Expression of AtWRI1 and AtDGAT1 during soybean embryo development influences oil and carbohydrate metabolism

Soybean oil is one of the most consumed vegetable oils worldwide. Genetic improvement of its concentration in seeds has been historically pursued due to its direct association with its market value. Engineering attempts aiming to increase soybean seed oil presented different degrees of success that varied with the genetic design and the specific variety considered. Understanding the embryo’s responses to the genetic modifications introduced, is a critical step to successful approaches. In this work, the metabolic and transcriptional responses to AtWRI1 and AtDGAT1 expression in soybean seeds were evaluated. AtWRI1 is a master regulator of fatty acid (FA) biosynthesis, and AtDGAT1 encodes an enzyme catalysing the final and rate-limiting step of triacylglycerides biosynthesis. The events expressing these genes in the embryo did not show an increase in total FA content, but they responded with changes in the oil and carbohydrate composition. Transcriptomic studies revealed a down-regulation of genes putatively encoding for oil body packaging proteins, and a strong induction of genes annotated as lipases and FA biosynthesis inhibitors. Novel putative AtWRI1 targets, presenting an AW-box in the upstream region of the genes, were identified by comparison with an event that harbours only AtWRI1. Lastly, targeted metabolomics analysis showed that carbon from sugar phosphates could be used for FA competing pathways, such as starch and cell wall polysaccharides, contributing to the restriction in oil accumulation. These results allowed the identification of key cellular processes that need to be considered to break the embryo’s natural restriction to uncontrolled seed lipid increase

Expression of \u3ci\u3eAtWRI1\u3c/i\u3e and \u3ci\u3eAtDGAT1\u3c/i\u3e during soybean embryo development influences oil and carbohydrate metabolism

Soybean oil is one of the most consumed vegetable oils worldwide. Genetic improvement of its concentration in seeds has been historically pursued due to its direct association with its market value. Engineering attempts aiming to increase soybean seed oil presented different degrees of success that varied with the genetic design and the specific variety considered. Understanding the embryo’s responses to the genetic modifications introduced, is a critical step to successful approaches. In this work, the metabolic and transcriptional responses to AtWRI1 and AtDGAT1 expression in soybean seeds were evaluated. AtWRI1 is a master regulator of fatty acid (FA) biosynthesis, and AtDGAT1 encodes an enzyme catalysing the final and rate-limiting step of triacylglycerides biosynthesis. The events expressing these genes in the embryo did not show an increase in total FA content, but they responded with changes in the oil and carbohydrate composition. Transcriptomic studies revealed a down-regulation of genes putatively encoding for oil body packaging proteins, and a strong induction of genes annotated as lipases and FA biosynthesis inhibitors. Novel putative AtWRI1 targets, presenting an AW-box in the upstream region of the genes, were identified by comparison with an event that harbours only AtWRI1. Lastly, targeted metabolomics analysis showed that carbon from sugar phosphates could be used for FA competing pathways, such as starch and cell wall polysaccharides, contributing to the restriction in oil accumulation. These results allowed the identification of key cellular processes that need to be considered to break the embryo’s natural restriction to uncontrolled seed lipid increase

Workflow for the Quantification of Soluble and Insoluble Carbohydrates in Soybean Seed

Soybean seed composition has a profound impact on its market value and commercial use as an important commodity. Increases in oil and protein content have been historically pursued by breeders and genetic engineers; consequently, rapid methods for their quantification are well established. The interest in complete carbohydrate profiles in mature seeds, on the other hand, has recently increased due to numerous attempts to redirect carbohydrates into oil and protein or to offer specialty seed with a specific sugar profile to meet animal nutritional requirements. In this work, a sequential protocol for quantifying reserve and structural carbohydrates in soybean seed was developed and validated. Through this procedure, the concentrations of soluble sugars, sugar alcohols, starch, hemicellulose, and crystalline cellulose can be determined in successive steps from the same starting material using colorimetric assays, LC–MS/MS, and GC–MS. The entire workflow was evaluated using internal standards to estimate the recovery efficiency. Finally, it was successfully applied to eight soybean genotypes harvested from two locations, and the resulting correlations of carbohydrate and oil or protein are presented. This methodology has the potential not only to guide soybean cultivar optimization processes but also to be expanded to other crops with only slight modifications

Microstructures and properties of direct laser sintered tungsten carbide (WC) particle reinforced Cu matrix composites with RE-Si-Fe addition : A comparative study

The poor wettability between ceramics and metals is a main obstacle in obtaining high-performance metal-matrix composites (MMCs) parts using direct metal laser sintering (DMLS). Rare earth (RE) elements, due to their unique physical and chemical properties, have high potential for improving laser processability of MMCs. In this work, a comparative study was performed to investigate the influence of RE-Si-Fe addition on microstructural features and mechanical properties of DMLS processed tungsten carbide (WC) particle reinforced Cu MMCs parts. It showed that by adding 3 wt% RE-Si-Fe, the WC reinforcing particles were refined, the particle dispersion state was homogenized, and the particle/matrix interfacial compatibility was enhanced. The RE-Si-Fe-containing WC/Cu MMCs parts possessed significantly elevated mechanical properties, i.e., densification level of 95.7%, microhardness of 417.6 HV, fracture strength of 201.8 MPa, and friction coefficient of 0.8. The metallurgical functions of the RE-Si-Fe additive for the improvement of DMLS quality of MMCs parts were discussed

Phosphorus availability of rock phosphates as compared with feed-grade phosphates for swine Disponibilidade de fósforo em fosfatos de rocha em comparação à de fosfatos bicálcicos para suínos

One hundred ninety two swine were used in a trial to assess the relative bioavailability of phosphorus (RBP) in six phosphate sources. Phosphates were three feed grade phosphates (FP), two made in Brasil, and one USA made, and three rock phosphate samples (RP) originated from two mines sites in Brasil, and one mine site in Israel. Levels of calcium, phosphorus and fluorine in RP were 29, 12 and 1.7% (RP source 1), 33, 14 and 1.4% (RP source 2), and 30, 14 and 3.6% (RP source 3), respectively. Pigs were fed a corn-soybean meal basal diet (18% CP, 0.95% Lys, 0.75% Ca, 0.37% P) or the basal diet with 0.15% P from a standard purified grade calcium phosphate (SP), or with 0.15% P from experimental FP or RP. Each diet was fed to six pen replicates of four pigs per pen for 35 days (14.4 to 39.9 kg). Weight gain (WG), feed/gain (FG), plasma P (PP), bone ash (BA), and breaking strength of metacarpals and metatarsals (BS-MM) and femurs (BS-F) were improved by phosphorus addition. However, performance and bone parameters were depressed by RP, as compared to FP dietary supplementation. WG, BA, BS-MM and BS-F were regressed to P added, and slope-ratios were calculated to assess RBP in the FP and RP sources. The average bioavailability of P in the FP and RP sources, relative to SP, were 89 and 49% (WG), 112 and 49% (BA), 78 and 28% (BS-MM), and 101 and 52% (BS-F), respectively. Low animal performance and bone strength related to toxicity should be expected if rock phosphates are used to feed pigs.<br>Cento e noventa e dois leitões foram usados em um experimento para avaliar a biodisponibilidade relativa do fósforo (RBP) em seis fontes fosfáticas. As fontes foram três fosfatos de uso em nutrição (FP), dois fabricados no Brasil e um nos Estados Unidos, e três amostras de fosfatos de rocha (RP), originados de duas minas brasileiras e uma mina situada em Israel. Os níveis de cálcio (Ca), fósforo (P) e flúor (F) nos fosfatos de rocha foram 29, 12 e 1,7% (fonte RP 1), 33, 14 e 1,4% (fonte RP 2) e 30, 14 e 3,6% (fonte RP 3), respectivamente. Os suínos foram alimentados com uma dieta basal de milho e farelo de soja (18% PB, 0,95% Lis, 0,75% Ca, 0,37% P) ou dieta basal contendo 0,15% P a partir de um fosfato bicálcico purificado padrão (SP), ou com 0,15% P a partir das fontes experimentais FP ou RP. Cada dieta foi fornecida a seis baias (replicatas) com quatro leitões durante 35 dias (14,4 a 39,9 kg). O ganho de peso (WG), a conversão alimentar (FG), o P plasmático (PP), as cinzas ósseas (BA) e a resistência óssea à quebra de metacarpos e metatarsos (BS-MM) e fêmures (BS-F) melhoraram com a adição de fósforo às dietas. Contudo, o desempenho e os parâmetros ósseos pioraram com o uso dos fosfatos de rocha, em comparação à suplementação dietética de FP. A análise de regressão dos dados de WG, BA, BS-MM e BS-F em relação ao P adicional foi realizada e foram calculadas slope-ratios para avaliar a RBP nas fontes de FP e RP. A biodisponibilidade média do P nas fontes FP e RP, relativas ao SP (com valor atribuído de 100), foram 89 e 49% (WG), 112 e 49% (BA), 78 e 28% (BS-MM), e 101 e 52% (BS-F), respectivamente. Baixos desempenho e resistência óssea relacionados à toxicidade devem ser esperados se fosfatos de rocha forem utilizados na alimentação de suínos