Effects of drought and the presence of the 1BL/1RS translocation on grain vitreosity, hardness and protein content in winter wheat

Grain texture is an important component of end-use quality in wheat. The effects of water availability on the components of texture; vitreosity, determined using a Light Transflectance meter (LTm), grain hardness measured using the single-kernel characterisation system (SKCS), and protein content, were studied in field experiments of winter wheat in the UK in 2001/2002 and 2002/2003. Experiments were grown on a drought prone soil and employed a mapping population of 46 doubled haploid (DH) lines and their parents, Beaver (+1BL/1RS, soft wheat) and Soissons (1B, hard wheat). The results showed that drought increased hardness in both seasons, but the effect was never sufficient to move a line from the soft class into the hard class. Puroindoline (PIN)-a:b peak height ratio explained ca. 78% of the variation in hardness, and drought also appeared to increase the amounts of PINs in the grain. Minor quantitative trait loci (QTLs) were found for hardness on chromosomes 2A, 2D, 3A and 6D, also associated with QTLs for PINs. Vitreosity also increased in response to drought in both seasons. Variation in vitreosity explained 7–11% of the overall variation in texture within a hardness class, with hardness increasing on average by 2.2 SKCS units for each 10% increase in the proportion of vitreous grains. The relationship between vitreosity and protein content was poor, despite the fact that protein content also increased in response to drought. Minor QTLs associated with both protein content and vitreosity were found on chromosomes 1B, 4D and 5D. A minor QTL for vitreosity was also found on chromosome 2D. However, there appeared to be no direct relationship between alleles at the Ha locus, the gene which controls the difference between hard and soft wheats, and vitreosity. A positive relationship between the presence of the 1BL/1RS translocation and the proportion of vitreous grains was identified, suggesting that vitreosity was strongly linked to changes in protein quality.     

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.     

Disease and pest resistance in grains of sorghum and millets

In this review available information on the mechanisms of resistance to insect pests and fungal pathogens in sorghum and millets is discussed. The primary source of resistance lies in the chemical and physical make up of the grain. Phenolic compounds such as ferulic acid and tannins present in some sorghums are potent inhibitors of pests and pathogens. Grain hardness is a major deterrent to infection and infestation in low tannin grains. The prolamins, the grain storage proteins of sorghum, are organized into protein bodies and provide a physical and a nutritional barrier since they are resistant to digestion by insect and fungal proteases. A plethora of proteins that belong to the ‘pathogenesis related protein’ group are distributed in various parts of the grain. Some of them are located in protein bodies. Notwithstanding, sorghum is still susceptible to insect pests and fungal pathogens. An understanding of the natural mechanisms of resistance in the grain is paramount for the development of durable resistance against pests and pathogens. The pyramiding of resistance genes and the development of transgenic lines based on this understanding are two sources of hope for the future protection of sorghum and millets.     

Genetic and Environmental Effects on Puroindoline-a and Puroindoline-b Content and their Relationship to Technological Properties in French Bread Wheats

Grain hardness (GHa) in hexaploid wheat (Triticum aestivum L.) is a major factor of end use quality. The variation of texture has been related to the Hardness locus, closely linked with the puroindoline-a (PIN-a) and puroindoline-b (PIN-b) genes. In order to study the role of puroindoline content in texture variation, the quantity of puroindolines was determined. Puroindoline-a (PIN-a) and puroindoline-b (PIN-b) content was determined and a total of 11 bread making parameters were obtained from 40 bread wheat cultivars grown in four experimental locations. The 11 parameters were significantly influenced by the genotype whereas location did not significantly affect PIN-a or PIN-b content and loaf volume. PIN-b and grain hardness displayed the highest heritability coefficients (both 0.88). PIN-a and PIN-b content were not correlated with grain protein content (Prot) and grain hardness in hard and soft wheat types. In soft samples PIN-(a+b) content was negatively correlated, with loaf volume in two locations. Multiple regression analyses, carried separately for soft and hard types, revealed that PIN-b content explained variation of dough strength (W) and loaf volume, however their influence was mostly significant in soft types. For each location, from 22 to 91% of the phenotypic variation of strength and loaf volume was explained by combining up to three flour traits. Protein content, PIN-b and the average score of high molecular weight glutenin subunits (HMW-GS) were frequently introduced by multiple regression (without GHa) as explanatory variables of strength and loaf volume. These results strengthened the significant role of PIN-b in breadmaking (loaf volume), and indicated that biochemical factors other than puroindolines are involved in the grain hardness variation.     

青海小麦籽粒硬度等位变异研究

为了解小麦品种籽粒硬度的遗传多样性,利用单粒谷物硬度测定、PCR扩增和核苷酸测序技术,分析了66份青海小麦品种籽粒硬度主效基因的等位变异。结果表明,青海小麦以硬质类型为主,比例达到47.0%,混合麦比例为19.7%,软质麦比例为33.3%。硬度基因有5种组合类型:野生型、Pina-D1a/PinbD1b、Pina-D1a/Pinb-D1c、Pina-D1a/Pinb-D1x和Pina-D1b/Pinb-D1a。野生型小麦类型比例最高,占59.09%,SKCS硬度指数平均为44.12,变化范围为12.75~84.89。突变类型的品种籽粒均为硬质。因此,在青海硬质小麦可以通过突变类型的分子标记进行选育,软质小麦选育需在利用硬度基因分子标记筛选的基础上进一步考察籽粒硬度性状的表现型。     

Mechanical properties of outer layers from near-isogenic lines of common wheat differing in hardness

The mechanical properties of the combined outer layers from near-isogenic wheats differing by hardness were determined. Results from traction tests showed significant differences between the isogenic lines, outer layers from grains of the soft type showing higher extensibility. Determination of the mechanical properties of the corresponding component tissues revealed significant differences between the isolated tissues from soft or hard wheat grains. It also allowed analysis of their respective contribution to the properties of the combined peripheral tissues using a simulation of their rupture as unseparated tissues. According to the results, if the component layers displayed similar maximum lineic force to rupture, the rupture of combined outer layers occurs when the least extensible individual tissue breaks. The major cell wall biochemical components of the combined outer layers and of their component tissues were analysed. The phenolic acid composition of soft wheat pericarp contained more ferulic acid in either monomeric or polymeric forms than the pericarp from hard wheat. Arabinoxylans in walls of the soft wheat pericarp appeared 1.6 times more cross-linked by ferulic acid dehydrodimers than walls of hard wheat. These differences in arabinoxylan cross-linking may explain the observed differences in pericarp mechanical properties.     

Effects of ozone treatment on the molecular properties of wheat grain proteins

Ozone is a powerful and highly reactive oxidizing agent, which has found increasing applications in the field of grain processing. However, in some cases, O₃ can potentially promote oxidation and/or degradation of the chemical constituents of grains. Experiments were carried out to evaluate the specific effects of gaseous ozone on the molecular properties of wheat grain proteins and their consequences on the bread-making quality of the resulting flours.Ozonation causes a significant reduction in the SDS solubility of the wheat prolamins, which can reasonably be attributed to conjugate effects of an increase in molecular dimensions and an increase in the compactness of the protein polymers initially present. In fact, our results demonstrate that this general reinforcement of the aggregative status of prolamins due to ozonation of wheat grains results from (i) the formation of new intermolecular S-S bonds, (ii) to a lesser extent, the formation of other types of intermolecular covalent cross-links (dityrosine cross-links) and finally, (iii) significant changes in secondary structure. By significantly affecting the molecular properties of wheat grain prolamins, ozone leads to profound changes in the rheological properties (i.e. increase in the tenacity and a great limitation of the extensibility) of the flours and/or doughs obtained.     

Characterisation of the Major Prolamins of Tef (Eragrostis tef) and Finger Millet (Eleusine coracana)

The major prolamins of tef (Eragrostis tef(Zucc.) Trotter) and finger millet (Eleusine coracanaL. Gaertn subsp.coracana) were purified and characterised by SDS–PAGE, amino acid analysis and N-terminal amino acid sequencing. These studies indicate that the major prolamins of tef and finger millet are similar to the α-prolamins of the Panicoideae (maize, sorghum andCoix), although they are classified in a separate sub-family of the Poaceae, the Chloridoideae.     

Effect of Genotype,Environment and Their Interaction on Quality Parameters of Wheat Breeding Lines of Diverse Grain Hardness

Abstract

Understanding the contribution of genotype, environment and genotype-by-environment interaction to wheat grain quality facilitates the selection for quality in breeding programs. Stability of grain quality characteristics is an important requirement in the baking industry. We assessed 24 winter wheat genotypes with different grain hardness in multienvironment trials at four locations and two levels of fertilization in each location. Grain samples were analyzed for hardness, protein and starch content, and wet gluten content, Zeleny sedimentation value, alveograph parameter (W) and hectoliter weight. All parameters were evaluated on whole grains using the near infrared transmittance technique. Differences between hard and soft genotypes appeared to be significant, apart from grain hardness, for protein content, Zeleny test and alveograph parameter. Genotype was found to have a major influence only on grain hardness; for protein content, wet gluten and Zeleny sedimentation value environment prevailed the influence of genotype, and for starch content, alveograph W parameter and hectoliter weight both sources of variation had similar importance. Genotype-by-environment interaction was of smaller size relative to genotype and environment in terms of all the studied quality parameters. Stable genotypes predominate the breeding lines studied. Response of unstable genotypes to environmental conditions was nonlinear in most cases.     

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.     

Genetic and environmental factors affecting grain texture in common wheat

Thirteen wheat cultivars grown in six locations were compared for kernel weight, protein content and grain texture, as determined by the Single Kernel Characterization System (SKCS). Moreover, puroindolines a (Pin-A) and b (Pin-B) bound to starch were quantified by densitometric scanning of A-PAGE fractionations. All cultivars shared allele Pina-D1a coding for wild-type Pin-A, and differed from each other in allele composition at Pinb-D1 coding for Pin-B. Cultivars with Pinb-D1a exhibited soft grain and high amounts of Pin-A and Pin-B compared to cultivars with Pinb-D1b or Pinb-D1d. Significant genetic variation for grain hardness and Pin-A level was detected in soft cultivars. The ratio between Pin-A and Pin-B levels in soft cultivars was approximately 6:5, whereas it varied between 9:5 and 10:1 in hard cultivars. Protein content was significantly correlated with Pin-B content (r=0.34) and SKCS value (r=0.36) in soft wheats. Significant correlations (0.68 and 0.73 for soft and hard wheats, respectively) were observed between Pin-A and Pin-B levels. Grain hardness was not correlated with puroindoline levels and Pin-A/Pin-B ratio in both textural classes. By contrast, kernel weight was found to act as a major environmental factor affecting grain texture in both soft and hard wheats.     

Identification of novel secaloindoline-a and secaloindoline-b alleles in CIMMYT hexaploid triticale lines

To characterize kernel hardness, an important trait in triticale breeding, and to identify secaloindoline alleles present in hexaploid triticale lines developed at International Maize and Wheat Improvement Center (CIMMYT) a total of 171 secondary hexaploid lines were analyzed for grain hardness using the Single Kernel Characterization System. They showed a large spectrum of kernel hardness types, from very soft to very hard, with values ranging from 8.6 to 84.9. The occurrence of starch granule-associated friabilin was studied in 30 lines, including 10 hard, 9 mixed and 11 soft genotypes. All soft lines displayed a high level of friabilin, whereas the hard lines showed almost no friabilin, indicating that friabilin is directly involved in the formation of grain texture in secondary hexaploid triticales. Two novel secaloindoline alleles were identified and designated as Sina-R1b and Sinb-R1c. Compared with SINAa, the deduced amino acid sequence of SINAb showed a Trp to Arg substitution at position 44. SINBc had a Gly to Ser substitution at position 78 and a Gly to Arg substitution at position 115, as well as a Cys insertion in the signal peptide, in comparison to SINBa. The novel alleles Sina-R1b and Sinb-R1c were detected in both the soft and hard triticale lines.     

Chemical imaging: the distribution of ions and molecules in developing and mature wheat grain

The ability to combine topographical and in situ chemical analysis of individual cereal grains, without recourse to fractionation, offers an opportunity to determine the distribution of functionally- and nutritionally-important components. Three such technologies are reviewed, including immunolocation using monoclonal antibodies specific for different types of wheat prolamins, secondary ion mass spectroscopy (SIMS) to detect the presence of inorganic elements such as sodium and sulphur, and infrared (including Raman and Fourier-transform infrared [FT-IR]) microspectroscopy to determine the distribution of biopolymers across the grain. Immunolabelling has shown that the distribution of prolamin proteins changes across the endosperm, with the outer endosperm containing a much greater proportion of prolamins than the inner endosperm. SIMS has shown, for the first time, the presence of Na⁺ in the phytin granules and that sulphur is enriched at the boundary between the starch granules and the protein matrix. Raman microspectroscopy has been used to investigate the distribution of proteins and the phenolic compound, ferulic acid, across the grain, whilst FT-IR has been used to define the microheterogeneity of arabinoxylans in endosperm cell walls. These methods highlight how in situ analysis can yield new insights into grain composition and how this may be altered by environmental conditions during grain development.     

Changes in the thermal and structural properties of maize starch during nixtamalization and tortilla-making processes as affected by grain hardness

The objective of this work was to evaluate the changes in the thermal and structural properties of maize starch during nixtamalization and the tortilla-making process and their relationship with grain hardness. Three maize types with varying hardness (hard, intermediate, soft) were processed by three nixtamalization processes (classic, traditional and ecological). Starch from the three maize types showed an A-type pattern and two endotherms corresponding to gelatinization and melting of the Type I amylose-lipid complexes. After cooking and steeping, in intermediate and soft grains the partial gelatinization and the annealing affected the starch properties and promoted the formation of amylose-lipid complexes. These effects were not observe in hard grains. The increase in melting enthalpy and the intensity of the peak 2θ∼20° from nixtamal to tortillas demonstrated the formation of amylose-lipid complexes. A third endotherm above 114 °C in some treatments of nixtamal and tortilla starch demonstrated the transformation of some amylose-lipid complexes in a most ordered structures (Type II complexes). The V-type polymorph structure found in native starch, nixtamal, and tortilla corresponds to a coexistence of Type I and Type II complexes. Formation of amylose-lipid complexes in tortillas had a partial effect on decreasing starch retrogradation (r = −0.47, P < 0.05).     

山西小麦品种籽粒硬度与主要品质性状研究

为明确山西省近十年育成小麦新品种的籽粒硬度和品质状况,利用单籽粒谷物特性测定仪(SKCS)、DA7200多功能近红外分析仪,对来自山西省近十年审定的56个小麦新品种的籽粒硬度、蛋白质含量、出粉率等指标进行了检测和分析。结果发现,山西省近十年审定的小麦品种中,硬质麦比例较高,为78%,混合型麦和软质麦比例较低,分别为12%和8%;硬度指数范围较宽,为16.33～78.93。经相关分析,小麦籽粒硬度指数与被测品质性状均呈正相关关系,其中,与出粉率、吸水率、最大拉伸阻力均呈极显著正相关(P0.01)。硬质麦的蛋白质含量、湿面筋含量、出粉率、沉降值等品质参数均显著高于混合型麦和软质麦。混合型麦的蛋白质含量、湿面筋含量、出粉率、沉降值等品质参数略高于软质麦。     

The co-operative interaction of puroindolines in wheat grain texture may involve the hydrophobic domain

The puroindoline genes are causatively associated with wheat grain hardness, a commercially significant property. The proteins puroindoline (PIN) A and B are both required in their wild-type (WT) to impart soft grain texture, and absence of/mutations in either/both PIN(s) results in hard wheat. However, there is no biochemical clarity yet that explains this interdependence. This work critically analyses the roles of the tryptophan-rich domain (TRD), the little-known hydrophobic domain (HD), and certain other residues, in the physical associations of PINs. Site-directed mutagenesis-PCR was used to delete the TRD or HD and introduce an Arg39Gly substitution in PINA. The PINB-D1c mutant (Leu60Pro) was also investigated. The yeast two-hybrid system was used to assess the protein–protein interactions (PPI) of proteins. The TRD deletion or Arg39Gly substitution in PINA did not adversely affect its PPI, while deletion of HD resulted in a significant reduction. No effect on PPI was observed for Leu60Pro PINB. The results of this expression system strongly suggest that the HD is essential (but not sufficient) in higher-order associations of PINs. We propose a two-event model that explains the co-operative action of the PINs and why mutations outside the TRD may alter grain texture.     

Protein bodies ontogeny and localization of prolamin components in the developing endosperm of wheat caryopses

During caryopsis development, prolamins are initially stored in individual protein bodies, then generate a protein matrix in the ripe caryopsis. The ontogeny of the protein bodies was analyzed by fluorescence and electron microscopy from 7 to 43 days after anthesis (dAA), a period of time from the cellularization of endosperm to its desiccation. A series of antibodies specific to each prolamin type (α/β-, γ-, ω-gliadins, low-molecular weight and high-molecular weight glutenin subunits) made it possible to localize and co-localize the different prolamins in organelles of endosperm cells at different developmental stages. Protein bodies containing prolamins were observed as early as 7 dAA. At the early developmental stages, protein bodies were spherical with diameters around 1–2 μm. Later, around 15 dAA, the PBs enlarged, and aggregation and/or coalescence were prominent at 21 dAA. From 33 dAA, individual PBs were no longer visible, but a protein matrix was confined in the space between starch granules. All prolamins were found in the same protein bodies, without any segregation according to their types. Immunochemical labelling of prolamins failed to reveal in TEM analyses any particular internal organization in protein bodies. Glutenin subunits and gliadins were observed in the Golgi apparatus at the early stages of endosperm development.     

Association of Puroindoline b-B2 variants with grain traits,yield components and flag leaf size in bread wheat (Triticum aestivum L.) varieties of the Yellow and Huai Valleys of China

A total of 169 wheat (Triticum aestivum L.) varieties (landraces and cultivars) were used to asses the relationship between Puroindoline D1 alleles and Puroindoline b-B2 variants and grain hardness, other grain traits, yield components, and flag leaf size. Results indicated that the average SKCS hardness of Pinb-B2v3 varieties was significantly greater than that of Pinb-B2v2 varieties within the soft Puroindoline D1 haplotype sub-group. Conversely, no statistically significant difference was obtained for SKCS hardness between varieties with the Pinb-B2v3 vs. Pinb-B2v2 alleles within the two hard Puroindoline D1 haplotypes (Pinb-D1b and Pinb-D1p sub-groups). Therefore, the Puroindoline b-B2 gene may have a bigger impact on soft wheat varieties than hard. Across all varieties, thousand-kernel weight, grain weight per spike, grain diameter, grain number per spike, flag leaf width and area of Pinb-B2v3 varieties were significantly greater than those possessing Pinb-B2v2. These results indicated that the Pinb-B2v3 allele was associated with preferable grain yield traits compared to the Pinb-B2v2 allele in bread wheat. This study provides evocative information for better understanding the molecular and genetic basis of wheat grain yield.     

Changes in common wheat grain milling behavior and tissue mechanical properties following ozone treatment

Ozone treatment (10 g/kg) of common wheat grains with a new patented process, Oxygreen^®, used before milling was found to significantly reduce (by 10–20%) the required energy at breaking stage whatever the grain hardness and without changes in the flour yield. Detailed study of each of the milling steps undertaken on a hard type cultivar showed that both the breaking and the reduction energy were decreased. Reduction of the coarse bran yield was also observed concomitantly with an increase in the yield of white shorts. Biochemical characterization of the milling fractions pointed out changes in technological flour properties as starch damage reduction, aleurone content enrichment and increase of insoluble glutenin polymers. Measurement of wheat grain tissue mechanical properties showed that ozone treatment leads to reduction of the aleurone layer extensibility and affects the local endosperm resistance to rupture. These data as well as the direct effect of ozone oxidation on biochemical compounds could explain the observed changes in milling energy, bran and shorts yield and flour composition.