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BACKGROUND: Cereals foods with a high content of dietary fibres or amylose have potential to lower postprandial glucose levels. Optimisation of cereal foods may improve management of type 2 diabetes (T2D). METHODS: We investigated the impact on 4 h postprandial glucose responses given as incremental area under curve (iAUC) of bread made of either 50% RNAi-based (genetically modified) amylose-only barley flour (AmOn) (and 50% wheat flour), 50% hulless barley flour (and 50% wheat flour) or 75% hulless barley (and 25% wheat flour) in subjects with T2D compared with 100% wheat flour bread. DESIGN: Twenty adults with T2D were randomly allocated to one of four breads at four separate visits. We measured fasting and 4 h postprandial responses of glucose, insulin, glucagon, triacylglycerol (TG), free fatty acids (FFA), glucagon-like peptide-1 (GLP-1) and gastric inhibitory polypeptide (GIP). Mixed model ANOVA was used to examine the differences. RESULTS: Bread made from 50% AmOn lowered the 4 h postprandial glucose by 34%, 27%, 23% (P < 0.05) compared with 100% wheat, 50% or 75% hulless barley, respectively. Bread made from 75% hulless barley reduced the postprandial glucose response (iAUC) by 11% (P < 0.05) compared to 100% wheat bread. Postprandial insulin responses (iAUC) were reduced for 50% AmOn compared with 100% wheat and 50% hulless barley and for 75% hulless compared to 50% hulless barley bread (P < 0.05). 4 h postprandial glucagon (tAUC) did not differ between the four bread types (P > 0.05). Lower postprandial GIP (iAUC) was observed after all barley breads compared to 100% wheat (P < 0.05), whereas no difference was seen in postprandial GLP-1. Postprandial TG and FFA (tAUC) were difficult to judge due to differences in fasting values. CONCLUSIONS: Bread made by replacing wheat flour with either 50% high-amylose or 75% hulless barley flour lowered postprandial glucose responses compared to 100% wheat bread indicating a beneficial impact on glucose regulation in T2D subjects. This trial was registered at clinicaltrials.gov as NCT04646746.
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Diabetes Mellitus Tipo 2 , Hordeum , Adulto , Humanos , Glucagón , Amilosa , Pan/análisis , Triticum/química , Glucemia , Harina , Péptido 1 Similar al Glucagón , Insulina , Glucosa , Polipéptido Inhibidor Gástrico , Grano Comestible , Periodo PosprandialRESUMEN
De novo domestication is a novel trend in plant genetics, where traits of wild or semi-wild species are changed by the use of modern precision breeding techniques so that they conform to modern cultivation. Out of more than 300,000 wild plant species, only a few were fully domesticated by humans in prehistory. Moreover, out of these few domesticated species, less than 10 species dominate world agricultural production by more than 80% today. Much of this limited diversity of crop exploitation by modern humans was defined early in prehistory at the emergence of sedentary agro-pastoral cultures that limited the number of crops evolving a favorable domestication syndrome. However, modern plant genetics have revealed the roadmaps of genetic changes that led to these domestication traits. Based on such observations, plant scientists are now taking steps towards using modern breeding technologies to explore the potential of de novo domestication of plant species that were neglected in the past. We suggest here that in this process of de novo domestication, the study of Late Paleolithic/Late Archaic and Early Neolithic/Early Formative exploration of wild plants and identification of neglected species can help identify the barriers towards domestication. Modern breeding technologies may then assist us to break these barriers in order to perform de novo domestication to increase the crop species diversity of modern agriculture.
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Starch with a high amylose (AM) content (high AM starch, HAS) has attracted increasing research attention due to its industrial application potential, such as functional foods and biodegradable packaging. In the past two decades, HAS structure, functionality, and applications have been the research hotspots. However, a review that comprehensively summarizes these areas is lacking, making it difficult for interested readers to keep track of past and recent advances. In this review, we highlight studies that benefited from rapidly developing techniques, and systematically review the structure, functionality, and applications of HAS. We particularly emphasize the relationships between HAS molecular structure and physicochemical properties.
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Amilosa , Almidón , Almidón/química , Amilosa/química , Estructura MolecularRESUMEN
We investigated the effects of drought stress (DS) on maize varieties with different amylose content (AC). In starches with AC of 33 %, DS increased the contents of amylopectin (AP) chains with a degree of polymerization (DP) > 36 and decreased the AP chains with DP ≤ 36, while the AC was unchanged. DS decreased the crystallinity, the thickness of both amorphous and crystalline lamellae, and average granular size. In contrast, the digestibility increased. For starches with AC of 45 %, DS increased the content of AP chains with DP > 24 and AC, while the contents of AP chains with DP ≤ 24 decreased. DS produced starch with thinner crystalline lamellae, thicker amorphous lamellae, more elongated and larger granules. The digestibility of the starches decreased. In starches with AC of 53 %, moderate DS led to similar structural and functional changes as found for starches with AC of 45 %. Finally, severe DS resulted in the decrease of AC.
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Amilopectina , Amilosa , Amilopectina/química , Amilosa/química , Sequías , Almidón/química , Zea mays/químicaRESUMEN
Nanoparticles are more promising than microcapsules as drug carriers because they can be absorbed directly by intestinal epithelial cells, significantly increasing the uptake and bioaccessibility of polyphenols. Our study aimed to use catechin (CC), epicatechin (EC) and proanthocyanidin (PAC) adsorption onto tapioca starch nanoparticles (TSNs), which were prepared by a physical method. These TSN loaded-polyphenols were subjected to adsorption kinetic, adsorption isotherm, adsorption capacity, antioxidant activity, and in vitro release analyses. The maximum adsorption capacities of TSNs for CC, EC and PACs were 179.39 mg g-1, 109.29 mg g-1 and 287.19 mg g-1, respectively. The adsorption dynamics and isotherms of polyphenols on TSNs conformed well to the pseudo-second-order kinetics and the Freundlich isotherms. Moreover, TSN loaded-polyphenols have low cytotoxicity and can continuously be released under stimulated gastric and intestinal conditions. Antioxidant activity tests showed that TSN loaded-polyphenols can remarkably scavenge DPPH free radicals; the IC50 of TSNs-CC, TSNs-EC and TSNs-PAC were 16.11, 16.59 and 16.93 µg mL-1, respectively.
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Catequina , Nanopartículas , Contaminantes Químicos del Agua , Adsorción , Antioxidantes , Catequina/química , Concentración de Iones de Hidrógeno , Cinética , Nanopartículas/química , Polifenoles/química , Almidón/químicaRESUMEN
This review systematically documents the major different strategies of generating high-amylose (HAS) starch mutants aiming at providing high resistant starch, by engineering the starch biosynthesis metabolic pathways. We identify three main strategies based on a new representation of the starch structure: 'the building block backbone model': i) suppression of starch synthases for reduction of amylopectin (AP) side-chains; ii) suppression of starch branching enzymes (SBEs) for production of AM-like materials; and iii) suppression of debranching enzymes to restrain the transformation from over-branched pre-AP to more ordered AP. From a biosynthetic perspective, AM generated through the second strategy can be classified into two types: i) normal AM synthesized mainly by regular expression of granule-bound starch synthases, and ii) modified linear AP chains (AM-like material) synthesized by starch synthases due to the suppression of starch branching enzymes. The application of new breeding technologies, especially CRISPR, in the breeding of HAS crops is also reviewed.
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Enzima Ramificadora de 1,4-alfa-Glucano , Almidón Sintasa , Enzima Ramificadora de 1,4-alfa-Glucano/genética , Enzima Ramificadora de 1,4-alfa-Glucano/metabolismo , Amilopectina/metabolismo , Amilosa/metabolismo , Vías Biosintéticas , Almidón/metabolismo , Almidón Sintasa/genética , Almidón Sintasa/metabolismoRESUMEN
Wheat grain nitrogen content displays large variations within different pearling fractions of grains because of radial gradients in the protein content. We identified how spatiotemporal mechanisms regulate this. The protein gradients emerged clearly at 19 days after anthesis, with the highest N content in aleurone and seed coat, followed by outer endosperm, whereas the lowest was in middle and inner endosperm. Laser microdissection, qRT-PCR and LC-MS were used to dissect tissue from aleurone, outer endosperm, middle endosperm, inner endosperm and transfer cells, measure gene expression and levels of free and protein-bound amino acids, respectively. The results showed that different FAA transportation pathways worked in parallel during grain filling stage while the grain protein gradient did not follow spatial expression of storage proteins. Additionally, two nitrogen (N) topdressing timings were conducted, either at the emergence of top third leaf (standard timing) or top first leaf (delayed timing), finding that delayed N topdressing enhanced both amino acids supply and protein synthesis capacity. The results provide insight into protein synthesis and amino acid transport pathways in endosperm and suggest targets for the enhancement of specialty pearled wheat with higher quality.
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Aminoácidos/metabolismo , Endospermo/química , Proteínas de Plantas/metabolismo , Semillas/química , Triticum/genética , Endospermo/crecimiento & desarrollo , Endospermo/metabolismo , Triticum/química , Triticum/metabolismoRESUMEN
The biosynthesis of starch granules in plant plastids is coordinated by the orchestrated action of transferases, hydrolases, and dikinases. These enzymes either contain starch-binding domain(s) themselves, or are dependent on direct interactions with co-factors containing starch-binding domains. As a means to competitively interfere with existing starch-protein interactions, we expressed the protein module Carbohydrate-Binding Motif 20 (CBM20), which has a very high affinity for starch, ectopically in barley plastids. This interference resulted in an increase in the number of starch granules in chloroplasts and in formation of compound starch granules in grain amyloplasts, which is unusual for barley. More importantly, we observed a photosystem-independent inhibition of CO2 fixation, with a subsequent reduced growth rate and lower accumulation of carbohydrates with effects throughout the metabolome, including lower accumulation of transient leaf starch. Our results demonstrate the importance of endogenous starch-protein interactions for controlling starch granule morphology and number, and plant growth, as substantiated by a metabolic link between starch-protein interactions and control of CO2 fixation in chloroplasts.
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Dióxido de Carbono/metabolismo , Hordeum/genética , Proteínas de Plantas/genética , Plastidios/metabolismo , Almidón/metabolismo , Ciclo del Carbono , Hordeum/metabolismo , Proteínas de Plantas/metabolismoRESUMEN
To investigate the effect of high atmospheric NO concentrations on crop plants and the role of phytoglobins under these conditions, we performed a long-term study on barley 'Golden Promise' wild type (WT), class 1 phytoglobin knockdown (HvPgb1.1-) and class 1 phytoglobin overexpression (HvPgb1.1+) lines. Plants were cultivated with nitrogen-free nutrient solution during the entire growth period and were fumigated with different NO concentration (ambient, 800, 1500, and 3000 ppb). Analysis of fresh weight, stem number, chlorophyll content, and effective quantum yield of PSII showed that NO fumigation promoted plant growth and tillering significantly in the HvPgb1.1+ line. After 80 d of NO fumigation, dry matter weight, spikes number, kernel number, and plant kernel weight were significantly increased in HvPgb1.1+ plants with increasing NO concentration. In contrast, yield decreased in WT and HvPgb1.1- plants the higher the NO level. Application of atmospheric 15NO and 15NO2 demonstrated NO specificity of phytoglobins. 15N from 15NO could be detected in RNA, DNA, and proteins of barley leaves and the 15N levels were significantly higher in HvPgb1.1+ plants in comparison with HvPgb1.1- and WT plants. Our results demonstrate that overexpression of phytoglobins allows plants to more efficiently use atmospheric NO as N source.
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Regulación de la Expresión Génica de las Plantas , Hordeum/genética , Óxido Nítrico/metabolismo , Proteínas de Plantas/genética , Hordeum/metabolismo , Proteínas de Plantas/metabolismoRESUMEN
Nitrogen fertilization regimes significantly affect both grain quality and yield. Wheat plants were subjected to different application timing of topdressed nitrogen at the emergence of the top fifth (TL5), top third (TL3) and top first leaf (TL1), respectively. The iTRAQ (isobaric tag for relative and absolute quantitation) technology was adopted to obtain the complete proteome of wheat flour and to identify the differentially expressed proteins (DEPs) as regulated by nitrogen topdressing timing. Collectively, 591 proteins into 17 functional categories in flour of mature grains were identified. In comparison to TL3, 50 and 63 DEPs were identified in TL5 and TL1, respectively. Nine of the DEPs commonly dependent on nitrogen topdressing timing are the γ-gliadins or high-molecular-weight glutenin subunits. Additionally, delaying nitrogen topdressing modified the grain hardness and allergic protein content. The results suggested that altering nitrogen topdressing timing is a potential strategy for pursuing targeted processing quality of wheat flour.
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Grano Comestible/efectos de los fármacos , Grano Comestible/metabolismo , Glútenes/metabolismo , Dureza/efectos de los fármacos , Nitrógeno/farmacología , Harina/análisis , Calidad de los Alimentos , Hojas de la Planta/metabolismo , Proteómica , Factores de TiempoRESUMEN
Plant starch is the main energy contributor to the human diet. Its biosynthesis is catalyzed and regulated by co-ordinated actions of several enzymes. Recently, a factor termed Protein Targeting to Starch 1 (PTST1) was identified as being required for correct granule-bound starch synthase (GBSS) localization and demonstrated to be crucial for amylose synthesis in Arabidopsis. However, the function of its homologous protein in storage tissues (e.g. endosperm) is unknown. We identified a PTST1 homolog in barley and it was found to contain a crucial coiled-coil domain and carbohydrate-binding module. We demonstrated the interaction between PTST1 and GBSS1 by fluorescence resonance energy transfer (FRET) in barley endosperm. By tagging PTST1 with the fluorophore mCherry, we observed that it is localized in the stroma of barley endosperm amyloplasts. PTST1 overexpression in endosperm increased endogenous gbss1a gene expression and amylose content. Gbss1a and ptst1 mutants were generated using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-related protein 9 (Cas9)-based targeted mutagenesis. Homozygous gbss1a mutants showed a waxy phenotype. Grains of ptst1 mutants did not accumulate any starch. These grains dried out during the desiccation stage and were unable to germinate, suggesting that PTST1 is essential for development of starchy endosperm and viable grains.
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Endospermo/crecimiento & desarrollo , Hordeum/metabolismo , Proteínas de Plantas/metabolismo , Almidón/biosíntesis , Cloroplastos/metabolismo , Grano Comestible/crecimiento & desarrollo , Endospermo/metabolismo , Hordeum/genética , Hordeum/crecimiento & desarrollo , Fenotipo , Hojas de la Planta/metabolismoRESUMEN
BACKGROUND: Nitrogen is one basic element of amino acids and grain protein in wheat. In field experiments, wheat plants were subjected to different timing of nitrogen topdressing treatments: at the stages of emergence of the top fifth leaf (TL5), top third leaf (TL3) and top first leaf (TL1) to test the regulatory effects of nitrogen topdressing timing on grain protein quality. The underlying mechanisms were elucidated by clarifying the relationship between proteolysis in vegetative organs and accumulation of amino acids in the endosperm cavity, conversion of amino acids, and storage protein synthesis in endosperm of wheat grain. RESULTS: Delayed nitrogen topdressing up-regulated gene expression related to nitrogen metabolism and protease synthesis in the flag leaf, followed by more free amino acids being transported to both the cavity and the endosperm from 7 days after anthesis (DAA) to 13 DAA in TL1. TL1 enhanced the conversion between free amino acids in endosperm and upregulated the expression of genes encoding high molecular weight (HMW) and low molecular weight (LMW) subunits and protein disulfide isomerases-like (PDIL) proteins, indicating that the synthesis and folding of glutenin were enhanched by delayed nitrogen topdressing. As a consequense, the content of glutenin macropolymers (GMP) and glutenin increased with delaying nitrogen topdressing. CONCLUSIONS: The results highlight the relationship between nitrogen remobilization and final grain protein production and suggest that the nitrogen remobilization processes could be a potential target for improving the quality of wheat grain. Additionally, specific gene expression related to nitrogen topdressing was identified, which conferred more detailed insights into underlying mechanism on the modification protein quality.
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Aminoácidos/metabolismo , Grano Comestible/metabolismo , Nitrógeno/metabolismo , Proteínas de Plantas/metabolismo , Triticum/metabolismo , Aminoácidos/análisis , Grano Comestible/química , Endospermo/química , Endospermo/metabolismo , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Nitrógeno/administración & dosificación , Hojas de la Planta/metabolismo , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
Grasses such as wheat and barley constitute some of the main crops currently cultivated worldwide. Cultivation of wild cereals such as Hordeum spontaneum (wild barley) and Triticum diccocoides (wild emmer) started in southwest Asia. Despite the many studies on the origins of agriculture and plant domestication, surprisingly few studies have discussed the importance of the nutritional quality of barley and emmer wheat in their evolution. A comparison of domesticated forms of these cereals with their wild progenitors suggests an evolution towards bigger grains with higher glycemic loads in the form of higher relative starch content with lower relative protein, fiber and mineral contents. In this work we hypothesize that in addition to the simple explanation that larger grains emerged under domestication, complex and indirect effects such as increased glycemic index and sweet taste should also be taken into consideration.
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Evolución Biológica , Dieta , Domesticación , Grano Comestible/química , Hordeum/química , Valor Nutritivo , Triticum/química , Agricultura , Productos Agrícolas , Índice Glucémico , GustoRESUMEN
Nitric oxide (NO) is an important signalling molecule that is involved in many different physiological processes in plants. Here, we report about a NO-fixing mechanism in Arabidopsis, which allows the fixation of atmospheric NO into nitrogen metabolism. We fumigated Arabidopsis plants cultivated in soil or as hydroponic cultures during the whole growing period with up to 3 ppmv of NO gas. Transcriptomic, proteomic and metabolomic analyses were used to identify non-symbiotic haemoglobin proteins as key components of the NO-fixing process. Overexpressing non-symbiotic haemoglobin 1 or 2 genes resulted in fourfold higher nitrate levels in these plants compared with NO-treated wild-type. Correspondingly, rosettes size and weight, vegetative shoot thickness and seed yield were 25, 40, 30, and 50% higher, respectively, than in wild-type plants. Fumigation with 250 ppbv 15 NO confirmed the importance of non-symbiotic haemoglobin 1 and 2 for the NO-fixation pathway, and we calculated a daily uptake for non-symbiotic haemoglobin 2 overexpressing plants of 250 mg N/kg dry weight. This mechanism is probably important under conditions with limited N supply via the soil. Moreover, the plant-based NO uptake lowers the concentration of insanitary atmospheric NOx, and in this context, NO-fixation can be beneficial to air quality.
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Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Hemoglobinas/metabolismo , Óxido Nítrico/metabolismo , Nitrógeno/farmacología , Simbiosis , Amoníaco/metabolismo , Arabidopsis/efectos de los fármacos , Arabidopsis/genética , Arabidopsis/crecimiento & desarrollo , Fumigación , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Nitratos/metabolismo , Óxido Nítrico/farmacología , Nitritos/metabolismo , Fenotipo , Hojas de la Planta/efectos de los fármacos , Hojas de la Planta/metabolismo , Propanoles/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , S-Nitrosotioles/metabolismoRESUMEN
Plants contain three classes of hemoglobin genes of which two, class 1 and class 2, have a structure similar to classical vertebrate globins. We investigated the effect of silencing the class 1 non-symbiotic hemoglobin gene, GLB1, and the effect of overexpression of GLB1 or the class 2 non-symbiotic hemoglobin gene, GLB2, in Arabidopsis thaliana. Lines with GLB1 silencing had a significant delay of bolting and after bolting, shoots reverted to the rosette vegetative phase by formation of aerial rosettes at lateral meristems. Lines with overexpression of GLB1 or GLB2 bolted earlier than wild type plants. By germinating the lines in a medium containing the nitric oxide (NO) donor, sodium nitroprusside (SNP), it was demonstrated that both GLB1 and GLB2 promote bolting by antagonizing the effect of NO, suggesting that non-symbiotic plant hemoglobin controls bolting by scavenging the floral transition signal molecule, NO. So far, NO scavenging has only been demonstrated for class 1 non-symbiotic hemoglobins. A direct assay in Arabidopsis leaf cells shows that GLB1 as well as the class 2 non-symbiotic hemoglobin, GLB2, scavenge NO in vivo. NO has also been demonstrated to be a growth stimulating signal with an optimum at low concentrations. It was observed that overexpression of either GLB1 or GLB2 shifts the optimum for NO growth stimulation to a higher concentration. In conclusion, we have found that expression of NO scavenging plant hemoglobin is involved in the control of bolting in Arabidopsis.