Chickpea (Cicer arietinum L.), a self-pollinated crop belonging to the subfamily Papilionaceae within the Fabaceae family, plays a significant role in global agriculture. Known for its adaptability to diverse climatic conditions, low production costs and contribution to nitrogen fixation, chickpea is a crucial legume in crop rotation systems. Despite its ability to thrive in a wide range of environments, chickpea cultivation faces challenges when exposed to low temperatures. As Cani (2009) highlighted, chickpea is not only vital for food security but also for sustaining agricultural ecosystems.

A critical aspect of chickpea production is seed quality, which directly affects both germination and storability. Several factors influence seed storage potential, including the pre-storage history, seed maturity and environmental conditions during pre- and post-harvest stages (Mahesha et al., 2001, Ahmad et al., 2014). Premature harvested seeds exhibit poor storage characteristics and reduced viability, making them less suitable for long-term storage (Rao et al., 2023). In contrast, seeds harvested at physiological maturity-when they reach peak viability and vigour-offer the best chance for successful storage. The timing of harvest, which is influenced by both genetic traits and environmental factors, plays a key role in ensuring high-quality seeds (Singh, 1995; Raghu et al., 2016).

Seed quality testing involves evaluating key attributes such as seed size, vigour, viability, germination rate and nutritional content, along with electrical conductivity (Ray, 2022). Furthermore, seed characteristics can be influenced by the specific location within the plant from which the seeds are harvested.

Given the importance of seed quality in ensuring optimal crop performance, the present study aims to investigate the impact of genotype selection and storage conditions on chickpea seed quality. This research seeks to assess how different chickpea genotypes respond to varying storage containers and environmental conditions. The rationale behind this study is to identify strategies that will enhance both seed quality and storage potential, contributing to improved chickpea production and ensuring its availability for both immediate use and long-term preservation. By understanding the relationship between genotype, seed maturation and storage conditions, this study will provide valuable insights into optimizing chickpea seed storage techniques for better agricultural sustainability.

Seeds from eight chickpea genotypes-Anuradha (V₁), Bidisha (V₂), Bidhan-1 (V₃), BG-3043 (V₄), Digvijay (V₅), GNG-2299 (V₆), ICCV-191609 (V₇) and ICCV-191611 (V₈)-were utilized. After harvest, seeds were cleaned thoroughly and sun-dried to approximately 8% moisture content. Seeds of each genotype were stored under ambient and refrigerated conditions in six types of storage containers: Cloth bags (T₁), aluminium foil pouches (T₂), brown paper packets (T₃), glass containers (T₄), 700-gauge polythene packets for ambient conditions (T₅) and 700-gauge polythene packets for refrigerated conditions (T₆, 4-5^oC). The study was set up using a two-factor complete randomized design (CRD) analysis with three replications. Laboratory experiments were conducted during April, 2022 to March, 2023 at the seed testing laboratory, Bidhan Chandra Krishi Viswavidyalaya, Nadia, West Bengal, India. Seed samples were collected at bi-monthly intervals over twelve months, resulting in six evaluations from each replication. For biochemical estimation, seeds were thoroughly cleaned, oven-dried and ground into fine powder, where 100 mg of seed powder per sample was taken for protein and carbohydrate analysis. Seed protein content was determined using Lowry et al., (1951), based on the reaction of peptide bonds with copper under alkaline conditions (Biuret reaction), followed by reduction of the Folin-Ciocalteu reagent by aromatic amino acids, producing a blue colour measured spectrophotometrically. Carbohydrate contents of the seeds were measured using Hedge (1962) method, involving hydrolysing carbohydrates to simple sugars, which react with anthrone reagent in acidic medium to form a green coloured complex. The intensity of the colour, read spectrophotometrically, is proportional to carbohydrate content. Electrical conductivity (EC), indicative of membrane stability, was measured to assess the physiological status of the seeds.

Seed protein content (mg g^-1) at different stages of storage
 
The data analysis (Table 1) reveals significant differences among genotypes at each storage interval, as indicated by the critical difference (CD at 0.01) values being consistently higher than the corresponding standard error of the mean (SEm). This confirms that the observed variation in performance across genotypes is statistically significant at 1% level. Notably, genotype V₅ maintained significantly higher values throughout all storage periods, indicating superior storability, while V₂ and V₄ consistently recorded the lowest values, signifying poor seed quality retention over time. The increasing CD values from 0.307 at 2 months to 0.954 at 12 months further suggest that genotype differences become more pronounced with prolonged storage, emphasizing the importance of selecting genotypes with stable performance over time for long-term seed viability.



The treatment-wise data (Table 2) demonstrates statistically significant differences in performance across all storage intervals, as the critical difference at 1% (CD/ 0.01) consistently exceeds the standard error of the mean (SEm), confirming the reliability of observed treatment effects. Among all treatments, T₆ consistently recorded the highest values throughout the 12-month storage period, significantly outperforming others and indicating superior effectiveness in maintaining seed quality. In contrast, T₁ showed the lowest performance at every interval, suggesting poor storability. The distinction among treatments becomes more evident over time, with increasing CD values from 0.266 at 2 months to 0.826 at 12 months, reinforcing that treatment effects on storability become more pronounced during extended storage. These findings highlight T6 as the most effective treatment for preserving seed quality over long-term storage.



The interaction effect between treatment and genotype (Table 3) shows statistically significant differences at 2 and 4 months after storage, as the critical difference at 1% (CD/0.01) exceeds the standard error of the mean (SEm) at these time points, indicating that the treatment-genotype combinations significantly influenced the trait during early storage. However, from 6 months onward, the differences become statistically non-significant (NS), suggesting that the effect of specific treatment-genotype combinations diminishes over time, likely due to progressive uniform deterioration across all combinations. Notably, the highest-performing combinations across time include T₆V₅, T₆V₈ and T₆V₁, all of which consistently retained superior values, whereas T₁V₂ and T₁V₄ exhibited the lowest values, especially by 12 months. These results emphasize that while certain treatment-genotype interactions initially confer storage advantages, their distinctiveness may wane with prolonged storage duration.


 
Seed carbohydrate content (mg g^-1) at different stages of storage
 
The genotype mean data (Table 4) across six storage durations reveals statistically significant differences among genotypes at all time points, as the critical difference at 1% (CD/0.01) consistently exceeds the standard error of the mean (SEm). This indicates that the genotypes differ significantly in their ability to maintain the trait under study-likely seed viability or vigour-over time. Genotypes V₅, V₈ and V₇ consistently maintained higher mean values, particularly V₈, which performed best at later stages (e.g., 12 months: 5.79), suggesting superior storability. In contrast, V₄ showed the lowest values throughout the storage duration, with a drastic decline to 3.34 at 12 months, indicating poor seed longevity. The significant differences highlight the importance of genotype selection for enhancing storage performance and long-term seed quality maintenance.



The treatment-wise analysis (Table 5) across storage durations demonstrates statistically significant differences at all time points, as the critical difference at 1% (CD/ 0.01) is consistently greater than the standard error of the mean (SEm), confirming the reliability of treatment effects. Among the treatments, T₆ consistently outperformed the others, maintaining the highest values from 2 to 12 months, with a notable value of 6.49 at the end of the storage period, indicating superior efficacy in preserving seed quality. Conversely, T₁ consistently recorded the lowest values, particularly after prolonged storage, suggesting it was the least effective treatment. The growing differences among treatments over time, as reflected in increasing CD values, emphasize the long-term impact of effective storage treatments, highlighting T6 as the most promising for sustaining seed viability and vigour over extended periods.



The analysis of the interaction effects between treatments and genotypes (Table 6) on seedling vigour index over a 12-month storage period reveals no statistically significant differences, as indicated by the absence of a critical difference (CD) at the 1% level across all storage intervals. Despite observable numerical variation in seedling vigour index values among the 48 treatment x genotype combinations, the lack of significance suggests that these differences are not statistically reliable and could be due to random variation. This indicates that the combined effect of treatment and genotype does not contribute meaningfully to variation in seedling vigour index under storage conditions, emphasizing the dominant role of main effects (treatment or genotype alone) rather than their interaction in determining storability traits.



Electrical conductivity (mS cm^-1 g^-1) of seeds at different stages of storage
 
The analysis of genotype means (Table 7) for electrical conductivity (EC) over a 12-month storage period indicates statistically significant differences among genotypes at each time point, as the calculated critical difference (CD) at the 1% level exceeds the Standard Error of Mean (SEm) throughout. Genotypes V₂, V₄ and V₆ consistently recorded higher EC values, indicating greater membrane deterioration and, consequently, reduced seed vigour during storage. In contrast, genotypes V₅, V₈ and V₇ maintained comparatively lower EC values, suggesting better seed membrane integrity and storability. The significant differences highlight the genetic variability in seed quality deterioration patterns, with EC values steadily increasing across storage durations for all genotypes, reflecting the typical aging-related loss of membrane stability.



The data on treatment means (Table 8) for electrical conductivity (EC) across storage durations reveals statistically significant differences among treatments at all time points, as the critical difference (CD) at the 1% level exceeds the corresponding standard error of mean (SEm). Notably, treatment T₁ consistently exhibited the highest EC values throughout the storage period, indicating the greatest level of seed membrane deterioration and reduced vigour. Conversely, treatments T₅ and T₆ showed significantly lower EC values, reflecting better membrane integrity and higher storability potential. Among them, T₆ emerged as the most effective in preserving seed quality, with the lowest EC across all durations. The consistent and significant variation underscores the influence of storage treatments on seed deterioration dynamics and highlights T₆ as the most promising treatment for maintaining seed quality during long-term storage.



The analysis of Treatment x Genotype interaction data for electrical conductivity (Table 9) over 12 months of storage reveals no statistically significant differences at the 1% level (CD > SEm) across all time intervals, as indicated by the non-significant (NS) critical difference values. This suggests that while variations in EC values exist among different genotype and treatment combinations, these variations are not substantial enough to be considered statistically significant. Therefore, the interaction effect between storage treatments and genotypes on seed membrane deterioration was not pronounced. The trend across the dataset still shows that certain combinations (such as T₁V_2, T₁V₄ and T₁V₆) consistently exhibited higher EC values, while others like T₆V₅, T₆V₈ and T₅V₅ maintained lower EC levels, indicating comparatively better seed storability. However, these observations remain descriptive and not statistically validated at the 1% level due to the lack of significant interaction effects.



Agarwal, (1987) noticed that protease activity was elevated in chickpea and mung bean seeds when they aged naturally or artificially. According to Ebone et al., (2019), the downregulation of antioxidant enzymes, including glutathione peroxidase (GPX), ascorbate peroxidase (APX), catalase (CAT) and superoxide dismutase (SOD), is the initial step in the aging process of seeds. The down-regulation and decrease in scavenging antioxidant activity resulted in the depression of the antioxidant enzyme system (Yin et al., 2014).  Reduced antioxidant activity can cause soluble protein degradation and decreased enzyme activity, as noted by Pukacka (2007), who also reported an increase in reactive oxygen species (ROS) during storage. Similar type of result was obtained in Bengal gram by Laxman et al., (2017) in terms of carbohydrate content, where they found that total carbohydrate decreased with storage time. Satasiya et al., (2020) and Chakraborty et al., (2024) in Chickpea also found similar type of result as well as corroborate with the findings.

The loss of leachate includes sugars, amino acids, fatty acids, proteins, enzymes and inorganic ions (K⁺, Ca⁺², Mg⁺², Na⁺ and Mn⁺²) and the test evaluates that the higher amount of ion as well as electrolyte leakage from the seeds resulting higher chances of degradation of seeds. Those genotypes and the containers measured higher amount of electrical conductivity are more prone to higher electrolyte leakage from the seeds compared to the seeds of other genotypes and containers. Beedi et al., (2018) observed similar type of result in kabuli chickpea. They further noted that, damage to the membrane system could be repaired and protected against such changes by primed treatment as indicated by low electrical conductivity of seed leachate and protective action of primed chemicals could presumably have extended the viability of seeds. Dias et al., (2004) also revealed that seed soaking in water effectively controlled the leakage of electrolytes, sugars and amino acids from the seeds.

Therefore, Digvijay (V₅) and ICCV-191611 (V₈) exhibited superior seed quality retention, while refrigerated storage (T₆) was most effective in preserving seed biochemical integrity, highlighting the importance of optimal genotype selection and storage conditions for maintaining seed viability and quality over time.

Dr. Kanu Murmu, Assistant Professor, Department of Agronomy and Dr. Raju Das, Assistant Professor, Department of Plant Pathology are duly acknowledged. This work was part of the Ph.D. program completed by Anish Choudhury and no funding was involved.

The authors declare that they have no conflict of interest.