Efficiency of isolated atmosphere packaging in maintaining the physiological quality of soybean seeds stored with different moisture contentsABSTRACT:
Maintaining the physiological quality of soybean (Glycine max L.) seeds after harvest is challenging due to their lipidrich composition, which accelerates deterioration under high moisture contents and inadequate packaging. This study evaluated seeds stored with initial moisture contents of 9%, 11%, and 13% in conventional big bags and ATI (isolated atmosphere) packaging over 225 days. Seed quality was strongly influenced by the interaction between moisture content, packaging type, and storage duration. Seeds stored at 9% moisture showed greater metabolic stability, maintaining germination above 95% and high vigor throughout the storage period. In contrast, seeds stored at 11% and especially 13% moisture exhibited accelerated deterioration, reflected in reduced vigor, increased formation of weak seedlings, and higher sensitivity to stress tests. ATI packaging minimized hygroscopic fluctuations and slowed degenerative processes, outperforming conventional packaging, particularly for seeds with higher moisture. Tetrazolium, accelerated aging, and emergence tests confirmed that combining lower initial moisture with an efficient hygroscopic barrier significantly extends physiological longevity. Overall, the results indicate that storing soybean seeds at 9% moisture in ATI packaging is the most effective strategy for preserving quality during prolonged storage, whereas moisture levels above 11% intensify deterioration even under controlled environmental conditions.
Index terms:
moisture content; packaging; seed quality
RESUMO:
A manutenção da qualidade fisiológica da soja (Glycine max L.) após a colheita é um desafio devido à sua composição lipídica, cuja deterioração é acelerada por teores elevados de água e por embalagens com baixa eficiência higroscópica. Este estudo avaliou sementes armazenadas com 9%, 11% e 13% de umidade inicial em big bags convencionais e com atmosfera modificada (ATI) ao longo de 225 dias. Os resultados mostraram que a qualidade fisiológica foi fortemente influenciada pela interação entre umidade, embalagem e tempo de armazenamento. Sementes com 9% mantiveram maior estabilidade metabólica, apresentando germinação superior a 95% e elevado vigor durante todo o período. Em contraste, teores de 11% e, principalmente, 13% aceleraram a deterioração, refletida em maior queda no vigor, aumento de plântulas fracas e maior sensibilidade aos testes de estresse. A embalagem ATI reduziu oscilações higroscópicas e retardou processos degenerativos, sendo mais eficiente que a convencional, sobretudo em sementes com maior teor de água. Testes de tetrazólio, envelhecimento acelerado e emergência confirmaram que a combinação entre menor umidade inicial e barreira higroscópica eficiente prolonga a longevidade fisiológica. Concluise que o teor de 9% associado à embalagem ATI é a estratégia mais eficaz para preservar a qualidade da soja em armazenamento prolongado, enquanto teores superiores a 11% intensificam a deterioração mesmo sob ambiente controlado.
Termos para indexação:
teor de água; embalagem; qualidade de sementes
INTRODUCTION
Soybean (Glycine max (L.) Merrill) has consolidated itself as the main crop of Brazilian agribusiness, with total production estimated at 166.28 million tons for the 2025/26 harvest, which represents a significant increase compared to the previous cycle. This performance was supported by a cultivated area of 47.40 million hectares (CONAB, 2026).
A seed of high physical, physiological, genetic, and sanitary quality is essential for agricultural success, as it ensures rapid and uniform emergence, especially under adverse conditions (Arachchige et al., 2024). However, soybean seeds have high susceptibility to postharvest deterioration due to their lipid-rich composition, initiating losses soon after physiological maturity. In tropical and subtropical environments, maintaining viability during storage becomes a major challenge, since management failures accelerate the degenerative process of seeds (Nardelli et al., 2025).
Proper storage acts as a regulator of seed basal metabolism, aiming to preserve vigor for as long as possible. The control of environmental variables, such as temperature and relative humidity, is essential to mitigate the respiratory rate. Uncontrolled environments expose the embryo to fluctuations that trigger the degradation of vital reserves, resulting in lots that perform poorly in the field (Marcos-Filho, 2015).
Complementary to the environment, the type of packaging used for storage plays a decisive protective role. The technological evolution of materials has allowed the development of packages that work as efficient hygroscopic barriers, capable of isolating the seed from atmospheric humidity variations, preventing moisture oscillations, and preserving the integrity of cell membranes and the stability of organelles (Capilheira et al., 2019). In this context, Oliveira et al. (2024) evaluated chickpea seeds and observed that the interaction between environment and storage directly influenced physiological quality.
The initial moisture content of the seed at the time of bagging is one of the variables with the greatest weight in longevity during storage, as moisture content is the regulator of the enzymatic activity and the speed of the internal biochemical reactions. Seeds stored with moisture content above the critical limit suffer an increase in respiration, which generates heat and creates an environment conducive to the proliferation of storage fungi, compromising physiological quality (Krzyzanowski et al., 2022).
Thus, the interaction between moisture, packaging and storage time defines the deterioration curve of each cultivar, with the loss of vigor being an event that precedes the loss of germination capacity, which makes the rigorous monitoring of these factors essential to avoid losses to the producer. Strategies that integrate technological packaging and initial moisture control are fundamental for the competitiveness of the soybean production chain (Marcos-Filho, 2015).
Mapiemfu-Lamare et al. (2026) evaluated the quality of soybean seeds stored under different packaging materials, temperatures, and genotypes, and demonstrated that the interaction between environment and packaging directly affects seed germination, viability, and vigor. In a complementary way, Coradi et al. (2020) also investigated different types of packaging in soybean storage and found that materials with greater barrier significantly reduce the loss of vigor over time.
Although recent literature advances in understanding the isolated effects of moisture, temperature, or packaging, there is still a clear gap: a lack of studies that simultaneously evaluate different initial moisture contents and different types of packaging over prolonged storage periods, especially with detailed physiological monitoring.
Thus, the objective of the present study was to fill this gap by analyzing, in a joint and temporally extensive way, how these factors modulate the deterioration and physiological performance of seeds over 225 days.
MATERIAL AND METHODS
The research was carried out in a soybean seed production company, located in the municipality of Campo Novo do Parecis, MT (-13.8177390, -57.6566850). A commercial lot of soybean seeds of the O800 I2X (NEO 800) cultivar, from the 2025/2026 harvest and from the aforementioned company, was used in the study.
A big bag of five million seeds from the lot previously selected after processing, with initial moisture content of 13%, was used. Subsequently, part of the seeds were dried to 11% and part to 9% with the aid of a Marcold Turbograin® in an adapted drying system, with drying air temperature and humidity of 30 °C and 50%, respectively. Soon after drying, the seeds were placed in the packages and stored in a cold chamber, with an average temperature of 12 °C and 60% relative humidity.
To conduct the experiment, the seeds were packed in small bags of 5 kg capacity, one conventional and one with Isolated Atmosphere (Atmosfera Isolada - ATI). The conventional bag was produced with braided polypropylene (PP) raffia fabric, with free passage of air and moisture, leaving the content exposed to the external environment and pests. The ATI bag, on the other hand, completely blocks the entry of oxygen and moisture, creating an isolated atmosphere.
The experimental design was completely randomized, in a 3 x 2 x 6 factorial scheme, with three replicates for each treatment. The treatments consisted of three initial storage moisture contents (9, 11 and 13%), two packages (conventional big bag and ATI) and six storage periods (0, 45, 90, 135, 180 and 225 days), respectively.
After each storage period, a 1 kg sample was collected from each 5 kg experimental unit. These samples were homogenized with the help of a soil homogenizer and sent to the company’s seed analysis laboratory.
The variables analyzed were:
Moisture content: evaluated using a Motomco® 999ESI moisture meter. The evaluations were carried out in triplicate, and the results were expressed as a percentage of moisture.
Germination: determined with four replicates, each consisting of four subsamples of 50 seeds, sown on paper towel substrate (Germitest®), previously moistened with distilled water, using 2.5 times the weight of the dry paper, and kept in a germinator at a temperature of 25 °C. The evaluations were carried out eight days after sowing, and the results were expressed as a percentage of normal seedlings, according to the Rules for Seed Testing (Brasil, 2025). Concomitant with this test, the seedlings were classified into strong and weak according to Krzyzanowski et al. (2020).
Tetrazolium test: performed using 100 seeds per replicate, divided into two subsamples of 50 seeds, which were pre-conditioned between paper for 16 hours at a temperature of 25 °C. After the time necessary for conditioning, the seeds were completely submerged in the solution of 0.075% tetrazolium salt, placed in an oven at a temperature of 40 °C and kept for a period of 2.5 hours, being subsequently washed and analyzed individually for viability and vigor, with the results expressed as percentage (França-Neto and Krzyzanowski, 2018).
Accelerated aging: conducted with 4 subsamples of 50 seeds per replicate, which were arranged in a single layer on a stainless steel screen inside plastic boxes (gerbox) containing 40 mL of distilled water at the bottom (Marcos-Filho, 2020). Subsequently, the boxes were transferred to a BOD germination chamber at 41 °C and kept for 48 hours. After this period, the seeds were subjected to the germination test, as previously described. The evaluation was carried out on the fifth day after sowing, computing the seedlings considered normal (Brasil, 2025). The results were expressed as a percentage of normal seedlings.
Seedling emergence: conducted with four replicates with four subsamples of 50 seeds, distributed in beds with dimensions of 5 meters long and 1 meter wide (5 x 1m), filled with soil collected from the A1 horizon of a Latossolo vermelho distrófico (Oxisol) and medium sand, in a 1:1 ratio. The evaluations were carried out seven days after sowing, computing the number of seedlings that emerged with a length equal to or greater than 3.0 cm above the soil layer. The results were expressed as a percentage of emerged seedlings.
The data were previously subjected to the Shapiro-Wilk normality test and the Bartlett homogeneity of variances test. Once the assumptions were met, the data were subjected to analysis of variance (ANOVA) by the F test at 5% probability level. For the qualitative factor (initial moisture and packaging), the means were compared by Tukey test at 5% probability level. For the quantitative factor (storage periods), the data were subjected to regression analysis, and the models were chosen based on the significance of the coefficients. The analyses were performed using the statistical software SISVAR version 5.6 (Ferreira, 2011).
RESULTS AND DISCUSSION
The analysis of variance indicated a significant triple interaction between the factors, PA × M × P (packaging x moisture x storage periods), only for the moisture content variable, indicating that the variation in seed moisture content over time is simultaneously influenced by the type of packaging and the initial moisture content. However, it was observed that the interaction between packaging and moisture (PA × M) was significant for the moisture content and vigor evaluated by the tetrazolium test (Table 1).
Analysis of variance of the variables moisture content (M), germination (GER), germination with classification of strong seedlings (GERS), viability in tetrazolium test (VIATZ), vigor in tetrazolium test (VIGTZ), 48-hour accelerated aging (48AA) and seedling emergence in bed (EB) for soybean seeds subjected to storage in conventional and ATI (Isolated Atmosphere) packaging, with different levels of initial seed moisture and storage periods.
In relation to the interaction between packaging and storage period (PA × P), there was a significant effect only for moisture content and emergence in bed with classification of strong seedlings (Table 1). On the other hand, the interaction between moisture and storage period (M × P) was significant for variables such as germination, 48-hour accelerated aging with classification of strong seedlings and emergence in bed with classification of strong seedlings (Table 1).
Regarding seed moisture content, it was found that, for all storage periods, seeds stored in conventional packaging (with initial moisture content of 13%) had values lower than those observed in seeds stored in ATI packaging with the same initial moisture, except for the period of 225 days (Table 2). In addition, there was no significant difference for the moisture content between the types of packaging in seeds with initial moisture contents of 9 and 11%, for all periods evaluated (Table 2).
At 45 days of storage, it was observed that the moisture content of seeds stored in both packages showed similar behavior to that of the initial time (0 days) (Table 2). However, this pattern was not maintained in the other periods, for which, regardless of the packaging, the seeds with initial moisture content of 11 and 13%, when compared, did not differ significantly in the final moisture content (Table 2).
A quadratic behavior was observed in Figure 1A for the moisture content of seeds stored in conventional packaging, with a reduction in the initial storage periods. This reduction continued until 41, 74 and 176 days, reaching minimum values of 8.90, 10.42 and 11.10% for seeds with initial moisture contents of 9, 11 and 13%, respectively. On the other hand, when the seeds were stored in ATI packaging, there was a linear decreasing behavior throughout the storage period (Figure 1B), indicating greater stability in the variation of the moisture content when compared to those stored in the conventional packaging, since the maximum moisture reduction was approximately 1 percentage point after 225 days at the initial moisture content of 13%.
Moisture content of soybean seeds subjected to storage in conventional (A) and ATI (B) packages, with different initial moisture contents and storage periods.
The dynamics observed in Figure 1 are directly associated with the role of respiration and lipid peroxidation in seed deterioration. At higher moisture contents, respiration increases, increasing the production of heat and ROS, which initiate reactions of oxidation of the unsaturated fatty acids present in the membranes (Krzyzanowski et al., 2022). This process, known as lipid peroxidation, is particularly critical in oilseeds such as soybeans, as it leads to membrane rupture, solute leakage, and loss of cell compartmentalization. The hygroscopic stability provided by ATI packaging reduces these metabolic oscillations, slowing the advance of oxidative deterioration and explaining the more uniform behavior of seeds throughout storage (Waterworth et al., 2024).
In general, the germination of soybean seeds during storage, as a function of the different initial moisture contents, remained above 95% throughout the storage period. At 45 days of storage, no significant differences were observed in the germination of seeds with different initial moisture contents (Table 3). In this context, Baributsa and Baoua (2022) highlight that there is a strong negative correlation between moisture content and germination of soybean seeds, indicating that the absence of increment in moisture prevents the intensification of deterioration, indicated by the reduction in seed moisture throughout storage (Figure 1).
The germination of soybean seeds throughout the storage period showed a trend of reduction for all the initial moisture contents evaluated (Figure 2), although this decrease occurred gradually and not very markedly. For seeds with initial moisture of 9%, a decreasing linear fit was observed, indicating low germination loss throughout storage. A similar behavior was observed in seeds with initial moisture of 11%, showing the highest rate of germination reduction compared to the other initial moisture contents (Figure 2). On the other hand, seeds with initial moisture content of 13% showed quadratic fit, with a reduction only in the final stages of storage (Figure 2).
Average germination of soybean seeds stored in different packages and subjected to storage with different initial moisture contents and storage periods.
Environments with humidity control contribute to maintaining the hygroscopic balance of seeds, avoiding variations that accelerate the loss of physiological quality. This is especially relevant for soybean seeds, whose deterioration is directly associated with fluctuations in moisture content during storage. For this reason, the combination of controlled environmental conditions and moisture content stability explains the behavior observed in Figure 2, in which germination remained high and showed a gradual reduction, even for seeds with initial moisture of 13%.
The results obtained converge with the findings of Nardelli et al. (2025), who demonstrated that the combination of initial moisture content and storage temperature is determinant for maintaining the physiological quality of soybean seeds. Even under controlled conditions, seeds with higher moisture content showed accelerated deterioration, with reduced vigor before germination drop, a pattern also observed in this study for seeds with 13% moisture. Thus, the data reinforce that, although the controlled environment is essential, the initial moisture content remains the main factor modulating seed longevity during storage.
According to the data in Table 4, soybean seeds showed high viability in the tetrazolium test regardless of the initial moisture content, with average percentages of 97, 98 and 96% at the initial moisture contents of 9, 11 and 13%, respectively. These results indicate that, within the range evaluated, small variations in seed moisture content do not drastically compromise the viability detected by the tetrazolium test.
The vigor of soybean seeds, evaluated by the tetrazolium test, 48-hour accelerated aging and seedling emergence in bed varied as a function of the combination between initial moisture content and type of packaging (Table 5). In the tetrazolium test, the means ranged from 95 to 97%, with no differences due to the initial moisture, because the differences between packages and moisture contents were small and with non-significant interactions.
By the 48-h accelerated aging test, it was observed that the soybean seeds maintained high percentages of vigor, with means ranging from 92 to 94% between the different levels of initial moisture and types of packaging. In addition, a significant difference was observed only for seeds stored with 9% moisture, with those stored in the ATI packaging showing greater vigor by the accelerated aging test (Table 5). Results observed by Capilheira et al. (2019) corroborate this trend, showing that the ATI packaging promotes greater stability of moisture content and contributes to the maintenance of vigor, especially in seeds with lower initial moisture. However, unlike the study conducted by Capilheira et al. (2019), which mainly evaluated the effect of the modified atmosphere, the present study advances by demonstrating that the effectiveness of the physical barrier of ATI is enhanced when associated with lower moisture contents, indicating a strategic interaction between adequate drying and choice of packaging.
In summary, the data from the 48-h accelerated aging test corroborate its efficacy as a sensitive test to identify high levels of vigor in soybean seeds (Table 5), demonstrating that small variations in the initial moisture content of soybean seeds significantly influenced the response to accelerated stress (França-Neto et al., 2025).
Regarding the results of seedling emergence in bed, high percentages of emergence were found for all treatments, ranging from 93 to 95% (Table 5). Significant differences were observed only in the treatment with initial moisture of 13%, in which seedlings grown from seeds stored in ATI packaging had a higher percentage of emergence (95%) compared to conventional packaging (93%). At the other moisture contents (9% and 11%), no statistically significant differences were observed between the types of packaging (Table 5).
When evaluating the percentage of strong seedlings in the 48-hour accelerated aging test at the beginning of storage (0 days), no significant difference was observed as a function of the initial moisture content of the seeds (Table 6). At 45, 90 and 135 days, there were also no significant differences between moisture contents, indicating maintenance of vigor regardless of the initial moisture and regardless of the packaging used. However, the moisture content of 13% led to significantly lower values at 180 and 225 days, with 84 and 79% in the ATI packaging, and at 225 days, with 82% in the conventional packaging, compared to the moisture contents of 9% and 11%, which did not differ from each other. This highlights that, over prolonged periods, higher moisture contents favor seed deterioration (Table 6).
Soybean seeds stored in conventional packaging, regardless of moisture, resulted in an average of 89% of strong seedlings. In the ATI packaging, there was an average of 91% of strong seedlings with 9% moisture, and the quality decreased as the moisture content increased (Table 6). This reinforces that packaging with an adequate barrier can attenuate the effects of moisture on vigor during storage.
In summary, the results showed that, within each storage period, the initial moisture content influenced the vigor of the seeds, with the highest moisture content (13%) being the most sensitive to deterioration in the long term (Table 6).
The physiological superiority of seeds stored with 9% moisture content is related to the fact that this content keeps the seed in a state of extremely reduced metabolism. Under conditions of low water availability, respiratory activity remains minimal, limiting the production of reactive oxygen species (ROS) and slowing down the initial deterioration processes. However, the deterioration is not restricted to the accumulation of ROS, which involves a coordinated sequence of cellular events that intensify as moisture increases (Waterworth et al., 2024).
As detailed by Krzyzanowski et al. (2022), the increase in moisture favors the partial reactivation of metabolism, increasing respiratory rate and the formation of free radicals capable of attacking essential components of cells. One of the first targets is the membrane system, whose destabilization results in greater electrical conductivity and solute leakage, which are classic indicators of loss of cellular integrity. In oilseeds such as soybeans, this process is aggravated by the high lipid content: the abundant presence of unsaturated fatty acids makes the membrane highly susceptible to lipid peroxidation, a central mechanism of deterioration. The oxidation of these lipids generates toxic compounds, such as malondialdehyde and other reactive aldehydes, which cause the structural collapse of organelles, especially mitochondria, reducing respiratory efficiency and the ability to synthesize ATP, elements that are directly associated with seed vigor (Stewart and Bewley, 1980).
In addition, recent studies show that oxidative deterioration also compromises the DNA repair system and genome stability, reducing the seed’s ability to reorganize its metabolism during germination. This cumulative damage affects the expression of genes involved in the synthesis of essential proteins, delays the resumption of growth and slows down the speed of emergence, characterizing a loss of vigor.
Thus, the 9% moisture content can be understood as a safe hygroscopic range, in which the seed remains stable and with less risk of oxidative damage throughout storage.
In general, the results regarding the percentage of strong seedlings, obtained from the 48-hour accelerated aging and emergence in bed tests (48AA and EB), showed a progressive reduction in the vigor of soybean seeds over the 225 days of storage (Figure 3). A decreasing linear fit was observed for the strong seedlings variable in 48AA for all moisture contents evaluated, indicating that physiological deterioration was continuous under these conditions (Figure 3A). The sharpest decrease was observed in seeds stored at 13% moisture, which showed a reduction rate of 2.9 percentage points in each period. On the other hand, seeds at 9% moisture content showed greater preservation of physiological potential, exhibiting the lowest degradation rate over the period (Figure 3A).
Strong seedlings from the 48-h accelerated aging test (A) and emergence in bed test (B) of soybean seeds subjected to storage with different initial moisture contents and storage periods.
A similar behavior was observed in the EB test with strong seedlings (Figure 3B) for the moisture contents of 11 and 13%, which also showed linear reductions with storage time, with coefficients of determination of 0.80 and 0.86, respectively. However, the lot stored at 9% moisture content was described by a quadratic model, maintaining emergence values above 90% and showing greater stability compared to treatments with higher moisture content. In general, the data confirm that the increase in the initial moisture content of the seeds accelerates the loss of vigor during storage, with the 9% moisture content being the most efficient in maintaining the physiological quality of soybeans for up to 225 days.
Souza et al. (2023) showed that environmental differences and variations within the bags themselves affect the moisture content and vigor of the seeds, especially in the most exposed layers. The results of this study reinforce this conclusion by demonstrating that ATI packaging minimizes these internal variations, creating a more uniform environment. This stability explains the higher percentage of strong seedlings observed in ATI over long storage periods, highlighting the importance of packaging with efficient barrier to reduce the negative effects of high moisture on vigor.
CONCLUSIONS
The maintenance of the physiological quality of soybean seeds during storage is dependent on the interaction between the initial moisture content and the efficiency of the protection provided by the packaging. Seeds stored at 9% initial moisture content have higher metabolic stability and lower deterioration rate over 225 days, regardless of the type of packaging used.
The use of isolated atmosphere (ATI) packaging is effective in mitigating hygroscopic fluctuations in the environment, preserving viability and vigor more efficiently than conventional packaging, especially in prolonged storage periods and with lower initial moisture in the seeds.
Initial moisture contents above 11% accelerate the loss of physiological quality with greater intensity, resulting in gradual reductions in seed germination and vigor.
Although the adoption of ATI packaging can still be conditioned by its cost, the advancement and greater diffusion of this technology tend to increase its accessibility and its potential for use as a tool for differentiation due to the quality. In addition to preserving the physiological potential, its greater resistance and ability to protect against contact with moisture can represent advantages during seed storage and transportation. In this context, its use may be promising for seeds of cultivars that are difficult to preserve, so further studies considering the interaction between packaging, genetic characteristics of cultivars, storage conditions and sanitary quality of the seeds are needed.
ACKNOWLEDGEMENTS
The authors are thankful for the financial support from the Brazilian agencies Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). Additionally, to André Capilheira, on behalf of the company Xpert Pack and Morena Sementes.
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