Puroindoline genotype,starch granule size distribution and milling quality of wheat

Genetically-diverse wheat samples from the Australian Winter Cereals Collection propagated in two environments were sequenced to identify puroindoline genotypes then the relationships between flour yield, genotype, starch granule size distribution and starch-bound puroindoline protein content were investigated. The Pina-D1a, Pinb-D1b genotype resulted in a higher average flour yield than either the Pina-D1b, Pinb-D1a or the Pina-D1a, Pinb-D1a but the ranges of flour yields for the three genotypes showed considerable overlap. For both hard wheat genotypes (Pina-D1a, Pinb-D1b or Pina-D1b, Pinb-D1a), a higher proportion of type A to type C starch granules was associated with higher flour yield and this relationship accounted for between 31% and 33% of the variation in flour yield. This result is consistent with previously reported findings for soft wheat. For the Pina-D1a, Pinb-D1b genotype, increased flour yield was also associated with a decrease in starch granule-bound puroindoline protein, which accounted for 31–35% of the variation in flour yield across the two environments. The combined effect of starch granule type and associated puroindoline content accounted for 68% of the variation in flour yield within the Pina-D1a, Pinb-D1b genotype.     

A novel hardness-related and starch granule-associated protein marker in wheat: LMW-GS-‘S’

Starch granule (SG)-associated proteins are involved in starch synthesis and the interaction between SGs and the endosperm protein matrix. In this study, SG proteins were sequentially extracted with the chaotropic reagent, urea from 1 M to 4 M, and then profiled using an integrated proteomic approach including one- and two-dimensional electrophoresis, mass spectrometry and antibody-based enzyme-linked immunosorbent assay (ELISA). The results demonstrated that the SG-associated proteins were dominated by granule-bound starch synthase (GBSS), gliadin, low molecular weight glutenin subunits (LMW-GS), serine protease inhibitors, α-amylase inhibitors and puroindolines. A protein with an apparent molecular mass of 50 kDa, expressed in cultivar hard wheat Kukri but not in soft wheat Triller was identified as a novel member of the ‘S’ group of LMW-GS, designated as LMW-GS-‘S’. Further characterization using a broad wheat population revealed that LMW-GS-‘S’ was selectively expressed in hard wheat cultivars while deleted in all soft wheats tested. Its relationship with hardness was confirmed by its expression in tetraploid durum wheats, which are among the hardest wheats around the world. Monoclonal antibody (MAb) F8-14E6 against LMW-GS-‘S’ was developed and used in an ELISA to screen 90 Glu allele-defined doubled haploid Janz/Kukri wheat lines. The allele that encodes LMW-GS-‘S’ was mapped to GluB3h (p < 0.001).     

Variation in polar lipid composition among near-isogenic wheat lines possessing different puroindoline haplotypes

The exact mechanism underlying wheat (Triticum aestivum L.) kernel hardness is unknown. Similar to puroindoline proteins, polar lipids are present on the surface of starch granules. The objective of this research was to determine the specific polar lipid species present on the surface of wheat starch from near-isogenic wheat lines that have different puroindoline haplotypes and endosperm hardness. Four near-isogenic wheat lines were used in this study, all derived from the soft cultivar Alpowa. Direct infusion tandem mass spectrometry was used to identify the lipid species in whole-meal, flour and starch samples. Endosperm hardness had no significant effect on the polar lipid contents in wheat whole-meal, a slight influence on the polar lipid contents of the flour fractions and a significant influence on the polar lipid composition of the polar lipids located on the surface of wheat starch. The greatest quantities of polar lipids on the starch-surface occurred when both puroindoline proteins were present in their wild-type form. Starch-surface polar lipid content dramatically decreased when one of the puroindoline proteins was null or if pin-B was in the mutated form. The least amount of polar lipids was present when pin-B was in its mutated form and pin-A was in its wild-type form.     

糯小麦籽粒淀粉粒度分布特征

为进一步加深对糯小麦淀粉特性的认识,选用2个糯小麦和2个普通非糯小麦品种为材料,利用激光粒度分析仪分析了糯小麦完熟籽粒中淀粉的粒度分布特征,及其与普通非糯小麦品种的差异。结果表明,糯小麦淀粉粒的粒径分布范围为1.0~43.7 μm,不同粒径淀粉粒的数目比例分布呈单峰曲线变化,体积和表面积分布均呈双峰曲线变化;糯小麦籽粒中存在2种类型淀粉粒,即A型大淀粉粒（直径≥10 μm）和B型小淀粉粒（直径<10 μm）,B型淀粉粒的数目、体积和表面积比例均高于A型淀粉粒。与普通非糯小麦相比,糯小麦籽粒中B型小淀粉粒的数目、体积和表面积比例较高,淀粉粒的平均粒径和中位粒径较大。糯小麦与普通非糯小麦不同粒径淀粉粒的数目、体积和表面积比例分布曲线存在差异。糯小麦B型淀粉粒处的峰高显著高于普通非糯小麦,A型淀粉粒处的峰值低于后者。糯小麦体积和表面积分布中B型淀粉的峰值粒径显著小于普通非糯小麦。     

小麦胚乳 A、B 型淀粉粒发育特征及黏度特性

利用扫描电镜对4个小麦品种不同发育时期籽粒自然断面和分离提纯的A、B型淀粉粒进行观察,并采用黏度糊化仪对小麦面粉和成熟小麦胚乳A、B型淀粉粒分别进行黏度测定.电镜扫描结果显示,小麦授粉3d有凝胶状物质产生于造粉体内壁,6d可明显看到部分造粉体被淀粉粒充满,此时淀粉粒雏形已现,9d淀粉粒体积增大,基本充满所有造粉体,12d前观察到的均为A型淀粉粒,15d有B型淀粉粒出现,18d以后造粉体被完全破坏降解,只留存有A、B型淀粉粒和各自表面的结合蛋白.这说明12d前是A型淀粉粒形成时期,后逐渐形成B型淀粉粒.黏度糊化测定结果表明,A、B型淀粉粒糊化后呈天蓝色,而面粉糊化后呈白色.其次,所测小麦样品糊化焓变和峰值黏度大小表现为B型淀粉粒＞A型淀粉粒＞小麦面粉.     

小麦籽粒淀粉粒粒级分布特征及其与淀粉理化特性关系研究进展

小麦籽粒淀粉粒粒级分布范围大,而且呈现梯度分布,这是小麦淀粉粒区别于其他植物淀粉粒的一个显著特征。不同粒径的淀粉粒在理化特性上有极大的差异,因此除直/支链淀粉比例之外,淀粉粒粒级分布也是影响淀粉品质的重要指标。本文综述了国内外在小麦籽粒淀粉粒粒级分布及其与淀粉品质特性关系方面的研究进展,提出应注意小麦籽粒不同粒级淀粉粒分布特征及其形成机理与调控途径等领域的研究,以促进优质小麦品种选育和调优栽培技术的建立。     

小麦淀粉粒结合蛋白多态性及蛋白表达差异的研究

为了解小麦淀粉粒结合蛋白(Starch granule proteins,SGPs)的多样性及与淀粉品质性状的关系,对来自中国、墨西哥和埃及的149份春小麦材料淀粉粒结合蛋白进行SDS-PAGE电泳分析,探讨了淀粉粒结合蛋白多态性及表达差异。结果表明,供试材料的SGP组成存在多态性,149份材料中分离了10种主要的SGPs和56种组合带谱,其中SGP-g和SGP-e是小麦中出现频率最高的淀粉粒结合蛋白(出现频率均在90%以上),而绝大多数小麦材料(92.6%)中包含了7～9条SGPs条带。材料的地理来源与SGPs蛋白谱带聚类的结果并不完全一致。     

小麦淀粉粒蛋白表达差异及其与淀粉糊化特性的关系

为探明小麦淀粉粒蛋白与淀粉糊化特性的关系,对河南省18个主栽小麦品种淀粉粒蛋白进行分离,并对淀粉粒蛋白类型、淀粉含量、糊化特性参数进行了研究.结果表明,18个小麦品种分离出了8种主要的淀粉粒蛋白,9种组合带谱,并以组合类型Ⅰ和组合类型Ⅶ所占比例最高,为55.56%,均表现为7条谱带.通过对淀粉粒蛋白条带数量以及谱带光密度总量和淀粉特性的相关性分析表明,淀粉粒蛋白光密度总量和直链淀粉含量、峰值黏度、低谷黏度、最终黏度呈显著负相关,表明淀粉粒蛋白表达量影响淀粉的糊化特性.同时选取6个小麦品种,分别采用盐、SDS、蛋白酶对淀粉进行处理后,淀粉峰值黏度显著提高,其中以盐处理提高幅度最大,且不同处理之间差异达显著水平.对处理后淀粉粒蛋白进行电泳分析表明,处理后的条带明显减少,说明处理后小麦淀粉粒蛋白含量下降.综合试验结果,不同小麦品种淀粉粒蛋白表达存在差异,同时淀粉粒蛋白表达量高低影响淀粉峰值黏度.     

小麦淀粉的粒度分布、组分及糊化特性对氮硫肥的响应

为明确氮硫肥对小麦淀粉粒度分布及主要理化特性的影响,以强筋小麦品种西农9718和中筋品种陕农138为材料,采用不同氮硫肥水平进行处理,系统分析了小麦A、B淀粉的粒度分布、淀粉组成、膨胀势及糊化特性对氮硫肥的响应。结果表明,强筋和中筋小麦品种的淀粉粒度分布基本一致。A淀粉粒体积分布占总淀粉的74.95%～75.43%,但淀粉粒数量远少于B淀粉粒。A淀粉的直链淀粉、峰值黏度、低谷黏度、崩解值、最终黏度以及回生值高于B淀粉,而B淀粉有更高的支链淀粉、膨胀势和糊化温度。不同氮硫肥处理改变了小麦淀粉的粒度分布,进而影响淀粉的组成和糊化特性。在施氮(230kg.hm-2)条件下,强筋品种西农9718和中筋品种陕农138的A、B淀粉分别在硫肥施用量为46和56kg.hm-2时有较低的直链淀粉含量、较高的膨胀势及较好的糊化特性。陕农138淀粉的粒度分布、组成及糊化特性对氮硫肥的响应比西农9718更敏感。适当的氮、硫肥配施有利于改善小麦的淀粉品质。     

Channels within soft wheat starch A- and B-type granules

The nature of channels within wheat starch granules was investigated using scanning electron and confocal laser scanning microscopy. A-type granules stained with 3-(4-carboxybenzoyl)quinoline-2-carboxaldehyde (CBQCA, protein-specific probe) revealed a network of radially oriented, channel-like protein structures similar to those previously reported. However, treatment of the same starch granules with methanolic merbromin (fluorescent dye) solution, which is used to highlight external granule surfaces (including those of channels) under non-swelling conditions, revealed few, if any, channels extending into the granule interior. This discrepancy suggested that channels within wheat starch granules were filled at least in part with protein. Removal of protein with protease facilitated greater access of methanolic merbromin to channels and/or cavities for both granule types. For A-type starch granules, relatively large channels were observed in the equatorial groove region, while finer channels originated from other regions of the granule. This work reports the first visualization of B-type granule channels, which most frequently occurred as less-defined voids (as opposed to the fine, discrete channels of A-type granules) extending to granule surfaces. Channels of A- and B-type starch granules appeared to facilitate transfer of chemical reagent into the granule matrix, though this effect was aided by granule swelling (hydration) and/or removal of channel-associated protein.     

Influence of environmental temperature during grain filling period on granule size distribution of rice starch and its relation to gelatinization properties

High temperature (HT) is the major environmental factor affecting grain starch properties of cooking rice cultivars. However, little information has been available on the effect of environmental temperature on the starch granule size distribution of rice grains. In this paper, five indica rice genotypes, including the wild type (9311) and its four mutants differing in amylose content (AC), were used to investigate the effect of environmental temperature on the starch granule size distribution, as well as its relation to AC and gelatinization properties of rice starch. Two temperature treatments (HT and NT) at filling stage were imposed to rice plants under the controlled temperature chambers. The result showed that HT increased the average diameter of starch granules and enhanced the proportion of large starch granules (LSG, D > 2.6 μm) by number, volume and surface area, respectively. However, influence of HT on GT and starch granule size distribution was relatively independent of their alteration in AC level for different rice genotypes. Therefore, HT-induced increase in the average diameter of starch granules and LSG percentage was strongly responsible for the higher starch gelatinization temperature and inferior cooked palatability of HT-ripening rice grains, which be not inherently associated with their varying AC level.     

Hydrolysis of granular corn starch with controlled pore size

Native corn (Zea mays L.) starch granules were hydrolyzed using glucoamylase at 50 °C for 1–8 h. The degree of hydrolysis over time was analyzed by the concentration of glucose released into solution. The pore sizes of hydrolyzed starch granules increased gradually with the degree of hydrolysis, as evidenced by scanning electron micrographs. It was deduced that every pore on the surface of granules was formed by hydrolysis of one enzyme molecule, so the size of pores distributed on the surface of starch granules was almost homogeneous for the same hydrolysis time. The specific surface area (S_BET), porosity, adsorptive capacity and mean pore radius of porous starch granules were determined to analyze the effect of digestion time on granule properties.     

高直链淀粉小麦西农836淀粉理化性质及消化特性分析

淀粉是人体摄入碳水化合物的主要来源,经消化后以葡萄糖的形式被人体吸收,其消化特性与人体健康密切相关。为探究小麦品种西农836淀粉理化性质与消化特性的关系,以商品粉金沙河、香雪、金龙鱼作为对照,测定其与西农836中直链淀粉含量、淀粉粒度和结晶度、糊化特性以及体外消化率的差异,并在小鼠体内模拟消化。结果表明,与金龙鱼、金沙河、香雪相比,西农836淀粉中直链淀粉含量和B型淀粉粒含量最高,与前3者差异显著（P<0.05）;西农836淀粉的峰值温度最高,峰值黏度和最终黏度最低;淀粉体外消化试验结果显示,西农836的快消化淀粉（RDS）、总消化淀粉（RDS+SDS）含量最低,抗性淀粉（RS）含量最高。小鼠体内消化试验结果表明,与金沙河相比,灌食西农836淀粉的小鼠血糖峰值较低,说明高直链淀粉小麦西农836有利于延缓消化和控制血糖升高。相关性分析结果显示,直链淀粉含量与抗性淀粉含量呈显著正相关（P<0.05）。     

Starch granule morphology in oat endosperm

Mature and developing oat (Avena sativa) grains were sectioned and image analysis methods used to estimate the starch granule-size distribution and morphology in endosperm cells. This showed that oat endosperm cells contain two types of starch granule: compound starch granules such as those seen in rice endosperm and in most other grasses; and simple granules similar to the B-type starch granules seen in the endosperm of Triticeae species such as wheat (Triticum aestivum). The simple granules in oats are similar in size and relative abundance to B-type granules in Triticeae suggesting that they may share a common evolutionary origin. However, there is a fundamental difference between oats and Triticeae in the timing of granule initiation during grain development. In Triticeae, the B-type granules initiate several days after the A-type granules whereas in oats, both the simple and compound granule types initiate at the same time, in early grain development.     

西藏冬青稞胚乳淀粉粒发育特征研究

为了解青稞胚乳淀粉及淀粉粒形成与生长特征,以西藏育成青稞品种冬青8号为材料,对籽粒发育过程中胚乳灌浆速度、淀粉积累量及淀粉粒的产生、二维面积、粒径变化进行了分析。结果表明,胚乳发育过程中,干物质和淀粉积累量呈S型曲线,两者呈正相关。花后5d,青稞胚乳中可零星见到粒径和二维面积较小的A型小淀粉粒;花后10~15d,A型小淀粉粒数量显著增大,但粒径和二维面积增加较慢;花后20d,B型小淀粉粒可能已经形成,与A型小淀粉粒组成一个复合群体;花后25d,形成了粒径10μm的A型淀粉粒、粒径5~10μm的B型淀粉粒和粒径5μm的C型淀粉粒;花后30~55d,不同粒径的淀粉粒数量和二维面积逐渐增加,其中,C型淀粉粒数量不断增加直至籽粒成熟。青稞成熟籽粒胚乳中,淀粉粒的粒径为1.45~44.76μm,二维面积为3.56~1 750.8μm2;A型淀粉粒形状为近圆形,表面光滑,个别表面附着有C型淀粉粒,B型淀粉粒形状有近圆形、椭圆形多种类型,C型淀粉粒形状比B型淀粉粒的变异更大。     

Starch isolation method impacts soft wheat (Triticum aestivum L. cv. Claire) starch puroindoline and lipid levels as well as its functional properties

Wheat (Triticum aestivum L.) kernel hardness is a major quality characteristic, which has been ascribed to the presence of puroindolines a and b. These proteins occur in higher levels at the surface of water-washed starch granules from soft wheat cultivars than at that of starch from hard wheat cultivars. In the present study, prime starch was isolated from flour from soft wheat (cultivar Claire) using a dough ball or batter based separation method. Starch isolated with the dough ball method contained lower levels of puroindolines, as well as of other starch granule associated proteins and lipids than that isolated with the batter method. Similar patterns of puroindoline and lipid levels after starch isolation can presumably be related to (polar) lipid binding by puroindolines. Both isolated starch fractions showed comparable differential scanning calorimetry thermograms, whereas higher levels of starch surface associated components restricted starch swelling. Necessary controls demonstrated that the observed differences did not arise from artefacts associated with hydration, fractionation or freeze-drying in the experimental protocols. Apparently, proteins and lipids at the starch granule surface impact water absorption and, as such, starch swelling, but they do not affect starch granule internal phenomena such as melting of the crystalline amylopectin chains.     

藁城8901与豫麦50籽粒淀粉粒的粒度分布特征

为研究小麦籽粒发育过程中胚乳淀粉粒形成与粒度分布特征,以小麦品种藁城8901与豫麦50为材料,研究了籽粒胚乳淀粉粒形成、生长与分布特征。结果表明,花后4d,小麦胚乳出现不同范围大小的淀粉粒,最大粒径8μm。花后7d,籽粒中淀粉粒增多增大,最大粒径20μm左右。花后10~14d,淀粉粒体积继续增大,并产生了一个新的小淀粉粒群体。花后17d,淀粉粒以体积增大为主。花后21d,淀粉粒最大粒径较成熟期变化较小。花后24d,小于0.6μm的淀粉粒数目急剧增加,大于0.6μm的淀粉粒数目占比则明显减少,表明这一时期又产生了一个新淀粉粒群体(后期形成的B型淀粉粒)。花后24~28d,小于0.6μm的淀粉粒数目仍不断增加,而直径较大的淀粉粒数目增加减少,表明籽粒中新淀粉粒的产生仍在继续,但以小粒径淀粉粒为主。花后28d至成熟期,最小粒径淀粉粒进一步生长,其他淀粉粒粒径变化相对较小。     

氮素对弱筋小麦宁麦9号淀粉形成的调节效应

为了给弱筋小麦保优栽培提供理论依据,选用长江中下游主推的优质弱筋小麦品种宁麦9号作为试验材料,研究氮素对弱筋小麦籽粒淀粉形成的调节效应。结果表明,氮肥运筹比例相同,减少施氮量,支链淀粉、总淀粉含量增加;施氮量相同,氮肥前移,支链淀粉、总淀粉含量增加;氮肥后移对直链淀粉含量的影响随施氮量的不同而不同。低施氮量下,随氮肥后移,直链淀粉含量呈上升趋势;高施氮量下则相反。因此弱筋小麦栽培应降低总氮肥施用量,控制中后期施氮量并使氮肥前移。强、弱势粒淀粉含量差异显著,强势粒淀粉积累量高于弱势粒。不同处理间淀粉粒形态结构存在差异。     

Interaction of granular maize starch with lysophosphatidylcholine evaluated by calorimetry, mechanical and microscopy analysis

In this study we evaluated the thermo-mechanical properties of maize starch pastes (80% wt/wt) under the effect of exogenous lysophosphatidylcholine (LPC) using differential scanning calorimetry (DSC), dynamic mechanical spectrometry (DMS), and scanning electron microscopy (SEM). Particular attention was paid to the development of the amylose-LPC inclusion complex. Results from SEM and DSC showed that with no exogenous LPC, granular maize starch developed the amylose network structure for starch gelling at 80–95 °C. In comparison, at 1.86 and 3.35% of LPC, heating up to 130 °C was needed to develop the three-dimensional network required for starch gelling. Results showed that at these LPC concentrations LPC interacted mainly with amylose within the starch granule. At concentrations ≥8.26% the LPC interacted with amylose both inside the granule and on the granule's surface. At such LPC concentrations heating to 130 °C did not fully develop the starch network structure for gelling. These results suggested that a higher thermal stability was achieved by starch granules because of LPC inclusion complex formation. DSC or DMS did not detect the development of this complex, probably because its formation took place below the onset of gelatinization under conditions of limited molecular mobility. Subsequently, a lower level of organization (i.e. complex in form I) was achieved than in the complex developed at high temperature and water excess (i.e. complex in form II). On the other hand, the changes in the starch granule structure observed by SEM as a function of the time–temperature variable were well described by the phase shift angle (δ) rheograms for starch pastes with and without addition of LPC.     

啤酒大麦与饲用大麦籽粒结构和淀粉粒的比较研究

为了探索大麦籽粒结构和淀粉粒与其用途的关系，利用扫描电子显微镜（SEM）和差示扫描量热分析仪（DSC）对啤酒大麦（港啤1号和扬农啤2号）与饲用大麦（扬饲麦1号）的籽粒结构和淀粉粒进行了比较研究。结果表明，3个品种表现相似的胚乳结构，但啤酒大麦的淀粉粒与蛋白质结合不紧密，饲用大麦的淀粉粒和蛋白质结合较紧密，且饲用大麦的蛋白质基质含量高于啤酒大麦。采用碱处理和差速沉淀法分离了籽粒胚乳大、中、小淀粉粒，虽然品种间淀粉粒的形态无明显差别，但大、中、小淀粉粒的大小和含量差异很大。DSC分析所得参数表明，大淀粉粒糊化时的起始温度（To）、峰值温度（Tp）和终结温度（n）高于相同品种的中、小淀粉粒，且糊化过程所需的热焓（△H）也较高。