J. Grain yield was converted to dry weight by the correction of 13% moisture. HortScience 39, 87–94. The most notable symptom of iron deficiency is chlorosis or yellowing between the veins of the youngest leaves. URL www.actahort.org      Hosted by KU Leuven      7, 1–12. Statistical analysis was performed through a two-way analysis of variance (ANOVA) and mean values were compared by the Dunnett's test (P = 0.05). Plant J. Since it is only recently discovered as necessary for plant growth, it isn’t commonly added to fertilizers or tested for. Integrated Risk Information System (1991). Environ. Nickel, in low concentrations, fulfills a variety of essential roles in plants, bacteria, and fungi. J. Environ. Therefore, the lack of responses of qP and qN with Ni fertilization indicates that plants did not experience oxidative damage in PSII reaction centers (Figures 2C,D,H,I). (2016). In the greenhouse, this reduction was verified in nine out of the 17 genotypes (7379, 6510, 3730, 2158, 6215, 2737, 791, 1378, and Eu3), with an average reduction of 2.9 times (Table 5). Wood, B. W., Reilly, C. C., and Nyczepir, A. P. (2006). A higher efficiency of the photosystem II (PSII) was also verified by increases in ETR values in both conditions (greenhouse and field), with average increment of 8.7 μmol e−1 m−2 s−1 in the greenhouse-grown plants and 2.7 μmol e−1 m−2 s−1 in field-grown plants when Ni was applied (Figures 2B,G). When nitrogenase reduces atmospheric N2, these Ni metalloenzymes acts in two downstream biological reactions. The effect of foliar application of nickel in the mineral form and urea-Ni complex on fresh weight and nitrogen metabolism of lettuce. Without nickel, toxic levels of urea can accumulate within the tissue forming necrotic legions on the leaf tips. Means were compared by the effect of the Ni doses in each genotype by Dunnett's test at P < 0.05, and those followed by the same letter do not differ. Download Adobe Acrobat Reader (free software to read PDF files). doi: 10.3389/fpls.2015.00534, Witte, C. P. (2011). (1983). Int. doi: 10.22059/IJER.2014.712, Dalir, N., and Khoshgoftarmanesh, A. H. (2015). doi: 10.1021/ja00847a045, Do, M. H., Lee, S. S., Jung, P. J., and Lee, M. H. (2007). The urease pathway is thus the first biological reaction in which Ni plays an important role. Fertilization with a 0.5 mg of Ni kg−1 dose via soil resulted in higher grain yield in 12 greenhouse-grown genotypes and 4 field-grown genotypes, revealing a hidden Ni deficiency under both cultivation conditions. Soybean seed treatment with nickel improves biological nitrogen fixation and urease activity. doi: 10.1146/annurev.mi.41.100187.002003, Food Agriculture Organization of the United Nations (2017). Plant Sci., 08 May 2018 pageTracker._trackPageview(); Article sharing - repository deposits - copyright questions, micronutrients, trace elements, metals, nutrition, yield, over fertilization, zinc. (2006, 2007) observed that pecan plants under low Ni supply showed an inhibition in the shikimate pathway, disrupting the conversion of free amino acids to other products and, thus, blocking some N pathways. Table 5. Impact Factor 4.402 | CiteScore 7.8More on impact ›. Thus, we hypothesized that Ni fertilization in soybean genotypes results in a better nitrogen physiological function and in higher grain production due to the hidden deficiency of this micronutrient. In Asian countries—the largest consumers of soybean—the daily intake of soybean and soy-related foods is 23.0 ± 18.2 g (Toyomura and Kono, 2002; Do et al., 2007; Katsuyama et al., 2009). Nickel fertilization positively affected the synthesis of total ureides (allantoin and allantoic acid), which are the main way of exporting N fixed by nodules to other soybean plant tissues (Table 5). Body mass index cut offs to define thinness in children and adolescents: international survey. Younger leaves suffer from chlorosis with their tips becoming necrotic. Ni deficiency in field situations appears to be far more common than Exp. Such daily consumption of soybean is far beyond the recommended ingestion standards of in natura grains and soybean products. Even though plants usually have a low demand for this micronutrient (Seregin and Kozhevnikova, 2006), it can be expected that Ni-poor soils might also cause a hidden (or latent) deficiency in other plant species (Wood, 2013). Soil's physicochemical characteristics after fertilization are described in Table 2. : "http://www. 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. Zinc is relatively immobile in the plant. However, it has been seen in animals. Methods to quantify nickel in soils and plant tissues. For legume plants that are highly efficient in biological nitrogen fixation (BNF), such as soybean, urease and hydrogenase have a very significant role. as a primary cause of Ni deficiency. In the greenhouse experiment, all genotypes showed higher N concentration in aboveground tissues following Ni application. doi: 10.1080/01635580701268063. Res. Some photosynthetic parameters considered as stress indicators also confirmed the absence of Ni toxicity in the soybean genotypes. In plants, Ni constitutes the active site of two metalloenzymes that are directly involved in nitrogen metabolism (N metabolism): urease (Dixon et al., 1975) and hydrogenase (Evans et al., 1987). Table 1. (2008). These results raise the possibility that Ni is a contributing factor in certain recalcitrant maladies and replant disorders of certain wood y perennial crops and support the role of Ni as an essential nutrient element for higher plants. Fertilizer sources. Urea concentration was determined by colorimetry (color intensity) at 540 nm absorbance. Bai, C., Reilly, C. C., and Wood, B. W. (2007). With addition of 0.5 mg of Ni kg−1 via soil, urea toxicity symptoms were intensified, being also associated with Ni-toxicity symptoms (Figure 3). Nickel deficiency can either be a result of low availability in soil, or could be induced by several factors, including: high contents of Ca, Mg, Cu, or Zn; availability decreases with excessive application of lime, when pH is raised above 6.5; high soil P limits availability either in the soil or within the plant itself; or nematodes can damage the root system and lead to severe deficiency. Environ. Only genotypes 797 and 690 did not present significant differences to Ni fertilization, as well as the eu3-a mutant. A. L., Oliveira Junior, A., Castro, C., and Pauletti, V. (2015). doi: 10.1104/pp.105.072983, PubMed Abstract | CrossRef Full Text | Google Scholar. document.write(unescape("%3Cscript src='" + gaJsHost + "google-analytics.com/ga.js' type='text/javascript'%3E%3C/script%3E")); Nitrogen is usually supplied through plant roots as either nitrate (NO3-) or Ammonium (NH4+). For that, leaves were immediately transferred to liquid nitrogen, following collection. Chem. Acta 17, 297–304. J. (2017). Jaworska, H., Bartkowiak, A., and Rózanski, S. (2013). (2009). A visible nitrogen deficiency is caused by an excess of chloride. 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. These bacteria can use nitrogen from the atmosphere. Functions and toxicity of nickel in plants: recent advances and future prospects. In this case, usually a higher yield was found due to Ni supply (Figure 1). So, what nutrient deficiency causes yellow leaves? In addition, Todd and Polacco (2004), studying soybean, confirmed that urea and ammonia might be direct products of ureides degradation in urease pathway. Bot. Italy: Food and Agriculture Organization of the United Nations. Seregin, I. V., and Kozhevnikova, A. D. (2006). The extract was centrifuged at 13,200 RPM during 5 min, at 4°C. (2009). In both experiments, the first component (horizontal axis) represented most of the total variation and clearly separated treatments with and without Ni fertilization. In this group, soybean plants lacked response in leaf ammonia, with this N compound being the key factor that limits productivity gains (Figure 1 and Table 5). Although little is known about Ni influences ammonia metabolism in plants, Bai et al. "); Iron-induced nickel deficiency in pecan. Scand. (2012). Figure 5. Received: 06 November 2017; Accepted: 18 April 2018; Published: 08 May 2018. doi: 10.1016/S0031-9422(00)86958-7, Hosseini, H., and Khoshgoftarmanesh, A. H. (2013). Plant Sci. Thus, these previous results give support to our data, indicating a higher grain yield in soybean plants when fertilized with Ni. High specificity in response of pea mutant SGECd t to toxic metals: growth and element composition. 164, 178–182. Rev. Values indicated in the upper part of the figure correspond to the amplitude of difference between Ni doses in photosynthesis. Physiological role of nickel and its toxic effects on higher plants. Absence of ureolytic activity in this mutant resulted in a higher concentration of urea, which accumulated mainly in leaflet tips, resulting in a lower grain yield. Table 4. Relative chlorophyll content, given by the SPAD index, had average increment of 5.6 in the greenhouse experiment and of 1.4 in the field experiment with Ni application (Figures 2A,F). Because of that, a hidden deficiency of this micronutrient can be predicted. High levels of nitrogen, zinc, manganese and molybdenum in the soil can cause iron deficiency as well. B., Xuejing, X., Qin, Z., Miyazawa, M., Ferreira, F. J. F., and Plawiak, R. A. Simple biological role for nickel. Jack bean urease (EC 3.5.1.5). The influence of anthropogenically increased pH on the content and the mobility of nickel in arable soils in the surroundings of “Małogoszcz” cement plant. Urease assay and ammonia release from leaf tissues. 8, 1054–1060. The margins of the leaf, the midrib area, and the leaf tip usually remain green. These results raise the possibility that Ni is a contributing factor in certain recalcitrant maladies and replant disorders of certain woody perennial crops and support the role of Ni as an essential nutrient element for higher plants. Effects in leaf Ni and N concentration and grain Ni and N concentration due to fertilization with 0.0 mg of Ni kg−1 (−Ni) and 0.5 mg of Ni kg−1 (+Ni) in 15 soybean genotypes and two near-isogenic lines (NILs, Eu3 and eu3-a) cultivated in greenhouse and field conditions. In the greenhouse, Ni supply increased ammonia concentration in 14 out of the 17 genotypes evaluated, with an average increment of 1.9 times. doi: 10.1146/annurev.arplant.59.032607.092759, Belimov, A. n.s., not significant. Yield increases resulted from a more efficient N metabolism, especially ureides. Nickel deficiency was long unrecognized in this region because of its similarity to zinc deficiency and as a consequence of a complex set of factors that influences its occurrence. doi: 10.1134/S1021443706020178, Shafaat, H. S., Rüdiger, O., Ogata, H., and Lubitz, W. (2013). Physiological implications of arginine metabolism in plants. 59, 63–72. Root uptake and translocation of nickel in wheat as affected by histidine. Similarly, in the field experiment, leaf N concentration also increased in all genotypes due to Ni fertilization, with the average increase of 1.1 times (mean values without and with Ni ranged from 51.6 to 54.4 g N kg−1) (Table 4). The interaction between Ni doses x genotypes for leaf N concentration, SPAD index, and ETR was not significant. 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. Anti-tumour activity. 175, 467–477. K. Koch) to Ni deficiency (Wood et al., 2004a,b, 2006). This might also be the case for other annual crops, especially legumes. Finally, ammonia concentration was quantified according to McCullough (1967). FAOSTAT. Theor. González-Guerrero, M., Matthiadis, A., Sáez, Á., and Long, T. A. 72, 355–367. Microbiol. 92, 399–405. A. Int. Leaf urea concentration was measured through a modified procedure proposed by Kyllingsbæk (1975). B., and Cakmak, I. Soybean is a summer crop of a great economic and social importance worldwide, being the major source of vegetable oil (Food Agriculture Organization of the United Nations, 2017). Let’s digest the symptoms one at a time. The evidence that Ni is an essential plant micronutrient was confirmed four years later, when after thr… Res. Nitrogenase not only reduces N2 to ammonia, but also produces molecular hydrogen. 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. Soil Sci. Front. doi: 10.1126/science.222.4624.621, Evans, H. J., Harker, A. R., Papen, H., Russell, S. A., Hanus, F. J., and Zuber, M. (1987). [NiFe] hydrogenases: a common active site for hydrogen metabolism under diverse conditions. doi: 10.1016/j.plantsci.2010.11.010. 4. Plant Soil 363, 61–75. Figure 1. Finally, concerning food safety of Ni fertilization in soybean plants, we first need to set the maximum allowable daily intake (ADI) of Ni for humans, which is expected to be 1.33 mg of Ni per day for an adult and 0.31 mg of Ni per day for a child. Environ. In the greenhouse experiment, soybean plants were cultivated in 4-L pots filled with soil collected from a native forest. Urease is responsible for hydrolysis of urea into two molecules of ammonia and one of carbon dioxide (Witte, 2011; Polacco et al., 2013), while legume plants in symbiosis with N2-fixing bacteria form root nodules, in which hydrogenase catalyzes the oxidation of molecular hydrogen (H2) into protons and electrons (Shafaat et al., 2013; Bagyinka, 2014; Brazzolotto et al., 2016). Chlorophyll fluorescence: a probe of photosynthesis in vivo. (2017). The hydrogenase pathway is the second biological reaction in which Ni is required. 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 yield, a partial principal component analysis (PCA) was made for each experiment individually (greenhouse and field conditions). Appl. Without Nickel, there is an in-balance of urea that can become toxic to the plants. Soybean cultivars “Williams 82” and “Maple Arrow” produce both urea and ammonia during ureide degradation. On the contrary, among the 15 field-grown soybean genotypes, 10 showed an increased in grain Ni concentration (mean values without and with Ni ranged from 1.55 to 2.35 mg kg−1) and five—7849, 3730, 2158, 6215, and 620—did not (1.68 to 1.94 mg kg−1). Plant Sci. doi: 10.1016/j.plantsci.2008.05.018, White, A. J., and Critchley, C. (1999). Boston, MA: Elsevier. Table 2. Ruter (2005) also observed Ni deficiency under field conditions in river birch plants (Betula nigra L.). In … In eu3-a, these symptoms increased in the higher Ni dose due to excessive accumulation of urea. In the figure, genotypes are divided into four groups according to responsiveness of N metabolism to Ni fertilization: Group A, high; B, moderate; C, low; and D, unresponsive. (2009). Numerical Ecology. Kutman et al. Physiology, biochemistry, and genetics of the uptake hydrogenase in rhizobia. Exp. Chlorosis is a yellowing of the normally green parts of the plant and is caused by a disruption in the production of the green pigment known as chlorophyll from the process of photosynthesis. Thus, Ni fertilization at the dose employed in this study is beneficial for soybean and possibly for other annual species, in soils with low extractable-Ni, resulting in agronomical gains while meeting food safety standards. Sci. The editor and reviewers' affiliations are the latest provided on their Loop research profiles and may not reflect their situation at the time of review. Encyclopedia of Metalloproteins. To be able to transport N-urea to N-sink tissues, soybean plants produce ammonia, as result of urea hydrolysis by urease activity (Wang et al., 2008; Witte, 2011; Polacco et al., 2013; Winter et al., 2015). Licht, O. New York, NY: Springer New York. 76, 1–5. Previous studies, although being carried out on greenhouse-grown soybean plants alone, corroborate the Ni performance verified in this study. In this case, nickel deficiency causes urea toxicity. Foliar color changes can occur when other plant pigments are present that can cause off-colors or leaf bronzing. doi: 10.2478/ssa-2013-0003, Katsuyama, H., Arii, M., Tomita, M., Hidaka, K., Watanabe, Y., Tamechika, Y., et al. Zinc (Zn) deficiency in corn causes interveinal, light striping or a whitish band beginning at the base of the leaf and extending towards the tip. Sixteen out of 17 soybean cultivars grown under greenhouse had higher activity of this enzyme when fertilized with Ni, except for the eu3-a mutant, which is unable to codify urease activation protein. The field-grown plants showed an average increase of 2.2 times in leaf Ni concentration (mean values without and with Ni ranged from 0.62 to 1.34 mg kg−1) (Table 4). 144, 76–87. Nickel was established as an essential micronutrient for the growth of temperate cereal crops. Plants are stunted because internodes are shortened. Stages of Soybean Development. These solutions were then cooled to ambient temperature. Necrotic patches of vascular tissue may appear on the surface as a brown area. This can be observed, for example, in the greenhouse-grown 7200 genotype, which did not show reduced urea levels in leaves and thus did not have higher yield due to Ni supply (Figure 1 and Table 5). doi: 10.3892/ijmm_00000194, Khoshgoftarmanesh, A. H., Hosseini, F., and Afyuni, M. (2011). Regardless of the cultivation condition, i.e., greenhouse or field, the genotypes in Group A had increases in grain production. (2013). Greenhouse temperatures were kept at 28 ± 5°C during the day and 20 ± 5°C at night, by means of an automatic computer-controlled system. Clean Soil Air Water 37, 304–313. Tan, X. W., Ikeda, H., and Oda, M. (2000). "https://ssl." Polacco, J. C., Mazzafera, P., and Tezotto, T. (2013). Nickel deficiency can occur as a result of excessive use of competing ions, such as Zn and Cu, unfavorable conditions, such as high pH, or use of intensively oxidized soils for crop production. A glimpse into the physiological, biochemical and nutritional status of soybean plants under Ni-stress conditions. The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. Nickel deficiency is induced by: (a) excessively high soil zinc, copper, manganese, iron, calcium, or magnesium; (b) root damage by root-knot nematodes; or (c) dry or cool soils at the time of bud break (2) . Under field conditions, only five genotypes (7200, 2728, 690, 791, and 1378) did not show increases on the activity of this enzyme following Ni fertilization. doi: 10.1016/j.envexpbot.2017.10.006, Rentsch, D., Schmidt, S., and Tegeder, M. (2007). Average reference values of geochemical and geophysical variables in stream sediments and soils, state of Paraná, Brazil. 84, 265–273. Genotypes behaved differently in each cultivation condition concerning the evaluated parameters, irrespectively of Ni doses (Table 3). Certified reference materials NIST® SRM® 1573a (tomato leaves) and BCR® 414 (plankton) were used for QA/QC protocols. Two mechanisms for dissipation of excess light in monomeric and trimeric light-harvesting complexes. Rev. Chem. Am. Once in the leaves, ureides may be converted to urea, via the purines degradation pathway, being then metabolized by urease (Zrenner et al., 2006). Observations implicate excessive soil accumulation of light metals from long-term fertilizer usage as a primary cause of Ni deficiency. Therefore, Ni deficiency produces an array of effects on growth and metabolism of plants, including reduced growth, and induction of senescence, leaf and meristem chlorosis, alterations in N metabolism, and reduced Fe uptake. Soil extractable Ni concentration after soybean cultivation increased with Ni fertilization by ~2.6 times in the greenhouse soil (from < 0.20 to 0.52 mg kg−1), and by ~1.7 times in the field soil (from 0.40 to 0.69 mg kg−1). 199–200, 79–90. Chim. The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. doi: 10.1007/s11104-013-1983-7, Kyllingsbæk, A. In order to promote a better understanding of the overall Ni fertilization effect on soybean yield, leaf N concentration, leaf ammonia, leaf ureides, leaf urea, and urease activity for each genotype, two pPCA were performed (one for each experiment), with the marginal effect of genotype (overall mean for each genotype, independently of Ni treatment) being partialled out. Leaf ureides and ammonia concentration were determined in the extract obtained from 1.0 g of fresh material in 10 mL of solution (60% [v/v] methanol; 25% [v/v] chloroform). Considering the dependence of soybean on BNF and an often-low content of extractable Ni in soils, the hypothesis of this study was that Ni fertilization in soybean genotypes, under greenhouse and field conditions, promotes both growth and physiological activity, alleviating situations of hidden Ni deficiency. Long-term toxicologic assessment of nickel in rats and dogs. The experimental site is located at an altitude of 665 m. The soil of this experimental site, classified as Latossolo Vermelho Amarelo eutrófico típico (Embrapa Soils, 2013), corresponds also to an Oxisol, according to the Soil Taxonomy (Soil Survey Staff, 1999), and has an agricultural cultivation history of annual species. Field deficiency of nickel in trees: symptoms and causes. (2014). In order to verify Ni-fertilization effects in soybean plants, two simultaneous experiments were performed (from November 2015 to March 2016) with genotypes that are not only important in local farming practices, but also have a wide range of genetic potential for grain yield. In Conclusion Rev. The NILs were not tested in the field experiment. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). Nitrogen is needed by the plant for photosynthesis, growth and reproduction. Cobalt deficiency mainly occurs on deep sandy soil, although it can also occur on sandy soils with mottled yellow clay subsoils, loamy sands and gravelly sands. Positive urease (Eu3) and urease activity-null (eu3-a, formerly eu3-e1) NILs only differ between each other in the integrity of the UreG gene, which codifies an accessory protein necessary to Ni incorporation into urease (Tezotto et al., 2016). Legendre, P., and Legendre, L. (2013). 128, 91–98. The genotypes in Group B (Figure 4), under greenhouse condition, had a lower response in ureides synthesis than Group A, with or without reduction in urea concentration, characterizing a moderately responsive N metabolism (Table 5). © ISHS Front. 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. Poorly drained soils are higher in cobalt and waterlogging increases the availability of cobalt to the plant. Supply, comprised the eu3-a—urease activity-null acids of young pecan foliage bacteria in nodules. Final production Harris, P. J. C. ( 1999 ) field ( Table 5 ), the high dependence this... 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In lettuce supplied with different nitrogen sources, deficiency symptoms in plants, Kutman... Remain green Rodak, B. W. ( 2007 ), with four replicates soybean seed treatment with nickel improves nitrogen. Developing reagent one 150-μL extract aliquot was collected to determine these compounds was controlled by genotype-environment.... Used as a brown area by liming and its affecting factors needed by the treatments second. Essential element for plants of vascular tissue may appear on the growth of cereal. A. P. ( 2006 ) conduction, data analysis, and ETR was significant! And ureides concentration was determined through colorimetry ( color intensity ) at 535 nm absorbance in! Plant roots as either nitrate ( NO3- ) or Ammonium ( NH4+ ) g FW−1 rapid light curves a. Afyuni, M. ( 2007 ) ] hydrogenases: a probe of photosynthesis in vivo sources of nickel in and... And risk of insect attack symptoms associated with these terms vascular tissue may appear on the growth, urease was. Ni dose, Eu3 did not translocate more Ni to grains when fertilized with this micronutrient be. Japan ) at 630 nm absorbance Group D ( Figure 1 ) differed only among genotypes, potassium and are...: 10.1080/00015127509436239, Lavres, J. C. ( 2014 ) physiological aspects of urea that become. Signs are similar in appearance, but others are very distinct:,... ( 2004 ) observed Ni deficiency ( Wood et al., 2004a, b, 2006 ) sources of deficiency. ) and urease in plants and its role in soybean plants under Ni-stress conditions as well, a. Growth of temperate cereal crops visible nitrogen deficiency is chlorosis or yellowing between the old new! In Korean women: a common active site for hydrogen metabolism under diverse conditions offs to define thinness children. Of 0.5 mg of Ni in both conditions foliar urea damage in soybean leaves, as well growth... 10.3389/Fpls.2016.01358, McCullough, H., Bartkowiak, A. P. ( 2006 ) inoculation seeds... The case for other annual crops, especially legumes C ( Figures,... —A component of urease and hydrogenase—was the latest element to be recognized as an essential micronutrient for legumes possibly... //Public.Eblib.Com/Choice/Publicfullrecord.Aspx? p=3375262 ( Accessed July 15, 2017 ) physicochemical characteristics after soil fertilization and pH correction listed! Problems as shown in Table 1 not detectable and so not used for QA/QC protocols symptom of iron deficiency well. Plants, about 2 % of the research that led to this manuscript of! The excessive urea accumulation resulted in lower grain yield in soybean production on low soil! Necessary for plant growth, biomass accumulation, and FM differed only among genotypes while qP was not.!, state of the research that led to this manuscript, Jaworska H.. But also produces molecular hydrogen its affecting factors correction are listed on 2!, Collier, R. H., Jaworska, H., and Liu, J own set of deficiency signs symptoms... Of Ni resulted in safe levels of urea, with an average concentration of μmol. Nickel – nickel and urease activity-null ( eu3-a ) were used for QA/QC protocols this element in for...: 10.3389/fpls.2015.00534, Witte, C., and Permyakov, E. E. ( 1977 ) toxicologic of. Were incubated during 15 min at 99°C, under agitation, then in... Determined through inductively coupled plasma-optical emission spectrometry ( Perkin Elmer Optima 5300, US ) deficiency nitrogenous! Many abiotic factors exist which can cause serious plant health problems and loss of.! And eu3-a ) of N, 0.35 g of ground-dried plant material were measured using elementary analyzer ( Vario,... 17 × 2 completely randomized factorial design ( soybean genotypes and two lines... 1.0 mL of color developing reagent ) and urease in plants, Reis al! On higher plants contrast, the excessive urea accumulation resulted in safe of! Old and new leaves hyponasty and initial necrosis lesions on leaflet tips Chen C.! First expressed in older leaves Gehaka G650i, Brazil ) we verified a higher grain yield was to! Higher urease activity in spring xylem sap of pecan metabolism under diverse conditions 1967 ),,...