不同灌水处理对强筋小麦谷蛋白大聚合体粒度分布和品质的影响

在田间条件下,以两个优质强筋小麦品种(藁城8901和济麦20)为供试材料,研究了不同灌水处理(全生育期不灌水、拔节期灌1次水、越冬期和拔节期灌2次水、越冬期、拔节期和灌浆期灌3次水,每次灌水量675 m³·hm^-2)对强筋小麦谷蛋白大聚合体含量与粒度分布、品质和产量的影响.结果表明: 两个小麦品种的面团形成时间、面团稳定时间、面包体积、籽粒产量、谷蛋白大聚合体含量以及体积加权平均粒径、表面积加权平均粒径、粒径>100 μm的体积百分比和表面积百分比均以灌2水处理最高.相关分析显示,两个小麦品种的面团形成时间、面团稳定时间和面包体积与粒径<10 μm和10～100 μm的谷蛋白大聚合体颗粒体积百分比呈显著负相关,而与粒径>100 μm的谷蛋白大聚合体颗粒体积百分比、体积加权平均粒径和表面积加权平均粒径呈显著正相关.水分供应过多或过少均不利于籽粒产量和品质的同步改善,灌溉水平可通过改变谷蛋白大聚合体粒度分布影响小麦籽粒品质.     

小麦谷蛋白亚基基因的PCR鉴定及其在品质改良中的应用

小麦谷蛋白是胚乳中的主要贮藏蛋白,对面包品质具有重要作用。因此,改变品种的谷蛋白等位基因组成是品质改良的主要内容,而谷蛋白亚基基因的选择手段是决定品质育种成效的关键。本文介绍了近年来发展起来的小麦谷蛋白亚基基因PCR分子鉴定技术,通过与传统SDS－PAGE方法的比较,总结了PCR技术应用于谷蛋白基因鉴定和品质改良计划的优越性及其研究进展,并讨论了分子标记技术在小麦品质改良计划中的应用前景及今后的研究方向。     

小麦谷蛋白亚基基因的PCR鉴定及其在品质改良中的应用

小麦谷蛋白是胚乳中的主要贮藏蛋白,对面包品质具有重要作用。因此,改变品种的谷蛋白等位基因组成是品质改良的主要内容,而谷蛋白亚基基因的选择手段是决定品质育种成效的关键。本文介绍了近年来发展起来的小麦谷蛋白亚基基因PCR分子鉴定技术,通过与传统SDSPAGE方法的比较,总结了PCR技术应用于谷蛋白基因鉴定和品质改良计划的优越性及其研究进展,并讨论了分子标记技术在小麦品质改良计划中的应用前景及今后的研究方向。     

谷蛋白聚合体大小分布与面粉揉面特性的初步研究

用单向一步SDS-PAGE方法分析表明小麦品种Suneca和Cook在麦谷蛋白5个亚基位点（Glu-B1,Glu-D1,Glu-A3,Glu-B3和Glu-D3)均含不同等位基因。选用Suneca×Cook的F4代群体中麦谷蛋白亚基位点均为纯合基因的60个系,研究麦谷蛋白基因型不同的株系间谷蛋白聚合体粒度大小分布（用SE-HPLC测定）和面粉揉面特性的变异。结果表明,不同的谷蛋白基因型,其谷蛋白聚合体粒度大小相对分布（用不溶谷蛋白聚合体占总谷蛋白聚合体含量的百分数表示,即UPP%）和面团形成时间（即揉面仪曲线图峰值的和面时间,简写PTM）均有显差异;面粉的揉面曲线形状与其UPP%值密切相关;UPP%与PTM呈极显正相关,与揉面仪曲线图峰高（PHM）呈显负相关;与面粉蛋白质含量（FP%）相比,UPP%对PTM和PHM的影响更大些,可作为育种早代品质性状选择一个指标。     

小麦高分子量麦谷蛋白亚基鉴定及其品质效应研究进展

高分子量谷蛋白亚基(HMW-GS,high molecular weight glutenin subunits)是小麦子粒贮藏蛋白的重要组成成分,其组成、搭配、表达水平及含量决定面团弹性和面包加工品质。本文主要介绍了小麦HMW-GS编码基因的克隆、分子特征、分子标记开发及其在小麦育种中的应用,并综述了不同HMW-GS与面粉加工品质之间的关系,以及HMW-GS基因遗传转化、微量配粉和突变体培育等方面的研究进展,分析了目前研究中存在的主要问题,认为通过分子标记辅助选择和转基因技术聚合优质亚基,培育优质面包小麦品种和明确各个HMW-GS基因的品质效应是今后的研究重点。     

150个小麦品种高分子量谷蛋白亚基组成与蛋白质含量和沉降值关系的研究

对150个不同小麦品种高分子量麦谷蛋白亚基与蛋白质含量、沉降值之间的关系进行了研究,结果表明:Glu-1三位点控制的亚基等位变异与品质性状关系密切.A1位点亚基出现频率高于N,对品质效应以1>N;B1位点7+9亚基对出现频率最高,其次为7+8,各亚基对蛋白质含量效应以8>17+18>7+8>13+19>7+9>14+15,对沉降值效应以8>13+19>7+8>7+9>14+15>7>17+18;D1位点2+12亚基对频率高于5+10,各亚基对品质效应以5+10>5+12>2+12.具有亚基1,8或7+8,5+10组合类型的小麦品种可望为品质较好的品种.     

西北春麦区小麦地方品种高分子量麦谷蛋白亚基组成分析

为了给品质改良提供基础材料,并了解西北春麦区小麦地方品种的遗传多样性,采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)方法,分析了493份小麦地方品种的高分子量麦谷蛋白亚基(HMW-GS)的组成.结果表明:在供试材料中,Glu-1位点共有26个等位基因,其中Glu-A1位点3个,Glu-B1位点9个,Glu-D1位点14个,亚基null、7+8、2+12在各自的位点上出现频率最高,分别达到了94.53%、92.92%、86.24%;亚基组成类型共有30种,主要为null/7+8/2+12,频率达79.76%;同时筛选出一些含有1、2*、13+16、14+15、5+10、1.5+10等优质亚基或亚基对的材料,可作为优质基因源;西北春麦区小麦地方品种间Glu-1位点的遗传多样性,以Glu-D1位点最高,其次是Glu-B1位点,Glu-A1位点最低.     

小麦高分子量麦谷蛋白亚基的遗传变异与小麦加工品质改良

高分子量麦谷蛋白亚基（HMW-GS）是小麦胚乳中一种具有多态性的蛋白质组分,在面团中它们可以通过相互之间或与低分子量麦谷蛋白亚基（LMw-Gs）之间形成二硫键来组成麦谷蛋白多聚体。由于其在小麦面粉加工所需的粘性和弹力方面具有极其重要的作用,过去几十年间在小麦加工品质相关蛋白研究方面的工作大多数集中在高分子量麦谷蛋白亚基上。近几年在高分子量麦谷蛋白亚基及其编码基因的鉴定、基因的遗传变异以及不同变异在小麦加工品质中的作用方面进行了大量研究。本文对近几年在HMW-GS领域的研究进展进行综述并且重点讨论HMW-GS的变异及其对小麦品质育种的重要意义。     

小麦高分子量谷蛋白亚基效应的比较研究

采用SDS-PAGE方法,通过对5个亲本间杂交获得的F2群体每一单株的F3籽粒样本及其亲本进行小麦高分子量谷蛋白亚基（HMW-GS）组成分析,并对每一F2单株上F3籽粒群体的高分子量谷蛋白亚基组成与其籽粒蛋白质含量、SDS-沉降值的关系进行研究,分析比较黄淮麦区出现频率较高的7个亚基或亚基对的品质效应。结果表明：小麦高分子量谷蛋白亚基组成不同群体间籽粒的蛋白质含量和SDS-沉降值基本达到显著或极显著水平。优质亚基表现为：1、7＋8、14＋15和5＋10亚基。因此,黄淮麦区小麦育种应加强对这些优质亚基的引入和利用,特别是对14＋15和5＋10亚基的引入和利用。     

Production and characterization of a transgenic bread wheat line over-expressing a low-molecular-weight glutenin subunit gene

The end-use properties, and thus the value, of wheat flours are determined to a large extent by the proteins that make up the polymeric network called gluten. Low molecular weight glutenin subunits (LMW-GS) are important components of gluten structure. Their relative amounts and/or the presence of specific components can influence dough visco-elasticity, a property that is correlated with the end-use properties of wheat flour. For these reasons, manipulation of gluten dough strength and elasticity is important. We are pursuing this goal by transforming the bread wheat cultivar Bobwhite with a LMW-GS gene driven by its own promoter. Particle bombardment of immature embryos produced several transgenic lines, one of which over-expressed the LMW-GS transgene. Southern blots confirmed that the transgene was integrated into the wheat genome, although segregation analyses showed that its expression was sometimes poorly transmitted to progeny. We have determined that the transgene-encoded LMW-GS accumulates to very high levels in seeds of this line, and that it is incorporated into the glutenin polymer, nearly doubling its overall amount. However, SDS sedimentation test values were lower from the transgenic material compared to a non transgenic flour. These results suggest that the widely accepted correlation between the amount of the glutenin polymers and flour technological properties might not be valid, depending on the components of the polymer.     

A 1B-coded low-molecular-weight glutenin subunit associated with quality in durum wheats shows strong similarity to a subunit present in some bread wheat cultivars

Received: 25 May 1999 / Accepted: 22 June 1999     

Identification of new low-molecular-weight glutenin subunit genes in wheat

To clarify the composition of low-molecular-weight glutenin subunits (LMW-GSs) in a soft wheat cultivar, we cloned and characterized LMW-GS genes from a cDNA library and genomic DNA in Norin 61. Based on alignment of the conserved N- and C- terminal domains of the deduced amino-acid sequences, these genes are classified into 12 groups. One of these groups (group 5), the corresponding gene of which has not been reported previously, contains two additional hydrophobic amino-acid clusters interrupting the N-terminal repetitive domain. Other groups (groups 11 and 12), which were not identified in other cultivars as a protein product, showed all eight cysteines in the C-terminal conserved domain. With specific primer sets for these groups it was revealed that Glu-D3 and Glu-A3 encoded the former and the latter, respectively. Both groups of genes were expressed in immature seeds. The presence of these groups of LMW-GSs may affect the dough strength of soft wheat. Received: 26 March 2001 / Accepted: 16 July 2001     

四川小麦地方品种AS1643中低分子量谷蛋白基因的克隆及其蛋白质的二级结构预测

根据小麦低分子量谷蛋白基因保守区序列设计引物P1/P2, 采用PCR法对四川小麦地方品种AS1643的基因组DNA进行扩增, 获得1条约900 bp的片段, 分离、纯化后连接到载体pMD18-T上, 对筛选阳性克隆测序, 获得1个低分子量谷蛋白基因LMW-AS1643(GenBank登录号: EF190322), 其编码区长度为909 bp, 可编码302个氨基酸残基组成的成熟蛋白。序列分析结果表明, LMW-AS1643具有典型的低分子量谷蛋白基因的基本结构, 其推导氨基酸序列与其它已知的LMW-GS相比, 最高相似性为93.40%。生物信息学分析表明, 在LMW-AS1643低分子量谷蛋白中, 无规则卷曲含量最高, 为67.90 %, 其次是a-螺旋, 占30.46 %, b-折叠含量最少, 为1.64 %。     

Variation and classification of B low-molecular-weight glutenin subunit alleles in durum wheat

 The B low-molecular-weight (LMW) glutenin subunit composition of a collection of 88 durum wheat cultivars was analyzed. Extensive variation has been found and 18 different patterns were detected. Each cultivar exhibited 4–8 subunits, and altogether 20 subunits of different mobility were identified. The genetic control of all these subunits was determined through the analysis of nine F₂ populations and one backcross. Five subunits were controlled at the Glu-A3 locus, 14 at Glu-B3 and 1 at Glu-B2. At the Glu-A3 locus each cultivar possessed from zero to three bands and eight alleles were identified. At the Glu-B3 locus each cultivar showed four or five bands and nine alleles were detected. Only one band was encoded by the Glu-B2 locus. A nomenclature for these alleles is proposed and the relationship between them and the commonly used LMW-model nomenclature is discussed. Received: 10 February 1997 / Accepted: 25 April 1997     

小麦高分子量谷蛋白亚基及其基因的研究进展

主要介绍了小麦高分子量谷蛋白亚基（HMW－GS）及其基因的研究进展情况,目前,转基因小麦的技术已经逐渐成熟,由于分子生物学领域分子标记技术的迅速发展,尤其是PCR技术的广泛应用,为实现外源优良储藏蛋白基因导入改良品种提供了可能,利用已知小麦品种的基因序列设计引物,从众多的未知小麦品种中扩增出新基因加以研究并做外源优质HMW－GS基因的转入已成为一种趋势。     

小麦高分子量谷蛋白亚基对加工品质影响的效应分析

分析了 2 50份小麦材料的高分子量谷蛋白亚基 (HMW- GS)组成以及其中 66份材料的加工品质及面条制作品质。回归分析表明 :HMW- GS与 1 0种加工品质性状均有显著的线性关系。不同亚基对综合品质效应的得分大小依次为 :Glu- Al,1 >2 * >null;Glu- Bl,1 4 +1 5>7+8>1 7+1 8>>7+9;Glu- Dl,5+1 0 >>2 +1 2 >4+1 2。不同基因位点对品质的贡献大小顺序为 :Glu- Dl>Glu- Al>Glu- Bl。首次提出了 HMW- GS综合品质评分系统     

高冰草中一种新型高分子量麦谷蛋白亚基编码序列的研究

高冰草(Agropyron elongatun)是普通小麦(Triticum aestivum)的近缘禾草,SDS-PAGE显示其所编码的麦谷蛋白亚基的类型较普通小麦更加丰富,是普通小麦品质改良的重要亲本之一。利用基因组PCR的方法从高冰草中克隆到一个新的高分子量麦谷蛋白亚基(HMW-GS)基因(AgeloG2)全编码序列,同源性分析表明：与普通小麦的1Dy12基因比较在少数位点发生了碱基替换和一处6碱基序列的缺失,同源性为99％;与普通小麦的1Dy10基因比较,该基因亦只有少数碱基的替换和两处18碱基序列的增加及一处6碱基序列的缺失,同源性为98％。从推导的编码序列分析,AgeloG2编码y型HMW—GS。综上分析,AgeloG2是一个新的高分子量麦谷蛋白y-型亚基基因。聚类分析结果显示,无论在基因序列还是推导的氨基酸序列上,小麦1Dy亚基与AgeloG2的同源性都高于与粗山羊来源的y型亚基的同源性。     

Single-seed PCR of LMW glutenin genes to distinguish between durum wheat cultivars with good and poor technological properties

Polymerase chain reaction (PCR) was used to amplify low-molecular-weight (LMW) glutenin sequences from genomic DNA extracted from a single germinating seed of several durum wheat genotypes. Electrophoretic analysis of PCR reactions showed the presence of amplified products characteristic of durum wheat cultivars with good and poor technological properties. This PCR-based approach is proposed as a very efficient and safe alternative to standard procedures for selecting durum wheat genotypes with good qualitative characteristics.     

Atomic force microscopy of a hybrid high-molecular-weight glutenin subunit from a transgenic hexaploid wheat

The high-molecular-weight glutenin subunits (HMW-GS) of wheat gluten in their native form are incorporated into an intermolecularly disulfide-linked, polymeric system that gives rise to the elasticity of wheat flour doughs. These protein subunits range in molecular weight from about 70 K-90 K and are made up of small N-terminal and C-terminal domains and a large central domain that consists of repeating sequences rich in glutamine, proline, and glycine. The cysteines involved in forming intra- and intermolecular disulfide bonds are found in, or close to, the N- and C-terminal domains. A model has been proposed in which the repeating sequence domain of the HMW-GS forms a rod-like beta-spiral with length near 50 nm and diameter near 2 nm. We have sought to examine this model by using noncontact atomic force microscopy (NCAFM) to image a hybrid HMW-GS in which the N-terminal domain of subunit Dy10 has replaced the N-terminal domain of subunit Dx5. This hybrid subunit, coded by a transgene overexpressed in transgenic wheat, has the unusual characteristic of forming, in vivo, not only polymeric forms, but also a monomer in which a single disulfide bond links the C-terminal domain to the N-terminal domain, replacing the two intermolecular disulfide bonds normally formed by the corresponding cysteine side chains. No such monomeric subunits have been observed in normal wheat lines, only polymeric forms. NCAFM of the native, unreduced 93 K monomer showed fibrils of varying lengths but a length of about 110 nm was particularly noticeable whereas the reduced form showed rod-like structures with a length of about 300 nm or greater. The 110 nm fibrils may represent the length of the disulfide-linked monomer, in which case they would not be in accord with the beta-spiral model, but would favor a more extended conformation for the polypeptide chain, possibly polyproline II.