Multi-method research strategy for understanding changes in barley grain protein composition and its relation to improved nutritional quality

In this study, we successfully expressed the active 1Ay subunit of Triticum urartu in barley by Agrobacterium-mediated transformation with a transformation efficiency of 19.9%. The results of SDS-PAGE revealed that the expressed proteins of 1Ay subunit were present at some grains of each of 46 original T0 plants, showing identical mobility to those of positive standards of T. urartu grain protein and bacteria expressional proteins. In the T2 generation, three homozygous lines, 2-28, 3-11, and 5-6, were identified. The results of scanning electron microscopy showed an increased amount of protein bodies in these transgenic lines. The main effects in the expression of the 1Ay subunits was a considerable increase in the glutenin content, but a decrease in the contents of gliadins while there were no effects in the contents of albumin, globulin and the total protein. We found that the gluten could not be washed out from the flour obtained from transgenic barley lines when using a Gluten index analyzer and a Farinograph indicating that the transgenic barley lines could not form dough. The lack of x-type HMW-GS and the reduction in number of subunit were inferred as the possible reasons for the inability to form gluten polymer.

Mixing properties and dough functionality of transgenic lines of a commercial wheat cultivar expressing the 1Ax1, 1Dx5 and 1Dy10 HMW glutenin subunit genes

As a primary food crop, maize is widely grown around the world. However, the deficiency of essential amino acids, such as lysine, tryptophan, and methionine, results in poor nutritional quality of maize. In addition, the protein concentration of maize declines with the increase in yield, which further reduces the nutritional quality. Here, the photosynthesis of leaves, grain amino acid composition, and stoichiometry of N and S are explored. The results show that N and S maintained the redox balance by increasing the content of glutathione in maize leaves, thereby enhancing the photosynthetic rate and maize yield. Simultaneously, the synergy of N and S increased the grain protein concentration and promoted amino acid balance by increasing the cysteine concentration in maize grains. The maize yield, grain protein concentration, and concentration of essential amino acids, such as lysine, tryptophan, and methionine, could be simultaneously increased in the N:S ratio range of 11.0 to 12.0. Overall, the synergy of N and S simultaneously improved the maize yield and nutritional quality by regulating the redox balance of maize leaves and the amino acids balance of grains, which provides a new theoretical basis and practical method for sustainable production of maize.

The main physiology and biochemistry metabolism including amino acids composition and gamma-aminobutyric acid (GABA) content of sorghum during 3-day germination period was investigated. Respiratory rate increased during germination. Protease activity, free amino acid and reducing sugar content increased with germination time. While moisture, protein, fat and starch content decreased, significantly. Meanwhile, amino acid composition redistributed and the relative content of essential amino acids increased except for leucine and valine. In addition, some nonessential amino acids content such as proline, glycine and histidine was largely enhanced by germination. GABA content increased about 3 folds after germination. Furthermore, combining tannin (the main anti-nutritional factor) was converted to be free and solvable. These results greatly suggest that germinated sorghum could be a functional food rich in GABA and other health-promoting nutrients.Â

The use of barley grains as bioreactors for high-level production of cellulase (1,4-beta-glucanase) was investigated. A hybrid cellulase gene, cel-hyb1, driven by the rice GluB-1 promoter was expressed specifically in developing endosperm. Codon usage optimisation of cel-hyb1 increased its expression in barley grains 527-fold and led to cellulase production of up to 1.5% of total grain protein. CEL-HYB1 enzyme in barley grains was highly stable during post-harvest storage. Selectable marker gene ( hph) was subsequently eliminated from transgenic lines through segregation of hph from synthetic cel-hyb1 ( syn.cel-hyb1) in T1 progeny, using a binary plasmid containing hph and syn.cel-hyb1 in separate T-DNAs. These data suggest that barley grains can potentially be used for the commercial production of cellulase.

Topicality. In the conditions of ever-increasing technogenic pressure on the environment, the study of the influence of In the context of the growing anthropogenic load on the environment, studying the influence of abiotic factors on the translocation of heavy metals in the soil-plant system is of great practical importance for the agricultural sector. In the southeastern industrial region, the environmental safety of crop production largely depends on the heavy metal accumulation in plants and their translocation depends on both soil properties and climatic factors. Purpose. To investigate the effect of agroclimatic conditions in the eastern part of the Northern Steppe of Ukraine on the content of heavy metal mobile forms in soil and their translocation into spring barley grain. Materials and methods. Research was conducted in the field crop rotation of the Donetsk State Agricultural Research Station of NAAS on the Stepovyk variety in 2018–2022 with using field, laboratory, mathematical and statistical methods. Results. The highest increase in the acid-soluble zinc content was observed under increasing soil moisture. Under dry conditions in 2020, the acid-soluble zinc content was 45.3 mg/kg, while under excessive moisture conditions in 2022 it increased by 8.5 mg/kg or 18.8 %. The content of all studied heavy metals in spring barley grain did not exceed the maximum permissible concentration (MPC) with increasing soil moisture. The content of the studied heavy metals in spring barley grain under increasing soil moisture did not exceed the maximum permissible concentration (MPC). With increasing moisture availability, there was a tendency to increased copper content (by 8.2–9.9 %). The zinc accumulation in spring barley grain increased from 24.2 mg/kg to 28.9 mg/kg, or by 19.4 % in the excessively wet year of 2022 compared to the dry year of 2020. The cadmium content increased by 0.02 mg/kg, which is 40 %. Conclusions. Hydrometeorological conditions affect the content of heavy metals mobile forms in the soil and their translocation into spring barley grain, which necessitates the implementation of methods to reduce the environmental risks of heavy metal contamination of plant products under conditions of excessive wetting and real anthropogenic load. Key words: spring barley, agroclimatic conditions, soil, grain, content of heavy metals

The article presents a literature review on the risks of the global problem of fodder protein and protein amino acids in the feeding of agricultural animals and poultry. Alternative ways of solving it are due to the production and use of non-traditional feed raw materials in feed diets. A number of scientists consider atypical agricultural biotechnological products obtained from insects to be one of the alternative sources of fodder protein and amino acids and see prospects in the development of their production. The most common subjects for study in this regard are crickets, locusts, silkworm larvae, flies, worms, etc. Their protein value, amino acid, fatty acid and mineral composition, the content of vitamins, and other biologically active substances are studied. The development of this direction is considered one of the ways to solve a number of urgent environmental and fodder problems on a biological basis. Рurpose of the research was to study the protein value and complete amino acid composition of flour from insects, namely flour from the larvae of the BSF fly, crickets and snails, and the methods of its determining: crude protein by the Kjeldahl method, digestible protein with the help of 0,2% pepsin, and the digestibility coefficient, and the principles of conducting researches, as well as the characteristics of methods for determining the content of replaceable and essential amino acids, including sulfur-containing, using the Kapel-105M capillary electrophoresis system. As a result of the research, it was found that cricket flour is the least valuable in terms of protein value (digestible protein and digestibility coefficient), and it is also poor in the content of nonessential and essential amino acids, compared to other studied types of insect flour. The best indicators of protein value were noted in the flour from BSF fly larvae in terms of the content of crude and digestible proteins, as well as the digestibility coefficient, which is 75.6%, which indicates its good digestibility by the bodies of animals and birds. The highest content of amino acids is contained in the flour from BSF fly larvae. Taking into account the above results of crude protein content, digestibility, and digestibility in this sample, amino acid digestibility will also be higher compared to other flour samples. In addition, flour from BSF fly larvae is rich in sulfur-containing amino acids. The content of tryptophan and histidine, compared to other types of raw materials, is the highest. The studied samples of insect flour contain a complete set of essential and nonessential amino acids, but BSF fly larvae flour and snail flour are more valuable for essential amino acids.

Protein and amino acid composition of the mantle of juvenile O ctopus vulgaris (Cuvier, 1797) during fasting for 27 days were determined. Average protein content of octopus mantle was of 711.19 ± 46.80 g kg−1 DW, and it decreased with increasing fasting days. The non-essential amino acids content was higher (486.18 ± 11.08 g kg−1 protein) than essential amino acids (425.82 ± 9.15 g kg−1 protein) at the start of the experiment (unstarved animals). The results suggest that the amino acid profile of the mantle where the most abundant amino acids are Arg, His, Lys, Gly, Leu and Pro could indicate a prolonged fasting condition (>20 days) or poor nutrition of O . vulgaris. This study supports the idea of using mantle for metabolic needs of starved O . vulgaris suggesting that the degradation pathway of amino acids to pyruvate and tricarboxylic acid cycle intermediates was favoured contrary to the degradation pathway of ketogenic amino acids. Special considerations should be taken concerning Thr, Ile, Ser, Ala, Asx (Asp, Asn), Glx (Glu, Gln) (because of their fast intake) and Lys and His (due to their stable contents) during a prolonged period of fasting.

BackgroundEthiopia has enormous potential for malt barley production. However, the local supply is insufficient to meet the demand for both the existing and emerging malt and beer factories. As a result, millions of dollars were being spent nationally to import malt. This study sought to underscore the factors influencing smallholder farmers' decision to participate in malt barley marketing and the level of participation in Central Ethiopia. A multi-stage sampling method was used to select representative sample households. Descriptive statistics and Heckman two-stage model were used to analyze the data.ResultsThe result of the probit model revealed that malt barley producer farmers' decision to participate in the malt barley grain market was determined by age and formal education level, malt barley farming experience, amount of malt barley grain produced, total livestock holding, distance from local cooperative and nearest market and household access to enough stable food; whereas, the ordinary least squares regression results indicated that malt barley producer farmers' level of participation in the malt barley grain market was determined by the formal education level of the household head, total livestock holding, and distance from the local cooperative.ConclusionsThe policy recommendation of the study were raising farmers' awareness of malt barley quality parameters, organize producer farmers in nearby agricultural cooperatives, creating suitable transportation system, attractive grain prices, alternative stable food crops and forages species. In addition, collaborating with young and educated people will enhance malt barley output and market supply.

Agronomic solutions such as supplemental post‐emergence N may increase yield of feed‐barley grain. Grain yield, grain N yield, and grain protein generally increased as post‐emergence N rate increased. Largest grain yield increases occurred in environments with >300 mm of growing season precipitation. Yields decreased when N was applied as urea ammonium nitrate in hot and low‐precipitation conditions. There were no adverse negative effects to maturity, lodging, or grain quality in ‘Amisk’ feed barley. Small yield gains and decreasing feed barley (Hordeum vulgare L.) acreage in Alberta may be addressed with agronomic solutions. Field experiments were conducted from 2014 to 2016 at 3 irrigated and 11 rain‐fed environments in Alberta, Canada, to determine the effects of supplemental post‐emergence N on ‘Amisk’ feed barley growth, grain yield, and grain quality. At the time of seeding, urea N was applied at rates suitable for targeting average yield potential in each environment. Just prior to BBCH 30 (beginning of stem elongation), this was supplemented with additional 0, 34, and 68 kg N ha−1 in the form of undiluted urea ammonium nitrate (UAN), and 34 kg N ha−1 with the urease inhibitor N‐(n‐butyl) thiophosphoric triamide (NBPT). In most environments, grain yield, grain N yield (grain yield × percentage grain N), and grain protein increased with post‐emergence N rate. The largest positive grain yield responses (up to 19%) usually occurred in environments with more than 300 mm of growing season precipitation. Grain yield and grain N yield were significantly reduced when post‐emergence N was applied in high temperatures with low precipitation. Post‐emergence N increased height, but did not markedly change lodging or days to maturity. The grain quality parameters of test weight, starch, and fiber had minor responses to post‐emergence N application. The addition of NBPT to post‐emergence N had little effect on any response variables. Supplemental post‐emergence N provided an agronomic solution for increasing feed barley grain yield, but consistent increases were limited to irrigated or high precipitation environments.

The formation of the protein complex during barley grain maturation is characterized by unequal synthesis of different protein fractions. In ten day-old grain the dominating protein is glutelin, followed by albumin and globulin. The content of prolamin is at this stage negligible. Particularly after the eighteenth day of maturation intensive synthesis of prolamins begins, which continues during the entire maturing period in almost the same ratio as total N accumulation in the grain. From the twenty-fourth day the amount of prolamins exceeds that of glutelins, and at full maturity 50% of the proteins are prolamins. The glutelin content increases absolutely, but relatively it decreases from 41 to 26%.The amino acid composition of flour from total grain as well as the amino acid composition of the individual proteins is significantly changed during maturation. Generally the changes are manifested in an increasing content of Glu and Pro and a decreasing content of Asp, Ala and Lys. These changes seem to occur as a result of an increasing representation of prolamin, which is characterized by large content of Glu and Pro and low content of Asp, Ala and Lys compared with other protein fractions.

The expansion of the production of products with specified functional indicators (cereals, bakery products, pasta, desserts, muesli, yogurts) and feed with a certain amino acid composition is the main task of the food industry and the feed industry, and naked oats have firmly occupied their own niche in these areas of processing. The purpose of the research is to establish the amino acid composition of oat proteins of the hulled oat variety Konkur and naked varieties Bekas and Baget, to assess the balance of essential amino acids in oat proteins relative to the «reference protein» (amino acid scale according to FAO / WHO data) in order to use grain as an additive in food and feed to increase the proportion of essential amino acids in them. The research was carried out during 2018-2022 at the experimental base of the Samara Research Institute of Agriculture. Grain samples of the varieties of filmy and naked oats Konkur, Bekas and Baget were used as experimental material. It was found that the highest proportion of essential amino acids was observed in varieties of naked oats 39,9-41,0 g/kg. The sum of critical amino acids (lysine, methionine, tryptophan) in naked oat varieties was 9,4-9,6 g/kg, which is 1,1-1,3 g/kg higher than in the filmy variety Konkur. According to studies, 1 g of Konkur variety protein contains 249,7 mg of essential amino acids, proteins of naked oat variety Bekas - 268,4 mg and Baget variety - 262,9 mg, which corresponds to 69,4 %, 74,6 % and 73,0 % relative to FAO/WHO recommended standards. The excess of some essential amino acids in naked oat varieties (amino acid score over 100 %) contributed to the imbalance in the ratio of essential amino acids to the «reference» values, while increasing the coefficient of difference in amino acid scores (KRAS protein of Konkur variety was 24,2 %, Bekas variety 33,9 %, varieties Baget 35,5 %), thereby contributing to a decrease in the biological value of the proteins of the varieties Bekas and Baget in relation to the protein of the filmy variety Konkur. Grain of naked oats with a high content of essential amino acids should be used as an ingredient in the production of food and feed with high nutritional value.

A cDNA clone encoding the gamma-zein protein of maize was expressed in developing grain of barley using the starchy endosperm cell-specific promoter from the wheat Glu-1D-1 (HMW subunit 1Dx5) gene. Seven transgenic lines were recovered from 226 bombarded immature embryos, of which two were sterile and four tetraploid, while five were shown to express the gamma-zein protein based on western blotting. Southern blot analysis showed the presence of between about three and twelve transgene insertions. Detailed comparative studies of five null and five homozygous transformed sub-lines from transgenic line A showed that gamma-zein accounted for over 4% of the total prolamin fraction, corresponding to about 1.9% of the total grain N. Comparison of the proteins present in the gel protein fraction demonstrated that the gamma-zein was incorporated into polymers, as in maize. However, there was no effect on grain hardness measured using the Perten Single Kernel Characterisation System or on the vitreousness measured by visual inspection. This contrasts with the situation in maize where a clear association with vitreousness has been reported.

Different effects of dietary β-hydroxy-β-methylbutyrate on composition of fatty acid and free amino acid, and fatty metabolism in the different muscles of broilers

A mathematical model of the diffusive transport of abscisic acid (ABA) and gibberellins (GAs) through the aleurone layer of barley (Hordeum vulgare L.) grain is presented. The model consists of two partial differential equations describing the accumulation of phytohormone in the apoplastic and symplasmic compartments of the aleurone layer, both spatially and temporally. The mathematical model contains the morphology of the barley grain and the physicochemical properties of the two phytohormones. A mathematical derivation of the accumulation ratios for the two phytohormones between the symplast and apoplast under equilibrium conditions resulted in different distribution mechanisms for GAs and ABA. A sensitivity analysis of the accumulation ratio for GAs indicated high sensitivity to the apoplastic pH and the membrane potential, whereas the accumulation ratio for ABA proved to be most sensitive to the pH difference between the apoplast and symplast. The diffusive transport time for GAs to the basal site of the aleurone layer as calculated with the mathematical model is within a physiologically plausible timescale according to experimental data from the literature. Abscisic acid cannot be transported by diffusion to the end of the aleurone layer as quickly as GAs, according to model simulations. Therefore, the functional role of ABA in germination is likely to be in the vicinity of the embryo.