RESUMO
Amaranth is a pseudocereal that contains between 50 and 60% starch, gluten-free protein, and essential amino acids. This study investigates the physicochemical changes in Amaranthus spp. grains, flour, isolated starch and nanocrystals during germination and malting. The moisture content increased from 8.9% to 41% over 2 h of soaking. The percentage of germination increased rapidly, reaching 96% after 60 h, a remarkable advantage over other cereals. The nutrient composition varied, including protein synthesis and lipid degradation. Lipid concentration decreased during malting, except for soaking, which increased by 62%. Scanning electron microscopy shows that germination does not cause morphological changes on the outer surface of the grains, while transmission electron microscopy indicates the presence of isolated nanocrystals with orthorhombic crystal structure confirmed by X-ray diffraction. The viscosity profile shows a decrease in peak viscosity. Therefore, amaranth is a potential pseudocereal that can be used as an additive in the production of fermented beverages.
Assuntos
Amaranthus , Farinha , Germinação , Nanopartículas , Amido , Amaranthus/química , Amaranthus/crescimento & desenvolvimento , Amaranthus/metabolismo , Farinha/análise , Amido/química , Amido/metabolismo , Nanopartículas/química , Viscosidade , Sementes/química , Sementes/crescimento & desenvolvimento , Sementes/metabolismo , Manipulação de AlimentosRESUMO
Water and nitrogen availability are two major environmental factors that can impair plant growth, and when combined, their effects on plant performance can be either intensified or reduced. The objective of this study was to analyze the influence of nitrogen availability on the responses of Amaranthus cruentus's metabolism to water stress. The plants were cultivated in plastic pots filled with vermiculite, kept under greenhouse conditions, and were watered three times a week with 70% of a full strength nitrogen-free Long Ashton solution, containing 1.97 or 9.88 kg N ha-1 as ammonium nitrate. Photosynthetic parameters were evaluated in planta, and leaves were harvested for chemical analysis of photosynthetic pigments, proline, and phenolic contents. Higher nitrogen supply increased the shoot dry matter, photosynthetic pigments, photosynthesis, stomatal conductance, transpiration, total leaf nitrogen, proline, nitrate, and ammonium but reduced the concentration of flavonoids and total phenols. Six days of water stress did not affect dry matter, photosynthetic pigments, leaf nitrogen, ammonium, or specialized metabolites but increased the proline under high nitrogen and negatively affected stomatal conductance, transpiration, photosynthesis, relative water content, instantaneous water use efficiency, and leaf nitrate. The negative effect was more pronounced under high nitrogen supply. The results show that the addition of a high amount of nitrogen made the physiological processes of plants more sensitive to water stress, indicating that the plant response to water restriction depends on the interaction between the different environmental stressors to which the plants are subjected.
Assuntos
Amaranthus , Compostos de Amônio , Amaranthus/metabolismo , Desidratação , Secas , Nitratos , Nitrogênio , Prolina/metabolismo , Estresse FisiológicoRESUMO
BACKGROUND: Amaranth is a plant naturally resistant to various types of stresses that produces seeds of excellent nutritional quality, so amaranth is a promising system for food production. Amaranth wild relatives have survived climate changes and grow under harsh conditions, however no studies about morphological and molecular characteristics of their seeds are known. Therefore, we carried out a detailed morphological and molecular characterization of wild species A. powellii and A. hybridus, and compared them with the cultivated amaranth species A. hypochondriacus (waxy and non-waxy seeds) and A. cruentus. RESULTS: Seed proteins were fractionated according to their polarity properties and were analysed in one-dimensional gel electrophoresis (1-DE) followed by nano-liquid chromatography coupled to tandem mass spectrometry (nLC-MS/MS). A total of 34 differentially accumulated protein bands were detected and 105 proteins were successfully identified. Late embryogenesis abundant proteins were detected as species-specific. Oleosins and oil bodies associated proteins were observed preferentially in A. cruentus. Different isoforms of the granule-bound starch synthase I, and several paralogs of 7S and 11S globulins were also identified. The in silico structural analysis from different isoforms of 11S globulins was carried out, including new types of 11S globulin not reported so far. CONCLUSIONS: The results provide novel information about 11S globulins and proteins related in seed protection, which could play important roles in the nutritional value and adaptive tolerance to stress in amaranth species.
Assuntos
Amaranthus/metabolismo , Proteínas de Plantas/metabolismo , Sementes/metabolismo , Cromatografia Líquida , Eletroforese , Globulinas/análise , Globulinas/isolamento & purificação , Globulinas/metabolismo , Proteínas de Plantas/análise , Proteínas de Plantas/isolamento & purificação , Sementes/química , Espectrometria de Massas em TandemRESUMO
Amaranthus palmeri S. Watson is probably the worst glyphosate-resistant (GR) weed worldwide. The EPSPS (5-enolpyruvylshikimate-3-phosphate-synthase) gene amplification has been reported as the major target-site-resistance (TSR) mechanism conferring resistance to glyphosate in this species. In this study, TSR and non-target-site-resistance (NTSR) mechanisms to glyphosate were characterized in a putative resistant A. palmeri population (GRP), harvested in a GR soybean crop from Argentina. Glyphosate resistance was confirmed for the GRP population by dose-response assays. No evidence of TSR mechanisms, as well as glyphosate metabolism, was found in this population. Moreover, a susceptible population (GSP) that absorbed about 10% more herbicide than the GRP population was evaluated at different periods after treatment. The GSP population translocated about 20% more glyphosate to the remainder of the shoots and roots at 96 h after treatment than the control, while the GRP population retained 62% of herbicide in the treated leaves. This is the first case of glyphosate resistance in A. palmeri involving exclusively NTSR mechanisms.
Assuntos
Amaranthus/metabolismo , Glycine max/efeitos dos fármacos , Glicina/análogos & derivados , Herbicidas/metabolismo , Plantas Daninhas/metabolismo , 3-Fosfoshikimato 1-Carboxiviniltransferase/genética , 3-Fosfoshikimato 1-Carboxiviniltransferase/metabolismo , Amaranthus/efeitos dos fármacos , Argentina , Transporte Biológico , Glicina/metabolismo , Glicina/farmacologia , Resistência a Herbicidas , Herbicidas/farmacologia , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Daninhas/efeitos dos fármacos , Glycine max/crescimento & desenvolvimento , GlifosatoRESUMO
Organic amaranth starch (Amaranthus caudatus) was studied after heat-moisture treatment (HMT) using different moisture contents and different times. The starch extracted by the aqueous method presented low lipid and protein content. After HMT, an increase in the thermal stability was identified. The onset and peak temperatures were higher with an increase in moisture content and the times used in the modification. The gelatinisation enthalpy varied due to the heterogeneity of the crystals formed after the structural reorganisation caused by HMT. The relative crystallinity was lower for the physically modified starches. An increase in the pasting temperature was accompanied by a decrease in the viscosity, setback and breakdown, which were proportional to the moisture and time used. The morphology of the HMT-modified samples was not altered; however, agglomerations were noted. Low levels of dispersion homogeneity and suspension stability were observed for the modified samples due to the strong presence of agglomerates.
Assuntos
Amido/análise , Água/química , Amaranthus/metabolismo , Varredura Diferencial de Calorimetria , Microscopia Eletrônica de Varredura , Tamanho da Partícula , Amido/química , Temperatura , Termogravimetria , Viscosidade , Difração de Raios XRESUMO
Tomato yellow leaf curl virus (TYLCV; genus Begomovirus; family Geminiviridae) infects mainly plants of the family Solanaceae, and the infection induces curling and chlorosis of leaves, dwarfing of the whole plant, and reduced fruit production. Alternatives for direct control of TYLCV and other geminiviruses have been reported, for example, the use of esterified whey proteins, peptide aptamer libraries or artificial zinc finger proteins. The two latter alternatives affect directly the replication of TYLCV as well as of other geminiviruses because the replication structures and sequences are highly conserved within this virus family. Because peptides and proteins offer a potential solution for virus replication control, in this study we show the isolation, biochemical characterization and antiviral activity of a peptide derived from globulins of amaranth seeds (Amaranthus hypochondriacus) that binds to the replication origin sequence (OriRep) of TYLCV and affects viral replication with a consequent reduction of disease symptoms in Nicotiana benthamiana. Aromatic peptides obtained from papain digests of extracted globulins and albumins of amaranth were tested by intrinsic fluorescent titration and localized surface resonance plasmon to analyze their binding affinity to OriRep of TYLCV. The peptide AmPep1 (molecular weight 2.076 KDa) showed the highest affinity value (Kdâ¯=â¯1.8â¯nM) for OriRep. This peptide shares a high amino acid similarity with a part of an amaranth 11S globulin, and the strong affinity of AmPep1 could be explained by the presence of tryptophan and lysine facilitating interaction with the secondary structure of OriRep. In order to evaluate the effect of the peptide on in vitro DNA synthesis, rolling circle amplification (RCA) was performed using as template DNA from plants infected with TYLCV or another begomovirus, pepper huasteco yellow vein virus (PHYVV), and adding AmPep1 peptide at different concentrations. The results showed a decrease in DNA synthesis of both viruses at increasing concentrations of AmPep1. To further confirm the antiviral activity of the peptide in vivo, AmPep1 was infiltrated into leaves of N. benthamiana plants previously infected with TYLCV. Plants treated with AmPep1 showed a significant decrease in virus titer compared with untreated N. benthamiana plants as well as reduced symptom progression due to the effect of AmPep1 curtailing TYLCV replication in the plant. The peptide also showed antiviral activity in plants infected with PHYVV. This is the first report, in which a peptide is directly used for DNA virus control in plants, supplied as exogenous application and without generation of transgenic lines.
Assuntos
Amaranthus/metabolismo , Begomovirus/genética , Globulinas/metabolismo , Nicotiana/virologia , Peptídeos/metabolismo , Origem de Replicação , Replicação Viral , Antivirais/farmacologia , Begomovirus/efeitos dos fármacos , Begomovirus/isolamento & purificação , Begomovirus/fisiologia , Sítios de Ligação , Produtos Agrícolas/efeitos dos fármacos , Produtos Agrícolas/virologia , Peptídeos/isolamento & purificação , Peptídeos/farmacologia , Extratos Vegetais/metabolismo , Nicotiana/efeitos dos fármacos , Carga Viral/efeitos dos fármacosRESUMO
Introdução: Doenças cardiovasculares constituem importante causa de morte em todo mundo e a hipercolesterolemia está diretamente relacionada como fator agravante desta morbidade. A dieta desempenha papel importante neste processo e alguns alimentos como o amaranto, especialmente sua proteína, tem mostrado capacidade de redução do colesterol plasmático. Estudos sugerem que este efeito está relacionado a peptídeos formados durante a digestão da sua proteína, os quais desempenham um papel importante na regulação e modulação do metabolismo lipídico. Os efeitos hipocolesterolêmicos, já observados, indicam o uso da proteína do amaranto como um composto bioativo direcionado para a promoção da saúde. Considerando que os efeitos hipocolesterolêmicos destes peptídeos são complexos e há diversas hipóteses formuladas, torna-se importante a realização de estudos visando avaliar a interação dos peptídeos na absorção intestinal do colesterol e da sua modulação genética. Objetivos: Verificar os efeitos do hidrolisado da farinha do grão de amaranto na absorção de colesterol e modulação de genes ABCA1, ABCG1, NPC1L1, AMPK, HMGR e SREBP-2em células Caco-2, e modulação dos genes ABCG8, HMGR, SREBP-2 e AMPKem enterócitos de hamsters. Metodologia: O amaranto foi triturado, sua farinha desengordurada e sua proteína isolada, com posterior digestão in vitro e filtração dos peptídeos. Três experimentos in vitro foram conduzidos com as células: permeação de hidrolisado, permeação de colesterol e de efeito sob a expressão gênica. No primeiro, o hidrolisado proteico de amaranto foi permeado em culturas celulares de Caco-2 no tempo de 2 horas. O permeato foi coletado e analisado por LC/MS/MS. No segundo, o hidrolisado de amaranto foi incorporado a micelas de colesterol e incubados em culturas celulares, nas concentrações de 1,0 mg/ml, e 3,0 mg/ml em tempos de 2h. Também em concentrações de 3,0 mg/ml foi adicionado albumina e caseína para efeito comparativo. O conteúdo de colesterol na porção apical e basolateral foi analisado em HPLC. O terceiro experimento foi avaliaçãoda exposição do hidrolisado, em concentrações de 0,5 mg/ml, 1,0 mg/ml e 3,0 mg/ml, em tempos de 2h e 12h. Após este período, foi realizada a extração de RNA total, avaliação de rendimento e integridade do material; medida quantitativa de expressão de RNAm por RT-PCR e quantificação relativa da expressão por ?CT dos genes ABCA1, ABCG1, ABCG8, NPC1L1, AMP1, HMGR e SREBP-2das células Caco-2 e tecido intestinal de hamsters, coletados em ensaios anteriores. Resultados: Na permeação de colesterol não houve diferença entre as concentrações dos hidrolisados proteicos e controle, porém o hidrolisado de amaranto em 1,0 mg/ml demonstrou uma tendência em diminuir a absorção de colesterol (p = 0,05). Na exposição das células Caco-2 aos peptídeos por 2h, houve uma diminuição nas concentrações de RNAm dos genes ABCA1, NPC1L1, AMPK, HMGR e SREBP-2 nas concentrações de 3,0 mg/ml. O tempo de exposição de 12h apresentou resultados semelhantes ao tempo de 2h. Somente a expressão gênica de ABCG8foi influenciada pelo isolado proteico de amaranto no experimento in vivo. Conclusão: A partir do exposto, podemos concluir que os peptídeos do grão de amaranto influenciam o metabolismo de colesterol por mecanismos genéticos. Portanto, torna-se uma alternativa a ser introduzida na dieta de indivíduos saudáveis e em pacientes com hipercolesterolemia, visando a prevenção de agravos e como estratégia de terapia adicional no controle dos níveis de LDL-c plasmático. Contudo, mais experimentos in vivo e em humanos são necessários para estabelecer a dose efetiva para consumo
Introduction: Cardiovascular diseases are an important cause of death worldwide and hypercholesterolemia is directly related as an aggravating factor of this morbidity. Diet plays an important role in this process and some foods such as amaranth, especially its protein, have shown ability to lower plasma cholesterol. Studies suggest that this effect is related to peptides formed during the digestion of their protein, which play an important role in the regulation and modulation of lipid metabolism. The hypocholesterolemic effects, already observed, indicate the use of amaranth protein as a bioactive compound aimed to promoting health. Considering that the hypocholesterolemic effects of these peptides are complex and there are several hypotheses formulated, it is important to carry out studies to evaluate the interaction of peptides in the intestinal absorption of cholesterol and its genetic modulation. Objectives: To verify the effects of amaranth grain flour hydrolyzate on cholesterol uptake and ABCA1, ABCG1, NPC1L1, AMPK, HMGR and SREBP-2 genes modulation in Caco-2 intestinal cells, and modulation of ABCG8, HMGR, SREBP-2 genes and AMPK in hamster intestinal cells. Methodology: Amaranth was crushed, the created flour was defatted and its protein isolated, with subsequent in vitro digestion and filtration of the peptides. Three in vitro experiments were conducted with the cells: hydrolyzate permeation, cholesterol permeation and genetic expression. In the first, the amaranth protein hydrolyzate was permeated in Caco-2 cell cultures in the time of 2 hours. The permeate was collected and analyzed by LC/MS/MS. In the second, the amaranth hydrolyzate was incorporated into cholesterol micelles and incubated in cell cultures at concentrations of 1.0 mg/ml and 3.0 mg/ml in times of 2 h. Also, at concentrations of 3.0 mg/ml albumin and casein were added for comparison. Cholesterol content in the apical and basolateral portion was analyzed by HPLC. The third experiment was to evaluate the exposure of the hydrolyzate at concentrations of 0.5 mg/ml, 1.0 mg/ml and 3.0 mg/ml, in times of 2 h and 12 h. After this period, the extraction of total RNA, evaluation of yield and integrity of the material was performed; quantitative measurement of mRNA expression by RT-PCR and relative quantification of ?CT expression of the ABCA1, ABCG1, ABCG8, NPC111, AMPK, HMGR and SREBP-2 genes from Caco-2 cells and hamster intestinal tissue, collected in previous assays, were finalized. Results: In cholesterol permeation there was no difference between the concentrations of the protein hydrolysates and control, but the amaranth hydrolyzate at 1.0 mg/ml showed a tendency to decrease the cholesterol absorption (p = 0.05). Exposure of Caco-2 cells to peptides for 2 h resulted in a decrease in ABCA1, NPC111, AMPK, HMGR and SREBP-2 mRNA levels at concentrations of 3.0 mg/ml. The exposure time of 12h presented results similar to the time of 2h. Only the gene expression of ABCG8 was influenced by the amaranth protein isolate in the in vivo experiment. Conclusion: From the above, we can conclude that amaranth peptides influence the metabolism of cholesterol by genetic mechanisms. Therefore, it becomes an alternative to be introduced in the diet of healthy individuals and in patients with hypercholesterolemia, aiming at the prevention of aggravations and as a strategy of additional therapy in the control of plasma LDL-c levels. However, more studies should bedone with animals and humans to define the dose-efficiency for diet.
Assuntos
Peptídeos , Expressão Gênica , Colesterol , Transporte Proteico , Amaranthus/metabolismo , Hipercolesterolemia , Técnicas In VitroRESUMO
The ability of different fruit by-products, okara, and amaranth flour, to support the growth of probiotic and non-probiotic strains was evaluated. The tests were conducted with three commercial starter cultures (Streptococcus thermophilus), ten probiotic strains (seven Lactobacillus spp. and three Bifidobacterium spp. strains), and two harmful bacteria representative of the intestinal microbiota (Escherichia coli and Clostridium perfringens). In vitro fermentability assays were performed using a modified MRS broth supplemented with different fruits (acerola, orange, passion fruit, and mango), and soy (okara) by-products or amaranth flour. Orange and passion-fruit by-products were the substrates that most promoted the growth of bacterial populations, including pathogenic strains. On the other hand, the acerola by-product was the substrate that showed the highest selectivity for beneficial bacteria, since the E. coli and Cl. perfringens populations were lower in the presence of this fruit by-product. Although the passion fruit by-product, okara, and amaranth stimulated the probiotic strains, the growth of the pathogenic strains studied was higher compared to other substrates. Different growth profiles were verified for each substrate when the different strains were compared. Although pure culture models do not reflect bacterial interaction in the host, this study reinforces the fact that the ability to metabolize different substrates is strain-dependent, and acerola, mango, and orange by-products are the substrates with the greatest potential to be used as prebiotic ingredients.
Assuntos
Amaranthus/química , Técnicas de Cultura de Células/métodos , Frutas/química , Glycine max/química , Probióticos , Agricultura , Amaranthus/metabolismo , Bactérias/química , Bactérias/crescimento & desenvolvimento , Bactérias/metabolismo , Farinha , Frutas/metabolismo , Resíduos Industriais , Probióticos/química , Probióticos/metabolismo , Glycine max/metabolismoRESUMO
Following the introduction of glyphosate-resistant (GR)-cotton crops in Mexico, farmers have relied upon glyphosate as being the only herbicide for in-season weed control. Continuous use of glyphosate within the same year and over multiple successive years has resulted in the selection of glyphosate resistance in Palmer amaranth (Amarantus palmeri). Dose-response assays confirmed resistance in seven different accessions. The resistance ratio based on GR50 values (50% growth reduction) varied between 12 and 83. At 1000 µM glyphosate, shikimic acid accumulation in the S-accession was 30- to 2-fold higher at compared to R-accessions. At 96 h after treatment, 35-44% and 61% of applied 14C-glyphosate was taken up by leaves of plants from R- and S-accessions, respectively. At this time, a significantly higher proportion of the glyphosate absorbed remained in the treated leaf of R-plants (55-69%) compared to S-plants (36%). Glyphosate metabolism was low and did not differ between resistant and susceptible plants. Glyphosate was differentially metabolized to AMPA and glyoxylate in plants of R- and S-accessions, although it was low in both accessions (<10%). There were differences in 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) enzyme activity by 50% (I50) between R- and S-accessions. However, no significant differences were found in the basal EPSPS activity (µmol inorganic phosphate µg-1 total soluble protein min-1) between R- and S-accessions. A point mutation Pro-106-Ser was evidenced in three accessions. The results confirmed the resistance of Palmer amaranth accessions to glyphosate collected from GR-cotton crops from Mexico. This is the first study demonstrating glyphosate-resistance in Palmer amaranth from Mexico.
Assuntos
Amaranthus/efeitos dos fármacos , Glicina/análogos & derivados , Herbicidas/farmacologia , Amaranthus/genética , Amaranthus/metabolismo , Glicina/farmacologia , Resistência a Herbicidas , Mutação , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Daninhas/genética , Ácido Chiquímico , GlifosatoRESUMO
Biogas digestates contain valuable nutrients but also have high water contents. Di-gestates were sampled from two different biogas facilities before and after solid-liquid separation and were analyzed with regard to their composition and phosphorus (P) fractions. Additionally, to investigate the P fertilizer effects of these digestates in comparison with undigested slurry or TripleSuper-P (TSP), they were applied in a pot experiment (6 kg soil per pot) in an amount corresponding to 200 mg P per pot in combination with various crops (amaranth, maize, maize + beans mixed cropping, sorghum). A separation of digestates resulted in higher P concentrations of the solid fraction in comparison with the liquid fraction. The proportion of the readily soluble P fractions (H2O-P, NaHCO3-P) to the total P was higher than 70 % in all digestates. The digestates increased P uptake of the tested crops and concentrations of bioavailable P in the soil to the same extent as highly soluble TSP. Activities of soil enzymes were lower after application of the digestates in comparison to unfermented slurry. The fertilizer management of digestates can be improved by a solid-liquid separation since the solid fraction showed a relatively high concentration of P resulting in a reduction in application doses required to meet the P demands of crops.