97份安徽省种植小麦品种HMW-GS组成及品质分析

为探究安徽小麦品种高分子量麦谷蛋白亚基（HMW-GS）组成及品质性状,对97份安徽省主要种植的小麦品种进行了HMW-GS组成及品质性状分析。结果表明,供试材料中共检测出10种HMW-GS亚基类型和19种亚基组合,在 Glu-A1位点检测出3种亚基类型,以1亚基（57.73%）和Null亚基 （40.21%）为主;在 Glu-B1位点共检测出3种亚基类型,以7+8亚基（52.58%）为主;在 Glu-D1位点共鉴定出4种亚基类型,优质亚基5+10占比最大,为45.36%;供试品种亚基组合品质得分在4~10分之间,10分的亚基组合1/7+8/5+10和1/17+18/5+10共19份材料（19.59%）,8分以下的共44份材料（45.36%）。对供试材料HMW-GS与品质性状进行相关分析发现, Glu-1的三个位点与硬度指数、溶剂保持力和吸水率相关达显著或极显著水平,对溶剂保持力影响表现为 Glu-A1> Glu-B1> Glu-D1;不同亚基组合对品质性状的影响存在差异,亚基组合为1/17+18/5+10的品种具有最高的硬度指数、4种溶剂保持力、吸水率、稳定时间和粉质质量指数,具有最低的L^*和W,该类品种有烟农19、泰农19、山农17和糯小麦1012;亚基组合为Null/7+9/4+12的品种具有最低的硬度指数、4种溶剂保持力、吸水率、稳定时间和粉质质量指数,具有最高的弱化度、L^*和W,该类品种有荃麦725、皖麦52和未来0818。     

SDS-PAGE与分子标记相结合分析宁夏小麦HMW-GS组成与变化特点

为了建立准确有效小麦HMW-GS的检测方法,提高优质小麦品种鉴定和筛选效率,以已知HMW-GS组成的16份小麦品种为对照,优化完善SDS-PAGE结合分子标记检测小麦HMW-GS的方法,并对103份宁夏小麦品种进行了验证和分析。结果表明,8.5%的分离胶可以有效区分除了 Glu-B1位点的7^*、7^OE与8^*亚基之外的其他亚基,分辨效果优良;SDS-PAGE结合 Bx7^OE、 Bx7^*/Bx7和 By8基因的分子标记可以准确鉴定小麦HMW-GS。在103份宁夏小麦品种中,发现15种HMW-GS和29种组合类型;首次在该地区小麦品种的 Glu-B1位点检测出了携带7^OE、7^*和8^*亚基的品种;1/17+18/5+10为当地小麦HMW-GS的优势亚基组合类型,占20%。自1970年至2010年,HMW-GS的优质亚基（1、17+18和5+10）的出现频率呈明显增长趋势;宁夏小麦的HMW-GS种类和优质亚基出现频率随品种更换呈增加趋势。综上所述,分离胶浓度为8.5% 的SDS-PAGE和 Bx7^OE、 Bx7^*/Bx7和 By8分子标记的方法可准确有效地检测小麦HMW-GS组成,此方法可用于优质小麦品种的鉴定和筛选。     

50份哈萨克斯坦小麦高分子量谷蛋白亚基解析

为发掘优良的小麦种质资源,利用SDS-PAGE技术分析了50份哈萨克斯坦小麦种质高分子量谷蛋白亚基组成及其品质得分。结果表明,50份供试材料中共检测到8种亚基类型,其中在 Glu-A1位点有2种亚基类型（Null,1）, Glu-B1位点有2种亚基类型（7＋8,7＋9）,在 Glu-D1位点有4种亚基类型（5＋10, 2＋10,2＋12,4＋12）。利用PCR分子标记对所得到的高分子量谷蛋白亚基进行验证,结果一致。在50份哈萨克斯坦小麦中,优质亚基1、7＋8、5＋10的出现频率较高,分别为34%、22%、56%。亚基组合Null/7＋9/5＋10在所有亚基组合类型中出现频率最高,为28%,品质得分为7分,而组合1/7＋8/5＋10出现频率仅为8%,品质得分为10分;供试材料品质得分大多为7、8分,平均为7.10分。出现频率较高的优质亚基可以作为改善现有品种亚基组成状况的亲本材料。     

黄淮南片小麦高分子量谷蛋白亚基组成及其与品质的关系

为了解小麦高分子量谷蛋白亚基(HMW-GS)组成与品质之间的关系,对177份黄淮南片地区小麦品种(系)的HMW-GS组成及其品质进行了检测。结果表明,177份材料在Glu-A1位点上有3种亚基类型(N、1、2*),在Glu-B1位点上有5种亚基类型(7+8、7+9、14+15、17+18、13+16),在Glu-D1位点上有3种亚基类型(2+12、5+10、5+12),1、14+15、5+10亚基的出现频率分别为73.45%、11.86%和48.02%。1、7+8、17+18、5+10亚基对蛋白质含量、湿面筋含量、峰值曲线面积、峰高、SDS沉淀值、8min带高、8min曲线面积、形成时间、稳定时间都具有较大的正向效应。共检测到19种不同的亚基组合类型,其中1/7+8/5+10亚基组合的小麦品种(系)各项品质指标均较优,其次是1/7+8/2+12亚基组合品种(系),N/7+9/2+12和N/14+15/5+10亚基组合类型小麦的品质较差。     

甘肃省冬小麦农家品种与改良品种的HMW GS变异及品质效应比较

为了解甘肃冬小麦农家品种和改良品种HMWGS变异及品质效应,为小麦品质改良和亲本选用提供依据,采用SDSPAGE法和近红外反射光谱法检测340份品种的HMWGS和其中252份的沉淀值、蛋白质和湿面筋含量。结果表明,340份品种在Glu1位点共有14种亚基变异,34种亚基组合形式。GluA1位点共有3种变异,亚基缺失（null)的频率最高,1和2^*亚基出现的频率改良品种（36.9%）比农家品种（12.0%）高;GluB1位点共有7种变异,7+8亚基出现的频率最高,改良品种比农家品种降低36.1个百分点;GluD1位点有4种亚基变异,2+12亚基出现频率最高,改良品种比农家品种降低11个百分点,5+10亚基的频率改良品种较农家品种提高17.6个百分点。含5+12亚基的农家品种及含14+15亚基的改良品种综合品质较优。从供试改良品种中筛选出在2个以上基因位点具有优质亚基的品种46个,其中10个品种在3个位点上都具有优质亚基。     

绵阳(绵麦)系列小麦品种高分子量谷蛋白亚基分析

为研究高分子量谷蛋白亚基(HMW-GS)在四川小麦品质育种中的价值,利用SDS-PAGE方法对43个绵阳(绵麦)系列小麦品种的HMW-GS进行了分析。结果共检测出7种HWM-GS类型和9种亚基组合类型。Glu-A1位点有2种亚基,null(N)占优势;Glu-B1位点有3种亚基,7+9与7+8占优势;Glu-D1位点有2种亚基,优质亚基5+10占优势。优势亚基组合类型为N、7+8、5+10。在优质亚基中,1亚基出现的频率为27.91%,7+8亚基出现的频率为44.19%,5+10亚基出现的频率高达65.12%。随时间变迁,优质亚基1和5+10呈上升趋势,而7+8呈下降趋势。优质亚基对蛋白质含量、湿面筋含量、稳定时间和沉降值等品质性状都有正向影响,尤其是优质亚基1或5+10的导入显著提高了沉降值。因此,在四川小麦品质育种中,可以通过聚合优质亚基1和5+10来改善沉降值等品质性状;同时还应引进优质亚基2*、17+18和14+15。     

甘肃小麦品种(系)HMW-GS遗传变异分析

为了了解甘肃省小麦品种HMW-GS的遗传变异和组成，为甘肃优质专用小麦品种筛选和品种改良提供依据,采用SDS-PAGE法，分析了254份甘肃省小麦材料(育成品种(系)和农家品种)Glu-1位点的HMW-GS变异，共检测到22种HMW-GS变异，Glu-A1位点3种，Glu-B1位点11种，Glu-D1位点8种。其中110个育成品种(系)的15种HMW-GS有27种亚基组合类型。Glu-A1位点有2种亚基，47．3％的品种该位点具有优质亚基；Glu-B1位点有7种亚基，76．4％的品种具优质亚基；Glu-D1位点有6种亚基，32．7％的品种具优质亚基。14．5％的品种(n：15)Glu-1位点具优质亚基。144份农家品种的18种HMW-GS共有29种亚基组合形式，Glu-A1位点有3种亚基,Glu-B1位点有9种亚基，Glu-D1位点有6种亚基，无优质亚基组合。     

云南省普通小麦育成品种(系)高分子量麦谷蛋白亚基组成分析

为深入了解云南省建国以来普通小麦育成品种(系)的高分子量麦谷蛋白亚基(HMW-GS)组成情况,利用SDS-PAGE电泳技术对152份云南省1950s以来普通小麦育成品种(系)HMW-GS组成和变异进行了分析。结果表明:(1)云南省普通小麦在Glu-A1位点具有N(56.58%)和1(43.42%)2种亚基类型,在Glu-B1位点具有7+8(42.11%)、7+9(34.87%)、6+8(0.66%)、14+15(7.24%)、17+18(13.16%)和13+16(1.97%)6种亚基类型,在Glu-D1位点具有2+10(5.26%)、2+12(54.61%)、5+10(24.34%)和5+12(15.79%)4种亚基类型;(2)云南省普通小麦HMW-GS组合类型比较丰富,共出现27种亚基组合类型,其中"N,7+8,2+12"、"N,7+9,2+12"、"1,7+8,2+12"与"1,7+9,5+10"较多,出现频率分别为20.39%、9.87%、7.89%和7.89%;(3)云南省各个时期育成品种(系)的HMW-GS品质评分基本维持在4.50左右,1990s以后育成的品种(系)中优质亚基5+10出现的频率随普通小麦育成时期的推移而逐渐增加。由此可见,云南省普通小麦的HMW-GS在Glu-A1、Glu-B1、Glu-D1位点上表现出丰富的多态性,共有12种HMWGS等位变异,包括13+16、2+10和5+12三种稀有亚基类型和27种亚基组合类型;对加工品质具有正效应的优质亚基17+18和5+10频率较小,缺乏优质亚基2*。因此,在云南省普通小麦的品质改良中应加强优质亚基2*、17+18和5+10引入及合理应用。     

1BL/1RS易位系HMW-GS组成与品质分析

为深入挖掘1BL/1RS易位系的品质育种潜力,对收集的300份1BL/1RS易位系的高分子量麦谷蛋白亚基(HMW-GS)组成及其品质性状进行了分析。在300份1BL/1RS易位系 Glu-1位点上共检出12种HMW-GS类型和27种HMW-GS组合,表明这些1BL/1RS易位系具有较高的遗传多样性。在 Glu-A1位点上,亚基Null和1亚基的出现频率分别为47.67%和52.33%; Glu-B1位点有7种等位变异,其中出现频率最高的为7+9亚基对(58.67%); Glu-D1位点有3种等位变异, 2+12亚基对为主要类型,出现频率为71.33%。亚基组合类型中,"Null/7+9/2+12"的出现频率最高(31. 67%)。 Glu-A1、 Glu-B1和 Glu-D1编码优质HMW-GS的比例分别为52.33%、38.34%和16.00%,其中31份1BL/1RS易位系在3个位点均编码优质HMW-GS类型。300份1BL/1RS易位系的品质变异范围较大,有9份材料具有较高的品质育种价值。     

小麦体细胞杂种渐渗系F6代的高分子量谷蛋白亚基组成与位点变异

为了发掘新的小麦高分子量谷蛋白亚基用于品质育种,测定了小麦/长穗偃麦草体细胞杂种F₆代共322个株系的高分子量谷蛋白亚基（HMWGS）组成,并进行了品质评分。结果表明,体细胞杂种株系在GluA1、GluB1和GluD1位点上的遗传变异率分别为0.52、0.58和0.46,一共出现27种不同的亚基组合形式,其中Null、7+9、2+12（同亲本小麦济南177）出现的频率最高,而2^*、7+8+9、2+5+12出现的频率最低;5+12出现的频率约为33.5%;在与优质可能相关的亚基组合当中,13+16出现的频率约为31.1%。由于5+12尚未有品质评分,因此部分株系无法给出分数。本研究表明,通过体细胞杂交可以产生大量的高分子量谷蛋白亚基/组合变异,这有助于今后小麦品质育种工作。     

高分子量麦谷蛋白亚基组成及其与小麦品质性状的关系分析

为进一步明确小麦高分子量麦谷蛋白亚基(HMW-GS)与小麦品质性状的关系,以黄淮麦区的127份小麦品种(系)为材料,利用SDS-PAGE技术、近红外谷物分析仪、粉质仪和拉伸仪等对其进行HMW-GS鉴定和品质检测。结果表明,参试材料在 Glu-A1、 Glu-B1和 Glu-D1 3个位点上分别检测到2(x1、x-null)、4(x7+y8、x7+y9、x14+y15、x17+y18)、2(x5+y10、x2+y12)种不同的亚基类型,其中x1、x7+y9、x5+y10在各自位点上出现的频率均最高,分别为70.1%、42.5%和51.2%;共发现有14种HMW-GS组合类型,其中1Ax1/1Bx7+1By8/1Dx5+1Dy10和1Ax1/1Bx7+1By9/1Dx2+1Dy12出现的频率较高,分别为18.9%和17.3%。1Ax1、1Bx7+1By8、1Bx17+1By18、Dx5+1Dy10亚基对蛋白质、沉降值、稳定时间、最大抗延阻力和拉伸面积等品质性状有显著的正向效应,而1Bx14+1By15亚基对除蛋白质和湿面筋以外的其他品质性状有负向效应。携带1Ax1/1Bx7+1By8/1Dx5+1Dy10品种(系)的被测品质性状显著高于携带其他组合类型的品种(系),其次是携带1Ax1/1Bx17+1By18/1Dx5+1Dy10的品种(系),而携带1Ax-null/1Bx7+1By9/1Dx2+1Dy12和1Ax-null/1Bx14+1By15/1Dx5+1Dy10品种(系)的各个品质性状显著低于携带其他组合类型的品种(系)。该结果可为进一步提高优质亚基的育种利用率和我国小麦品质的遗传改良提供参考依据。     

Stacking HMW-GS transgenes in bread wheat: Combining subunit 1Dy10 gives improved mixing properties and dough functionality

We have determined the technological properties of four lines containing combinations of three HMW-GS transgenes, encoding HMW-GS 1Ax1, 1Dx5 and 1Dy10. These lines were produced by conventional crossing of three single transgenic lines of the bread wheat cultivar Anza that contains the endogenous HMW-GS pairs 1Dx2 + 1Dy12 and 1Bx7* + 1By8 and is null for the Glu-A1 locus. Consequently, the total number of HMW-GS ranged from 4 in the control line Anza to 7 in line T618 which contains all three HMW-GS transgenes. The lines were studied over two years using a range of widely used grain and dough testing methods. All lines with transgenic subunits showed higher levels of glutenin proteins than the Anza control, and these differences were highly significant for lines T616, T617 and T618, containing, respectively, the transgenes encoding HMW-GS 1Ax1 and 1Dy10, 1Dx5 and 1Dy10 and 1Ax1, 1Dx5 and 1Dy10. These increases in glutenin levels are compensated by lower levels of gliadins present in transgenic lines. These changes affected the ratio of polymeric to monomeric gluten proteins (poly:mono), the ratio of HMW-GS to LMW-GS (HMW:LMW) and the contents of individual 1Ax, 1Bx, 1By, 1Dx and 1Dy subunits. Transgenic lines expressing subunit 1Dy10 together with x-type subunits (T616, T617 and T618) were superior to line T606, which had only increases in x-type subunits. In particular, the combination of transgenic subunits 1Dx5 and 1Dy10 (line T617) gave better dough rheological properties than the other combinations of transgenic subunits. For example, dough development time and stability were increased by 3.5-fold and 8.5-fold, respectively, while the mixing tolerance index (MTI) was decreased by 3.3-fold in line T617 with respect to the control line. Alveograph analyses showed that all four transgenic combinations had increased P values compared to the Anza control but subunit 1Dx5 greatly reduced the extensibility (L). These results show that stacking HMW-GS transgenes by conventional crossing is a valid strategy for the improvement of wheat quality, with different effects being related to the different HMW-GS combinations.     

Mixing properties and dough functionality of transgenic lines of a commercial wheat cultivar expressing the 1Ax1, 1Dx5 and 1Dy10 HMW glutenin subunit genes

In this work we report the effects of the HMW-GS 1Ax1, 1Dx5 and 1Dy10 on the breadmaking quality of the bread wheat cultivar Anza that contains the HMW-GS pairs 1Dx2 + 1Dy12 and 1Bx7* + 1By8, and is null for the Glu-A1 locus. This allows the characterization of individual subunits 1Dx5 and 1Dy10 in the absence of subunit 1Dx5, and the interactions between these subunits and subunits 1Dx2 and 1Dy12 to be determined. Three transgenic lines termed T580, T581 and T590, containing, respectively, the HMW-GS 1Ax1, 1Dx5 and 1Dy10 were characterized over 3 years using a range of widely-used grain and dough testing methods. The transgenic subunits 1Ax1, 1Dx5 and 1Dy10 accounted for 25.2%, 20.3% and 17.9%, respectively, of the total HMW-GS in the three transgenic lines. Although lines T581 and T590 expressed similar levels of subunits 1Dx5 and 1Dy10 they had different effects on other aspects of protein composition, including changes in the ratios of glutenin/gliadin, of HMW/LMW-GS, the 1Dx2/1Dy12, the x-type/y-type HMW-GS and the proportions of high molecular mass glutenin polymers. In contrast, lines transformed to express subunits 1Ax1 and 1Dx5 showed similar changes in protein composition, with higher protein contents and decreased ratios of glutenin/gliadin and 1Dx2/1Dy12. In addition, both transgenic lines showed similar increases in the ratio of x-type/y-type subunits compared to the control line. The transgenic lines were analysed using Farinograph, Mixograph and Alveograph. This confirmed that the expression of all three subunits resulted in increased dough strength (and hence breadmaking quality) of the cultivar Anza. A beneficial effect of subunit 1Dx5 has not been reported previously, transgenic wheat lines expressing this subunit giving overstrong dough unsuitable for breadmaking. However, the expression of subunit 1Dy10 had a greater effect on breadmaking quality than subunits 1Ax1 and 1Dx5. The Farinograph parameters such as dough stability and peak time were increased by 9.2-fold and 2.4-fold, respectively, in line T590 (expressing 1Dy10) with respect to the control line. Similarly, the Mixograph mixing time was increased by four-fold and the resistance breakdown decreased by two-fold in line T590 compared with the control line. The Alveograph W value was also increased by 2.7-fold in line T590 compared to the control line. These transgenic lines are of value for studying the contribution of specific HMW-GS to wheat flour functional properties.     

小麦高分子量谷蛋白亚基基因分子育种研究进展

为给小麦品质改良工作者提供通过分子生物技术优化HMW-GS组成方面的全面信息,综述了HMW-GS的基因克隆、分子标记以及基因工程改良三个方面近年来的国内外研究进展.迄今为止, 被克隆和测序的HMW-GS基因已有20多个,即1Ax1、1Ax2*、1Ax2*B、1Ay1、1Bx7、1Bx9、1Bx17、1Dx2、1Dx5、1Bx20、1By8、1By9、1Dy10、1Dy12、1AxNull、1Bx14、 1Bx23、1Dx2.2、1Dx2.1、1Dy10.1、1Dy12t等,还不断有新的基因被发现和克隆.克隆方法可概括为两种:一种是以HMW-GS克隆作为探针筛选cDNA或基因组DNA文库, 从而获得所需的靶基因序列, 然后再选择合适的载体进行克隆测序;另一种则是采用PCR技术.HMW-GS基因的分子标记方法主要有RFLP法、PCR法和SNP(单核苷酸多态性)法.目前已有研究者通过基因工程方法将部分外源HMW-GS基因导入小麦,有效地改善了受体品种的加工品质.     

Incorporation of high-molecular-weight glutenin subunits into doughs using 2 gram mixograph and extensigraphs

To study the contributions of high-molecular-weight glutenin subunits (HMW-GS) to the gluten macropolymer and dough properties, wheat HMW-GS (x- and y-types) are synthesized in a bacterial expression system. These subunits are then purified and used to supplement dough mixing and extensigraph experiments through dough partial reduction and reoxidation to allow these exogenously added HMW-GS to incorporate into gluten polymers. Detailed results are given for seven mixing and two extension parameters. HMW-GS synthesized in bacteria behaved similarly under these conditions to the same HMW-GS extracted from wheat flour. These experiments initially focused on the HMW-GS of the D-genome of hexaploid wheat encoded at the Glu-D1 locus; e.g. the Dx2, Dx5, Dy10, and Dy12 subunits. Experiments used five different flours and results are shown to be consistent when normalized to results from Dx5. The incorporation of Dx-type subunits into the gluten disulfide bonded network has greater effects on dough parameters than incorporation of Dy-type subunits. When Glu-D1 x- and y-type subunits are incorporated together, there are synergistic effects greater than those with either subunit type alone. This synergistic effect was greatest with approximately equal amounts of Dx- and Dy-type subunits - implying a 1:1 stoichiometric relationship.     

甘肃冬小麦品种(系)面粉品质性状相关基因分析

为了解甘肃省近20年育成的106份冬小麦品种(系)中加工品质性状相关基因分布情况,用22个分子标记对供试材料的HMW-GS、LMW-GS、面粉色泽及籽粒硬度等品质性状相关基因进行了分析。结果发现,供试品种(系)的HMW-GS相关基因中,在Glu-A1位点检测到34份品种(系)含有AxNull,频率为32.08%;在Glu-B1位点检测到Bx7+By8和Bx14+By15共2种基因组合,分别占17.92%和25.47%;在Glu-D1位点检测到11份品种(系)含有Dx5+Dy10,占10.38%。对LMW-GS鉴定结果显示,29份品种(系)含Glu-A3d基因,分布频率为27.36%。HMW-GS和LMW-GS亚基组合中,含有4个、3个和2个位点优质亚基基因组合的品种(系)分别占0.94%、8.49%和3.77%。对面粉色泽相关基因Ppo-A1、Ppo-D1、Psy-A1、Lox-B1和TaPod-A1位点的检测发现,优异等位变异占比分别为39.62%、50.94%、31.13%、30.19%和38.68%。对籽粒硬度相关基因检测发现,在Pina、Pinb和Pinb-2等位变异位点的检测...     

Pasting properties of transgenic lines of a commercial bread wheat expressing combinations of HMW glutenin subunit genes

Seven transgenic lines of a commercial wheat (Triticum aestivum L.) cultivar expressing transgenic subunits 1Ax1, 1Dx5 and 1Dy10, alone or in combination have been developed. Pasting properties were determined in these transgenic lines using a Rapid Visco Analyser (RVA) in order to determine the possible impact of HMW-GS transgene expression on the starch properties. Expression of the HMW-GS transgenes increased the proportions of the corresponding 1Ax, 1Dx and 1Dy subunits affecting significantly the ratios of HMW-GS:LMW-GS and x-type:y-type HMW-GS. Starch granule size distribution varied significantly among all transgenic lines, with the Anza control and transgenic line T616 (expressing subunits 1Ax1 and 1Dy10) showing the highest and the lowest percentage of B granules, respectively. All transgenic lines increased the water-binding capacities (WBC) at 25 °C and 90 °C. Line T606 (expressing subunits 1Ax1 and 1Dx5) and line T590 (expressing subunit 1Dy10) showed the lowest and the highest values for peak viscosity, respectively. Notably, lines expressing only transgenic x-type subunits (T580, T581 and T606), with high ratios of x-type:y-type HMW-GS, had low peak viscosities, final viscosities and breakdown viscosities. Line T590 had the highest breakdown viscosity while lines T606 and T581 had the lowest.     

氮肥运筹对小麦HMW GS形成与积累的影响

为给小麦优质栽培提供理论依据,以多穗型品种031085和大穗型品种山农91为试验材料,利用SDSPAGE电泳等分析技术,研究了氮肥运筹对不同类型小麦品种高分子量谷蛋白亚基（HMWGS）形成的影响。结果表明,小麦HMWGS出现的时间多在花后10~15 d,且多穗品种相对早于大穗品种;氮肥运筹对HMWGS出现时间影响不大;亚基组合1Bx7+1By9出现时间早于亚基1Dx2+1Dy12;不同施氮处理下,开花后各小麦品种HMWGS亚基的积累过程均呈三次曲线增长;各亚基积累量所占百分含量随时间变化较为平稳,受施氮影响较小,y型亚基百分含量高于x型亚基。