Serine hydroxymethyltransferase participates in the synthesis of cysteine-rich storage proteins in rice seed.

The low level of cysteine-rich proteins (lcrp) mutation indicates a decrease in cysteine-rich (CysR) prolamines, α-globulin, and glutelin. To identify the causing factor of lcrp mutation, to elucidate its function, and to elucidate the role of CysR proteins in the formation of protein bodies (PBs), lcrp mutant was analyzed. A linkage map of the LCRP gene was constructed and genomic DNA sequencing of a predicted gene within the mapped region demonstrated that LCRP encodes a serine hydroxymethyltransferase, which participates in glycine-serine interconversion of one-carbon metabolism in the sulfur assimilation pathway. The levels of l-Ser, Gly, and Met in the sulfur assimilation pathway in the lcrp seeds increased significantly compared to that in the wildtype (WT). As the lcrp mutation influences the growth of shoot and root, the effects of the addition to the medium of amino acids and other compounds on the sulfur assimilation pathway were studied. Electron-lucent PBs surrounded by ribosome-attached membranes were observed accumulating cysteine-poor prolamines in the lcrp seeds. Additionally, glutelin-containing PBs were smaller and distorted in the lcrp seeds compared to those in the WT. These analyses of PBs in the lcrp seeds suggest that cysteine-rich proteins play an important role in the formation of PBs in rice.

Grain chalkiness traits is affected by the synthesis and dynamic accumulation of the storage protein in rice.

BACKGROUOND: Grain chalkiness lowers the market value of rice. Alleviating grain chalkiness is the most challenging issue in many rice-producing areas of the world. Nitrogen (N) metabolism has received increasing attention as a result of its relationship with grain chalkiness, although little information is available on the mechanism of N-induced grain chalk. RESULTS: A highly chalky rice variety OM052 was used to explore the protein synthesis and its accumulation in the grain exposed to N topdressing (N+) at the panicle initiation stage and a control (N-). The results showed that chalky kernels were stimulated by the N+ treatment and more prone to occur on the top and primary rachis. The grain protein content was increased because of the increased average and maximum rates of protein accumulation during grain filling, which was related to the enhanced activities of glutamine synthetase, glutamate synthase, glutamic oxalo-acetic transaminase and glutamate pyruvate transaminase under the N+ treatment. The activities of these enzymes at 15 days after flowering (DAF) were notably positively correlated with grain chalky traits and protein content. CONCLUSION: N topdressing regulates the synthesis and accumulation of the protein by affecting the key enzymes, especially at 15 DAF, which is attributed to grain chalkiness in rice. © 2021 Society of Chemical Industry.

Design of a Seed-Specific Chimeric Promoter with a Modified Expression Profile to Improve Seed Oil Content.

Increasing the yield of plant oil is an important objective to meet the demand for sustainable resources and energy. Some attempts to enhance the expression of genes involved in oil synthesis in seeds have succeeded in increasing oil content. In many cases, the promoters of seed-storage protein genes have been used as seed-specific promoters. However, conventional promoters are developmentally regulated and their expression periods are limited. We constructed a chimeric promoter that starts to express in the early stage of seed development, and high-level expression is retained until the later stage by connecting the promoters of the biotin carboxyl carrier protein 2 (BCCP2) gene encoding the BCCP2 subunit of acetyl-CoA carboxylase and the fatty acid elongase 1 (FAE1) gene from Arabidopsis. The constructed promoter was ligated upstream of the TAG1 gene encoding diacylglycerol acyltransferase 1 and introduced into Arabidopsis. Seeds from transgenic plants carrying AtTAG1 under the control of the chimeric promoter showed increased oil content (up by 18⁻73%) compared with wild-type seeds. The novel expression profile of the chimeric promoter showed that this could be a promising strategy to manipulate the content of seed-storage oils and other compounds.

Buried treasure: biosynthesis, structures and applications of cyclic peptides hidden in seed storage albumins.

Covering: 1999 up to the end of 2017The small cyclic peptide SunFlower Trypsin Inhibitor-1 (SFTI-1) from sunflower seeds is the prototypic member of a novel family of natural products. The biosynthesis of these peptides is intriguing as their gene-encoded peptide backbone emerges from a precursor protein that also contains a seed storage albumin. The peptide sequence is cleaved out from the precursor and cyclised by the albumin-maturing enzymatic machinery. Three-dimensional solution NMR structures of a number of these peptides, and of the intact precursor protein preproalbumin with SFTI-1, have now been elucidated. Furthermore, the evolution of the family has been described and a detailed understanding of the biosynthetic steps, which are necessary to produce cyclic SFTI-1, is emerging. Macrocyclisation provides peptide stability and thus represents a key strategy in peptide drug development. Consequently the constrained structure of SFTI-1 has been explored as a template for protein engineering, for tuning selectivity towards clinically relevant proteases and for grafting in sequences with completely novel functions. Here we review the discovery of the SFTI-1 peptide family, their evolution, biosynthetic origin, and structural features, as well as highlight the potential applications of this unique class of natural products.

Global transcriptome analysis and identification of genes involved in nutrients accumulation during seed development of rice tartary buckwheat (Fagopyrum Tararicum).

Tartary buckwheat seeds are rich in various nutrients, such as storage proteins, starch, and flavonoids. To get a good knowledge of the transcriptome dynamics and gene regulatory mechanism during the process of seed development and nutrients accumulation, we performed a comprehensive global transcriptome analysis using rice tartary buckwheat seeds at different development stages, namely pre-filling stage, filling stage, and mature stage. 24 819 expressed genes, including 108 specifically expressed genes, and 11 676 differentially expressed genes (DEGs) were identified. qRT-PCR analysis was performed on 34 DEGs to validate the transcriptome data, and a good consistence was obtained. Based on their expression patterns, the identified DEGs were classified to eight clusters, and the enriched GO items in each cluster were analyzed. In addition, 633 DEGs related to plant hormones were identified. Furthermore, genes in the biosynthesis pathway of nutrients accumulation were analyzed, including 10, 20, and 23 DEGs corresponding to the biosynthesis of seed storage proteins, flavonoids, and starch, respectively. This is the first transcriptome analysis during seed development of tartary buckwheat. It would provide us a comprehensive understanding of the complex transcriptome dynamics during seed development and gene regulatory mechanism of nutrients accumulation.

Overview on the mechanisms of coffee germination and fermentation and their significance for coffee and coffee beverage quality.

Quality of coffee is a complex trait and is influenced by physical and sensory parameters. A complex succession of transformations during the processing of seeds to roasted coffee will inevitably influence the in-cup attributes of coffee. Germination and fermentation of the beans are two bioprocesses that take place during post-harvest treatment, and may lead to significant modifications of coffee attributes. The aim of this review is to address the current knowledge of dynamics of these two processes and their significance for bean modifications and coffee quality. The first part of this review gives an overview of coffee germination and its influence on coffee chemistry and quality. The germination process initiates while these non-orthodox seeds are still inside the cherry. This process is asynchronous and the evolution of germination depends on how the beans are processed. A range of metabolic reactions takes place during germination and can influence the carbohydrate, protein, and lipid composition of the beans. The second part of this review focuses on the microbiota associated with the beans during post-harvesting, exploring its effects on coffee quality and safety. The microbiota associated with the coffee cherries and beans comprise several bacterial, yeast, and fungal species and affects the processing from cherries to coffee beans. Indigenous bacteria and yeasts play a role in the degradation of pulp/mucilage, and their metabolism can affect the sensory attributes of coffee. On the other hand, the fungal population occurring during post-harvest and storage negatively affects coffee quality, especially regarding spoilage, off-tastes, and mycotoxin production.

Identification and Assessment of the Potential Allergenicity of 7S Vicilins in Olive (Olea europaea L.) Seeds.

Olive seeds, which are a raw material of interest, have been reported to contain 11S seed storage proteins (SSPs). However, the presence of SSPs such as 7S vicilins has not been studied. In this study, following a search in the olive seed transcriptome, 58 sequences corresponding to 7S vicilins were retrieved. A partial sequence was amplified by PCR from olive seed cDNA and subjected to phylogenetic analysis with other sequences. Structural analysis showed that olive 7S vicilin contains 9 α-helixes and 22 ß-sheets. Additionally, 3D structural analysis displayed good superimposition with vicilin models generated from Pistacia and Sesamum. In order to assess potential allergenicity, T and B epitopes present in these proteins were identified by bioinformatic approaches. Different motifs were observed among the species, as well as some species-specific motifs. Finally, expression analysis of vicilins was carried out in protein extracts obtained from seeds of different species, including the olive. Noticeable bands were observed for all species in the 15-75 kDa MW interval, which were compatible with vicilins. The reactivity of the extracts to sera from patients allergic to nuts was also analysed. The findings with regard to the potential use of olive seed as food are discussed.

Tailoring Grain Storage Reserves for a Healthier Rice Diet and its Comparative Status with Other Cereals.

A global rise of diet-related noncommunicable diseases calls for a focus on diet-based nutritional intervention across the entire socioeconomic consumer spectrum. We review recent reports in the area of healthier rice aimed at developing rice grains with improved dietary fiber compositions (increased amounts of nonstarch polysaccharides and resistant starch), and less digestible starch (higher amylose and phospholipid complex in the endosperm) resulting in reduced glycemic impact upon grain consumption. We furthermore elaborate on the interconnections of elevated amounts of protein and a balanced composition of essential amino acids. The importance of a nutritious aleurone layer and its role in lipid storage and micronutrient composition is discussed briefly in the context of brown rice benefits. We identify gene targets for precision breeding that will facilitate the production of rice grains and rice-based products to mitigate the impact of nutrition-related preventable diseases.

Higher endogenous methionine in transgenic Arabidopsis seeds affects the composition of storage proteins and lipids.

Previous in vitro studies demonstrate that exogenous application of the sulfur-containing amino acid methionine into cultured soybean cotyledons and seedlings reduces the level of methionine-poor storage proteins and elevates those that are methionine-rich. However, the effect of higher endogenous methionine in seeds on the composition of storage products in vivo is not studied yet. We have recently produced transgenic Arabidopsis seeds having significantly higher levels of methionine. In the present work we used these seeds as a model system and profiled them for changes in the abundances of 12S-globulins and 2S-albumins, the two major groups of storage proteins, using 2D-gels and MALDI-MS detection. The findings suggest that higher methionine affects from a certain threshold the accumulation of several subunits of 12S-globulins and 2S-albumins, regardless of their methionine contents, resulting in higher total protein contents. The mRNA abundances of most of the genes encoding these proteins were either correlated or not correlated with the abundances of these proteins, implying that methionine may regulate storage proteins at both transcriptional and post-transcriptional levels. The elevations in total protein contents resulted in reduction of total lipids and altered the fatty acid composition. Altogether, the data provide new insights into the regulatory roles of elevated methionine levels on seed composition.

Fluctuation of Dof1/Dof2 expression ratio under the influence of varying nitrogen and light conditions: involvement in differential regulation of nitrogen metabolism in two genotypes of finger millet (Eleusine coracana L.).

In order to gain insights into the mechanism of high nitrogen use efficiency (NUE) of finger millet (FM) the role of Dof2 transcription factor (TF), which is a repressor of genes involved in C/N metabolism was investigated. The partial cDNA fragment of EcDof2 (912-bp; GenBank acc. no. KF261117) was isolated and characterized from finger millet (FM) that showed 63% and 58% homology with Dof2 of Zea mays at nucleotide and protein level, respectively. Its expression studies were carried out along with the activator EcDof1 in two genotypes (GE3885, high protein genotype (HPG); GE1437, low protein genotype (LPG)) of FM differing in grain protein contents (13.8% and 6.2%) showed that EcDof2 is expressed in both shoot and root tissues with significantly (p≤0.05) higher expression in the roots. The diurnal expression of both EcDof1 and EcDof2 in shoots was differential having different time of peak expression indicating a differential response to diurnal condition. Under continuous dark conditions, expression of EcDof1 and EcDof2 oscillated in both the genotypes whereas on illumination, the fold expression of EcDof1 was higher as compared to EcDof2. Under increasing nitrate concentration, EcDof2 expression increases in roots and shoots of LPG while it remains unchanged in HPG. However, the EcDof1 expression was found to increase in both genotypes. Further, time kinetics studies under single nitrate concentration revealed that EcDof2 was repressed in the roots of both genotypes whereas EcDof1 oscillated with time. The EcDof1/EcDof2 ratio measured showed differential response under different light and nitrogen conditions. It was higher in the roots of HPG indicating higher activation of genes involved in N uptake and assimilation resulting in high grain protein accumulation. The results indicate that both light and nitrogen concentration influence Dof1 and Dof2 expression and suggests a complex pattern of regulation of genes influenced by these plant specific TFs. In nutshell, the Dof1/Dof2 ratio can serve as an index for measuring the N responsiveness and NUE of crops and can be further validated by Dof2 knock down approach.

Seed-specific expression of a lysine-rich protein gene, GhLRP, from cotton significantly increases the lysine content in maize seeds.

Maize seed storage proteins are a major source of human and livestock consumption. However, these proteins have poor nutritional value, because they are deficient in lysine and tryptophan. Much research has been done to elevate the lysine content by reducing zein content or regulating the activities of key enzymes in lysine metabolism. Using the naturally lysine-rich protein genes, sb401 and SBgLR, from potato, we previously increased the lysine and protein contents of maize seeds. Here, we examined another natural lysine-rich protein gene, GhLRP, from cotton, which increased the lysine content of transgenic maize seeds at levels varying from 16.2% to 65.0% relative to the wild-type. The total protein content was not distinctly different, except in the six transgenic lines. The lipid and starch levels did not differ substantially in Gossypium hirsutum L. lysine-rich protein (GhLRP) transgenic kernels when compared to wild-type. The agronomic characteristics of all the transgenic maize were also normal. GhLRP is a high-lysine protein candidate gene for increasing the lysine content of maize. This study provided a valuable model system for improving maize lysine content.

Differences between easy- and difficult-to-mill chickpea (Cicer arietinum L.) genotypes. Part II: protein, lipid and mineral composition.

BACKGROUND: Part I introduced the concept of easy- and difficult-to-mill chickpea genotypes, the broad chemical composition of their seed fractions and proposed mechanistic explanations for physical differences consistent with observed variation in milling ease. Part II continues this research by delving deeper into the amino acid, fatty acid and mineral components. RESULTS: No association between fatty acid composition and ease of milling was observed. However, particular amino acids and mineral elements were identified that further support roles of lectins, pectins and mineral-facilitated binding in the adhesion of chickpea seed coat and cotyledons. CONCLUSION: These differences suggest underlying mechanisms that could be exploited by breeding programmes to improve milling performance. This study shows that the content and composition of amino acids, fatty acids and minerals within different chickpea tissues vary with seed type (desi and kabuli) and within desi genotypes in ways that are consistent with physical explanations of how seed structure and properties relate to milling behaviour.

Differences between easy- and difficult-to-mill chickpea (Cicer arietinum L.) genotypes. Part I: broad chemical composition.

BACKGROUND: Ease of milling is an important quality trait for chickpeas (Cicer arietinum L.) and involves two separate processes: removal of the seed coat and splitting of cotyledons. Four chickpea genotypes (two desi types, one kabuli type and one interspecific hybrid with 'wild' C. echinospermum parentage) of differing ease of milling were examined to identify associated seed composition differences in the seed coat, cotyledons and their junctions (abaxial and adaxial). RESULTS: Several components in different fractions were associated with ease of milling chickpea seeds: primarily soluble and insoluble non-starch polysaccharides (including pectins) and protein at the seed coat and cotyledon junctions, and the lignin content of the seed coat. CONCLUSION: This study shows that the chemical composition of chickpea does vary with seed type (desi and kabuli) and within desi genotypes in ways that are consistent with physical explanations of how seed structure and properties relate to milling behaviour.

Differential expression and elution behavior of basic 7S globulin among cultivars under hot water treatment of soybean seeds.

Basic 7S globulin (Bg7S), which accumulates in mature soybean (Glycine max) seeds, is an extracellular matrix protein. A large amount of Bg7S is synthesized de novo and is eluted from soybean seeds when immersed in 50-60°C water (hot water treatment, HWT). However, the Bg7S elution mechanism remains unclear. Under HWT, the seeds probably undergo heat stress and flooding stress. To obtain fundamental knowledge related to how Bg7S is eluted from hot-water-treated seeds, this study compared Bg7S elution among soybean cultivars having different flooding tolerance during pre-germination. The amounts of Bg7S eluted from seeds varied significantly among cultivars. Elution was suppressed by seed coats regarded as preventing the leakage of seed contents by rapid water imbibition. Furthermore, Bg7S expression levels differed among cultivars, although the difference did not result from any variation in Bg7S promoter sequences. However, the expression levels of Bg7S under HWT were not associated with the flooding tolerance level. Immunoelectron microscopy revealed that the Bg7S accumulated in the intercellular space of hot-water-treated seeds. Plasma membrane shrinkage was observed. The main proteins eluted from seeds under HWT were located in the extracellular space. This study clarified the mechanism of Bg7S elution from seeds under HWT.

Chemical compounds related to nutraceutical and industrial qualities of non-transgenic soybean genotypes.

BACKGROUND: Information about the chemical profile of soybean seed is valuable for breeding programs aimed at obtaining value-added products to meet the demands of niche markets. The objective of this study was to determine seed composition of non-transgenic soybean genotypes with specialty characters in different environments of Argentina. RESULTS: Protein and oil contents ranged from 396 to 424 g kg⁻¹ and from 210 to 226 g kg⁻¹, respectively. Oleic and linolenic acid ratio, the general indicator of oil quality, varied from 2.7 to 3.8. The oil contained high levels of total tocopherols (1429-1558 mg kg⁻¹) and the meal exhibited high levels of total isoflavones (2.91-4.62 mg g⁻¹). The biplot showed that oleic, linoleic and linolenic acids, γ-, δ- and total tocopherols, genistin, malonyl daidzin and genistin, acetyl daidzin and glycitin and total isoflavones allowed the greatest discrimination among the genotypes studied. CONCLUSION: Different chemical profiles of each non-transgenic genotype analyzed were established and, therefore, their identity was defined. These results are important for breeders who intend to obtain new genotypes with improved meal and oil quality, as well as for processors and exporters, who could use them directly as raw material for soyfood processing for nutraceutical purposes.

Effect of nitrogen fertilization and cover cropping systems on sorghum grain characteristics.

Cover crop treatments and nitrogen (N) fertilization rates were investigated for their impact on sorghum grain quality attributes. Sorghum was planted in field plots treated with differing cover cropping systems and fertilization rates. The size (weight and diameter) and hardness of the kernels were influenced by both the cover crop and N rates. The protein content increased as the N rate increased and also with the addition of cover crops to the system. The protein digestibility values and starch granule size distributions were not affected by N rate or the cover cropping treatments. Soil properties were tested to determine relationships with grain quality attributes. The utilization of cover crops appears to increase the protein content without causing a deleterious effect on protein digestibility. The end-product quality is not hampered by the use of beneficial cropping systems necessary for sustainable agriculture.

Effects of soybean protein composition and processing conditions on silken tofu properties.

BACKGROUND: Texture and water holding are important for silken tofu manufacturers and are affected by many factors, including soybean variety and processing conditions. In this study we evaluated these two key quality attributes of silken tofu produced from two soybean varieties - Bunya and Cowrie - using a soak or a dry processing method with glucono-δ-lactone or nigari as coagulant at concentrations of 1.5-5.0 g kg⁻¹. RESULTS: The soak method produced substantially firmer silken tofu with either coagulant. The optimum coagulant concentration to achieve maximum hardness was 2.5-3.0 g kg⁻¹. At 3.0 g kg⁻¹, Bunya produced silken tofu with firmer texture for each of the coagulant-method combinations and lower water loss, an indicator for water holding capacity, for the soak method. The two varieties differed significantly in the composition of major protein subunits for the seed and soymilk. Bunya had no 11SA4, less 11S and more 7S and therefore a lower 11S/7S ratio. CONCLUSION: Both variety and processing conditions affect textural properties and water loss of silken tofu. The absence of 11SA4 and subsequent protein subunit compositions are important in determining these two key quality attributes. Variety Bunya produces silken tofu with firmer texture and lower water loss.

Ammonium as sole N source improves grain quality in wheat.

BACKGROUND: The skilful handling of N fertilizer, including N source type and its timing, is necessary to obtain maximum profitability in wheat crops in terms of production and quality. Studies on grain yield and quality with ammonium as sole N source have not yet been conducted. The aim of this study was to evaluate the effect of N source management (nitrate vs. ammonium), and splitting it into two or three amendments during the wheat life cycle, on grain yield and quality under irrigated conditions. RESULTS: This experiment demonstrates that Cezanne wheat plants growing with ammonium as exclusive N source are able to achieve the same yield as plants growing with nitrate and that individual wheat plants grown in irrigated pots can efficiently use late N applied in GS37. Ammonium nutrition increased both types of grain reserve proteins (gliadins and glutenins) and also increased the ratio gli/glu with respect to nitrate nutrition. The splitting of the N rate enhanced the ammonium effect on grain protein composition. CONCLUSIONS: The application of ammonium N source, especially when split into three amendments, has an analogous effect on grain protein content and composition to applications at a higher N rate, leading to higher N use efficiency.

Interactive effect of supplemental ultraviolet B and elevated ozone on seed yield and oil quality of two cultivars of linseed (Linum usitatissimum L.) carried out in open top chambers.

BACKGROUND: Current scenarios of global climate change predict a significant increase in ultraviolet B (UV-B) and tropospheric ozone (O3) in the near future. Both UV-B and O3 can have detrimental effects on the productivity and yield quality of important agricultural crops. The present study was conducted to investigate the individual and interactive effects of supplemental UV-B (sUV-B) (ambient + 7.2 kJ m⁻² day⁻¹) and O3 (ambient + 10 ppb) on the yield and oil quality of two cultivars of linseed (Linum usitatissimum L.). RESULTS: The mean monthly ambient O3 concentration varied from 27.7 to 59.0 ppb during the experimental period. O3 affected fruit formation, while sUV-B was mainly responsible for ovule abortion. Seed sugar and protein contents showed maximum reduction in O3-treated plants, while mineral nutrient levels were most affected by sUV-B + O3 treatment. Rancid oil of low nutritional quality and containing long-chain fatty acids was favoured along with a decrease in oil content. CONCLUSION: sUV-B and O3 individually as well as in combination caused deterioration of the yield and quality of oil and seeds of linseed. However, the individual effect of O3 was more damaging than the effect of sUV-B or sUV-B + O3, and cultivar T-397 performed better than Padmini.

Proteins in oats; their synthesis and changes during germination: a review.

Oats (Avena sativa L.) are distinct among cereals due to their considerably higher protein concentration. At the same time oats possess a protein quality of high nutritional value and a special protein composition. Most cereals like wheat, barley, and rye have a high percentage of prolamins, the alcohol-soluble fraction, which usually contains most of the storage proteins, but oats are an exception. Their major storage proteins belong to the salt-water soluble globulin fraction, whereas oats prolamins are a minor component. During oats groat development, most obvious is the fairly linear increase in the globulin fraction. Oats globulins share structural features with the 11 S globulins of legumes and other dicots. Amino acid composition of oats is superior to that of other cereals due to the higher amount of limiting amino acids like lysine and threonine. During germination, total amino acid analysis revealed an increase in essential amino acids like lysine and tryptophan, which leads to an increased nutritional value of germinated oats. Oats protein products including globulin have been chemically modified by various methods to improve their functional properties.