陕西省部分小麦栽培品种高分子量麦谷蛋白亚基组成及其与品质关系

本研究以36个陕西省主要栽培品种为材料,利用SDS-PAGE方法分析了高分子量麦谷蛋白亚基组成,并对其SDS沉淀值进行了测定,分析了高分子量麦谷蛋白亚基组成与代表面包烘烤品质的SDS沉淀值间的关系.结果表明:Glu-1位点存在着广泛的等位基因变异,在所分析的36个品种中,Glu-A1位点亚基1出现频率最高,为63.9%;Glu-B1位点等位基因变异最丰富,其中亚基7 8和7 9出现频率最高,其次为亚基14 15;Glu-D1位点亚基组成以2 12为主;对各位点品质评分与SDS沉淀值的简单相关分析结果表明各位点品质评分与SDS沉淀值均存在着显著或极显著相关关系,各位点影响大小依次为Glu-D1＞Glu-A1＞Glu-B1;Glu-1品质评分与SDS沉淀值极显著正相关.     

Disulphide structure of high-molecular-weight (HMW-) gliadins as affected by terminators

This study aimed at elucidating SS-bonds of HMW-gliadins (HGL) from wheat with the focus on terminators of glutenin polymerisation. HGL from wheat flour extracts non-treated or treated with the S-alkylation reagent N-ethylmaleinimide (NEMI) were compared. HGL from wheat flour Akteur were isolated, hydrolysed with thermolysin and the resulting peptides pre-separated by gel permeation chromatography and analysed by liquid chromatography/mass-spectrometry using alternating electron transfer dissociation/collision-induced dissociation. Altogether, 22 and 28 SS-peptides from samples without and with NEMI treatment, respectively, were identified. Twenty-six peptides included standard SS-bonds of α- and γ-gliadins, high-molecular-weight and low-molecular-weight glutenin subunits. Eleven SS-bonds were identified for the first time. Fifteen peptides unique to HGL contained cysteine residues from gliadins with an odd number of cysteines (ω5-, α- and γ-gliadins). Thus, gliadins with an odd number of cysteines, glutathione and cysteine had acted as terminators of glutenin polymerisation. Decisive differences between samples without and with NEMI treatment were not obvious showing that the termination of polymerisation was already completed in the flour. The two HGL samples, however, were different in the majority of ten peptides that included disulphide-linked low-molecular-weight (LMW) thiols such as glutathione and cysteine with the former being enriched in the non-treated HGL-sample.     

中国小麦品种资源Glu-1位点组成概况及遗传多样性分析

 分析了 5 12 9份中国小麦初选核心种质样品HMW GS的组成情况 ,其中地方品种 345 9份、育成品种(系 ) 16 70份。这些材料作为初级核心种质基本代表了保存在国家长期库中的普通小麦种质资源的遗传多样性 ,覆盖了中国小麦栽培的 10大生态区。总体来看 ,在Glu A1、Glu B1和Glu D13个位点上的主要等位变异分别为null、7+8和 2 +12。育成品种中 1、7+9、14 +15、5 +10和 5 +12亚基 (对 )的频率比地方品种有很大的提高。在Glu 1位点上 ,地方品种与育成品种的遗传丰富度差异甚微 ,但育成品种的遗传离散度指数却显著高于地方品种。在 3个位点中 ,Glu B1位点的多样性最丰富 ,其次为Glu D1位点 ,Glu A1位点的多样性最差。从生态区来讲 ,地方品种变异类型最丰富的 3个大区是黄淮冬麦区、西北春麦区和西南冬麦区 ;选育品种最丰富的 4个大区是西南冬麦区、黄淮冬麦区、长江中下游冬麦区和北部冬麦区。由于广泛的引种、杂交、选择以及亲本选配中的偏爱 ,造成许多生态区遗传离散度指数高低与遗传丰富度出现相矛盾的现象 ,这点在长江中下游冬麦区材料中表现尤为突出。育成品种与地方品种间遗传分化系数分析表明 ,现代引种和杂交育种使我国小麦品种“群体”遗传组成和结构发生了质的变化。     

我国春麦区部分小麦品种品质状况分析

1999年将 38份春小麦品种 (品系 )种植于内蒙古呼和浩特 ,对其磨粉品质和面包烘烤品质进行了评价。结果表明 ,我国春小麦品种的面包烘烤品质较差 ,不同地区品种间品质差异较大 ,辽宁和内蒙品种的磨粉品质和面包烘烤品质优于其它地区的品种。回归分析表明 ,蛋白质含量和单位蛋白质含量的面包体积决定了面包体积总变异的 99.9% ,硬度、沉淀值和吸水率对面包总分有重要作用。品种的出粉率主要取决于 1心槽路出粉率 ,2心和 1皮槽路出粉率对出粉率贡献也较大。高分子量麦谷蛋白亚基 (HMW GS)Glu A1和Glu D1位点的等位变异与品质性状密切相关。面包烘烤品质的改良应在分析HMW GS的基础上 ,将硬度作为选择指标之一 ,适当提高蛋白质含量 ,重点加强对沉淀值的选择     

Glu A1和 Glu D1位点高分子量谷蛋白亚基共同缺失对弱筋小麦品质的影响

为了研究高分子量谷蛋白亚基(HMWGS)缺失对小麦品质的影响,以 GluA1和 GluD1位点HMWGS共同缺失材料2GS041410作供体亲本,以弱筋小麦品种扬麦13和扬麦18作轮回亲本进行回交,构建不同遗传背景的BC¹F³和BC²F³群体,测定受体、供体亲本的品质,以及回交群体BC¹F³和BC²F³中 GluA1和 GluD1位点HMWGS共同缺失纯合单株及两位点均正常表达纯合单株的品质。结果表明,供体2GS041410具有较低的SDS沉降值、较短的面团形成时间和稳定时间;在BC¹F³和BC²F³中, GluA1和 GluD1位点HMWGS共同缺失对蛋白含量影响不显著,但可显著或极显著降低SDS沉降值和水溶剂保持力(SRC)。 GluA1和 GluD1位点HMWGS双缺失在弱筋小麦品质育种中具有一定的应用价值。     

烟草高分子量核DNA提取方法比较

高分子量(HMW)核DNA的制备是构建BAC文库关键环节之一。为构建高质量的烟草BAC文库,作者研究了一次选择法和二次选择法提取烟草高分子量核DNA的效果。研究结果表明:虽然二次选择法得到的高分子量核DNA浓度较低,但此法省略了对细胞核连续3￣5次的漂洗,而且得到含DNA的凝胶块(plugs)数量是用一次选择法的3倍,因此得到高分子量DNA的总量更高;同时二次选择法得到的plugs纯度更高,排除了酶切时细胞杂质对HMWDNA的干扰,此外在节约材料等方面也优于一次选择法。     

Construction and characterization of a BAC library for sunflower (Helianthus annuus L.)

A bacterial artificial chromosome (BAC) library was constructed using the sunflower (Helianthus annuus L.) restorer line RHA325, which carries the restorer gene Rf1 and the Pl2-gene conferring resistance to downy mildew. High molecular weight DNA was prepared from nuclei using leaf material from two-week old seedlings. The library was constructed using the HindIII site of pBeloBAC11. The current BAC library comprises 104,736 clones. The insert size of the clones varied between 20 and 270 kb, with an average insert size of 60 kb. The whole 1.9× sunflower BAC library was spotted in duplicate on four high-density filters, each carrying 55,296 clones. The content of organellar DNA, which was estimated by colony hybridisation against the mitochondrial probe coxI and the chloroplast probe rbcL, proved to be less than 0.03 and 0.1%, respectively. BAC pools, allowing PCR-based screening, were made and used to identify positive BAC clones for the markers OP-K13_454, closely linked to the restorer gene Rf1. The PCR-based screening was verified by the results obtained for this marker by colony hybridisation.     

Duplication of the Bx7 high-molecular-weight glutenin subunit gene in bread wheat (Triticum aestivum L.) cultivar'Red River 68'

SDS-PAGE analysis of seed proteins of the cultivar‘Red River 68’showed a considerably higher staining intensity of the band corresponding to HMW-GS Bx7 relative to the equivalent band in the cultivars‘Chinese Spring’and‘Cheyenne’. Southern blots of restriction enzyme fragments from DNA of these three cultivars were analyzed densitometrically to reveal that the band corresponding to the Bx7 gene of‘Red River 68’had a double staining intensity compared to the equivalent bands from the other two cultivars, which indicates that in‘Red River 68’a duplication of the Bx7 gene has occurred. Although the possibility of the gene copy being a pseudogene was not ruled out, the greater amount of protein corresponding to Bx7 in‘Red River 68’most likely is in accord with an increase in active gene number. SDSPAGE analysis of the proteins showed also that the mobility of Bx7 in‘Cheyenne’was slightly different from the mobilities of the Bx7 subunits of‘Red River 68’and‘Chinese Spring’. The same difference was observed at the gene level by PCR amplification of the genes encoding these subunits.     

西藏普通小麦地方品种特有高分子量谷蛋白亚基组合“Tibetan Dx5*+Tibetan Dy10”中Tibetan Dy10亚基基因测序与分析

 【目的】鉴定在西藏小麦地方品种中发现的特有高分子量谷蛋白亚基组合“Tibetan Dx5*+Tibetan Dy10”中的Tibetan Dy10亚基是否与普通小麦Dy10亚基为同一亚基。【方法】利用SDS-PAGE分析和Tibetan Dy10亚基基因的克隆和测序。【结果】表明Tibetan Dy10亚基与普通小麦中Dy10亚基广泛存在的Dx5+Dy10组合形式中的Dy10亚基的分子序列非常相似，但分别在2个六肽中的1个氨基酸部位发生替换，第335位的甘氨酸（G）和第451位的谷氨酰氨（Q）在Tibetan Dy10 中均被替换为精氨酸（R）。【结论】Tibetan Dy10与普通小麦中常见的Dy10亚基基因的DNA序列存在微小差异，属于Dy10位点的一个新变异。