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"PROLAMINE"
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Genetic Variation Analysis of Avenin Components in the Population of F2 Common Oat Hybrids
by
Renev, Evgeny P.
,
Eremin, Dmitry I.
,
Lyubimova, Anna V.
in
Avena sativa L
,
avenin-coding locus
,
Barley
2024
The use of avenins as biochemical markers successfully complements the use of molecular markers in oat breeding. Currently, the genes controlling the synthesis of oat prolamins are insufficiently studied. The purpose of the work was to study the genetic variation of avenin components in populations of F2 common oat hybrids and to describe new allelic variants of component blocks. The avenins component of F2 grain in 19 hybrid oat populations was studied using the native electrophoresis method. Cultivars with new combinations of avenin components were used as parental genotypes to produce hybrids. The protein separation was conducted in vertical plates of 13.2% polyacrylamide gel. The number of avenin components in the spectra of cultivars varied from 8 to 12. The observed ratio of the grain number that compose the phenotypic classes for allele pairs at each of the loci corresponded to the theoretically expected one for codominant monohybrid inheritance. Our results confirm the assumption that avenin synthesis is controlled by three independent gene clusters located on three chromosomes. In the course of the studies, hybrid combinations were not identified in the spectra of which avenin components were manifested that were absent in both parents. The prolamin component blocks in oat are formed by 2–5 components, are characterized by high stability, and are inherited unchanged. Fifteen new allelic variants of blocks of components of the avenin electrophoretic spectrum have been identified: six for the Avn A locus, six for the Avn B locus, and three for the Avn C locus. This expands the possibilities of using prolamins as biochemical markers of economically valuable oat traits and certification of new cultivars and valuable breeding lines.
Journal Article
Down-Regulation of Rice Glutelin by CRISPR-Cas9 Gene Editing Decreases Carbohydrate Content and Grain Weight and Modulates Synthesis of Seed Storage Proteins during Seed Maturation
2023
The glutelins are a family of abundant plant proteins comprised of four glutelin subfamilies (GluA, GluB, GluC, and GluD) encoded by 15 genes. In this study, expression of subsets of rice glutelins were suppressed using CRISPR-Cas9 gene-editing technology to generate three transgenic rice variant lines, GluA1, GluB2, and GluC1. Suppression of the targeted glutelin genes was confirmed by SDS-PAGE, Western blot, and q-RT-PCR. Transgenic rice variants GluA1, GluB2, and GluC1 showed reduced amylose and starch content, increased prolamine content, reduced grain weight, and irregularly shaped protein aggregates/protein bodies in mature seeds. Targeted transcriptional profiling of immature seeds was performed with a focus on genes associated with grain quality, starch content, and grain weight, and the results were analyzed using the Pearson correlation test (requiring correlation coefficient absolute value ≥ 0.7 for significance). Significantly up- or down-regulated genes were associated with gene ontology (GO) and KEGG pathway functional annotations related to RNA processing (spliceosomal RNAs, group II catalytic introns, small nucleolar RNAs, microRNAs), as well as protein translation (transfer RNA, ribosomal RNA and other ribosome and translation factors). These results suggest that rice glutelin genes may interact during seed development with genes that regulate synthesis of starch and seed storage proteins and modulate their expression via post-transcriptional and translational mechanisms.
Journal Article
Isolation and characterization of gluten protein types from wheat, rye, barley and oats for use as reference materials
by
Koehler, Peter
,
Scherf, Katharina Anne
,
Lexhaller, Barbara
in
Alcohol
,
Allergies
,
Amino acids
2017
Gluten proteins from wheat, rye, barley and, in rare cases, oats, are responsible for triggering hypersensitivity reactions such as celiac disease, non-celiac gluten sensitivity and wheat allergy. Well-defined reference materials (RM) are essential for clinical studies, diagnostics, elucidation of disease mechanisms and food analyses to ensure the safety of gluten-free foods. Various RM are currently used, but a thorough characterization of the gluten source, content and composition is often missing. However, this characterization is essential due to the complexity and heterogeneity of gluten to avoid ambiguous results caused by differences in the RM used. A comprehensive strategy to isolate gluten protein fractions and gluten protein types (GPT) from wheat, rye, barley and oat flours was developed to obtain well-defined RM for clinical assays and gluten-free compliance testing. All isolated GPT (ω5-gliadins, ω1,2-gliadins, α-gliadins, γ-gliadins and high- and low-molecular-weight glutenin subunits from wheat, ω-secalins, γ-75k-secalins, γ-40k-secalins and high-molecular-weight secalins from rye, C-hordeins, γ-hordeins, B-hordeins and D-hordeins from barley and avenins from oats) were fully characterized using analytical reversed-phase high-performance liquid chromatography (RP-HPLC), sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), N-terminal sequencing, electrospray-ionization quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF-MS) and untargeted LC-MS/MS of chymotryptic hydrolyzates of the single GPT. Taken together, the analytical methods confirmed that all GPT were reproducibly isolated in high purity from the flours and were suitable to be used as RM, e.g., for calibration of LC-MS/MS methods or enzyme-linked immunosorbent assays (ELISAs).
Journal Article
Maize-A potential source of human nutrition and health: A review
2016
Maize or corn (Zea mays L.) is an important cereal crop of the world. It is a source of nutrition as well as phytochemical compounds. Phytochemicals play an important role in preventing chronic diseases. It contains various major phytochemicals such as carotenoids, phenolic compounds, and phytosterols. It is believed to have potential anti-HIV activity due to the presence of Galanthus nivalis agglutinin (GNA) lectin or GNA-maize. A tablespoon of maize oil satisfies the requirements for essential fatty acids for a healthy child or adult. Decoction of maize silk, roots, leaves, and cob are used for bladder problems, nausea, vomiting, and stomach complaints. Zein an alcohol-soluble prolamine found in maize endosperm has unique novel applications in pharmaceutical and nutraceutical areas. Resistant starch (RS) from maize reduces the risk of cecal cancer, atherosclerosis, and obesity-related complications. This review presents a detailed view on the nutritional and potential health benefits of maize.
Journal Article
Nitrogen Fertilizer Regulated Grain Storage Protein Synthesis and Reduced Chalkiness of Rice Under Actual Field Warming
2021
Our previous study has shown that nitrogen plays an important role in dealing with significantly increased chalkiness caused by elevated temperature. However, the role of nitrogen metabolites has not been given sufficient attention, and its regulatory mechanism is not clear. This study investigated the effects of high temperature and nitrogen fertilizer on the synthesis of grain storage protein and further explored the quality mechanism under the actual scenario of field warming. Results showed that increased temperature and nitrogen fertilizer could affect the activities of nitrogen metabolism enzymes, namely, glutamate synthetase, glutamine synthetase, glutamic pyruvic transaminase, and glutamic oxaloacetic transaminase, and the expressions of storage protein synthesis factor genes, namely, GluA and GluB , and subfamily genes, namely, pro14, BiP1 , and PDIL1 , which co-induced the changes of storage protein synthesis in rice grains. Furthermore, the increased temperature changed the balance of grain storage substances which may lead to the significantly increased chalky rate (197.67%) and chalkiness (532.92%). Moreover, there was a significant negative correlation between prolamin content and chalkiness, indicating that nitrogen fertilizer might regulate the formation of chalkiness by affecting the synthesis of prolamin. Results suggested that nitrogen application could regulate the related core factors involved in nitrogen metabolism pathways, which, in turn, affects the changes in the storage protein components in the grain and further affects quality. Therefore, as a conventional cultivation measure, nitrogen application would have a certain value in future rice production in response to climate warming.
Journal Article
Extraction, Composition, Functionality, and Utilization of Brewer’s Spent Grain Protein in Food Formulations
by
Moran, Galo Chuchuca
,
Grossmann, Lutz
,
Devnani, Bhanu
in
alternative proteins
,
Amino acids
,
Barley
2023
In recent years, brewer’s spent grain (BSG) has gained attention as a plant-based protein source because it occurs in large quantities as a by-product of beer brewing. BSG can contribute to future food requirements and support the development of a circular economy. In light of the dynamic developments in this area, this review aims to understand the proteins present in BSG, and the effect of extraction techniques and conditions on the composition, physicochemical, and techno-functional properties of the obtained protein extracts. The water-insoluble hordeins and glutelins form the major protein fractions in BSG. Depending on the beer brewing process, the extraction technique, and conditions, the BSG protein isolates predominantly contain B, C, and ϒ hordeins, and exhibit a broad molecular weight distribution ranging between <5 kDa and >250 kDa. While the BSG isolates obtained through chemical extraction methods seem promising to obtain gelled food products, physical and enzymatic modifications of BSG proteins through ultrasound and proteolytic hydrolysis offer an effective way to produce soluble and functional protein isolates with good emulsifying and foaming capabilities. Specifically tailored protein extracts to suit different applications can thus be obtained from BSG, highlighting that it is a highly valuable protein source.
Journal Article
Quality of winter wheat in relation to heat and drought shock after anthesis
by
Li, Z.,CSIRO Food Future Flagship, North Ryde (Australia)
,
Balla, K.,Agricultural Research Inst. of the Hungarian Academy of Sciences, Martonvasar (Hungary)
,
Rakszegi, M.,Agricultural Research Inst. of the Hungarian Academy of Sciences, Martonvasar (Hungary)
in
ABSORCION
,
ABSORPTION
,
AGUA
2011
This study investigated the effect of high temperature and drought (during grain-filling) on the quality and components yield of five winter wheat varieties. Drought and drought + heat were found to have a much greater influence on the yield and quality than heat stress alone. Averaged over the varieties, the yield losses were 57% after drought, 76% after drought + heat, and only 31% after heat stresses. The reductions in the unextractable polymeric protein fraction and glutenin-to-gliadin ratio indicated a poorer grain yield quality, despite the higher protein content. Quality deterioration was observed after drought or drought + heat, while high temperatures alone resulted in no change or in a better ratio of protein components. A significant negative correlation was observed between starch granule size and relative protein content after drought.
Journal Article
Genetic Basis of Variation in Rice Seed Storage Protein (Albumin, Globulin, Prolamin, and Glutelin) Content Revealed by Genome-Wide Association Analysis
2018
Rice seed storage protein (SSP) is an important source of nutrition and energy. Understanding the genetic basis of SSP content and mining favorable alleles that control it will be helpful for breeding new improved cultivars. An association analysis for SSP content was performed to identify underlying genes using 527 diverse
accessions grown in two environments. We identified more than 107 associations for five different traits, including the contents of albumin (Alb), globulin (Glo), prolamin (Pro), glutelin (Glu), and total SSP (Total). A total of 28 associations were located at previously reported QTLs or intervals. A lead SNP sf0709447538, associated for Glu content in the
subpopulation in 2015, was further validated in near isogenic lines NIL(Zhenshan97) and NIL(Delong208), and the Glu phenotype had significantly difference between two NILs. The association region could be target for map-based cloning of the candidate genes. There were 13 associations in regions close to grain-quality-related genes; five lead single nucleotide polymorphisms (SNPs) were located less than 20 kb upstream from grain-quality-related genes (
,
,
,
, and,
). Several starch-metabolism-related genes (
,
,
,
, and
) were also associated with SSP content. We identified favorable alleles of functional candidate genes, such as
,
,
, and other four candidate genes by haplotype analysis and expression pattern. Genotypes of
and
with higher Pro were not identified in
and exhibited much higher expression levels in
group. The lead SNP sf0601764762, repeatedly detected for Alb content in 2 years in the whole association population, was located in the
locus that controls the synthesis of amylose. And Alb content was significantly and negatively correlated with amylose content and the level of 2.3 kb
pre-mRNA examined in this study. The associations or candidate genes identified would provide new insights into the genetic basis of SSP content that will help in developing rice cultivars with improved grain nutritional quality through marker-assisted breeding.
Journal Article
Characterization and Relative Quantitation of Wheat, Rye, and Barley Gluten Protein Types by Liquid Chromatography–Tandem Mass Spectrometry
by
Scherf, Katharina A.
,
Colgrave, Michelle L.
,
Lexhaller, Barbara
in
Allergies
,
allergy
,
Amino acids
2019
The consumption of wheat, rye, and barley may cause adverse reactions to wheat such as celiac disease, non-celiac gluten/wheat sensitivity, or wheat allergy. The storage proteins (gluten) are known as major triggers, but also other functional protein groups such as α-amylase/trypsin-inhibitors or enzymes are possibly harmful for people suffering of adverse reactions to wheat. Gluten is widely used as a collective term for the complex protein mixture of wheat, rye or barley and can be subdivided into the following gluten protein types (GPTs): α-gliadins, γ-gliadins, ω5-gliadins, ω1,2-gliadins, high- and low-molecular-weight glutenin subunits of wheat, ω-secalins, high-molecular-weight secalins, γ-75k-secalins and γ-40k-secalins of rye, and C-hordeins, γ-hordeins, B-hordeins, and D-hordeins of barley. GPTs isolated from the flours are useful as reference materials for clinical studies, diagnostics or in food analyses and to elucidate disease mechanisms. A combined strategy of protein separation according to solubility followed by preparative reversed-phase high-performance liquid chromatography was employed to purify the GPTs according to hydrophobicity. Due to the heterogeneity of gluten proteins and their partly polymeric nature, it is a challenge to obtain highly purified GPTs with only one protein group. Therefore, it is essential to characterize and identify the proteins and their proportions in each GPT. In this study, the complexity of gluten from wheat, rye, and barley was demonstrated by identification of the individual proteins employing an undirected proteomics strategy involving liquid chromatography-tandem mass spectrometry of tryptic and chymotryptic hydrolysates of the GPTs. Different protein groups were obtained and the relative composition of the GPTs was revealed. Multiple reaction monitoring liquid chromatography-tandem mass spectrometry was used for the relative quantitation of the most abundant gluten proteins. These analyses also allowed the identification of known wheat allergens and celiac disease-active peptides. Combined with functional assays, these findings may shed light on the mechanisms of gluten/wheat-related disorders and may be useful to characterize reference materials for analytical or diagnostic assays more precisely.
Journal Article
Gene Duplication and Evolution Dynamics in the Homeologous Regions Harboring Multiple Prolamin and Resistance Gene Families in Hexaploid Wheat
2018
Improving end-use quality and disease resistance are important goals in wheat breeding. The genetic loci controlling these traits are highly complex, consisting of large families of prolamin and resistance genes with members present in all three homeologous A, B, and D genomes in hexaploid bread wheat. Here, orthologous regions harboring both prolamin and resistance gene loci were reconstructed and compared to understand gene duplication and evolution in different wheat genomes. Comparison of the two orthologous D regions from the hexaploid wheat Chinese Spring and the diploid progenitor
revealed their considerable difference due to the presence of five large structural variations with sizes ranging from 100 kb to 2 Mb. As a result, 44% of the
and 71% of the Chinese Spring sequences in the analyzed regions, including 79 genes, are not shared. Gene rearrangement events, including differential gene duplication and deletion in the A, B, and D regions, have resulted in considerable erosion of gene collinearity in the analyzed regions, suggesting rapid evolution of prolamin and resistance gene families after the separation of the three wheat genomes. We hypothesize that this fast evolution is attributed to the co-evolution of the two gene families dispersed within a high recombination region. The identification of a full set of prolamin genes facilitated transcriptome profiling and revealed that the A genome contributes the least to prolamin expression because of its smaller number of expressed intact genes and their low expression levels, while the B and D genomes contribute similarly.
Journal Article