Physiological quality of wheat seeds influenced by nitrogen sources and rates in soils with low organic matter contentABSTRACT
The production of high-quality wheat seeds in soils with low soil organic matter (SOM) content requires the use of nitrogen (N) fertilization techniques, since different types of N fertilizers have different impacts on crop uptake and development. The present study aimed to evaluate the quality of wheat seeds from modern cultivars, grown in soils with low SOM content, under different ammoniacal and nitric N fertilization rates in Southern Brazil. Seeds were produced in three locations (Capão do Leão, Pelotas and Rio Grande, RS, Brazil), during two crop seasons (2021/22 and 2022/23), under N rates of 0, 40, 80, 120, and 160 kg·ha-1 of N, supplied with urea (46% N) and ammonium nitrate (AN) (27% N). Germination test, dry weight and length of aerial part and root of seedlings, accelerated aging test (AAT), emergence in seedbed, and electrical conductivity (EC) of seeds were evaluated. It was observed that seed germination presented acceptable values, with rates of viable seeds varying from 86 to 99%, except location 3, which presented values below 80% germination at the rate of 160 kg·ha-1 of N. A reduction in seed vigor was observed with the use of AN. Increasing N rates reduced germination in AAT and increased seed EC. EC has a negative correlation with all parameters indicating good physiological seed quality. It was concluded that excessive rates of N negatively impact the physiological potential of wheat seeds grown in soils with low SOM contents.
Key words
Triticum aestivum
; urea; ammonium nitrate; nitrogen efficiency
INTRODUCTION
Wheat (Triticum aestivum L.) is among the most cultivated cereals in the world, occupying the largest planting area (USDA 2023). In Brazil, wheat occupies an area of around 3 million hectares and 8.9 million tons in the 2023/24 crop season, with the largest concentration in the states of the southern region. However, national wheat production does not meet the internal demand for the cereal, requiring the import of around 5.8 million tons annually (CONAB 2024).
Currently, wheat cultivation has demonstrated better productive performance, due to the use of high-yielding cultivars, associated with the use of N fertilization (Bazzo et al. 2020). N-based fertilizers account for about 70% of the total chemical fertilizers used worldwide (Li et al. 2009, Wang et al. 2016, Pasa et al. 2024), However, their efficiency is low, presenting values around 30 to 50% of the applied N (Li et al. 2009, Drury et al. 2017). Low nitrogen use efficiency is the result of N loss processes, such as nitrate (NO3-) leaching, ammonia (NH3), and nitrous oxide (N2O) emissions (Ren et al. 2023, Santos et al. 2023). The low nitrogen use efficiency associated with soils in southern Brazil, which have low SOM content (Carlos et al. 2022), result in low N availability, limiting crop development.
The physiological quality of the seed is influenced by several aspects. One of them is mineral nutrition, in which a well-nourished plant provides higher yield and seed quality (Prando et al. 2012). According to Bazzo et al. (2021), adequate nutrient availability directly affects seed storage structures, chemical composition and embryo formation, influencing the physiological performance of seeds. Among nutrients, N is the most required for good seed formation (Marinho et al. 2022). Nitrogen availability directly interferes with the attributes reported above, in addition to the metabolic functions that impact the initial development of the embryo during germination (Kolchinski and Schuch 2004).
Nitrogen is uptake by the roots in the form of NO3- and ammonium (NH4+), through various physiological mechanisms according to the form absorbed, and the form in which N is found in greater concentration is influenced by temperature, precipitation, and mineralization and nitrification rate (Jackson and Bloom 1990). In the absence of N application, it is through the mineralization process that N is converted from SOM to NH4+, and to NO3- by the nitrification process. However, these processes are less intense at low temperatures, due to the low activity of microorganisms (Beck 1983, Bouwmeester et al. 1985, Pasa et al. 2024). This fact, associated with low SOM content in determined soils, results in a limitation of the natural availability of N, requiring an adequate balance in the supplementation of the nutrient. The use of fertilizers based on NO3- and NH4+ is an alternative, as they provide a more balanced availability of N throughout the crop cycle, improving the source: sink ratio, resulting in higher protein content (Pasa et al. 2024). According to Carvalho and Nakagawa (1988) and Olivoto et al. (2017), there is a positive correlation between the increase in protein content and seed vigor, showing that the use of N sources that increase protein content is essential to produce seeds with high physiological potential.
In this context, there are few studies that aimed to evaluate the physiological quality of wheat seeds in modern cultivars associated with fertilization and plant nutrition with N fertilizer sources in soils with low SOM. In this sense, the objective of this study was to evaluate the quality of wheat seeds of modern cultivars, grown in soils with low organic matter content, under different ammoniacal and nitric nitrogen fertilization rates in southern Brazil.
MATERIALS AND METHODS
Seeds were collected over a period of two years (2021/22 and 2022/23) in the municipalities of Capão do Leão, RS (location 1) (31°48’07,9”S and 52°30’17,9”W, altitude 45 m), Pelotas, RS (location 2) (31°31’50,7”S and 52°14’10,8”W, altitude 26.7 m), and Rio Grande, RS (location 3) (32°12’30,9”S and 52°32’09,7”W, altitude de 6.6 m). The soil from locations 1 and 2 are classified as Luvisol, with SOM contents of 2 and 2.1%, respectively, and from location 3 is Planosols, with 1.2% of SOM (WRB 2014). The cultivars used were Tbio Audaz, Tbio Astro, and Tbio Toruk, respectively, in locations 1, 2 and 3. The Tbio Audaz was used because it has an early cycle and is similar to the Tbio Toruk, but with a medium cycle, but both are demanding in N. Tbio Astro is characterized by high resistance to lodging and has a super early cycle. For a better understanding of the results, from this point onwards a coding similar to that used by Pasa et al. (2024) was adopted, location and year being referred as L (1, 2, 3) and Y (2021, 2), respectively. For instance, for study 1 in the year 2021 the coding L1-Y2021 was used.
The study was conducted in all locations under field conditions (Table 1). In previously prepared areas, under both fallow with summer grasses, and after wheat cultivation, soybeans were cultivated in order to represent the crop used in the region, without the presence of irrigation. The design used was randomized blocks with four replications, with locations 1 and 3 using a 2 × 3 + 1 factorial (2 N sources × 3 N rates + 1 control) and location 2 using a 2 × 4 + 1 factorial (2 N sources × 4 N rates + 1 control). The N rates were 0, 40, 80 and 120 in L1-Y2021, 0, 40, 80 and 160 in L1-Y2022 and L3-Y2022, and in L2-Y2022, 0, 40, 80, 120 and 160 kg·ha-1 of N. The N rates were split into two applications at phenological growth stages (GS), GS 21 and GS 31 (Zadoks 1985), each one with 50% of the N rate. Ammonium nitrate (27% N, Yarabela) and urea (46% N, common urea) were used as N sources, and 200 kg·ha-1 of formulated fertilizer 5 – 20 – 20 (N – P2O5 – K2O) was used as starter fertilizer, applied in the sowing furrow. The experimental plots consisted of nine rows with a spacing of 0.17 cm by 4 m in length, with the seven central rows of 3 m in length being used as the useful area of the plot.
Planting, harvest, average temperature, and precipitation data from the three cultivation locations.
The seeds were harvested after they reached physiological maturity. After harvesting, the seeds were dried in a stationary dryer at the temperature of 40°C until they reached a moisture content of 13%, with moisture content monitored using a John Deere GT 5300 grain moisture tester. After drying, the seed samples were stored for four months in a cold chamber (15.6 ± 2°C), with relative humidity around 40 ± 5%, until the physiological quality analyses of the seeds were carried out.
Physiological seed analyses
The germination test was performed with 200 seeds per replicate, divided into four subsamples of 50 seeds, mounted on germitest paper rolls moistened with distilled water equivalent to 2.5 times the weight of the paper, being placed in a germinator at 20°C. The first count was performed four days after the test was set up, and the final count was performed on the eighth day. The seedlings were identified as normal, abnormal and dead seedlings, and represented as a percentage (Brasil 2009).
Along with the germination test, four replicates of 20 seeds were set up, placed on the upper third of the germitest paper sheet, to analyze the length and mass of seedlings. After the fifth day, the seedling length was evaluated, with the aid of a graduated ruler and the results expressed in cm (Nakagawa 1999). The seedling dry weight was measured after the seedling length assessment, in which they were placed in paper bags and taken to an oven for 72 hours, at 65°C and forced air circulation, until reaching a constant weight. After that, a precision scale (0.0001 g) was used to calculate the dry weight. The results were expressed in mg per seedling.
The accelerated aging test (AAT) was performed in biochemical oxygen demand (BOD) at 43°C for 48 hours, and the seeds were placed in gerbox-type boxes, homogeneously on the metal screen, and with 40 mL of saturated saline solution (Pedroso et al. 2010), composed of 11 grams of NaCl per 100 mL of distilled water, inside the boxes. Subsequently, 200 seeds were used per replicate, divided into four subsamples of 50 seeds and subjected to the standard germination test, for four days.
Seedling emergence was carried out according to Prando et al. (2012) in beds containing soil as substrate. Sowing consisted of four replicates of 50 seeds, carried out at the depth of 3 cm, with a distance between rows of 10 cm and humidity maintained with irrigation whenever necessary. The results were expressed as a percentage, and the evaluation was carried out on the 14th day after the test was implemented, consisting of the manual counting of seedlings emerged in each replicate.
Electrical conductivity (EC) analysis was performed by weighing four subsamples of 50 seeds from each treatment repetition, which were weighed in a precision scale (0.0001 g) and then placed in plastic cups with 75 mL of deionized water. As a standardization of the test, two controls containing only 75 mL of deionized water were used. The samples were taken for 24 h to a BOD germinator at the temperature of 25°C, and after this period the reading was performed with the aid of a bench conductivity meter (Vieira and Krzyzanowski 1999).
Statistical analysis
The data were subjected to the F-test, with the aid of R software (R Core Team 2020), and, when analysis of variance (ANOVA) was significant for the treatments, comparison of means (Tukey’s test) and polynomial regression were performed for qualitative and quantitative data, respectively, with support of the ExpDes package (Ferreira et al. 2013). The locations were considered as a factor only to run a three-way interaction and verify whether there were significant differences between the locations for each parameter, and ,when significant, data for each location were presented separately. Pearson’s correlation was performed with p < 0.05 and with the aid of the R packages corrplot (Wei and Simko 2021) and Hmisc (Harrell Jr. and Harrell Jr. 2019).
RESULTS AND DISCUSSION
The germination normal seedlings (GNS) test (Table 2) showed interaction between factors in L3-Y2022, with seed germination superiority with urea of 12.6 and 11.8% in relation to AN, at the respective rates of 40 and 80 kg·ha-1 of N. In the other locations and years evaluated, no difference was observed between the N sources, only the simple effect of the N rate was observed, with a decreasing linear behavior in the GNS in relation to the increase in N rates, except for L1-Y2021 (Table 2).
Germination of normal seedlings (GNS) from the germination test of wheat seeds produced with different N rates using urea and ammonium nitrate (AN).
The absence of variation in the germination test is attributed by Bazzo et al. (2021) and Marinho et al. (2022), because seeds with high vigor have reserves that correctly supply the embryo. It is described that high vigor seeds undergo less variation with variation of N rates (Bazzo et al. 2021). However, in soils with low SOM levels, the variation in N rates directly impacts the crop, due to the relatively low SOM N mineralization rates (Chen et al. 2015, Ray et al. 2020) and low N availability, restricting crop development and yields.
The decrease in germination with increasing N rates may be the result of excessive N rates leading to a higher incidence of diseases such as blast (Magnaporthe oryzae Triticum) (Silva et al. 2019) and cell toxicity (Elhanafi et al. 2019). However, the germination percentage remained satisfactory up to the rate of 120 kg·ha-1 of N in all locations, which is reported by Alves et al. (2017), who showed that the application of up to 120 kg·ha-1 of N is beneficial for the wheat crop. Pasa et al. (2024) also observed that the most significant increase in the protein content in wheat grains of modern cultivars occurred up to the rate of 120 kg·ha-1 of N. Protein content has a direct correlation with the improvement of the physiological potential of seeds (Carvalho and Nakagawa 1988, Olivoto et al. 2017), but, in the present study, no improvement in quality was observed with the increase in N rates. However, except for the rate of 160 kg·ha-1 of N in L3-Y2022, all other locations and rates evaluated showed a germination percentage above 80%, which is required by Brazilian legislation as a minimum value for commercialization of wheat seeds (Brasil 2013).
Regarding the seedling root length (SRL), a simple effect of the N source was observed in L2-Y2022 (Table 3), with superiority when AN was used in relation to urea. The effect of the N rate was observed in L2-Y2022 and L3-Y2022, with a decreasing linear behavior in relation to the increase in N rates. In a study conducted by Marinho et al. (2022), a quadratic response was observed in relation to N rates on the length of wheat seedlings. The dry weight of the seedling root did not show any significant effect under the conditions evaluated. The length of the aerial part of the seedlings (LAPS) showed interaction between the factors in L1-Y2021 and L3-Y2022, with AN being superior in L1-Y2021 (Table 4) in relation to urea at the rate of 40 kg·ha-1 of N, and in L3-Y2022 (Table 3) urea was superior in relation to AN at a N rate of 160 kg·ha-1 of N. These results disagree with those found by Prando et al. (2012), who did not observe significant differences in the rates of 0, 40, 80 and 120 kg·ha-1 of N. The variation observed between locations can be attributed to the genotype, since the effect of N fertilization can vary between cultivars (Benincasa et al. 2022, Pasa et al. 2024), due to distinct characteristics of each genotype, which can alter the N absorption, assimilation and conversion (Luo et al. 2020). According to Gonçalves et al. (2020), the choice of genotype is essential to minimize the effects of the genotype × environment interaction, and provide more favorable conditions for the production of quality seeds.
Seedling root length (SRL) and length of the aerial part of the seedling (LAPS), dry weight of the seedling root (DSR), and dry weight of the aerial part of the seedling (DAPS) of wheat seeds subjected to different rates of N with the use of urea and ammonium nitrate (AN), at location L2-Y2022 and L3-Y2022.
Seedling root length (SRL) and length of the aerial part of the seedling (LAPS), dry weight of the seedling root (DSR), and dry weight of the aerial part of the seedling (DAPS) of wheat seeds subjected to different rates of N with the use of urea and ammonium nitrate (AN), at location L1-Y2021 and L1-Y2022.
The first count of germinated seeds (Fig. 1) from the germination test showed interaction between the factors in L1-Y2021 (Fig. 1a) and L3-Y2022 (Fig. 1c). In L1-Y2022, superiority of germinated seeds was observed with the use of AN only at the rate of 160 kg·ha-1 of N, as well as a decreasing linear behavior with the increase in N rates with the use of urea (Fig. 1a). In L1-Y2022 and L2-Y2022 (Fig. 1b), the increase in rates showed a decreasing linear behavior with the increase in N rates, regardless the N source used. In L3-Y2022, a higher rate of germinated seeds was observed with the use of urea at rates of 40, 80 and 160 kg·ha-1 of N, in relation to AN (Fig. 1c). Seed vigor is directly affected by N fertilization, as higher N availability contributes to more intense responses on the seed protein content and assimilation of reserves (Marcos Filho 2015).
Emergence of normal seedlings in the first count of germination test of wheat seeds subjected to different N rates, with urea and ammonium nitrate, in (a) L1-Y2021, (b) L1-Y2022, L2-Y2022 and (c) L3-Y2022. Asterisks at the top of the graph indicate significant difference by Tukey’s test (p < 0.05).
The accelerated aging test showed a simple effect of the N source in L1-Y2021 (Fig. 2), with a higher rate of germinated seeds with the use of urea. However, it was observed that seed vigor was reduced with increasing N rates (Fig. 2). Excessive rates of N can lead to lodging (Galetto et al. 2017) and diseases, such as blast (Magnaporthe oryzae Triticum) (Silva et al. 2019), thus reducing the physiological quality of the seed. The accelerated aging test allows to determine the physiological quality of the seed after storage, indicating possible poor formation of embryos and reserves, reducing seed germination (Do Lago and Martins 1998).
Percentage of emergence of normal seedlings of wheat seeds subjected to the accelerated aging test (AAT), produced with different N rates, supplemented with urea and ammonium nitrate, in L1-Y202(1-2) and L2-Y2022.
The evaluation of seedlings emergence in the seedbed (SES) showed a significant interaction between N rate and N source only in L1-Y2021 (Table 5), with a higher seedling emergence ratio at rates of 40 and 80 kg·ha-1 of N with the use of urea in relation to AN. This fact may be related to the differences observed in AAT and EC in L1-Y2021, indicating that the seeds produced with urea in this year and location have a better physiological potential than those produced with AN. Seeds from L1-Y2022 and L2-Y2022 did not show significant differences. Similarly, Bazzo et al. (2018), in white oats (Avena sativa L.), did not observe SES variation with the increase in N rates.
Percentage of seedlings emerged in a seedbed from wheat seeds subjected to different rates of N with the use of urea and ammonium nitrate (AN), from locations L1-Y202(1-2) and L2-Y2022.
Prando et al. (2012) observed an increase in EC with the higher N application rate, corroborating the results found in the present study, which, in both locations evaluated, presented higher EC with the highest N rates (Fig. 3). An interaction was observed between the source and N rate factors in L2-Y2022 (Fig. 3b), with a higher EC index when AN was used in relation to urea, with superiority of 24.6, 31.1 and 30.6%, respectively, at rates of 80, 120 and 160 kg·ha-1 of N. This increase with the use of AN may be associated with the higher protein content in wheat grains observed by Pasa et al. (2024), increasing the concentration of solutes that can migrate into the water during the test.
Electric conductivity (EC) of wheat seeds of (a) L1-Y202(1-2) and of (b) L2-Y2022, resulting from difference N rates with the use of urea and ammonium nitrate. Asterisks at the top of the graph indicate significant difference by Tukey’s test (p < 0.05).
However, it is observed that EC presents a high negative correlation with all evaluated parameters of seed physiological quality (Fig. 4), that is, the higher the EC of the seeds, the lower their physiological quality. This occurs because, according to Abreu et al. (2011) and Prado et al. (2019), the increase in EC is associated with greater degradation of the membranes, reducing the germination potential of the seeds, increasing the amount of nutrients and solutions leached into the water, contributing to higher EC. The observed positive correlation of LAPS with SRL and normal seedlings in the first count of the seed germination test (FCGT), and of FCGT with normal seedlings in the second count of the seed germination test and AAT, highlights the direct influence that seed vigor has on seed physiological quality, since, according to Finch-Savage and Bassel (2016), the germination rate and seedling growth, called vigor characteristics, are interconnected and play a critical role in crop establishment.
Pearson’s correlation between variables in wheat seeds with difference N rates, with the use of urea and ammonium nitrate, in two crop seasons and two locations in Southern Brazil*.
CONCLUSION
Nitrogen rates higher than 120 kg·ha-1 in Planosols reduced seed germination of modern wheat cultivars to unacceptable levels when grown in soils with low organic matter content. In Luvisols, seed germination was maintained with rates of up to 160 kg·ha-1 of N.
Seedling emergence in the Tbio Audaz bed was reduced with increasing N rates, but Tbio Astro did not show this behavior, indicating that the response to nitrogen fertilization depends on the genotype. Increasing N rates increases the EC of wheat seeds, as does the use of ammonium nitrate, presenting negative effects on the physiological quality of seeds in relation to the use of urea.
The physiological quality of seeds is affected by soil and climate conditions, with variations in quality and response to nitrogen fertilization according to the genotype and type of soil cultivation. In this context, higher N rates in soils with low organic matter content contribute to maintaining the physiological quality of seeds. However, when cultivation is carried out in soils with low organic matter content, the type of nitrogen source interferes with the physiological quality of seeds.
ACKNOWLEDGMENTS
The authors would like to thank all the students and staff for their contributions in the development of this research. The authors would also like to thank the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).
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How to cite: Pasa, E. H., Pasa, M. S., Ferreira, J. P., Weinert, C., Vargas, V. L., Otero, A. S., Martinez, F. P., Pedó, T. and Carlos, F. S. (2025). Physiological quality of wheat seeds influenced by nitrogen sources and rates in soils with low organic matter content. Bragantia, 84, e20240178. https://doi.org/10.1590/1678-4499.20240178
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FUNDING
Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorFinance code 001
DATA AVAILABILITY STATEMENT
Data will be available from the corresponding author on reasonable request.
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SRL: wheat seedling root length; LAPS: length of the aerial part of the wheat seedling; DSR: dry weight of the wheat seedling root; DAPS: dry weight of the aerial part of the wheat seedling; FCGT: normal seedlings in the first count of the seed germination test; SCGT: normal seedlings in the second count of the seed germination test; AAT: normal seedlings from the accelerated aging test; ETS: seedlings emerged in the seedbed emergence test; EC: electrical conductivity test on wheat seeds; *cells with “X” are not statistically correlated (p < 0.05).
