunits,LMW-GS)组成、SDS沉降值和和面图指标,分析了麦谷蛋白等位亚基及其相互作用对品质的影响。结果表明:该群体麦谷蛋白组成仅在Glu-A1(1亚基或N亚基)与Glu-B1(14 15亚基或17 18亚基)位点有差异。Glu-A1位点1亚基的SDS沉降值显著高于N亚基,1亚基的峰高、7 min尾高显著大于N亚基,而在和面时间、7 min带宽以及衰落角(耐揉性),两亚基间差异不显著。Glu-B1位点亚基间SDS沉降值17 18>14 15/17 18>14 15,和面时间、7 min带宽两个指标17 18亚基显著高于14 15亚基,衰落角显著小于14 15亚基,峰高和7 min尾高差异不显著。对Glu-A1和Glu-B1两位点互作,除7 min带宽互作达5%显著水平外,其它四个指标均未达显著水平。1亚基相对于N亚基,17 18亚基相对于14 15亚基,虽以SDS沉降值为标准其效应相当,但两者却作用于面团的不同性能。     

Effects of amino acid substitutions outside an antigenic site on protein binding to monoclonal antibodies of predetermined specificity obtained by peptide immunization: demonstration with region 15-22 (antigenic site 1) of myoglobin.

Monoclonal antibodies (mAbs) of predetermined specificity were prepared by immunizing with a free (i.e., not conjugated to any carrier) synthetic peptide representing region 15–22 (site 1) of sperm whale myoglobin (SpMb). The cross-reactions of Mb variants with three mAbs were studied in order to determine whether such interactions are influenced by substitutions outsde the site. Finback whale Mb, which has no substitutions within region 15–22, showed lower cross-reactivity and relative binding affinity than the reference antigen, SpMb. Bottle-nose Atlantic dolphin myoglobin (BdMb) and badger myoglobin (BgMb), although they have identical substitutions within region 15–22 (Ala-15 to Gly and Val-21 to Leu), showed very different binding properties. The cross-reaction of BdMb was quite comparable to that of SpMb, while that of BgMb was much lower. Since the two proteins have identical structures in regions 15–22, the differences in their cross-reactivities are readily attributed to the effects of substitutions outside this region. Another pair of myoglobins, horse myoglobins (HsMb) and chicken myoglobin (ChMb), also have two identical substitutions (Ala-15 to Gly and Val-21 to Ile) within region 15–22, but possessed different cross-reactivity. The results indicate that the reaction of mAbs, whose specificity is precisely known and predetermined by the immunizing free peptide, can be markedly affected by substitutions outside the indicated binding region on the protein.