Microbiolization of Arachis hypogaea L. seedsAbstract
Peanut (Arachis hypogaea L.) is one of the most widely cultivated and consumed oilseed crops globally, with seed quality being a critical factor for successful cultivation and productivity. Among the main phytopathogenic agents associated with seeds, fungi stand out due to their frequency and the extent of damage they can cause, ranging from the sowing period to post-harvest. In this context, the present study aimed to evaluate the effects of commercial biological products on the sanitary and physiological quality of peanut seeds. The experiment was conducted in the laboratory and greenhouse of the Phytopathology Laboratory at CCA/UFPB, Areia–PB, Brazil. The treatments included: Vacciplant®, Shocker®, Tricho Turbo®, Auin®, Ecotrich®, Captana®, and a control using sterilized distilled water (SDW). The following parameters were evaluated: fungal incidence, seed moisture content, germination, first count, germination and emergence speed indices, as well as seedling growth and dry mass. All treatments reduced the incidence of Penicillium sp., Fusarium sp., Chaetomium sp., and Aspergillus sp. The highest germination speed index (GSI) was observed with Ecotrich® (87.29). The products Ecotrich®, Auin®, and Captana® promoted greater shoot and root growth, while Shocker®, Auin®, and Captana® were most effective in increasing shoot dry mass. All treatments outperformed the control in seedling emergence, with Captana® being the most effective. The results indicate that biological products are a promising alternative for peanut seed treatment, with positive effects on seed health, germination, and vigor.
Keywords:
peanut; biological control; parasitism; seed pathology
Resumo
O amendoim (Arachis hypogaea L.) é uma das principais oleaginosas cultivadas e consumidas mundialmente, sendo a qualidade das sementes um fator determinante para o sucesso do cultivo e da produtividade. Os fungos se destacam entre os principais agentes fitopatogênicos associados às sementes, tanto pela frequência quanto pelos danos que podem causar, desde o período de semeadura até a pós-colheita. Nesse contexto, o presente trabalho teve como objetivo avaliar os efeitos de produtos biológicos comerciais sobre a qualidade sanitária e fisiológica de sementes de amendoim. O experimento foi conduzido em laboratório e casa de vegetação do Laboratório de Fitopatologia do CCA/UFPB, Areia–PB. Os tratamentos consistiram em: Vacciplant®, Shocker®, Tricho Turbo®, Auin®, Ecotrich®, Captana® e um controle com água destilada esterilizada (ADE). Foram avaliadas a incidência de fungos, teor de água, germinação, primeira contagem, índices de velocidade de germinação e emergência, além do crescimento e massa seca das plântulas. Todos os tratamentos reduziram a incidência de Penicillium sp., Fusarium sp., Chaetomium sp. e Aspergillus sp.. O maior índice de velocidade de germinação (IVG) foi observado com Ecotrich® (87,29). Os produtos Ecotrich®, Auin® e Captana® promoveram maior crescimento da parte aérea e radicular, enquanto Shocker®, Auin® e Captana® se destacaram na massa seca da parte aérea. Todos os tratamentos superaram o controle na emergência de plântulas, sendo Captana® o mais eficiente. Os resultados indicam que os produtos biológicos representam uma alternativa promissora para o tratamento de sementes de amendoim, com efeitos positivos na sanidade, germinação e vigor das sementes.
Palavras-chave:
amendoim; controle biológico; parasitismo; patologia de sementes
1. Introduction
Peanut (Arachis hypogaea L.), a member of the Fabaceae family, is among the most widely produced and consumed oilseed crops worldwide, with significant social and economic importance. In the Northeast region of Brazil, it is cultivated primarily as a subsistence crop by smallholder farmers (Saita and Pandolfi, 2019), standing out for its drought tolerance, which is attributed to morphological and physiological traits that allow its cultivation under rainfed conditions or intercropped with other crops (Biai et al., 2021). According to data from the National Supply Company (Conab), Brazilian peanut production for the 2024/25 season is estimated at approximately 1.15 million tons, representing a 60.3% increase compared to the previous harvest of about 733.7 thousand tons (CONAB, 2025).
Peanut cultivation can be economically viable when appropriate technologies are adopted (Gabriel et al., 2025). In newly cultivated areas, seeds represent one of the main production costs, as they often exhibit low quality due to the crop’s susceptibility to various pests (Machado et al., 2022; Gabriel et al., 2025), which can lead to plant mortality and significant yield losses.
Given this scenario, seed quality plays a crucial role in ensuring proper plant establishment and the production of healthy seeds, as well as the development of vigorous plants. Among the microorganisms that infect peanuts, fungi are the most prevalent. These fungi are associated with a reduction in seed physiological quality, and they can cause lesions on plants during germination and/or storage, in addition to producing mycotoxins that are toxic and harmful to human and animal health (Krahn et al., 2020; Cavalcante and Oliveira, 2022).
Phytopathogenic fungi are also responsible for causing diseases in peanut plants, compromising both yield and grain quality. Among the main pathogens reported in the literature are Cercospora sp., Puccinia arachidis, Pseudocercospora personata, Sphaceloma arachidis, Rhizopus sp., Fusarium sp., Sclerotium rolfsii, Phoma arachidicola, Leptosphaerulina crassiasca, and Colletotrichum arachidis, which may be associated with a wide variety of symptoms such as leaf spots, stem and pod rots, wilting, and even premature plant death (Almeida et al., 2019; Devechio et al., 2021).
Due to the potential negative effects of agrochemical use on agricultural production and the environment, the use of biological products for disease control in plants has become increasingly common. Antagonistic microorganisms are considered environmentally sustainable and non-harmful, aiding in the selection and manipulation of efficient and economically feasible management systems (Gawai, 2018).
The use of Bacillus sp. CHEP5 for disease control in peanuts has shown promising results, with a significant reduction in the incidence and severity of smut caused by Thecaphora frezzii, suggesting that the biological agent has the potential to induce plant defense mechanisms, thereby contributing to the phytosanitary protection of the crop (Figueredo et al., 2024). Additionally, the GA1 strain of Bacillus velezensis was effective in reducing stem rot (white mold or southern blight) caused by Athelia rolfsii, both through direct antagonism and by stimulating systemic resistance in the host peanut plants (Alleluya et al., 2023).
When testing the effect of four soil-isolated antagonists, comprising three Aspergillus species (A. flavus, A. fumigatus, and A. niger) and one Trichoderma viride species, it was observed that Trichoderma viride exhibited the greatest antagonistic potential, being capable of inhibiting the growth of rice pathogenic fungi with high inhibition rates, particularly against Fusarium fujikuroi, with inhibition reaching up to 87.15% (Sultana et al., 2025).
Thus the objective of this study was to evaluate the effects of commercial biological products on the sanitary and physiological quality of peanut seeds.
2. Materials and Methods
2.1. Seed source
Seeds of the peanut cultivar BR1 were obtained from smallholder farmers in the municipality of Areia, Paraíba (PB), Brazil. The seeds were transported to the Phytopathology Laboratory at the Federal University of Paraíba (UFPB), Campus II, where they were manually processed and surface disinfected with 1% sodium hypochlorite for 3 minutes.
2.2. Control of pathogens associated with Arachis hypogaea seeds using biological products
Peanut seeds were subjected to the following treatments: T1: Vacciplant® (0.66 mL per 100 mL); T2: Shocker® (0.19 g per 100 mL); T3: Tricho Turbo® (0.33 mL per 100 mL); T4: Auin® (0.68 mL per 100 mL); T5: Ecotrich® (0.83 g per 100 mL); T6: Control (sterilized distilled water – SDW); T7: Captan® (240 g per 100 kg of seeds) (Table 1).
2.3. Sanitary assessment of seeds and pathogen identification
Seed health assessment and pathogen identification were performed through the incubation method using filter paper (Brasil, 2009). A total of 200 seeds per treatment were used, arranged in 20 replicates of 10 seeds each. Seeds were immersed for five minutes in their respective treatments. For the fungicide application, 200 seeds were weighed, and the appropriate amount of fungicide was applied by direct contact based on seed weight.
Treated seeds were placed in Petri dishes containing two layers of sterile filter paper moistened with sterilized distilled water (SDW) and incubated at 25 ± 2 °C under a 12-hour photoperiod for seven days. Fungal identification was conducted using optical microscopy, comparing vegetative and reproductive structures with descriptions found in the literature (Seifert et al., 2011). Results were expressed as the percentage of infected seeds.
2.4. Evaluation of seed physiological qualitymoisture content
Seed moisture content was determined based on fresh weight (Brasil, 2009), with results expressed as percentages. The analysis was conducted using four replicates of 25 seeds, which were incubated in an oven at 105 ± 3 °C for 24 hours and weighed using an analytical precision balance.
2.5. Germination test
The germination test was carried out with 200 seeds divided into four replicates of 50 seeds. Seeds were placed between two Germitest® paper sheets, covered with a third, and arranged in roll format. Rolls were moistened with SDW in an amount equivalent to 2.5 times their dry weight. The rolls were placed in transparent plastic bags to prevent water loss through evaporation and incubated in a germination chamber (BOD) at 25 ± 2 °C with a 12-hour photoperiod. Germinated seeds were counted every 24 hours up to the 10th day, with evaluations including hard and dead seeds (Brasil, 2009).
The first count was conducted simultaneously with the germination test, recording the number of germinated seeds on the 5th day after sowing (Brasil, 2009). Germination percentage (%G) was calculated using the formula: %G = (N2/N1) × 100, where %G = germinated seedling percentage, N2 = number of germinated seedlings, and N1 = number of seeds sown. The germination speed index (GSI) was calculated by recording daily germinated seed counts, using the formula proposed by Maguire (1962).
Shoot and root lengths were measured after the germination test using a millimeter-scale ruler, with results expressed in centimeters. Seedling shoots and roots were dried in an oven at 65 °C for 48 hours and weighed on a precision analytical balance (0.0001 g). Results were expressed in grams per seedling.
2.6. Seedling emergence test
The emergence test was conducted in a greenhouse using 200 seeds per treatment, divided into four replicates of 50 seeds, sown in polystyrene trays filled with sterile substrate. Seedlings were irrigated and emergence was recorded every 24 hours. The first count was performed on the 5th day after sowing, corresponding to the number of emerged seedlings (Brasil, 2009). Emergence percentage (%E) was calculated based on the percentage of emerged seedlings.
The emergence speed index (ESI) was calculated by recording the number of emerged seedlings daily until stabilization, which occurred on the 7th day, using the formula proposed by Maguire (1962).
Shoot and root lengths of emerged seedlings were measured with a millimeter-scale ruler, and results were expressed in centimeters. Shoots and roots were then oven-dried at 65 °C for 48 hours and weighed using an analytical precision balance (0.0001 g). Results were expressed in grams per seedling.
2.7. Experimental design and statistical analysis
The experiment followed a completely randomized design, with seven treatments. Data were subjected to analysis of variance (ANOVA), and means were compared using the Scott-Knott test at a 5% significance level. Fungal incidence data were previously transformed using √(x + 1), as described by Bartlett (1947). Statistical analyses were performed using the R software (R Core Team, 2024).
3. Results
Table 2 presents the results of the analysis of variance. For fungal genera, treatments had a significant effect on all pathogens evaluated, with Fusarium showing the highest mean square value (9.6467) (p ≤ 0.01). Aspergillus, Chaetomium, and Penicillium also showed significant responses (p ≤ 0.05), with mean square values of 1.6285; 0.29876; and 0.238404, respectively.
Summary of the analysis of variance for Aspergillus sp., Chaetomium sp., Fusarium sp., Penicillium sp., germination percentage (GP), first germination count (FGC), germination speed index (GSI), emergence (EM), emergence speed index (ESI), shoot length (SL), root length (RL), shoot dry mass (SDM), and root dry mass (RDM) from germination and emergence tests.
Regarding germination, treatments significantly affected the first germination count (FGC), with a mean square of 32.409, germination percentage (GP), germination speed index (GSI), and shoot length in the germination test (SL-G), all significant at p ≤ 0.05. Root length (RL-G), shoot dry mass (SDM-G), and root dry mass (RDM-G) also showed significant differences among treatments, with RL-G showing a mean square of 0.07696.
Seedling emergence (EM) was influenced by the treatments (p ≤ 0.05). Emergence percentage (EP), emergence speed index (ESI), shoot length (SL-E), and root length (RL-E) also differed among treatments. Shoot dry mass (SDM-E) was significant at the 5% level, whereas root dry mass (RDM-E) showed no significant difference among treatments.
The moisture content of BR1 peanut seeds was 8.57%. Four fungal genera were identified in the seeds: Penicillium, Fusarium, Chaetomium, and Aspergillus (Figure 1).
Fungal incidence in peanut (Arachis hypogaea L.) seeds treated with biological products. T1 – Vacciplant® (0.66 mL 100 mL−1); T2 – Shocker® (0.19 g 100 mL−1); T3 – Tricho-Turbo® (0.33 mL 100 mL−1); T4 – Auin® (0.68 mL 100 mL−1); T5 – Ecotrich® (0.83 g 100 mL−1); T6 – Control (sterilized distilled water – SDW); T7 – Captan® (240 g 100 kg−1). Means followed by the same letter in the row do not differ significantly according to the Scott-Knott test (p ≤ 0.05).
Fusarium sp. was the most prevalent genus, with an incidence of 61% in BR1 peanut seeds. However, all treatments reduced its incidence compared to the control, showing effectiveness equivalent to that of the fungicide.
All treatments, except Ecotrich®, reduced the incidence of Aspergillus sp. relative to the control. On the other hand, Vacciplant®, Tricho-Turbo®, and Auin® were not effective in controlling Penicillium sp., showing no significant difference from the control, whereas the remaining treatments significantly reduced the incidence of this pathogen.
Regarding the genus Chaetomium, only Auin® did not differ significantly from the control, while all other treatments were effective in reducing its incidence in the seeds.
Table 3 presents the effects of treatments on the physiological quality of peanut (Arachis hypogaea L.) seeds. The treatments Tricho-Turbo®, Auin®, Ecotrich®, and Captan® showed germination rates (GP ≥ 94%) and first germination counts (FGC) comparable to the control, indicating that these treatments did not impair seed germinative capacity.
Physiological quality of peanut (Arachis hypogaea L.) seeds treated with biological products, evaluated by germination percentage (GP), first germination count (FGC), germination speed index (GSI), shoot length (SL) and root length (RL), shoot dry mass (SDM), and root dry mass (RDM).
The highest germination speed index (GSI) was observed with Ecotrich® (87.29), indicating faster and more vigorous germination. This value was significantly higher than those of the other treatments, suggesting a greater potential to enhance germination. Regarding growth parameters, Ecotrich® again stood out, with the highest values for shoot length (4.98 cm) and root length (10.25 cm), exceeding nearly all other treatments and not differing significantly from Auin® (RL 10.16 cm) and the fungicide (RL 9.38 cm).
The treatments Shocker®, Auin®, and Captan® showed significantly higher shoot dry mass compared to the other treatments. Concerning root dry mass, Vacciplant® and Captan® performed better, differing significantly from the other treatments.
The treatment with Captan® was the most effective for emergence parameters, with 100% emergence (EM), 89% in the first emergence count (FEC), and an emergence speed index (ESI) of 44.02, significantly outperforming all other treatments (Table 4). However, all treatments were superior to the control, positively influencing seedling emergence, which suggests that biological products are promising alternatives to chemical treatments.
Quality of peanut (Arachis hypogaea L.) seeds treated with biological products, evaluated through emergence test parameters: emergence percentage (EM), first emergence count (FEC), emergence speed index (ESI), shoot length (SL), root length (RL), shoot dry mass (SDM), and root dry mass (RDM).
Shocker® and Auin® also showed high emergence percentages, both reaching 93%, and exhibited higher ESI values compared to the control and most other treatments, with values of 38.71 and 39.89, respectively. These treatments also demonstrated good performance in the first emergence count, with 73% for Shocker® and 80% for Auin®, indicating a positive influence of the products on early seedling development.
No significant differences were observed among treatments for shoot length (SL), root length (RL), and shoot dry mass (SDM). However, for root dry mass (RDM), the control and Vacciplant® showed greater biomass accumulation, surpassing the other treatments.
4. Discussion
Low seed moisture content is crucial for ensuring the physiological (Oliveira et al., 2022) and sanitary quality of peanut seeds, directly influencing viability, pathogen resistance, and germination performance during health testing (Menegaes and Nunes, 2021). Oilseeds such as peanuts must be stored with a moisture content between 4% and 9% to preserve their physiological quality and prevent deterioration (Azeredo et al., 2005), indicating that the moisture level of the seeds used in this study was adequate.
Fusarium sp. is one of the main pathogens responsible for root rot in peanuts and occurs in most regions where the crop is grown. In cases of high incidence, it can reduce yields by more than 20% (Sun et al., 2023). The primary methods for controlling this pathogen are chemical and biological strategies. Aspergillus and Penicillium are storage fungi that cause deterioration, increase respiratory rates, reduce germination, and can induce the production of mycotoxins harmful to humans and animals (Prestes et al., 2019). High rates of these pathogens were found in the sanity test, but the results demonstrated high efficiency in reducing infestation through the products used (Figure 1).
Biological control has become one of the most commonly used practices in peanut cultivation due to its beneficials effects, lower cost, and reduced environmental impact (Du and Li, 2024). The results obtained for the control of Fusarium sp. using Shocker®, which contains Trichoderma and Bacillus species, demonstrate the efficacy of antagonistic microorganisms in suppressing harmful crop pathogens.
Products such as Shocker®, Tricho-Turbo®, and Ecotrich® play an important role as biocontrol agents against various pathogens, as they are formulated with Trichoderma spp (Figure 1). This saprophytic fungus can colonize a variety of pathosystems, benefiting soil organisms and controlling plant diseases through several mechanisms, including the production of antifungal compounds, competition with pathogenic fungi, mycoparasitism, and the activation of host plant defenses (Ahmad et al., 2024; Al-Shuaibi et al., 2024).
Bacillus sp.-based products like Shocker® are widely used in biological control for their strong antimicrobial activity against plant pathogens and for stimulating plant growth and development (Karačić et al., 2024). These bacteria inhibit the growth of phytopathogens by producing antimicrobial compounds and extracellular hydrolytic enzymes, competing for resources, and activating systemic resistance in host plants (Miljaković et al., 2024). Seed treatment with Bacillus sp. can suppress pathogenic fungal infections and protect seedlings during emergence and early development (Chen et al., 2019).
Vacciplant® is a product derived from marine algae and functions as a biostimulant, rich in polysaccharides, oligosaccharides, peptides, proteins, and phytohormones. These components act as resistance elicitors in plants through salicylic acid and jasmonic acid signaling pathways, regulation of reactive oxygen species homeostasis, and activation of defense-related genes and enzymes (Akhyani et al., 2024). This can be observed in the quality health (Figure 1) and physiological (Table 3 and 4) results, in which the product had a high influence on the reduction of pathogens associated with peanut seeds and on plant physiology.
Biological treatments based on Trichoderma spp. and Beauveria bassiana stood out for their ability to increase root length. Chagas et al. (2017) emphasized the importance of Trichoderma spp. in promoting plant growth, with more than 60% increases in biomass accumulation in crops such as soybean (Glycine max), cowpea (Vigna unguiculata), rice (Oryza sativa), and maize (Zea mays). Similarly, Jaber and Enkerli (2016) observed that seed treatment with B. bassiana significantly improved nearly all growth parameters in fava bean (Vicia faba).
These microorganisms produce antimicrobial compounds that inhibit pathogen growth, reducing disease incidence in plants (Alvarez et al., 2019). In addition, they positively affect seed germination, root development, and overall plant growth (Jacques et al., 2021).
Shocker® is a biological fungicide in wettable powder form, while Auin® is an emulsifiable concentrate insecticide, both offering ease of handling and application and achieving results comparable to the chemical fungicide used. However, the use of chemical control can negatively affect microbial activity in the soil (Barbero et al., 2023). Therefore, biological seed treatment presents a promising alternative for improving productivity and sustainability in agriculture.
Illa et al. (2019) demonstrated that antagonistic agents such as Trichoderma harzianum and Bacillus subtilis had positive effects on peanut seedling emergence. In controlled conditions, the application of these biological agents, whether individually or in combination, resulted in an increased percentage of emergence at 28 days after sowing. In field conditions, this increase was approximately 37%, and biological treatments also led to greater plant biomass, higher yields, larger seed size, and reduced disease incidence. Similar results were observed in the emergence of peanut seeds treated with biological products in this study (Table 4).
Thus, antagonistic microorganisms significantly improved emergence and early development of peanut plants, contributing to more efficient production. The effectiveness of the microbial species used in seed microbiolization may vary depending on the environmental conditions under which the experiments are conducted (Dalzotto et al., 2020).
5. Conclusions
Biological products were effective in reducing seed-borne pathogens in peanut seeds. The use of Trichoderma sp., Bacillus sp. and Beauveria sp. positively influenced germination and emergence speed indices, as well a root system development and seedling dry mass accumulation.
Data Availability Statement
The entire data set that supports the results of this study was published in the article itself.
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Edited by
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Editor:
Takako Matsumura Tundisi
