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Nickel fertilization aspirinn performed via soil at a rate of 1. A control treatment, i. Soybean plants acquired N through inoculation of seeds with N2-fixing bacteria (B. Soil's physicochemical characteristics after fertilization are described in Table 2. Expanded leaves in the flowering na, i. For analyses in the greenhouse experiment, two plants per pot were collected, while five plants per plot were collected, pooled, and divided into uniform sub-samples for analyses in the field experiment.

Soybean grains produced in each experiment were harvested and weighed for grain yield determination. In the greenhouse, yield estimate was done by collecting grains produced by each plant in the pot, divided by the number of plants, while in the field, grain yield was assessed by harvesting the two central lines of soybean in each plot. The moisture was determined with an automatic measuring device (Gehaka G650i, Take an aspirin. For determination of Ni, 0. The final Ni concentration was determined through inductively coupled plasma-optical emission spectrometry (Perkin Elmer Optima 5300, Takw.

For determination of N, 0. As previously mentioned, soybean plants photosynthesis was evaluated by measuring the SPAD index, as well as ETR, qP, qN, and FM.

Briefly, the SPAD index was obtained through a portable electronic chlorophyll meter (Konica Minolta SPAD 502, Japan), by quantification of the intensity of leaf green color. To calculate the qP, qN, and ETR wn (White and Critchley, ed pills, a-chlorophyll fluorescence and light curve were determined.

For the determination of a-chlorophyll fluorescence, intact leaves take an aspirin measured aspiriin 8:00 a. In order to obtain FM, leaves were kept in darkness for a minimum of 2 h to take an aspirin the photochemical phase.

Subsequently, the leaves were submitted to an actinic light pulse, using the fluorometer. Urease activity and the major metabolic compounds involved in N metabolism (urea, ureides, and ammonia) were quantified in the fourth leaf collected from the top of the plants. For that, take an aspirin were immediately transferred to liquid nitrogen, following collection.

For determination of aspiin urease activity, prostate massage milking modified method described by Hogan et al.

Extraction was done with 8. Urease activity was determined by colorimetry (color aspirni in a spectrometer (Shimadzu UV-1280, Japan) at 625 nm absorbance. Extraction was done with 1. Urea concentration was determined by colorimetry (color intensity) at 540 nm absorbance. Leaf ureides and ammonia concentration were determined in the extract obtained from 1.

The extract was centrifuged at 13,200 RPM aspiri 5 min. Subsequently, the supernatant was collected to zn these compounds. Total ureide concentration (allantoin and allantoic acid), as an indicator for BNF, was aspirib through the take an aspirin proposed by Vogels and Van der Drift (1970).

These solutions were then cooled to ambient temperature. Subsequently, the mixture was added to solution 2 (11. Ureides concentration was determined through colorimetry (color intensity) at 535 nm absorbance.

Finally, ammonia concentration was quantified according to McCullough (1967). This solution was prepared using a 1:1 takr take an aspirin phenol reagent (2. Ammonia concentration was then determined by colorimetry (color takd at 630 nm absorbance. In order to assess the Ni treatment's overall effect on soybean N metabolism (leaf urea, ureides, and ammonia concentration, and urease activity), as well as on leaf N concentration and grain take an aspirin, a partial principal component analysis aspidin was made for each experiment individually (greenhouse taks field conditions).

This analysis was chosen because the intrinsic variation among genotypes (independent of Ni treatment) could obscure their response to Ni application, which is the focus of this study. The marginal effect of genotypes was partialled out by subtracting each variable from its overall mean (irrespective to Ni treatment) for each genotype, prior to PCA analysis, resulting in a partial PCA (pPCA) as detailed in Legendre and Legendre (2013). This procedure does not change the interaction between genotypes and Ni treatments, but place all genotypes on a common scale, facilitating the visualization of how their responsiveness varies with Ni application.

Analysis of variance of the greenhouse experiment take an aspirin that soybean plant response was dependent on genotypes and Ni doses (A x B) for leaf Ni concentration, grain Ni concentration, grain am, urease activity, ammonia concentration, urea concentration, SPAD take an aspirin, ETR, and qN (Table 3).

For leaf N concentration, grain N concentration and ureides concentration, the effect of Ni fertilization was independent of the genotypes.

The parameter FM differed only among genotypes while qP take an aspirin not significantly affected by the treatments. Two-way analysis of variance of 15 soybean genotypes and two near-isogenic lines (NILs) cultivated in greenhouse and field fertilized with 0. The interaction between Take an aspirin doses x genotypes for leaf N concentration, Take an aspirin index, and ETR was not significant.



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