Microstructure of hard and soft kernels of barley

Kernel hardness, an important quality trait of cereal grains, is known to influence pearling properties and malting quality of barley. To understand the endosperm micro-structural features of kernels and their relationship to kernel hardness, endosperms of three hard and three soft hulled spring barley lines based on single kernel characterization system hardness index were observed under light (LM) and scanning electron (SEM) microscopy. Under LM, endosperm cell wall of the three hard kernel lines was significantly thicker than that of the three soft kernel lines. Hard and soft lines showed differences in the degree of starch-protein association and continuity of protein matrix under the SEM. Hard kernel lines with a continuous protein matrix exhibited greater starch-protein adhesion than the soft kernel lines, suggesting that starch-protein binding may be one of the factors influencing barley kernel hardness. SEM of flour particles of soft kernel lines showed numerous well defined individual A and B-type starch granules, while, flour of hard kernel lines mostly showed small flour aggregates with few individual starch granules.     

小麦籽粒发育时期Puroindolines蛋白与硬度的关系

为探讨Puroindolines蛋白的表达特点与籽粒硬度的关系,采用改进的SDS-PAGE凝胶分析了不同硬度小麦品种的籽粒在各个发育时期Puroindolines蛋白的表达.结果表明,不同硬度的小麦籽粒中总Puroindolines(PinA和PinB)蛋白的表达量差异不大,但与胚乳淀粉颗粒结合的Puroindolines蛋白量差异非常明显:在籽粒发育的不同时期,软质小麦籽粒淀粉粒表面结合的Puroindolines蛋白量显著高于硬质小麦;基因型同为野生型但硬度有差异的品种,籽粒较软的材料其淀粉粒表面结合的Puroindolines蛋白量也明显高于较硬的材料,说明该蛋白的结合特性是决定籽粒硬度的直接原因.结果还表明,胚乳中水溶性戊聚糖与籽粒硬度关系密切.     

High Resolution Imaging of Starch Granule Surfaces by Atomic Force Microscopy

High resolution imaging of the surfaces of starch granules from two different botanical sources has been performed using two complementary techniques: low voltage scanning electron microscopy (LVSEM) and atomic force microscopy (AFM). LVSEM provided superior images ofuncoatedgranules than possible by conventional scanning electron microscopy, and these images were used to validate the features revealed at higher resolution by AFM. The AFM images demonstrated that, although intra-sample variation exists, the surfaces of wheat and potato starch granules possess substantially different topographies. Potato starch had many protrusions (50–300 nm in diameter), above a flatter surface, which contained structures in the order of 10–50 nm. Wheat starch had far fewer protrusions and generally had a smoother surface made up of 10–50 nm structures. The 10 to 300 nm structures are believed to be carbohydrate in nature and correspond to ‘blocklet’ structures, comprising groups of amylopectin side-chain clusters presenting at the granule surface. We conclude therefore that near-molecular resolution topography of the starch granule surface has been revealed, which has allowed further insight into starch granule structure and molecular organisation.     

Study of kernel structure of high-amylose and wild-type rice by X-ray microtomography and SEM

Rice kernels of a transgenic high-amylose rice line and its wild type were examined by a high-resolution X-ray microtomography (XMT) and an environmental scanning electron microscope. Two-dimensional (2-D) cross-sectional images and 3-D objects from XMT were reconstructed and analyzed to elucidate their structural features. The lack of two isoforms of starch branching enzyme, termed SBEI and SBEIIb in high-amylose rice (HAR), resulted in a distinctly different grain inner density than wild-type rice (WTR). HAR had smaller, elongated starch granules with air spaces inside the kernels resulting in an opaque grain, whereas WTR had a tight endosperm with little air space and polygonal starch granules. XMT allowed a full 3-D characterization of the rice kernel structure and revealed that air space distribution was not uniform in the HAR kernel.     

Scanning Electron Microscopic Study of Dough and Chapati from Gluten-reconstituted Good and Poor Quality Flour

Hard and soft wheat flours, which were used in the study, resulted in good and poor quality chapatis respectively. Gluten was isolated and interchanged among the two whole wheat flours and studied by scanning electron microscopy for its influence on structural characteristics of dough and its relation to chapati-making quality. Microscopic observations clearly indicated that larger gluten strands covered starch granules in hard wheat flour dough, while gluten was short and starch granules exposed in dough prepared from soft wheat flour. Greater film forming ability of gluten in hard wheat flour dough manifested in long and bulky starch strands interwoven with protein matrix in its chapati crumb. Higher moisture retention and starch gelatinization as a consequence of greater film forming ability of gluten in hard wheat flour resulted in pliable and soft textured chapati.     

Hardness Distribution and Endosperm Structure on Polishing Characteristics of Brewer’s Rice Kernels

Abstract

This study was designed to determine the effects of the hardness distribution and the endosperm structure on the polishing characteristics of brewer’s rice kernels. We used four brewer’s rice cultivars, Kairyo-omachi, Hattan-nishiki No. 1, Senbon-nishiki and Yamada-nishiki. The broken kernel ratios in Kairyo-omachi and Hattan-nishiki No. 1 were significantly higher than those in Senbon-nishiki and Yamada-nishiki. Vickers hardness (VH) values in white-core tissues in kernels differed among varieties, which were significantly lower in Kairyo-omachi and Hattan-nishiki No. 1. However, no varietal differences were observed in VH values in the peripheral translucent tissues surrounding the white-core tissues. The tissues along the dorsoventral axis were softer than those along the longitudinal axis of the kernels. The tissues on the ventral side were softer than those on the dorsal side. Scanning electron microscopy (SEM) observations revealed the presence of closely arranged compound starch granules and few varietal differences in the peripheral translucent tissues surrounding the white-core tissues. However, as compared with Yamada-nishiki and Senbon-nishiki, in Hattan-nishiki No. 1 and Kairyo-omachi, the starch granules were loosely packed and the airspaces between the starch granules were more numerous in the white-core tissues. A higher number of airspaces and less starch were present in the endosperm cells along the dorsoventral axis when compared with along the longitudinal axis and on the ventral side than on the dorsal side. The present study showed that polishing characteristics are closely related with the endosperm structure, which is characterized as the density of starch granules.     

Endosperm Structure of White-Belly and White-Core Rice Grains Shown by Scanning Electron Microscopy

Abstract:

White-belly and white-core are the major two types of grain chalkiness in japonica rice. This study aims to compare the morphological features of white-belly and white-core using a scanning electron microcope (SEM). A japonica rice cultivar Wuyujing3 and its mutants were used as materials. Nearly 1000 SEM images were observed, and 12 representative photos were selected. SEM images showed contrasting differences between white-belly and white-core in endosperm microstructure including the shape of endosperm cell, the size distribution of starch granules, and the amount of protein bodies. White-belly and white-core also varied markedly in morphological features of the cracked compound starch granules. Our findings should help to advance our understanding of the multi-faceted nature of grain chalkiness from the perspective of starch and protein accumulation, and should be of value for future work on rice grain chalkiness.     

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.     

Investigation of the fracture mode for hard and soft wheat endosperm using the loading–unloading bending test

Investigation of the fracture mode for hard and soft wheat endosperm was aimed at gaining a better understanding of the fragmentation process. Fracture mechanical characterization was based on the three-point bending test which enables stable crack propagation to take place in small rectangular pieces of wheat endosperm. The crack length can be measured in situ by using an optical microscope with light illumination from the side of the specimen or from the back of the specimen. Two new techniques were developed and used to estimate the fracture toughness of wheat endosperm, a geometric approach and a compliance method. The geometric approach gave average fracture toughness values of 53.10 and 27.0 J m⁻² for hard and soft endosperm, respectively. Fracture toughness estimated using the compliance method gave values of 49.9 and 29.7 J m⁻² for hard and soft endosperm, respectively. Compressive properties of the endosperm in three mutually perpendicular axes revealed that the hard and soft endosperms are isotropic composites. Scanning electron microscopy (SEM) observation of the fracture surfaces and the energy–time curves of loading–unloading cycles revealed that there was a plastic flow during crack propagation for both the hard and soft endosperms, and confirmed that the fracture mode is significantly related to the adhesion level between starch granules and the protein matrix.     

Effects of drought on the morphological and physicochemical characteristics of starch granules in different elite wheat varieties

In this study, scanning electron microscopy (SEM) revealed the formation of pits and pores on the surfaces of starch granules in response to drought stress, with substantially more pronounced effects in the ordinary yield potential wheat cv. Xindong 23 than the excellent yield potential wheat cv. Xindong 20. Drought induced a significant reduction in starch granule sizes in both wheat varieties, though the reduction observed in Xindong 23 was six times more pronounced than that observed for Xindong 20. Amyloglucosidase and α-amylase treatment of starch from wheat grown in drought conditions released significantly more reducing sugars compared with samples from irrigated controls. SEM and confocal laser scanning microscopy (CLSM) revealed that starch granules from the two wheat varieties grown under drought conditions had substantially increased fluorescence after treatment with proteolytic enzymes and staining with methanolic merbromin and 3-(4-carboxybenzoyl) quinoline-2-carboxaldehyde dyes. Analysis of pasting properties showed significant increases of peak viscosity, trough viscosity, break down, and setback following drought stresses. Furthermore, drought induced a significant reduction in the water binding capacity and increased damage to starch only in Xindong 23. These results provide insight into the potential mechanisms through which drought influences the ultrastructures and physicochemical properties of starch in wheat.     

Volume, texture, and molecular mechanism behind the collapse of bread made with different levels of hard waxy wheat flours

Physico-chemical properties of bread baked by partially replacing normal wheat (Triticum aestivum L.) flour (15, 30, and 45%) with two hard waxy wheat flours were investigated. Substitution with waxy wheat flour resulted in higher loaf volume and softer loaves. However, substitution at >30% resulted in excessive post-bake shrinkage and a ‘key-hole’ shape with an open crumb structure. Bread crumb microstructure indicated a loss of starch granule rigidity and fusing of starch granules. The cells in the interior of the bread did not become gas-continuous and as a result, shrunk as the loaf cooled. Soluble starch content was significantly higher in bread crumb containing waxy wheat flour than in control bread. Debranching studies indicated that the soluble starch in bread made with 30-45% hard waxy wheat flour was mostly amylopectin. Incorporation of waxy wheat flour resulted in softer bread immediately after baking but did not retard staling upon storage.     

Difference in the Physical Properties of White-Core and Non-White-Core Kernels of the Rice Varieties for Sake Brewing is Unrelated to Starch Properties

Abstract

This study was designed to determine whether or not the difference in the physical properties between white-core and non-white-core kernels of the rice varieties for sake brewing is associated with their starch properties. We used two rice cultivars for sake brewing, Senbon-nishiki and Yamada-nishiki, from three different plots in Hiroshima prefecture. Hardness values of kernels were significantly higher in non-white-core than in white-core kernels in both varieties. Vickers hardness (VH) values were lowest at the center of the kernel in both types of kernels. VH values of white-core tissues of white-core kernels were significantly lower than those of corresponding tissues of non-white-core kernels. No significant differences were observed between the two types of kernels in VH values of the surrounding translucent tissues and in the starch properties (amylose content, pasting properties analyzed using a rapid viscoanalyzer and gelatinization properties analyzed using a differential scanning calorimetry). These results suggest that the difference in physical properties between the two types of kernels of the rice varieties for sake brewing are associated with the difference in structure of endosperm cells and not in starch properties.     

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。     

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。     

Transformation of Pinb-D1x to soft wheat produces hard wheat kernel texture

Kernel hardness - a key quality trait of common wheat (Triticum aestivum L.) - is mainly conditioned by the Pina and Pinb genes. Mutation or deletion of Pina or Pinb increases kernel hardness, resulting in a hard wheat kernel texture. Here, Pinb-D1x gene was cloned from a hard wheat landrace Kashibaipi and transformed into a soft wheat cultivar Yangmai19 to assess its effect on kernel hardness and flour properties. PCR, RT-PCR and Western blot data confirmed the successful transformation and overexpression of Pinb-D1x gene in transgenic offsprings. The data of single kernel characterization system and scanning electron microscopy revealed that the introduction of Pinb-D1x in soft wheat increased the kernel hardness significantly and changed the internal structure of the kernel. Similarly, transgenic lines exhibited hard wheat like flour properties; flour whiteness and pasting temperature were significantly reduced in the transgenic lines, while the total protein content, damaged starch content, and compound parameter in the Mixograph tests (PT × TW value) showed a significant increase over the wildtype. The results showed that the transformation of the Pinb variants is a powerful strategy to alter the kernel hardness and flour properties in wheat breeding.     

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。     

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.     

小麦胚乳A、B型淀粉粒分离纯化方法研究

为了能够准确研究小麦籽粒胚乳中A、B型淀粉粒的形成机理、化学组分及理化特性,对前人分离提纯A、B型淀粉粒的方法进行改进,提出了适于小麦A、B型淀粉粒分离提纯的常量法和微量法。用4个不同淀粉含量类型的小麦品种籽粒对此方法进行验证,结果表明4个品种A、B型淀粉粒在常量法中的分离纯度分别达到79.46%~85.23%和90.27%~96.43%,在微量法中的分离纯度分别达到92.68%~96.70%和96.48%~99.00%。在常量法和微量法中,A型淀粉粒的分离纯度与品种淀粉含量成正相关, B型淀粉粒分离纯度与品种淀粉含量成负相关;而A、B型淀粉粒在两种方法中的分离产率与品种淀粉含量没有显著关系。     

On Predicting Roller Milling Performance V: Effect of Moisture Content on the Particle Size Distribution from First Break Milling of Wheat

Extended breakage functions incorporating both single kernel size and moisture content were determined for First Break milling of hard and soft wheats under a sharp-to-sharp disposition. A moisture correction function was constructed by milling narrow size fractions of wheat tempered to different moisture contents, and subtracting the breakage function at a base moisture content of 16% from that at other moisture contents. The effect of adding moisture was to change an initially inverted U-shaped distribution at low moisture contents to a linear distribution at 16% moisture, then to a U-shaped distribution at higher moisture contents, reminiscent of the particle size distributions produced by dull-to-dull milling. The extended breakage functions were used to predict milling of unseparated feed samples at different roll gaps and moisture contents. In addition, mixtures of hard wheat at 14% moisture and soft wheat at 20% moisture, mixed in different proportions, were milled and the resulting particle size distributions compared with predictions. Excellent predictions were obtained in all cases. This confirms the independent breakage of kernels during First Break milling, and demonstrates the potential of the breakage function approach for interpreting single kernel data in terms of predictions of milling performance.