Polymerisation of gluten proteins in developing wheat grain as affected by desiccation

The breadmaking quality of wheat is affected by the composition of gluten proteins and the polymerisation of subunits that are synthesised and accumulated in developing wheat grain. The biological mechanisms and time course of these events during grain development are documented, but not widely confirmed. Therefore, the aim of this study was to monitor the accumulation of gluten protein subunits and the size distribution of protein aggregates during grain development. The effect of desiccation on the polymerisation of gluten proteins and the functional properties of gluten were also studied. The results showed that the size of glutenin polymers remained consistently low until yellow ripeness (YR), while it increased during grain desiccation after YR. Hence, this polymerisation process was presumed to be initiated by desiccation. A similar polymerisation event was also observed when premature grains were dried artificially. The composition of gluten proteins, the ratios of glutenin to gliadin and high molecular weight-glutenin subunits to low molecular weight-glutenin subunits, in premature grain after artificial desiccation showed close association with the size of glutenin polymers in artificially dried grain. Functional properties of gluten in these samples were also associated with polymer size after artificial desiccation.     

Kernel vitreousness and protein content: Relationship,interaction and synergistic effects on durum wheat quality

Four sets of durum samples were used in this study to further understand the interrelationships among hard vitreous kernels (HVK), protein content, and pigment concentration, with a focus on the interaction and synergistic effects of protein content and vitreousness on durum quality. HVK level increases with higher protein content in the range of 9.5–12.5%, but this relationship is less evident in durum samples with high protein content (12.5–14.5%). Both protein content and kernel vitreousness can significantly affect durum milling quality. White starchy kernels (WSK) in low protein durum have a very detrimental impact on milling and pasta processing quality, but high protein content can mitigate the adverse impact of WSK on durum quality. Although protein content plays a dominant role, higher HVK might contribute positively to pasta firmness. There was no significant difference in yellow pigment content between HVK and WSK. However, pigment loss from semolina to dough was higher for WSK than HVK. Despite the difference in protein content, HVK and WSK have little difference in gluten strength. The monomeric protein was preferentially accumulated in HVK. The glutenin proteins of HVK and WSK were similar in the ratios of 1Bx/1By and HMW/LMW-GS.     

Influence of low soil nitrogen and phosphorus on gluten polymeric and monomeric protein distribution in two high quality spring wheat cultivars

The effect of low levels of nitrogen, phosphorus and a combination of the two on the distribution of polymeric and monomeric proteins in two high quality spring bread wheat cultivars was investigated for two consecutive seasons. Size exclusion-high performance liquid chromatography (SE-HPLC) was used to determine the quantity and relationships of monomeric and polymeric proteins, and their relationship with flour protein content (FPC) and SDS sedimentation volume (SDSS). The low nitrogen and combined low nitrogen and low phosphorus treatments had a much larger effect on the protein fractions than the low phosphorus treatment alone. The SDS-soluble large monomeric protein fraction and the percentage SDS-insoluble monomeric proteins, were significantly increased under low nitrogen and a combination of low nitrogen and low phosphorus treatments. The percentage SDS-insoluble large and total polymeric proteins was significantly reduced under low nitrogen and a combination of low nitrogen and phosphorus treatments. The SDS-soluble and -insoluble small polymeric proteins were significantly increased under both low nitrogen and a combination of low nitrogen and low phosphorus treatments. The low nitrogen treatment consistently caused the lowest FPC and SDSS values. Under low nitrogen conditions, there was a significant positive correlation between the SDS-soluble gliadins and SDSS, and FPC.     

Impact of ethanol,succinic acid,and the combination thereof at levels produced during sponge fermentation on hard wheat,soft wheat,and durum wheat farinograph rheology

The sponge and dough mixing process is one of the most common in the world, yet the mechanistic understanding of this process has yet to be sufficiently explored. In this study, aqueous solutions of ethanol, succinic acid, and their combination were prepared at concentrations intended to replicate fermentation times of 3, 4 and 6 h. These solutions were added to a farinograph mixer to make dough using hard wheat, soft wheat, and durum wheat flour. The results indicate that these yeast metabolites (ethanol, succinic acid) impact the mixing resistance, peak mixing value, and dough mixing stability in each of the flour types, likely primarily affected by the ratios of gliadin to glutenin and LMW glutenin in each flour type. Results suggest a stabilizing non-covalent interaction imparted by gliadin at peak mixing time, a stabilizing effect of HMW glutenin during break down, and synergistic effects of ethanol and succinic acid that leads to a faster rate of breakdown in later stages of mixing. It also suggests an increase in mixing resistance when acidulants are added to durum wheat dough. Taken together, this study adds new insights on the sponge and dough mixing process in a way that has not previously been conducted.     

反相超高效液相色谱分析小麦麦谷蛋白组分研究

为利用反相超高效液相色谱(RP-UPLC)分离、定量分析小麦麦谷蛋白组分，研究了流动相比例、洗脱时间、柱温、流速和进样量对色谱分离结果的影响，并利用新建立的分离麦谷蛋白亚基的方法分析了基因型及环境对麦谷蛋白组分的影响。结果表明，最优的洗脱程序为79%的流动相A(含0.08%三氟乙酸的超纯水)和21%的流动相B(含0.08%三氟乙酸的乙腈溶液)洗脱5 min,在25 min内，流动相A逐渐降低至48%,流动相B逐渐增加至52%,最后维持5 min,柱温、流速和进样量分别为55℃、0.5 mL·min^-1和16μL,该方法可以精确分离至少11种高分子量麦谷蛋白亚基(HMW-GS)。单个HMW-GS的相对含量和HMW-GS与低分子量麦谷蛋白亚基(LMW-GS)的比值均受基因型和环境的影响，且基因型的影响大于环境。     

Glu-B1位点亚基色谱鉴定及7OE对面团强度的影响

准准确鉴定高分子量谷蛋白亚基（HMW-GS）及探讨其对面团特性的影响是小麦品质研究的重要内容。用聚丙烯酰胺凝胶电泳（SDS-PAGE）和反相高效液相色谱（RP-HPLC）对62份品种（系）的HMW-GS组成进行鉴定。结果表明，结合SDS-PAGE和RP-HPLC两种方法可以对Glu-B1位点，尤其是Glu-B1（7+8）进行有效鉴定。选用13份姊妹系为材料，使用RP-HPLC和凝胶色谱（SE-HPLC）方法，研究了Glu-B1al（7^OE+8*）以及贮藏蛋白组分含量对面团强度的影响。结果表明，Glu-B1al（7^OE+8*）可显著提高HWM-GS总量和面团强度，7OE可作为优质亚基用于强筋小麦育种。相关性分析表明，谷蛋白总量、HWM-GS、LMW-GS以及x-HMW含量与最大抗阻力呈1%显著正相关，相关系数分别为0.76、0.76、0.77和0.72。SDS-不溶性谷蛋白大聚体（UPP）占谷蛋白聚合体总量的百分比与揉面仪峰值时间、粉质仪形成时间和稳定时间以及最大抗阻呈1%或0.1%显著正相关，相关系数分别为0.77、0.90、0.89和0.87。醇溶蛋白与谷蛋白含量的比值与揉面仪峰值时间呈1%显著负相关，相关系数为－0.69；与粉质仪稳定时间和最大抗阻呈5%显著负相关，相关系数分别为－0.58和－0.64。HPLC可有效分离和量化HWM-GS，并作为小麦品质育种有效的辅助手段。     

Glu-1位点缺失对小麦麦谷蛋白聚合体粒度分布及面团特性的影响

以Glu-1位点正常和部分缺失的小麦品系为材料,探讨HMW-GS和LMW-GS组成与谷蛋白聚合体粒度分布和面团特性的关系,为利用HMW-GS缺失系改良小麦品质提供理论依据。在20个供试硬白冬麦品系中,1个品系为Glu-A1位点缺失,5个品系为Glu-D1缺失,3个品系为Glu-A1和Glu-D1双缺失。所有品系的蛋白质含量皆较高(13.39%~14.12%),品系间无显著差异,缺失系与非缺失系间也无显著差异。Glu-1位点缺失显著降低了高分子量谷蛋白/低分子量谷蛋白比(HMW/LMW)、不溶性谷蛋白大聚体的含量和百分比。谷蛋白/醇溶蛋白比(GLU/GLI)在基因型间变幅较小,且在缺失系和非缺失系间无显著差异。Glu-1位点缺失显著降低了面团弹性,但显著提高了面团的延展性。部分Glu-1位点缺失系仍具有较高的面团强度和突出的延展性,谷蛋白聚合体粒度分布和面团特性受谷蛋白亚基组成和表达量的共同影响。研究结果表明,利用Glu-1位点亚基缺失可能是改善面筋延展性,提高食品加工品质的方法之一。     

Limited but specific variations of seed storage proteins in Japanese common wheat (Triticum aestivum L.)

The electrophoretic banding patterns ofgliadin in common wheat lines derived fromJapan were determined byacid-polyacrylamide gel electrophoresis. For the 107 wheat lines used in our study,27 different patterns were identified, 13corresponding to the -gliadin, 8 tothe , -gliadin and 6 to the-gliadin. The gliadin patterns ofJapanese wheat cultivars and landracesgreatly differed from the patterns of wheatlines from other countries, and thevariation seen in wheat lines from Japanwas limited to 46 patterns. Sevencollection or breeding areas in Japanshowed different frequencies in theirgliadin patterns. Combining the gliadinpatterns with high molecular weightglutenin subunit compositions, 67combinations were observed. One gliadinpattern consisting of -gliadinpattern F, , -gliadinpattern H and -gliadin pattern Dwas frequently found in many Japanese wheatlines, though the other patterns werelimited to only one or two wheat lines.     

High molecular weight glutenin subunit composition of Chinese bread wheats

Summary The endosperm storage proteins of 205 Chinese bread wheat cultivars and advanced lines were fractionated by SDS-PAGE to determine their high molecular weight (HMW) glutenin subunit composition. Seventeen alleles were identified: three at Glu-A1, eight at Glu-B1, and six at Glu-D1. The most common alleles were Null, 1, 7+8, 7+9, and 2+12. The results indicate that wheats from different regions differ in their frequencies of HMW glutenin subunits, however, none of the subunits could be related to specific environments. The glutenin quality scores of Chinese wheats ranged from 3 to 10, with an average of 6.7. Increasing quality scores have implications in improving steam-bread making quality for Chinese consumers. On the basis of HMW glutenin subunit composition, Chinese wheats are close to European wheats, especially Italian wheats because several Italian introductions are widely distributed in the pedigrees of Chinese wheat.     

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.