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This study investigated the quality of anaerobic (AnE) and oxic/anoxic (O/A) effluents from a continuous-feed structured-bed hybrid baffled reactor (SBHBR) treating dairy wastewater impacts on lettuce and cucumber germination. While sustainable technologies like SBHBR have successfully removed organic matter and total nitrogen from dairy wastewater, residual concentrations may still represent a risk to water resources. Therefore, phytotoxicity bioassays were conducted with lettuce and cucumber seeds in contact with effluent during early stages to evaluate the potential implications of dairy wastewater reuse in agriculture. The study also explored the potential of SBHBR technology in promoting water resource preservation and creating a sustainable energy and nutrient cycling system. The physicochemical parameters of both effluents were characterized, and the phytotoxicity was evaluated by measuring the germination index (GI), root length (RL), the number of germinated seeds (SG), and epicotyl elongation (EE) for both lettuce and cucumber. The study revealed that the O/A effluent demonstrated lower phytotoxicity than the AnE effluent. The mean results indicate that the O/A zone wastewater was more conducive to cucumber germination than the AnE zone. Moreover, a positive influence of organic matter in the effluent on root growth and epicotyl elongation in cucumber, as well as the presence of nitrogen on the germination index, in both plant species. These findings emphasize the importance of considering effluent characteristics for suitable irrigation, highlighting SBHBR's potential as an effective solution for treating and reusing dairy wastewater in agriculture. This approach helps conserve water resources and promote a sustainable energy and nutrient cycling system.

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Our objective was to evaluate the effects of feeding 3-nitrooxypropanol (3-NOP; Bovaer, DSM Nutritional Products) at two levels on methane emissions, nitrogen balance, and performance by feedlot cattle. In experiment 1, a total of 138 Nellore bulls (initial body weight, 360 ± 37.3 kg) were housed in pens (27 pens with either 4 or 5 bulls per pen) and fed a high-concentrate diet for 96 d, containing 1) no addition of 3-NOP (control), 2) inclusion of 3-NOP at 100 mg/kg dry matter (DM), and 3) inclusion of 3-NOP at 150 mg/kg DM. No adverse effects of 3-NOP were observed on DM intake (DMI), animal performance, and gain:feed (P > 0.05). In addition, there was no effect (P > 0.05) of 3-NOP on carcass characteristics (subcutaneous fat thickness and rib eye area). In experiment 2, 24 bulls (initial BW, 366 ± 39.6 kg) housed in 12 pens (2 bulls/pen) from experiment 1 were used for CH4 measurements and nitrogen balance. Irrespective of the level, 3-NOP consistently decreased (P < 0.001) animals' CH4 emissions (g/d; ~49.3%), CH4 yield (CH4/DMI; ~40.7%) and CH4 intensity (CH4/average daily gain; ~38.6%). Moreover, 3-NOP significantly reduced the gross energy intake lost as CH4 by 42.5% (P < 0.001). The N retention: N intake ratio was not affected by 3-NOP (P = 0.19). We conclude that feeding 3-NOP is an effective strategy to reduce methane emissions, with no impairment on feedlot cattle performance.

During fiber digestion in the rumen, enteric methane is produced. Methane is a potent greenhouse gas. Recently several studies have focused on developing synthetic compounds and their utilization as specific inhibitors of methanogenesis. 3-Nitrooxypropanol is a structural compound that can help to mitigate CH4 emissions. The objective of this study was to evaluate the effects of feeding 3-nitrooxypropanol (3-NOP; Bovaer, DSM Nutritional Products) at two levels on methane emissions, nitrogen balance, and performance by feedlot cattle. No effect of 3-NOP on animal performance and N balance was found. However, regarding CH4 production 3-NOP consistently decreased (P < 0.001) animals' CH4 emissions (g/d; ~49.3%), methane yield (CH4/dry matter intake; ~40.7%), and CH4 intensity (CH4/average daily gain; ~38.6%). This study provides information on the potential role of 3-NOP on reducing CH4 emissions from feedlot cattle without reducing animal performance.

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The Gram-positive bacteria lactic acid bacteria (LAB) are used in the food industry but are also known for inhibiting certain food spoilage microorganisms, especially fungi. Sources of nitrogen (N) for culture media are generally organic and expensive. Many attempts have been made to formulate economical culture media with alternative N sources obtained from agricultural and industrial byproducts. This study describes the design and optimization of an inexpensive culture medium for Lactiplantibacillus plantarum (formerly Lactobacillus plantarum) MZ809351 strain B31. The culture medium was optimized using statistical experimental designs to identify the factors with the most significant effects on biomass concentration to reduce the overall cost, aiming to obtain a biomass concentration similar to that obtained with the reference LAB culture medium (de Man, Rogosa and Sharpe; MRS). Sodium acetate and magnesium sulfate were the most significant factors (p < 0.005), and their contents were reduced by 22 % and 40 %, respectively, without affecting biomass concentration. Malt germ extract (MGE) was used as an alternative nitrogen source to replace meat extract (ME) and proteose peptone (PP). Through these experiments, the composition of a culture medium that is less expensive than MRS broth was defined, which produced a biomass concentration (3.8 g/L) similar to that obtained with MRS medium. The inhibitory effects of two LAB strains isolated from the Ivory Coast and Mexico on the growth and production of ochratoxin A (OTA) in an ochratoxigenic fungus was tested. The minimum cellular concentration of the LAB to prevent the development of Aspergillus carbonarius Ac 089 and the production of OTA was determined in a model assay in Petri dishes. The conditions to inhibit the germination of A. carbonarius Ac 089 and the production of OTA were found. Using the optimized medium and a ratio of 2 × 104 LAB/spore (1 × 108 CFU/mL) strain B7 (L. plantarum MZ809351) and 2 × 103 LAB/spore (1 × 107 CFU/mL) strain B31 (L. plantarum MN922335) completely inhibited the growth of the fungus. A ratio of 2 × 105 LAB/spore (1 × 109 CFU/mL) was required to inhibit OTA production with strains B7 and B31. This study indicates the potential of cultivating LAB in an optimized and inexpensive culture medium for use as a biological control agent against ochratoxigenic fungi in food.

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Silicon (Si) may be involved in the modification of C:N:P stoichiometry and in physiological processes, increasing sorghum growth and grain production. The objective was to evaluate the effect of Si supply on C:N:P:Si stoichiometry, physiological response, growth, and grain production of sorghum. The experiment was carried out in pots with four concentrations of Si: 0; 1.2; 2.4; and 3.6 mmol L-1 in a completely randomized design, with six replicates. Physiological attributes and dark green color index were measured and grain and biomass production were determined. Posteriorly, the plant material was ground to determine silicon (Si), carbon (C), nitrogen (N), and phosphorus (P) contents in order to analyze C:N:P:Si stoichiometry. C:Si and C:N ratios decreased at all Si concentrations applied (1.2, 2.4, and 3.6 mmol L-1) and in all plant parts studied, being lower at 3.6 mmol L-1. The lowest C:P ratios of leaves and roots were observed at 3.6 mmol L-1 Si and the lowest C:P ratio of stems was observed at 1.2 mmol L-1 Si. Si concentrations were not significant for the N:P ratio of leaves. The highest N:P ratio of stems was observed at 3.6 mmol L-1, while the lowest N:P ratio of roots was observed at 2.4 and 3.6 mmol L-1. Regardless of photosynthetic parameters, the application of 1.2 mmol L-1 Si enhanced photosynthetic rate. The application of 2.4 and 3.6 mmol L-1 enhanced stomatal conductance and dark green color index. The mass of 1000 grains was not influenced by Si applications, while Si applications at all concentrations studied (1.2, 2.4, and 3.6 mmol L-1) enhanced shoot and total dry matter, not affecting root dry matter and grain production. In conclusion, Si supply modifies C:N:P:Si stoichiometry and increases physiologic parameters, growth, development, and grain production in sorghum.

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The productivity of rainforests growing on highly weathered tropical soils is expected to be limited by phosphorus availability1. Yet, controlled fertilization experiments have been unable to demonstrate a dominant role for phosphorus in controlling tropical forest net primary productivity. Recent syntheses have demonstrated that responses to nitrogen addition are as large as to phosphorus2, and adaptations to low phosphorus availability appear to enable net primary productivity to be maintained across major soil phosphorus gradients3. Thus, the extent to which phosphorus availability limits tropical forest productivity is highly uncertain. The majority of the Amazonia, however, is characterized by soils that are more depleted in phosphorus than those in which most tropical fertilization experiments have taken place2. Thus, we established a phosphorus, nitrogen and base cation addition experiment in an old growth Amazon rainforest, with a low soil phosphorus content that is representative of approximately 60% of the Amazon basin. Here we show that net primary productivity increased exclusively with phosphorus addition. After 2 years, strong responses were observed in fine root (+29%) and canopy productivity (+19%), but not stem growth. The direct evidence of phosphorus limitation of net primary productivity suggests that phosphorus availability may restrict Amazon forest responses to CO2 fertilization4, with major implications for future carbon sequestration and forest resilience to climate change.

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This study aimed to evaluate the effects of untreated pig manure from diets incorporating growth-promoting supplements (antibiotics and Zn oxide) on the survival and reproduction of Eisenia andrei earthworms. The tested manures were obtained from four different groups of pigs fed with four different diets: CS, a diet based on corn and soymeal; TR, a diet based on corn, soymeal, and ground wheat (15%); CSa, a diet based on corn and soymeal + 100 ppm of doxycycline + 50 ppm of colistin + 2500 ppm of Zn oxide; and TRa, a diet based on corn, soymeal, and ground wheat (15%) + 100 ppm of doxycycline + 50 ppm of colistin + 2500 ppm of Zn oxide. The study used two soils representative of the Southern region of Brazil (Oxisol and Entisol). In general, there were no significant differences between the different manures tested in each soil. However, there were differences in the toxicity manure on E. andrei between the soils, and the magnitude of this effect was dependent on the applied dose. In Oxisol, LC50 values were higher than 80 m3 ha-1, and EC50 varied from 9 to 27 m3 ha-1. In Entisol, the LC50 values were below the lowest dose tested (< 25 m3 ha-1), and EC50 remained around 5 m3 ha-1. It may be possible that the effects observed were attributed to an excess of nitrogen, copper, and zinc, promoted by the addition of the untreated manure and how these factors interacted with soil type.

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The study aimed to evaluate the effect of powdered coconut water-based diluent (ACP-101c) associated with extra virgin coconut oil (CO) as an external cryoprotectant in the conservation of cryopreserved buck sperm. For cryopreservation, the ejaculates from four bucks were pooled and divided into three aliquots and diluted at 37°C for treatments T1 (ACP-101c + 2.5% egg yolk +7% glycerol), T2 (ACP-101c + 2.5% CO +7% glycerol), and T3 (ACP-101c + 5% CO +7% glycerol). Then, the samples were packaged and cooled at a rate of 1.07°C/min decrease. Upon reaching 4°C, the samples were stored in a refrigerator at 4°C for 30 minutes for stabilization. After this period, the straws were frozen in nitrogen vapor for 15 minutes and then immersed and stored in liquid nitrogen at -196°C. After thawing, the samples were evaluated for sperm kinetics, plasma membrane integrity, acrosomal integrity, membrane functionality, mitochondrial activity (MA), and sperm morphology. In this study, no statistically significant differences were observed between the three treatments regarding the kinetic parameters (p > 0.05; Table 1). However, in relation to the velocities, a reduction was observed beyond the expected. There were no statistically significant differences between the diluents T1, T2, and T3 for the three velocities (curvilinear velocity [VCL], linear velocity [VSL], average path velocity [VAP]). Furthermore, no statistically significant differences were observed (p > 0.05) among treatments regarding the evaluation of membrane integrity, the functional membrane, MA, and sperm morphology after thawing. In conclusion, the use of CO in concentrations of 2.5% and 5.0% is effective in maintaining goat sperm quality, presenting itself as an alternative diluent for international programs of artificial insemination and embryo transfers. [Table: see text].

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The objective of this study was to quantify the effect of potassium (K) supply on osmotic adjustment and drought avoidance mechanisms of Eucalyptus seedlings growing under short-term water stress. The effects of K supply on plant growth, nutritional status, leaf gas exchange parameters, leaf water potential (Ψw), leaf area (LA), stomatal density (SD), leaf carbon (C) and nitrogen (N) isotopic compositions (δ13C and δ15N ‰) and leaf C/N ratio under polyethylene glycol (PEG)-induced water deficit were measured. Under both control (non-PEG) and osmotic stress (+PEG) conditions, K supply increased plant growth, boosting dry matter yield with decreased C/N leaf ratio and δ15N ‰ values. The +PEG significantly reduced LA, plant growth, dry matter yield, Ψw, number of stomata per plant and leaf gas exchange, relative to non-PEG condition. Potassium supply alleviated osmotic-induced alterations in Eucalyptus seedlings by better regulating leaf development as well as SD, thus improving the rate of leaf gas exchange parameters, mesophyll conductance to CO2 (lower δ13C ‰ values) and water use efficiency (WUE). Consequently, K-supplied plants under drought better acclimated to osmotic stress than K-deficient plants, which in turn induced lower CO2 assimilation and dry matter yield, as well as higher leaf δ13C ‰ and δ15N ‰ values. In conclusion, management practices should seek to optimize K-nutrition to improve WUE, photosynthesis-related parameters and plant growth under water deficit conditions.

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BACKGROUND: Nitrogen (N) and phosphorus (P) are macronutrients essential for crop growth and productivity. In cultivated fields, N and P levels are rarely sufficient, contributing to the gap between realized and potential production. Fertilizer application increases nutrient availability, but is not available to all farmers, nor are current rates of application sustainable or environmentally desirable. Transcriptomic studies of cereal crops have revealed dramatic responses to either low N or low P single stress treatments. In the field, however, levels of both N and P may be suboptimal. The interaction between N and P starvation responses remains to be fully characterized. RESULTS: We characterized growth and root and leaf transcriptomes of young maize plants under nutrient replete, low N, low P or combined low NP conditions. We identified 1555 genes to respond to our nutrient treatments, in one or both tissues. A large group of genes, including many classical P starvation response genes, were regulated antagonistically between low N and P conditions. An additional experiment over a range of N availability indicated that a mild reduction in N levels was sufficient to repress the low P induction of P starvation genes. Although expression of P transporter genes was repressed under low N or low NP, we confirmed earlier reports of P hyper accumulation under N limitation. CONCLUSIONS: Transcriptional responses to low N or P were distinct, with few genes responding in a similar way to the two single stress treatments. In combined NP stress, the low N response dominated, and the P starvation response was largely suppressed. A mild reduction in N availability was sufficient to repress the induction of P starvation associated genes. We conclude that activation of the transcriptional response to P starvation in maize is contingent on N availability.

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The objective of this work was to evaluate the effect of nitrogen (N) fertilization and the biological nitrogen fixation efficiency in soybean crops grown in unfavorable environments (high temperature, low fertility, and areas with sandy soil). Six field experiments were conducted between 2015 and 2018. Two experiments were performed per year. They were conducted in two separate areas. One was previously covered with degraded pasture (post-pasture area). The other was previously used to plant soybean (post-soybean crop area). The treatments consisted of inoculations with N-fixing bacteria (NFB) (0, 4, 8, and 12 doses ha-1) and N fertilization with rates of 0, 25, 50, and 100 kg ha-1. N fertilization and inoculation with NFB increased soil N, though the levels were still low. Among the tested groups, those with the application of eight doses of inoculant recorded the highest grain yields in post-soybean areas. They showed 10% (237 kg ha-1) and 15% (336 kg ha-1) higher grain yields when compared to crops treated without inoculant and crops with four doses of inoculant, respectively. N fertilization with 25 and 100 kg ha-1 decreased the root nodules of soybean plants grown in the post-soybean and post-pasture areas, respectively. Soybean crops grown on degraded pasture areas also showed good response to N fertilization (50 kg ha-1) when combined with NFB inoculation (12 doses ha-1). These showed grain yields 22% (439 kg ha-1) higher than those of plants treated with just 12 doses of inoculant and no N fertilization.

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Several factors have the potential to influence microalgae growth. In the present study, nitrogen concentration and light intensity were evaluated in order to obtain high biomass production and high phycoerythrin accumulation from Porphyridium purpureum. The range of nitrogen concentrations evaluated in the culture medium was 0.075-0.450 g L-1 and light intensities ranged between 30 and 100 µmol m-2 s-1. Surprisingly, low nitrogen concentration and high light intensity resulted in high biomass yield and phycoerythrin accumulation. Thus, the best biomass productivity (0.386 g L-1 d-1) and biomass yield (5.403 g L-1) were achieved with NaNO3 at 0.075 g L-1 and 100 µmol m-2 s-1. In addition, phycoerythrin production was improved to obtain a concentration of 14.66 mg L-1 (2.71 mg g-1 of phycoerythrin over dry weight). The results of the present study indicate that it is possible to significantly improve biomass and pigment production in Porphyridium purpureum by limiting nitrogen concentration and light intensity.

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ß-fructofuranosidase (invertase) and ß-D-fructosyltransferase (FTase) are enzymes used in industrial processes to hydrolyze sucrose aiming to produce inverted sugar syrup or fructooligosaccharides. In this work, a black Aspergillus sp. PC-4 was selected among six filamentous fungi isolated from canned peach syrup which were initially screened for invertase production. Cultivations with pure carbon sources showed that invertase and FTase were produced from glucose and sucrose, but high levels were also obtained from raffinose and inulin. Pineapple crown was the best complex carbon source for invertase (6.71 U/mL after 3 days of cultivation) and FTase production (14.60 U/mL after 5 days of cultivation). Yeast extract and ammonium chloride nitrogen sources provided higher production of invertase (6.80 U/mL and 6.30 U/mL, respectively), whereas ammonium nitrate and soybean protein were the best nitrogen sources for FTase production (24.00 U/mL and 24.90 U/mL, respectively). Fermentation parameters for invertase using yeast extract were Y P/S = 536.85 U/g and P P = 1.49 U/g/h. FTase production showed values of Y P/S = 2,627.93 U/g and P P = 4.4 U/h using soybean protein. The screening for best culture conditions showed an increase of invertase production values by 5.10-fold after 96 h cultivation compared to initial experiments (fungi bioprospection), while FTase production increased by 14.60-fold (44.40 U/mL) after 168 h cultivation. A. carbonarius PC-4 is a new promising strain for invertase and FTase production from low cost carbon sources, whose synthesized enzymes are suitable for the production of inverted sugar, fructose syrups, and fructooligosaccharides.

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This study focused on evaluating whether the injection of CO2, which is associated with the use of thermoelectric fly ashes and a reduced supply of nitrogen, affects the production of intracellular carbohydrates from Spirulina. For this purpose, the addition of 0.25 g L-1 of NaNO3, along with a 10% (v v-1) of CO2 injection, a flow rate of 0.3 vvm for 1 or 5 min, as well as 0, 120 and 160 ppm of fly ashes, was studied. The assays with 120 ppm of fly ashes presented the best kinetic parameters and CO2 biofixation rate, regardless of the CO2 injection time. Meanwhile, the experiments with 120 and 160 ppm of fly ash and CO2 injection for 1 min presented 63.3 and 61.0% (w w-1) of carbohydrates, respectively. Thus, this study represents an important strategy to increase the accumulation of carbohydrates in Spirulina, with potential application in the production of bioethanol.

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Agricultural intensification offers potential to grow more food while reducing the conversion of native ecosystems to croplands. However, intensification also risks environmental degradation through emissions of the greenhouse gas nitrous oxide (N2O) and nitrate leaching to ground and surface waters. Intensively-managed croplands and nitrogen (N) fertilizer use are expanding rapidly in tropical regions. We quantified fertilizer responses of maize yield, N2O emissions, and N leaching in an Amazon soybean-maize double-cropping system on deep, highly-weathered soils in Mato Grosso, Brazil. Application of N fertilizer above 80 kg N ha-1 yr-1 increased maize yield and N2O emissions only slightly. Unlike experiences in temperate regions, leached nitrate accumulated in deep soils with increased fertilizer and conversion to cropping at N fertilization rates >80 kg N ha-1, which exceeded maize demand. This raises new questions about the capacity of tropical agricultural soils to store nitrogen, which may determine when and how much nitrogen impacts surface waters.

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Improved understanding of the nutritional ecology of arbuscular mycorrhizal (AM) fungi is important in understanding how tropical forests maintain high productivity on low-fertility soils. Relatively little is known about how AM fungi will respond to changes in nutrient inputs in tropical forests, which hampers our ability to assess how forest productivity will be influenced by anthropogenic change. Here we assessed the influence of long-term inorganic and organic nutrient additions and nutrient depletion on AM fungi, using two adjacent experiments in a lowland tropical forest in Panama. We characterised AM fungal communities in soil and roots using 454-pyrosequencing, and quantified AM fungal abundance using microscopy and a lipid biomarker. Phosphorus and nitrogen addition reduced the abundance of AM fungi to a similar extent, but affected community composition in different ways. Nutrient depletion (removal of leaf litter) had a pronounced effect on AM fungal community composition, affecting nearly as many OTUs as phosphorus addition. The addition of nutrients in organic form (leaf litter) had little effect on any AM fungal parameter. Soil AM fungal communities responded more strongly to changes in nutrient availability than communities in roots. This suggests that the 'dual niches' of AM fungi in soil versus roots are structured to different degrees by abiotic environmental filters, and biotic filters imposed by the plant host. Our findings indicate that AM fungal communities are fine-tuned to nutrient regimes, and support future studies aiming to link AM fungal community dynamics with ecosystem function.

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Despite the great diversity of plant growth-promoting bacteria (PGPB) with potential to partially replace the use of N fertilisers in agriculture, few PGPB have been explored for the production of commercial inoculants, reinforcing the importance of identifying positive plant-bacteria interactions. Aiming to better understand the influence of PGPB inoculation in plant development, two PGPB species with distant phylogenetic relationship were inoculated in maize. Maize seeds were inoculated with Bacillus sp. or Azospirillum brasilense. After germination, the plants were subjected to two N treatments: full (N+) and limiting (N-) N supply. Then, anatomical, biometric and physiological analyses were performed. Both PGPB species modified the anatomical pattern of roots, as verified by the higher metaxylem vessel element (MVE) number. Bacillus sp. also increased the MVE area in maize roots. Under N+ conditions, both PGPB decreased leaf protein content and led to development of shorter roots; however, Bacillus sp. increased root and shoot dry weight, whereas A. brasilense increased photosynthesis rate and leaf nitrate content. In plants subjected to N limitation (N-), photosynthesis rate and photosystem II efficiency increased in maize inoculated with Bacillus sp., whilst A. brasilense contained higher ammonium, amino acids and total soluble sugars in leaves, compared to the control. Plant developmental and metabolical patterns were switched by the inoculation, regardless of the inoculant bacterium used, producing similar as well as distinct modifications to the parameters studied. These results indicate that even non-diazotrophic inoculant strains can improve the plant N status as result of the morpho-anatomical and physiological modifications produced by the PGPB.

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The relationship between leaf photosynthesis and nitrogen is a critical production function for ecosystem functioning. Cultivated species have been studied in terms of this relationship, focusing on improving nitrogen (N) use, while wild species have been studied to evaluate leaf evolutionary patterns. A comprehensive comparison of cultivated vs wild species for this relevant function is currently lacking. We hypothesize that cultivated species show increased carbon assimilation per unit leaf N area compared with wild species as associated with artificial selection for resource-acquisition traits. We compiled published data on light-saturated photosynthesis (Amax ) and leaf nitrogen (LNarea ) for cultivated and wild species. The relationship between Amax and LNarea was evaluated using a frontier analysis (90th percentile) to benchmark the biological limit of nitrogen use for photosynthesis. Carbon assimilation in relation to leaf N was not consistently higher in cultivated species; out of 14 cultivated species, only wheat, rice, maize and sorghum showed higher ability to use N for photosynthesis compared with wild species. Results indicate that cultivated species have not surpassed the biological limit on nitrogen use observed for wild species. Future increases in photosynthesis based on natural variation need to be assisted by bioengineering of key enzymes to increase crop productivity.

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BACKGROUND: Maize breeding programs focus on the development of hybrid varieties and the cultivation of landrace materials is discouraged; however, they are a valuable source of genes and their conservation is advisable. OBJECTIVE: Analyzing some stress indicators during cryopreservation of maize landrace seeds. MATERIALS AND METHODS: Seeds of 35 accessions of landrace maize were collected in two regions of Costa Rica and cryopreserved by direct immersion in liquid nitrogen (LN). Membrane integrity, germination of seeds and DNA methylation in tissues were analyzed 5, 7 and 9 days after rewarming. RESULTS: Germination of landrace maize seeds was near 100 % for most accessions. No statistically significant differences in germination were observed between non-cryopreserved controls and seeds stored in LN for 1 h or 1 year. Membrane integrity, number of leaves and root and shoot length of plantlets were similar after cryostorage of seeds for 1 h and 1 year. A short delay in growth of cryostored seed compared to non-frozen controls was observed. Changes in the proportion of DNA methylation were noted from 0 to day 9 in the organs studied depending on the germination stage and cryopreservation treatment. CONCLUSION: It may be inferred that many of the methylated genes were related to growth and development. In addition, a cryobank of maize landraces from two regions of Costa Rica was established.

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The objective of this study was to examine the effects of the type and intensity of nutritional stress, and of the statistical treatment of the data, on the genotype x environment (G x E) interaction for tropical maize (Zea mays). For this purpose, 39 hybrid combinations were evaluated under low- and high-nitrogen and -phosphorus availability. The plants were harvested at the V6 stage, and the shoot dry mass was estimated. The variance components and genetic values were assessed using the restricted maximum likelihood/best linear unbiased prediction method, and subsequently analyzed using the GGE biplot method. We observed differences in the performances of the hybrids depending on both the type and intensity of nutritional stress. The results of relationship between environments depended on whether genotypic values or phenotypic means were used. The selection of tropical maize genotypes against nutritional stress should be performed for each nutrient availability level within each type of nutritional stress. The use of phenotypic means for this purpose provides greater reliability than do genotypic values for the analysis of the G x E interaction using GGE biplot.

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This experiment evaluated forage production, sward structure, stocking rate, weight gain per area and nutritive value of forage as grazed by beef heifers on Alexandergrass (Urochloa plantaginea (Link) Hitch) pasture fertilized with nitrogen (N): 0; 100; 200 or 300 kg of N/ha. The experiment was a completely randomized design following a repeated measurement arrangement. The experimental animals were Angus heifers with initial age and weight of 15 months and 241.5±5 kg, respectively. The grazing method was continuous, with put-and-take stocking. N utilization, regardless of the level, increase by 25% the daily forage accumulation rate and the weight gain per area by 23%. The level of 97.2 kg N/ha leads to a higher leaf blade mass and increases by 20% the leaf:stem ratio. Alterations in sward structure changes the nutritive value of forage as grazed. The utilization of 112.7 kg of N/ha allows the highest stocking rate (2049.8 kg of BW/ha), equivalent to 7.5 heifers per hectare.

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