Storability assessment of polymer coated and biologically treated maize seeds in different packaging systems

Introduction

Maize (Zea mays L.), is widely recognized as the "queen of cereals", is one the most adaptable and versatile cereal crops cultivated worldwide. Its high productivity, diverse end uses, and economic importance make it indispensable global food and feed. In India, maize ranks third among cereal crops after wheat and rice. It is cultivated year-round in multiple forms such as sweet corn, baby corn, green cobs, and popcorn. In India maize production reached 42.28 million tonnes, the government has set up long term target to increase maize production to 86 million tonnes by 2047 (IBEF, 2024) [25] . Despite this, the national productivity (~3.5 t ha⁻¹) still trails behind the global average, although states like West Bengal, Telangana, and Andhra Pradesh reports near-optimal yields (Livemint, 2024) [33] . Ensuring sustained high quality seed supply is essential for meeting this growing demand. As seed longevity of maize is influenced by several factors like moisture content, temperature, relative humidity, storage conditions, fungal growth, and insect pests. Proper seed storage methodologies, as controlling temperature, relative humidity, and seed moisture, can significantly slows the seed deterioration. The proper storage practices like maintaining low seed moisture, using effective packaging materials and minimizing thermal fluctuations can significantly delay seed deterioration. Seed coating has emerged as an important strategy to enhance seed performance during storage by forming a protective barrier and enabling the controlled delivery of active agents. Chemical seed coatings containing fungicides and insecticides are widely used to suppress seed and soil borne pathogens and reduce early seedling mortality (Dhiman et al., 2023) [20] . In recent years, biological seed coating an eco-friendly approach has gained attention. It involves applying a thin film of biodegradable polymer embedded with biological agents on the seed surface, thereby reducing chemical inputs and promoting sustainable agriculture. Biological coatings typically encapsulate plant growth-promoting microorganisms within biodegradable polymers to form a uniform and stable layer. Beneficial microbes such as Trichoderma spp., Pseudomonas fluorescens, Bacillus subtilis, and Rhizobium spp. exhibit strong antagonistic activity against pathogens, enhance nutrient availability, improve seedling vigour, and increase tolerance to abiotic stresses (Mastouri et al., 2010; Zhang et al., 2023) [36,59] . Polymer coatings also contribute to seed longevity by regulating moisture absorption, stabilizing applied chemicals or biological agents, and providing a physical barrier against environmental stress. Recent studies indicate that polymer-coated maize seeds maintain higher germination, vigour, and seedling biomass during extended ambient storage compared with uncoated seeds (Moharana et al., 2020; Badiger et al., 2024) [37,11] . Furthermore, research on natural polymers such as chitosan, super absorbent polymers (SAPs), and nano-biopolymer blends demonstrates improved seedling performance and stress tolerance, highlighting emerging opportunities for climate-resilient seedcoating technologies (Behboud et al., 2024; Ondreickova et al., 2023) [16,40] . Recent studies have shown that biological and polymer-based coatings effectively maintain microbial viability, promote early crop establishment (Priyadharshini et al., 2022; Artemyeva et al., 2023) [43,9] . Importantly, commercial polymers used in the seed industry have demonstrated high compatibility (96 -100%) with beneficial microbes, confirming their suitability for biological seed coating applications (Arpitha et al., 2024) [8] . In addition to seed treatments, packaging materials also play a crucial role in extending seed longevity by preventing insect infestation. Conventional packaging materials such as jute and polypropylene woven bags are widely used for storing the seeds. Although they are used for grain storage the combined use with polymer or biological seed coatings remains insufficiently evaluated for maize seeds. Despite substantial progress, limited research has focused on integrating biological and chemical seed coatings with different packaging systems to evaluate their combined influence on seed storability, particularly in both inbred and hybrid maize lines under tropical ambient conditions. Such information is essential for developing comprehensive storage strategies that ensure high seed quality, reduce post-harvest losses, and support India's expanding maize seed industry. Therefore, the present study was undertaken to assess the effect of chemical, biological, and untreated seed coatings in combination with different packaging materials on seed longevity, vigour, health, and overall storability of maize inbred and hybrid lines under ambient storage conditions.

Materials and Methods

Freshly harvested seeds of maize genotypes two inbreds (BML 7, PFSR 3) and two hybrids KNMH 4010131, KNMH 4010141) were collected from the Agricultural Research Station (ARS) at PJTAU, Karimnagar. The seeds were stored under ambient conditions with initially recorded germination rate of 94%.

Seed coating: Biological agents like Trichoderma viride and

Bacillus megaterium were applied at 2g kg⁻¹ of seed, along with a bio-friendly polymer at 6g kg⁻¹, fungicide metalaxyl at 2g kg⁻¹, and insecticide thiamethoxam at 3g kg⁻¹ and colourants. The required quantities of each coating material were weighed and mixed into a slurry in a glass beaker. This slurry was then uniformly applied to the 2000g maize seeds per treatment by thorough mixing in a polythene cover. The coated seeds were shade dried for 2 hours, after drying the treated seeds were divided into two replications of 1000g each and packed into jute bags and polypropylene woven sac bags. The seed quality parameters were assessed bi-monthly for the coated inbred and hybrid maize seeds are as follows:

Treatment details Experiment 1

The germinated seedlings were evaluated for seedling parameters,

Seedling length (cm): Ten normal seedlings of 7 th day old were selected the shoot and root length was measured from the cotyledon to the tip of the primary root and shoot, the respective values were recorded in cm.

Seedling dry weight:

The seedlings measured for length were kept in an oven at 60 o C for 48 hrs. Then the seedling dry weight was measured, and the mean values were expressed in grams (g).

Vigour index I & II:

It was calculated as per the formula given by (Abdul-Baki & Anderson, 1973) [3] .

Vigour index I =

Vigour index II = Germination% × Seedling dry weight (g).

Field Emergence (%): Field emergence percentage was recorded to assess the establishment potential of maize seeds under field conditions. Seeds from each treatment/replication were sown in rows at a uniform depth and spacing. The field was maintained under recommended agronomic practices. The number of seedlings that successfully emerged was counted at a specified interval (usually 10-14 days after sowing). Field Emergence (%) was calculated using the formula:

Insect infestation (%): Insect infestation was assessed using a representative 20 g seed sample from each replication. The sample was carefully examined, and insect-damaged, bored, or perforated kernels were separated from the healthy ones. The number or weight of infested kernels was recorded. Insect Infestation (%) was then calculated using:

Seed Infection (%): Seed health status was assessed using the Standard Blotter technique as described by ISTA (2023) [26] . For each treatment, 400 seeds (100 seeds per replication) were placed equidistantly on moistened, three-layer blotter paper inside sterile Petri plates. The plates were incubated at 22 ± 2°C under alternating 12 h light and 12 h dark cycles for 7 days. After incubation, seeds were examined under a stereobinocular microscope for the presence of fungal pathogens. Infected seeds showing mycelial growth, sporulation, or characteristic pathogen structures were counted. Seed Infection (%) was calculated as:

Statistical analysis: The experiment was conducted in a Completely Randomized Design (Factorial) with three replications. Data were analysed using ANOVA as described by Gomez and Gomez (1984) [23] . Standard Error of Difference (SED) and Critical Difference (CD) at 5% probability were calculated for comparison of means. Percentage data were transformed to arcsine values before analysis (Snedecor & Cochran, 1956) [53] .

Results

The study evaluated the longevity of maize inbred lines and hybrid seeds using seeds with 95% germination rate initially (Table 1 andTable 2). Among the hybrids, KNMH 4010131 showed better storability, with 81% germination rate compared to 56% in KNMH 4010141. Seeds treated with combined formulation of Thiamethoxam + Trichoderma viride + Bacillus megaterium + biofriendly polymer + colourant of CI recorded a germination percentage of 75%. In inbred lines, treatment T 9 (PFSR 3 seeds treated with Thiamethoxam, Trichoderma viride, Bacillus megaterium, biofriendly polymer, and colourant of CI, and stored in polypropylene woven sacs) recorded germination rate of 76%, followed by T 7 with 71%, compared to 65% for untreated seeds (Figure 1and Figure 2). This treatment resulted in a 13.3% improvement in germination, compared to a 4.41% improvement with chemical treatment alone. The best treatment, labelled T 3 , involved KNMH 4010131 seeds treated with Thiamethoxam, Trichoderma viride, Bacillus megaterium, biofriendly polymer, and colourant of CI, and stored in polypropylene woven sacks. This treatment achieved an 88% germination rate, followed by T 4 with 84%.

Similarly, seed coating with biological agents was found to improve seedling length. Over an eight-month storage period, the seedling length of maize hybrids decreased from 30.38 cm to 23.15 cm. However, among the hybrids, KNMH 4010131 exhibited highest seedling length of 23.95 cm compared to 22.22 cm for KNMH 4010141. Treating seeds with a combination of chemicals, biological agents, biofriendly polymer, and colourant of CI resulted in7.82% improvement in seedling length, while chemical treatment alone resulted in a 5% improvement with storage in polypropylene woven sac bags.

The mean seedling dry weight decreased from 865 mg to 418 mg over the storage period. After eight months, seeds stored in polypropylene woven sack bags recorded the highest improvement in seedling dry weight (5.35%) compared to those stored in jute bags. Among the hybrids, KNMH 4010131 demonstrated superior storability, maintaining a higher seedling dry weight of 526 mg compared to 332 mg for KNMH 4010141.

Field emergence (%) declined from 79% to 51% over the storage period in inbreds and hybrids. At the end of ageing the seed treatment with chemical and biological agents, biofriendly polymer, and colourant recorded the highest improvement in field emergence (16.4%) of maize inbred lines, followed by chemical seed treatment (11.5%) while, among the hybrids, KNMH 4010131 showed better storability, achieving a 70% field emergence rate, compared to 46% for KNMH 4010141 compared to untreated control. The combination of chemical and biological seed treatments, along with biofriendly polymers and polypropylene woven sac bags, was the best method for maintaining seed quality and improving field emergence in maize hybrid lines. This approach led to field emergence improvements of 48.3% for KNMH 4010141 and 17.1% for KNMH 4010131.

The mean insect infestation (%) increased gradually from 0% to 2.39% over the storage period. After eight months, maize inbred lines stored in polypropylene woven sack bags showed the highest percentage reduction in seed infection (5.75%) compared to seeds stored in jute bags. Among hybrids treatments T 1 to T 14 maintained high storage potential and

Discussion

Seed quality deterioration during storage is a natural and inevitable process driven by multiple physiological, biochemical, and environmental factors. In the present study, a progressive decline in germination, seedling vigour, seedling biomass, and field emergence was observed across maize hybrids and inbred lines over eight months of ambient storage, consistent with earlier reports in maize and other field crops (Shobha et al., 2015; Ozturk et al., 2006; Singh et al., 2020) [50,41,52] . However, the magnitude of deterioration differed significantly among genotypes, seed treatments, and packaging materials.

Germination declined from an initial mean of 87% to 69% after eight months, reflecting typical ageing-induced reductions reported in soybean, groundnut, sunflower, and maize (Rasheed et al., 2020; Davila-Olivas et al., 2021) [44,18] . The hybrid KNMH 4010131 showed superior storability compared to KNMH 4010141, highlighting genotype-dependent seed longevity a feature widely documented in maize and rice (Ghosh et al., 2018; Prasanna et al., 2021) [22,42] . Seeds treated with a combination of chemical protectants (Thiamethoxam, Metalaxyl), biological agents (Trichoderma viride, Bacillus megaterium), biofriendly polymer, and colourant of CI retained significantly higher germination percentage than untreated or chemically treated seeds. [6,35,49,1] . Recent studies further confirmed the role of polymer-enhanced seed coatings in maintaining hydration balance and reducing membrane damage, thereby prolonging seed viability (Waqas et al., 2023; Moreno-Perez et al., 2022) [56,38] . Polypropylene woven sacks were consistently superior to jute bags, likely due to their semi-permeable nature that restricts moisture ingress, maintains lower equilibrated moisture content, and prevents insect movement these findings are in agreement with Mutige et al. (2013) [39] , Sastry et al. (2007) [47] , and more recent reports by Singh and Kaur (2021) [51] and Teklu et al.

(2022) [54] . Thus, packaging material significantly influenced germination and longevity. Seedling length showed a steady decline during ageing, aligning with earlier studies linking reduced early growth to DNA degradation, chromosomal aberrations, and failure of enzyme activation during germination (Kapoor et al., 2010; Kibinza et al., 2020) [28,31] . Ageing-induced impairment in enzymatic pathways required for early seedling development is well documented, and recent evidence suggests that oxidative stress accelerates mitochondrial disfunction during storage (Bailly, 2020) [12] . Treatments involving integrated seed coatings (chemical + biological + polymer) significantly improved seedling length during storage. Trichoderma viride and Bacillus megaterium are known to promote early root -shoot growth by producing phytohormones, solubilizing nutrients, and mitigating oxidative stress, explaining the superior performance of treated seeds (Ameen et al., 2019; Tiwari et al., 2022) [5,55] . Comparable improvements have been reported in maize, wheat, and soybean seedling growth when coated with beneficial microbes and polymers (Bargaz et al., 2021; Zhang et al., 2020) [14,57] . A marked decline in seedling dry weight from 865 mg initially to 418 mg after eight months was observed in untreated and jutestored seeds. This reduction agrees with earlier reports that attribute seedling dry matter loss to restricted nutrient mobilization, weakening of endosperm reserves, and exhaustion of metabolic substrates during ageing (Khatun et al., 2009; Bedi et al., 2020) [30,15] . Recent studies confirm that oxidative injury to membranes and storage proteins accelerates reserve depletion during storage (Chen et al., 2021; Hasan et al., 2023) [17,24] . In contrast, seeds treated with combined biological and chemical agents, along with polymer coatings, maintained higher dry weights due to reduced deterioration, improved enzyme activity, and enhanced seed metabolism. [29,4,13] . The hybrid KNMH 4010131 again showed superior storability, confirming genetic influences on emergence potential and seed vigour under stress (Prasanna et al., 2021) [42] . Untreated seeds and those stored in jute bags recorded the highest insect infestation, primarily by rice moth consistent with reports from pigeon pea, chickpea, maize, and stored pulses (Rathod et al., 2018; Devi & Mohan, 2020) [45,46,19] . [7,48,10] .

Recent studies further demonstrate that polymer-fungicide combinations enhance long-term protection by controlling storage pathogens and sealing micro fissures on the seed coat (Zhang et al., 2021; Abdelhamid et al., 2023) [58,2] . The present study highlights that integrating chemical protectants + biological agents + biofriendly polymer + Colourant of CI, combined with polypropylene woven sack storage, provides the most effective strategy for maintaining maize seed quality during ambient storage. This integrated approach minimized deterioration, controlled insect infestation and seed infection, and maximized germination, vigour, biomass, and field emergence in both hybrid and inbred lines. Genotypes differed in storage potential, with KNMH 4010131 consistently displaying higher storability across all parameters, implying inherent physiological resilience a trend supported by recent genetic and physiological studies on maize seed longevity (Li et al., 2022; Jha & Singh, 2023) [32,27] . Overall, the findings reinforce that integrated seed coating technologies combined with appropriate packaging are critical for ensuring seed longevity under ambient storage conditions, especially in tropical environments.

Conclusion

The seed storage methodology involving treatment with a combination of chemicals, biological agents, biofriendly polymer, colourant of CI, and storage in polypropylene woven sac bags, showed the highest improvement in field emergence, seedling vigour index I, and seedling vigour index II in maize hybrid lines. Among all treatments, the integrated seed coating consisting of chemical protectants (Thiamethoxam, Metalaxyl), biological agents (Trichoderma viride, Bacillus megaterium), biofriendly polymer, and Colourant of CI combined with polypropylene woven sack packaging proved most effective in maintaining seed viability and vigour during the eight-month storage period. This treatment significantly improved germination, seedling growth, seedling dry weight, field emergence, and seed health while achieving complete control of insect infestation and substantial reduction in seed infection.Hybrid KNMH 4010131 consistently exhibited superior storability compared to KNMH 4010141, indicating inherent genetic advantages. In inbred lines, PFSR 3 seeds treated with the integrated coating and stored in polypropylene sacks recorded the highest retention of seed quality. Overall, the findings highlight that integrated seed treatment technologies combined with suitable semi-permeable packaging materials are essential for safeguarding seed quality under ambient storage, particularly in tropical and subtropical climates. The approach offers a practical, scalable, and cost-effective solution for enhancing seed longevity, improving field performance, and ensuring high-quality seed supply for maize growers.

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