Moisture content and storage temperature on the physical and physiological quality of soybean seeds treated with phytosanitary productsABSTRACT:
Phytosanitary seed treatment (ST) of soybeans is fundamental to guaranteeing the establishment of the crop’s stand, for control of diseases and pests, provided it is conducted correctly. A key factor in this process is the moisture content of the soybean seeds at the time of ST, which is also related to their preservation during storage. The aim of this study was to assess how the moisture content of the seeds at the moment of ST affects their physical and physiological quality during storage at different temperatures, with the aim of identifying the ideal moisture content values for ST. Seeds from the same lot were adjusted to moisture contents of 7%, 9%, 11%, 13% and 15%, treated with phytosanitary products and stored at temperatures of 10 °C, 20 °C, 30 °C and alternating 20-30 °C. Physical and physiological quality was assessed after 0, 45, 90 and 135 days, by the germination, emergence, tetrazolium and moisture content tests. Storage at 10 °C was the most efficient for maintaining quality, while higher temperatures accelerated deterioration. Seeds with a moisture content higher than 13% show greater loss of quality, whereas seeds with less than 9% moisture are more susceptible to mechanical damage during ST processing and imbibitional injury. Moisture content of 11% is the most recommended for soybean ST.
Index Terms:
Glycine max L.; industrial seed treatment; seed safety; water content
RESUMO:
O tratamento químico de sementes (TS) de soja é fundamental para garantir o estabelecimento do estande da cultura, desde que realizado de forma correta. Um fator chave nesse processo é o teor de água das sementes no momento do TS, que também está diretamente relacionado à sua conservação durante o armazenamento. Objetivou-se com este estudo avaliar como o teor de água das sementes de soja no momento do TS afeta sua qualidade física e fisiológica durante o armazenamento em diferentes temperaturas, buscando identificar os valores ideais de teor de água para o TS. Sementes de um mesmo lote foram ajustadas para teores de água de 7%, 9%, 11%, 13% e 15%, tratadas com produtos fitossanitários e armazenadas em temperaturas de 10 °C, 20 °C, 30 °C e alternada 20-30 °C. A qualidade física e fisiológica foi avaliada após 0, 45, 90 e 135 dias, pelos testes de germinação, emergência, tetrazolio e teor de água. O armazenamento a 10 °C foi o mais eficiente para manter a qualidade, enquanto temperaturas superiores aceleraram a deterioração. Em sementes com teor de água superior a 13% ocorre maior perda de qualidade, enquanto sementes com menos de 9% são mais suscetíveis a danos mecânicos no processo de TS e embebição. O teor de água de 11% é o mais recomendado para o TS de soja.
Termos para indexação:
Glycine max L.; tratamento industrial de sementes; seed safety; grau de umidade
INTRODUCTION
During seed production, several factors are critical to ensure high quality, including post-harvest care. Variations in seed moisture content and storage conditions directly influence the metabolic activity and physiological quality of seeds (Carvalho et al., 2022; Moraes et al., 2022), which can compromise the initial establishment of crops (Reis et al., 2022).
In Brazil, where the production and storage of soybean seeds are concentrated in tropical regions, the control of temperature and moisture content during storage has become increasingly relevant, due to the challenging environmental conditions of these areas (Virgolino et al., 2016; Carvalho et al., 2022). This care is even more critical with the growing practice of phytosanitary seed treatment (ST), which when carried out in advance, requires adequate storage, with low temperature and relative humidity, to preserve the physiological quality of the seeds until sowing (Reis et al., 2023; Rocha et al., 2025).
Industrial seed treatment (IST) has been widely used as a strategy to protect seed lots against pathogens and initial pests, in addition to ensuring adequate stand (Reis et al., 2023). However, for IST to be effective, it is essential that it does not compromise the physiological quality of the seeds, especially after prolonged storage after the process (Rocha et al., 2025). This makes the storage environment a key factor in preserving the viability of treated seeds, as storage carried out under conditions of high relative humidity (RH) and high temperatures accelerates the seed deterioration process and consequently reduces the quality of the batch (Toni et al., 2024).
The quality of a seed batch depends on physical, physiological, genetic, and sanitary attributes (Sundareswaran et al., 2023). Among these, moisture content, as a physical attribute, stands out for its impact on the metabolic processes of seeds and for increasing susceptibility to mechanical damage during harvesting, processing, and ST (Dias et al., 2018). Although this factor is extremely important, the understanding of its direct influence on ST is still incipient, especially in relation to the consequences on physiological quality after treatment and storage.
This is due to the fact that seeds with low moisture content are more susceptible to mechanical damage during processing. According to Carvalho et al. (2016) and Coppo et al. (2022), this damage is a key factor in the loss of quality, which occurs when the contact of seeds with rigid surfaces causes breaks and cracks in the seed, which in turn reduce the physiological potential of soybean seeds and increase sensitivity to the phytotoxic effect of phytosanitary treatments.
On the other hand, when using seeds with high moisture content in ST, latent mechanical damage can occur, which, together with the high moisture content, increases seed respiration, which evolves during storage and accelerates the deterioration process (Dias et al., 2018; Abati et al., 2021).
Adding these possible types of damage that can occur in ST, storage conditions in turn can accelerate or delay the deterioration of treated seeds (Moraes et al., 2022; Rocha et al., 2025), directly influencing their storability after phytosanitary treatment (seed safety) .
Therefore, the aim of the study was to evaluate how the moisture content at the time of phytosanitary treatment affects the physical and physiological quality of soybean seeds stored at different temperatures, in order to establish ideal values of water content for this operation.
MATERIAL AND METHODS
Soybean seeds of the cultivar Brasmax FOCO IPRO, with an initial moisture content of 11%, were used in the study. To achieve different moisture contents, the seeds were subjected to wetting or drying procedures, following specific protocols for each situation:
Drying: carried out in a longitudinal stationary experimental dryer, adjusted to a temperature of 35 ± 2 °C and an air flow of 23 m³ min-1. Seed moisture content was continuously monitored by weighing samples until reaching the desired value.
Wetting: carried out in a Mangelsdorf-type germinator, adjusted to a temperature of 25 ± 2 °C, with the seeds arranged in a single layer inside woven polyethylene mesh packages. Moisture content was also monitored by means of constant weighing of the samples, until reaching the desired value.
After the wetting or drying protocols, the seeds remained in kraft paper bags, covered by a polyethylene plastic bag, for 12 hours to stabilize the moisture content in their mass. After this period, batches with portions of seeds with moisture contents of 7%, 9%, 11%, 13% and 15% were obtained. The different moisture contents were confirmed using the method of drying in the oven at 105 °C for 24 hours (Brasil, 2025).
The batches were weighed and separated into two-kilogram portions for further treatment. The treatment was performed using the Momesso Arktos Laboratório L5K machine, with the objective of simulating IST in batches. All seeds were treated with the Fortenza® Duo recipe, which contains fungicides and insecticides, according to the active ingredients described in Table 1. In addition, polymer (BioCroma® Red by BioGrow, 100 mL.100 kg-1 of seeds) and finishing powder (Biogloss by BioGrow, 200 g.100 kg-1 of seeds) were added.
Active ingredients and commercial products used in the phytosanitary treatment of soybean seeds.
After treatment, the seeds were packed in kraft paper bags, which were drawn and identified for analysis after each storage period, 0, 45, 90 and 135 days. Storage was carried out in BOD (Biochemical Oxygen Demand) chambers, with temperature control, and kept under the following conditions: 10 °C, 20 °C, 30 °C and a variation of 20-30 °C (alternating for 12 hours), without the presence of light. During storage, there was no control of relative humidity, which was directly dependent on the environmental conditions.
Throughout the storage period, seed quality was evaluated by means of the following tests:
Moisture content: with three replications of 5 g ± 0.5 g of seeds, the moisture content was determined by the method of drying in the oven at 105 ± 3 °C for 24 hours, according to the methodology of Brasil (2025), and the values were expressed as percentages.
Germination in rolled paper (RP): four replications of 50 seeds sown on paper substrate, arranged in rolls and moistened with distilled water (equivalent to 2.5 times the weight of the paper). The rolls were kept in a Mangelsdorf germinator at 25 ± 2 °C. Normal seedlings were counted eight days after sowing, according to Brasil (2025), and the results were expressed as a percentage.
Germination in rolled paper with vermiculite (RP+V): four replications of 50 seeds per treatment were sown in paper rolls moistened with distilled water (3 times the weight of the paper). A thin layer of moistened thin vermiculite was added to the wet paper (two sheets), 100 mL per repetition (ratio of 1 g of vermiculite to 1 mL of water). The rolls were kept in a germinator at 25 ± 2 °C, and normal seedling counts were made at eight days after sowing and expressed as a percentage (Carvalho et al., 2024).
Seedling emergence in controlled conditions (SET): the substrate used was a mixture of sand and soil in a 2:1 ratio, placed in plastic trays. The substrate was irrigated to 60% of the water retention capacity at sowing and uniformly throughout the test. Four replications of 50 seeds were used, and the trays were kept in an oven at 25 ± 2 °C with an alternating regime of 12 hours of light and dark. Seedling emergence count was carried out eight days after sowing and expressed as a percentage (Krzyzanowski et al., 2020).
To evaluate the vigor and mechanical damage of the seeds, with the tetrazolium test, the test was carried out at 0, 90 and 135 days:
Tetrazolium test (TZ): Four replications of 25 seeds were placed between sheets of germination paper moistened with distilled water and preconditioned for 16 hours in a germinator at 25 ± 2 °C. At the end of this period, the seeds were transferred to 50mL plastic cups, fully immersed in tetrazolium solution (0.075%) and kept at 35 ± 2 °C in a BOD chamber until staining, a period of approximately 120 minutes. After staining and washing under running water, the seeds were classified for vigor (sum of levels 1 to 3) and mechanical damage (levels 1 to 8), according to the methodology of França-Neto and Krzyzanowski (2022). The results were expressed as a percentage.
The experimental design was completely randomized, with four replications, in a 5 × 4 × 4 factorial scheme, involving five seed moisture contents, four storage temperatures, and four evaluation times throughout the storage period, except for the tetrazolium test, which was used three storage times. The collected data were subjected to analysis of variance using the F-test (α < 0.05). When significant differences were identified, the means were compared using Tukey's test (α < 0.05) or, alternatively, by polynomial regression analyses, opting for the significant model that had the highest coefficient of determination and with a biologically coherent relationship. All statistical analyses were performed using RStudio software, with the aid of the ExpDes.pt package (R Core Team, 2022).
RESULTS AND DISCUSSION
In the evaluation of moisture content, a significant interaction was identified between the three factors analyzed (moisture content, storage period and temperature). Immediately after ST (0 day), a significant difference was observed only between the moisture contents of the seeds at each storage temperature (Figure 1 and Table 2). This result was expected, since the seeds were evaluated soon after ST, before any influence of the different temperature conditions during storage on their hygroscopic equilibrium.
Percentage variation of moisture contents in soybean seeds during storage, considering different initial moisture contents and storage temperatures
From 45 days on, differences began to emerge between treatments, depending on the storage temperature and on the initial moisture content of the seeds, as they tend to enter hygroscopic equilibrium. At temperatures of 30 °C and 20-30 °C, all seed moisture contents stabilized, with no significant differences between them, regardless of the initial moisture contents, indicating that the seeds reached hygroscopic equilibrium with average values of 6.6% and 8.1%, respectively. This stability at 45 days was not observed at the temperature of 10 °C, at which seeds with lower moisture contents (7%, 9% and 11%) continued with lower values when compared to seeds with higher initial contents (13% and 15%). At the temperature of 20 °C, only the seeds with initial content of 7% differed and remained below the others at 45 days (Table 2).
This indicates that high temperatures accelerate the process of water loss from seeds during storage, when there is no control of relative air humidity, leading to faster stabilization and lower moisture contents. This behavior can be attributed to the inverse relationship between temperature and relative humidity, which directly influences the moisture content of the seeds in their hygroscopic equilibrium (Marcos-Filho, 2015).
The moisture content of soybean seeds is important in storage and quality preservation until the commercialization period. It is worth pointing out that the moisture content of the seeds should be as uniform as possible, because even with its uniformity, it tends to be reduced during storage, until commercialization, when seeds are stored in regions with low relative humidity (Amaral et al., 2018).
In general, the storage temperature influenced the moisture content of the seeds. As shown in Figure 1, the seeds tended to a specific hygroscopic equilibrium for each temperature, at levels inversely proportional to the temperature. In an environment at 30 °C, this trend was more pronounced, with a rapid reduction and equalization of the moisture content for all batches with distinct initial moisture contents. From 90 days on, the moisture content in this environment stabilized in the narrow range between 6.29% and 6.58% (Table 2), the lowest values observed in the experiment. In contrast, at 10 °C, the seeds maintained the highest moisture contents, with the moisture curves remaining closer to the initial values and tending to an equilibrium at higher levels, above 10.6% at 90 days and 11.5% after 135 days. Intermediate behavior was observed at the temperatures of 20 °C and 20-30 °C.
The decrease in temperature in closed environments reduces the air’s ability to retain water vapor. When the absolute humidity is kept constant, such as inside a sealed BOD, there is an increase in relative humidity (Dalpasquale, 2016). Seeds, due to their hygroscopic nature, respond to RH variations by absorbing or releasing water vapor until they reach hygroscopic equilibrium. When RH rises and the seed moisture content is below the new hygroscopic equilibrium, moisture absorption occurs. Conversely, the increase in temperature reduces relative humidity and, consequently, hygroscopic equilibrium, favoring the drying process (Ziegler et al., 2016). The results obtained highlight the inverse relationship between temperature and RH, and consequently the influence on the moisture content of the seeds in hygroscopic equilibrium, reiterating the importance not only of temperature but also of the relative humidity of the environment in seed storage.
Storage at 10 °C resulted in a greater oscillation between treatments, reflecting the influence of low temperature in a closed environment (BOD) on the possible increase in relative humidity and consequently in the moisture content of seeds. A linear increase in moisture content was observed in the seeds with initial contents of 7%, while for the other treatments (9%, 11%, 13% and 15%), there were oscillations in the moisture content, due to the hygroscopic equilibrium over 135 days, with values between 13% and 11% (Figure 1 and Table 2).
For seeds stored at temperatures of 20, 30 and 20-30 °C, stability of the values was observed at 45 days among all samples, which indicates the hygroscopic equilibrium. The approximate moisture contents were 9%, 6.5% and 7.5%, respectively, in each environment. These values remained consistent throughout the storage period, with no further relevant differences between the treatments in relation to moisture content (Figure 1).
Under conditions of constant relative humidity, soybean seeds reach lower moisture contents as the temperature and storage time increase (ASAE, 2021). This behavior was corroborated by the results obtained, as the increase in storage temperature resulted in a progressive reduction in the moisture content of the seeds.
The low moisture content in the seeds, specifically 7% and 9%, resulted in lower germination after phytosanitary treatment (day 0) (Table 3). For Marcos-Filho (2015), seeds with moisture content below 11% are more vulnerable when exposed to substrates with high water availability, such as in germination tests in the laboratory compared to field conditions during sowing, due to imbibition injury. In this context, seeds with moisture contents above 15% do not show significant imbibition injury (Toledo et al., 2010). These results highlight the importance of premoistening for soybean seeds with moisture contents below 11%, as recommended by Toledo et al. (2010), especially for those treated with phytosanitary products.
Throughout the storage period, seeds with high moisture content (15%) lost their physiological quality more quickly, even after reaching hygroscopic equilibrium (Table 3). The damage was progressively intensifying, so that, at 45 days, the seeds showed germination around 78%, a value that is below the minimum standard required for the commercialization of soybean seeds in Brazil. These results suggest that the storage of seeds with high initial moisture contents considerably compromises germination, even after stabilization of moisture content below 9% after 45 days, indicating that the detrimental effects of the high initial moisture content in the seeds, even for only a short period, the first quartile of the storage period, are enough to accelerate the deterioration.
When analyzing the storage temperatures, it was found that at 10 °C there was no significant reduction in germination. At temperatures of 20 °C and 20-30 °C, there were reductions of 4 and 8 percentage points over the 135 days of storage, respectively. At 45 days of storage at 30 °C, with a decrease of 8 percentage points, germination was already below the minimum standard required for commercialization in Brazil (Table 4).
These results reinforce that the temperature of 10 °C is the most suitable to preserve the germination of soybean seeds, corroborating that storage at low temperatures (< 20 °C) not only reduces the speed of physiological deterioration, but also limits the development of pests and pathogens, prolonging the viability of the seeds (Dadlani et al., 2023). Similarly, Abati et al. (2020) observed that the increase in the volume of slurry in IST and the prolongation of storage resulted in a reduction in the quality of soybean seeds, an effect that was mitigated when seeds were stored at 10 °C and 50% relative humidity.
When comparing the factors temperature and storage period (Table 5), it was observed that, regardless of the moisture contents, germination decreased with the increase in temperature. This was more evident in batches with high moisture contents (13% and 15%), due to the combination of adverse conditions.
Subsequent studies have indicated that the humidity of the storage environment is a more determining factor for seed longevity than temperature. When seeds are dried to a relatively low moisture content, their storage capacity increases, even at higher temperatures, such as above 25 °C. However, at temperatures above 25 °C, seeds with moisture content above 12-14% showed greater physiological and sanitary deterioration, highlighting the need for strict moisture control to prolong seed viability (Carvalho et al., 2016; Dadlani et al., 2023; Toni et al., 2024).
In the germination test in rolled paper plus vermiculite (RP+V), it was observed that, at the storage temperature of 10 °C, there were no significant differences throughout the storage period between the initial moisture content treatments; however, at 20°C at 90 days there was a difference, with lower values for 13% and 15% (Table 6). This result was different from that obtained in the test of germination in rolled paper (RP), in which seeds with moisture contents of 7% and 9% had lower germination rates (Tables 3 and 5). The use of RP+V seemed to minimize the imbibition injury due to the larger contact area with the substrate, which retains more water, simulating conditions closer to those found in the field.
Additionally, when comparing the means of germination between the two tests, it was observed that the RP+V methodology tends to better express the germination potential. This may be related to the ability of the vermiculite substrate to attenuate the phytotoxic effect, probably due to its capacity to adsorb phytosanitary products, which is consistent with what has been reported by Carvalho et al. (2024) and Reis et al. (2025).
For seeds with moisture content of 15%, a reduction in the percentage of normal seedlings was observed when they were stored at higher temperatures, such as 20 °C, 30 °C and 20-30 °C (Table 6). This behavior is in line with the results obtained in RP, where high contents of initial moisture, combined with high temperatures, favored seed deterioration.
In the regression graphs for RP+V, it was observed that seeds stored at 10 °C showed germination percentages above 85%, regardless of the initial moisture content (Figure 2). Consistently, Mavaieie et al. (2019) also reported that, under climate-controlled storage conditions, such as in a cold chamber (10 °C and 50% relative humidity), the physiological quality of treated soybean seeds was maintained for up to eight months, with high values of vigor at the end of the period. These findings corroborate the importance of a controlled storage environment to preserve the physiological quality of the treated seeds.
Percentage variation of germination in soybean seeds during storage, considering different initial moisture contents and storage temperatures, by the paper roll + vermiculite test
At high temperatures, such as 20 °C, 30 °C and 20-30 °C, a marked reduction in quality was observed, especially in batches with higher moisture content, throughout the storage period (Figure 2). These results show the importance of controlling the temperature and initial moisture content of the seeds to maintain quality during storage.
In the emergence evaluation, the results differed from those observed in the RP and RP+V tests, showing that, at 0 days, there was no significant difference between the seed moisture contents for seedling emergence (Table 7). This fact demonstrates that, under conditions similar to those of the field, such as the direct contact of the seeds with the soil, the imbibition process occurs more gradually. This reduces the immediate proximity of the phytosanitary product to the seeds and reduces imbibition injury, especially in soybean seeds with low moisture contents (Marcos-Filho, 2015). This information reinforces the importance of choosing the most appropriate methodology for evaluating seed quality.
Throughout storage at 10 °C, up to 90 days, no significant differences were observed in seedling emergence values between the different initial moisture contents. However, at 135 days, seeds with moisture contents of 13% and 15% led to lower emergence values compared to the others (Table 7). This reduction can be explained by the high moisture content, which accelerates hydrolytic reactions and facilitates the increase in concentrations of sugars and fatty acids, favoring the deterioration process (Marcos-Filho, 2015).
At higher temperatures (20, 30 and 20-30 °C), a significant reduction in emergence values was observed, especially for seeds with moisture contents of 13% and 15% at 45 days of storage, and this trend was maintained in subsequent periods (Table 7). Dadlani et al. (2023) also highlighted that the increase in temperature increases the respiratory rate of seeds, intensified by the moisture content, and when this content exceeds 14%, respiration is accelerated, leading to further deterioration.
In addition, seeds with initial moisture contents of 7%, 9% and 11% showed better emergence results at all temperatures tested (Figure 3). On the other hand, seeds with moisture contents of 13% and 15% had greater deterioration, with more intense losses at temperatures of 20, 30 and 20-30 °C. These observations indicate that storage under conditions of high temperature and humidity favors the loss of physiological quality in the first 45 days.
Percentage variation of emergence in soybean seeds during storage, considering different initial moisture contents and storage temperatures.
In the evaluation of vigor by the tetrazolium test, seeds with moisture contents of 7% and 9% showed lower results compared to the other treatments, at temperatures of 10, 20 and 30 °C (Table 8).
This can be explained by the increased susceptibility of these seeds to imbibition injury, which has been discussed previously. This damage occurs mainly in seeds with low moisture content, during rehydration, compromising vigor and the evaluation by tetrazolium. From 90 days of storage, a reduction in seed vigor was observed at all temperatures and moisture contents, when compared to the initial period (0 day) (Table 9).
Seed deterioration is an inevitable process and occurs due to the gradual loss of viability and vigor, especially under less favorable storage conditions. High temperatures and high moisture contents accelerate the deterioration process, which can be explained by the higher metabolic activity of seeds under these conditions, leading to greater degradation of cellular components and compromising the integrity of seed membranes (Dadlani et al., 2023).
In the tetrazolium test, it was found that the reduction of seed moisture content led to an increase in the incidence of mechanical damage, which was more prevalent in seeds with moisture contents of 7% and 9%, regardless of the storage temperatures evaluated (Table 10). Seeds with higher intensity of mechanical damage are more susceptible to the phytotoxic effects of ST, which negatively affect their quality (Coppo et al., 2022). This higher susceptibility may explain the low vigor observed in seeds with moisture content below 9%, since soybean seeds with low moisture content are more negatively impacted by ST (Dias et al., 2018).
In addition, when comparing the moisture contents as a function of storage, seeds with moisture contents of 7% and 9% showed the highest percentages of mechanical damage for all storage periods. It was also observed that seeds with higher initial moisture contents (11%, 13%, and 15%) showed an increase in the incidence of mechanical damage during storage, especially at higher temperatures. This can be explained by the intensification of latent mechanical damage, which became more evident with time and adverse storage conditions (Abati et al., 2021).
These results reinforce the importance of adequate seed storage practices, especially with environments with lower temperatures, which help to preserve quality for longer periods. Storage at higher temperatures can accelerate deterioration and increase susceptibility and the advance of latent mechanical damage.
In addition, accurate knowledge of the moisture content of seeds at the time of treatment is essential to prevent mechanical damage, which occurs more frequently in seeds with low moisture contents. This damage not only affects vigor, but also compromises germination after treatment and during storage.
These practices ensure that the quality of seeds is maintained, from treatment to planting, preserving their germination potential and vigor until cultivation.
CONCLUSIONS
The initial moisture content of soybean seeds has a significant impact on quality during storage. Soybean seeds with initial moisture content higher than 13% show greater loss of quality during storage.
Soybean seed batches with moisture contents below 9% at the time of treatment are more likely to suffer mechanical damage and imbibition injury, especially when immediately subjected to the germination test, compromising their quality.
Moisture content of around 11% is the most suitable for phytosanitary treatment, as it avoids mechanical damage and preserves the quality of seeds during storage.
Storage at 10 °C is ideal for maintaining the physiological quality of treated soybean seeds for up to 135 days. In contrast, high temperatures, especially 30 °C, accelerate seed deterioration.
ACKNOWLEDGMENTS
This study was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico - CNPq (Process: 314408/2021-5), the Fundação de Amparo à Pesquisa do Estado de Minas Gerais - FAPEMIG (Process: APQ-01869-18), and the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES (Finance code: 001). Special thanks to Seedcare Institute Syngenta Brazil.
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Data availability
Additional data will be made available by the authors upon reasonable request.
Publication Dates
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Publication in this collection
01 Sept 2025 -
Date of issue
2025
History
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Received
17 Oct 2024 -
Accepted
01 July 2025
