Optimization of pre-sowing treatments to break seed dormancy in wild finger millet (Eleusine africana L) for enhanced conservation and utilization of genetic resources

Optimization of pre-sowing treatments to break seed dormancy in wild finger millet (Eleusine africana L) for enhanced conservation and utilization of genetic resources | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Optimization of pre-sowing treatments to break seed dormancy in wild finger millet (Eleusine africana L) for enhanced conservation and utilization of genetic resources Ovais Peerzada, Vetriventhan Mani, Kommineni Jagadeesh, Kuldeep Singh This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7526433/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 11 Dec, 2025 Read the published version in Genetic Resources and Crop Evolution → Version 1 posted 7 You are reading this latest preprint version Abstract Crop wild relatives of finger millet, a part of its genetic resources, are key to exploring the genetic potential of the crop for several climate-resilient and nutritional traits. Seed dormancy in wild species accessions poses a significant challenge to the conservation, evaluation, distribution, and their utilization in pre-breeding activities. In the present study, we tested 12 treatments to break dormancy and enhance germination in two wild accessions (IE 8414 and IE 8416). Germination of these two accessions under control conditions was 0%. Among the treatments tested, Thiourea at 0.5% for 16 hrs was the most effective treatment, achieving germination rates of 91.00% in IE 8414 and 95.25% in IE 8416. Potassium nitrate (KNO₃) at 0.2% for 16 hrs also showed significant efficacy, with 50.75% germination in IE 8414 and 70.75% in IE 8416. Gibberellic acid (GA₃) at 500 ppm for 2 hrs induced 51.00% germination in IE 8416, while < 1% in IE 8414. Chemical treatments have been more effective, and while other treatments such as ethrel, cold stratification, and water immersion, were largely ineffective, with germination rates remaining below 5%. These findings highlight the potential of Thiourea and KNO₃ as effective tools for breaking dormancy in wild Eleusine accessions. Wild relatives genetic conservation climate resilience dormancy and germination Figures Figure 1 Figure 2 Figure 3 Introduction Finger millet is one of the most important millets among dry land crops with an allotetraploid nature (2n = 4x = 36) and C4 photosynthetic machinery. Its cultivation has been spread across several countries i.e., India, Ethiopia, Nepal, Uganda, Tanzania, Kenya, Zimbabwe, Zambia, Malawi, Eritrea, Mozambique, Rwanda, which accounts for around 20% of global millet area and 26% of global millet production (Gebreyohannes et al. 2024 ). India is one of the largest global producers of finger millet, with its cultivation spread across 1.162 million ha and with production of 1.692 million tons and productivity of 1456 Kg/ha ( https://www.indiastat.com/ ). In addition, it possesses a better nutritional profile with superiority in several nutrients (Backiyalakshmi et al. 2023 ). In the current scenario of the intensified impact of climate change, genetic resources, specifically the crop wild species, hold the key to addressing the spiraling challenges. Crop wild relatives are the reservoir of several desirable alleles for various traits(Dida et al. 2021 ) and play a crucial role in breeding for traits such as drought tolerance, pest resistance, and enhanced nutritional profiles (Hilu and Dewet 1976 ). However, seed dormancy has been identified as one of the major hurdles among the wild species and needs to be addressed. Seed dormancy is the inability of the seed to germinate, though the embryo is intact and viable. While dormancy enhances survival in natural ecosystems, it poses a significant challenge in ex-situ conservation, where rapid and uniform germination is essential for conservation, regeneration, and breeding programs. It has been a major hurdle for the freshly harvested seeds for their immediate use, which has enhanced the need for standardization of several pre-sowing seed treatments for overcoming it. Seed dormancy is a complex trait, affected by multiple genes and environmental factors (Graeber et al. 2014 ; Lu et al. 2018 ) and an important component of plant adaptation (Donohue et al. 2005 ; Huang et al. 2015 ). It is regulated by the antagonistic hormones, abscisic acid (ABA) and gibberellic acid (GA) (Finkelstein et al. 2008 ; Shu et al. 2016 e et al. 2017 ). Moreover, seed dormancy is also considered an undesirable trait by farmers, affecting crop establishment. So, the dormancy-breaking pre-treatments have been found most useful in most of the crops, i.e., rice, wheat, maize, sorghum, pearl millet, chickpea, pigeon pea and many other crops (Burton 1969 ; Sahu et al. 1993 ; Matus-Cádiz and Hucl 2003 ; Shanmugavalli et al. 2007 ; Khadraji and Qaddoury 2023 ; Lamichaney et al. 2023 ). Compared to cultivated, the strong dormancy mechanisms present in wild genotypes often result in poor germination under standard conditions, complicating their conservation and utilization (Long et al., 2015). The study conducted by (Kannababu et al. 2025 ) reported several genes regulating seed vigour related traits in finger millet. The impact of seed dormancy breaking treatments on the expression of genes regulating seed vigour and related traits can elucidate further mechanisms involved. Currently, ICRISAT genebank holds a global finger millet collection of 7536 accessions collected from 25 countries including 206 wild germplasm accessions ( https://genebank.icrisat.org/ ). Seed viability testing, typically conducted through germination tests, is a critical component of routine genebank operations. To ensure that germination results accurately reflect true seed viability, it is essential to apply an effective dormancy-breaking treatment, but limited information regarding the effective pre-sowing seed treatments and standardized protocol to overcome seed dormancy in wild species of finger millet is limiting their efficient conservation as well as exploration of their potential in crop improvement programs. Hence, the current study was initiated to unveil the impact of various pre-seed treatments to develop a more efficient treatment for routine use in viability testing for conservation and also to enhance the utility of crop wild relatives of finger millet. Materials and methods Plant material The current study was carried out during the year 2023-24 at ICRISAT, Genebank, Patancheru, Hyderabad. The seeds belonging to the two wild finger millet accessions (IE 8414 and IE 8416) were sown in the glass house at ICRISAT in the month of October 2023. The seeds were grown in 12-inch pots in a mixture of red soil, sand and vermicompost in the ratio of 3:2:1. At the time of sowing, 2 g DAP was added to the mixture, and 2 g of urea was applied after 25 days of sowing. After thinning, only five seedlings were maintained in each pot, and the seeds from both accessions were harvested at maturity. The seeds were carefully harvested, threshed, and after harvesting, a sample of seeds from the accessions was taken in grip seal bags to determine the initial seed moisture content, which was 16.2% (Oven dry method). Seeds were dried in the sun (max. 30 ° C, min. 22 ° C) to bring the moisture content down to 12.6% and then transferred to the genebank drying room maintained at 15°C and 15% RH for further drying. The seeds were dried to a uniform moisture content (7.1%) before sampling for germination testing. The seeds were taken out from the drying room and kept at room temperature (25 ± 2 0 C) for saturation before being immersed in different chemical solutions. Experimental design and treatments The current experiment was carried out in a factorial completely randomized design with genotypes and treatments as two different factors and four replications. The combinations of genotype × treatments were applied randomly across replications. Table 1 Different kinds of pre-sowing seed treatments used for breaking seed dormancy S. No Treatment Concentration/Condition 1 Control Normal Incubation (25 0 C for 10 days) 2 Cold stratification 4°C for 7 days 3 Cold stratification 4°C for 15 days 4 Hydropriming Soaking in distilled water for 17 hr 5 Hydropriming Soaking in distilled water for 24 hr 6 Chemical treatment Potassium nitrate (KNO₃) at 0.2% (16 hr) 7 Chemical treatment Potassium nitrate (KNO₃) at 0.5% (16 hr) 8 Chemical treatment Ethrel 25 ppm (16 hr) 9 Chemical treatment Ethrel 50 ppm (16 hr) 10 Chemical treatment Thiourea 0.5% (16 hr) 11 Chemical treatment Thiourea 1% (16 hr) 12 Chemical treatment Gibberellic acid (GA₃) at 500 ppm (2 hr) 13 Chemical treatment Gibberellic acid (GA₃) at 1000 ppm (2 hr) Seed Viability and dormancy evaluation Before applying dormancy-breaking treatments, the seed viability of both the wild accessions was assessed using the tetrazolium (TZ) test following ISTA guidelines (ISTA 2021 ). For each accession, 100 seeds were preconditioned by soaking in distilled water at room temperature (23 ± 2°C) for 16 hours. The seeds were then longitudinally and carefully dissected under a magnifying lens to expose the embryo. Dissected seeds were immersed in a 1% tetrazolium chloride solution and incubated in the dark at 25°C for two hours. Following staining, seeds were rinsed twice with distilled water. Seeds were carefully examined under a Tagarno magnifying microscope for evaluation. Seeds were considered viable if the embryo was uniformly stained red. The viability percentage for each accession was calculated as the proportion of seeds with completely stained embryos relative to the total number of seeds tested. For dormancy evaluation, uniform and intact seeds were selected from the available seed, and a total of 400 seeds were randomly selected from them and divided into four replications of 100 seeds each. The seeds were then subjected to different kinds of pre-sowing seed treatments, including cold stratification and hydropriming with exposure for different number of days and variable periods, respectively. Moreover, the treatments include exposure to chemicals like potassium nitrate (KNO₃), thiourea, and also hormones like ethrel and gibberellic acid at different concentrations and periods ( Table 1 ). Following the treatments, after the soaking period was over, the seeds were taken out and washed thoroughly with distilled water before placing them on petri dishes for germination. Germination testing was performed at the seed laboratory of ICRISAT genebank, following the International Seed Testing Association rules (ISTA 2021 ) using the "Top of Paper" method, where seeds were evenly spaced on moistened germination sheets in petri dishes. The dishes were covered and incubated in a growth chamber under controlled conditions (25 ± 1°C temperature and 85–90% relative humidity). Germination was recorded on the tenth day by counting normal seedlings, and the germination percentage was calculated as Germination percentage (%) = (Number of germinated seeds / Total seeds) * 100 Statistical analysis Germination percentage data were arcsine square root transformed before statistical analysis to stabilize the variance. The data were analyzed based on the linear model in which genotype, treatment, genotype × treatment, and replication are considered fixed factors using R software (version 4.2.2) and the packages emmeans and multcompView. The interaction (Genotype × Treatment) and main effect mean (treatment and genotype) were calculated. Further, post-hoc analysis was performed using the Duncan multiple range test (DMRT) (α = 0.05). Results and Discussion Embryos of seeds from both wild finger millet accessions exhibited a uniform red coloration upon staining with 1% tetrazolium chloride (TZ) solution, indicating that all tested seeds were viable. The viability percentage recorded was 100% for the accession IE 8416, while it was 99% for the accession IE 8414. The complete and consistent staining of the embryo tissues suggests that cellular respiration was active, confirming the physiological integrity and viability of the seeds across both accessions. The differential impact of various dormancy-breaking treatments imposed on wild accessions of finger millet was evident ( Table 2 ). Highly significant variation in germination rates among different treatments (p < 0.001) and between accessions (p < 0.001), with a significant genotype × treatment interaction effect (p < 0.001), was observed. The control groups of both E. africana accessions (IE 8414 and IE 8416) demonstrated absolute germination failure (0% ± 0.00), confirming the persistence of primary dormancy under standard conditions (25 0 C for 10 days) ( Table 3 ). In addition, a significant difference in germination (%) among the two wild genotypes has also been observed. Table 2 Analysis of variance (ANOVA) for the germination percentage of Eleusine africana genotypes under various treatments Source of Variation DF Sum Sq Mean Sq F-Value Pr(> F) Accession 1 5835 5835 558.27 < 2e-16*** Treatment 12 47798 3983 381.1 < 2e-16*** Accession* treatment 12 6138 512 48.94 < 2e-16*** Residuals 78 815 10 - - Total 103 60,586 - - - Note: Significance codes: *** indicates P < 0.001 Genotype*Treatment means Among the various pre-sowing treatments ( Table 3 ) , similar to the control, no germination was observed in the case of ethrel under both concentrations (50 ppm and 25 ppm) for a period of 16 hr in both accessions. Though cold stratification of 4°C for 7 days and 15 days has induced germination in both accessions, the germination percentage was lower than in other treatments, and differential response was observed in the two accessions. Moreover, water immersion treatments (17 and 24 hr) have not been very effective and resulted in poor germination (ranged between 2.38–2.88%) in the case of both accessions. GA 3 has a significant positive impact on germination, but it was limited only to one of the two varieties i.e., effective in IE 8416 with germination around 50% under both the concentrations i.e., 500 ppm (2 hr) and 1000 ppm (2 hr). No significant difference in germination was observed in different concentrations of gibberellic acid i.e., 500 ppm for 2 hr and 1000 ppm for 2 hr. Similar to GA 3 , KNO 3 treatment for a period of 16 hr also had a differential impact on germination in two accessions, with a greater positive impact on IE 8416. In addition, it also varied based on concentration (0.2% and 0.5%), which was not observed in the case of GA 3 . The increase in the concentration of KNO₃ resulted in a significant decrease in germination and it was much more drastic in the case of IE 8414 (50.25–0.25%). Apart from all the treatments, thiourea is the chemical treatment that has shown the highest germination percentage (> 90%) in both the accessions under 0.5% for 16 hr while further increase in concentration (1% for 16 hrs) resulted in reduction in germination (~ 30%) . Table 3 Grouping of genotypes*treatment means of germination% based on Duncan multiple range test Duncan's Multiple Range Test _ Treatments Accession Treatment Mean ± SEM groups IE 8416 Cold stratification 4° C 15 days 10.25 ± 1.49 gh IE 8414 Cold stratification 4° C 15 days 0.25 ± 0.25 i IE 8416 Cold stratification 4° C (7 days) 13.50 ± 0.65 g IE 8414 Cold stratification 4° C (7 days) 0.50 ± 0.29 i IE 8414 Control (25 0 for 10 days) 0.00 ± 0.00 i IE 8416 Control (25 0 for 10 days) 0.00 ± 0.00 i IE 8414 Ethrel - 50 ppm(16 hr) 0.00 ± 0.00 i IE 8416 Ethrel - 50 ppm (16 hr) 0.00 ± 0.00 i IE 8414 Ethrel − 25 ppm (16 hr) 0.00 ± 0.00 i IE 8416 Ethrel − 25 ppm (16 hr) 0.00 ± 0.00 i IE 8416 GA₃ − 500 ppm (2 hr) 51.00 ± 4.74 cd IE 8414 GA₃ − 500 ppm (2 hr) 0.75 ± 0.48 i IE 8416 GA₃- 1000 ppm (2 hr) 46.25 ± 3.90 d IE 8414 GA₃- 1000 ppm (2 hr) 3.00 ± 1.08 i IE 8416 KNO₃ − 0.2% (16 hr) 70.75 ± 2.14 b IE 8414 KNO₃ − 0.2% (16 hr) 50.75 ± 0.48 cd IE 8416 KNO₃ − 0.5% (16 hr) 53.75 ± 4.07 c IE 8414 KNO₃ − 0.5% (16 hr) 0.25 ± 0.25 i IE 8416 Thiourea − 0.5% 95.25 ± 0.25 a IE 8414 Thiourea − 0.5% 91.00 ± 0.58 a IE 8416 Thiourea − 1% 36.25 ± 2.39 e IE 8414 Thiourea − 1% 27.50 ± 2.84 f IE 8416 Water immersion (17 hr) 4.50 ± 1.19 hi IE 8414 Water immersion (17 hr) 0.25 ± 0.25 i IE 8416 Water immersion (24 hr) 3.75 ± 0.75 hi IE 8414 Water immersion (24 hr) 2.00 ± 1.68 i Treatment means The means of germination % for various treatments ( Table 4 ) and grouping based on significant differences ( Fig. 2 ) can provide an overview of the impact of various treatments on finger millet wilds. No significant difference was observed among the treatments, control, ethrel (25 ppm for 16 hr and 50 ppm for 16 hr) and water immersion (17 hrs and 24 hrs) and these are not effective in breaking seed dormancy. The cold stratification treatment of 4°C for 7 days and 15 days treatments are also similar and had minimal impact on enhancing germination. No significant increase in germination percentage when the concentration of GA 3 was increased from 500 ppm to 1000 ppm (24.63% and 25.88%). In the case of KNO 3 for a period of 16 hr, a lower concentration of 0.2% shown a greater positive impact than a higher concentration (0. 5%) with germination of 60.75% and 27% respectively. Finally, thiourea (0.5%) for a period of 16 hr is the chemical treatment that has been most effective and enhanced germination levels of > 90%, but a drastic reduction in was witnessed when its concentration was increased to 1%. Table 4 Treatment grouping based on Germination % using Duncan multiple range test (DMRT) Treatment Mean Group Thiourea − 0.5% (16 hr) 93.13 a KnO₃ − 0.2% (16 hr) 60.75 b Thiourea − 1% (16 hr) 31.88 c GA₃- 1000ppm (2 hr) 24.63 d GA₃ − 500ppm (2 hr) 25.88 d KnO₃ − 0.5% (16 hr) 27.00 d Cold stratification 4° C 7 days 7.00 e Cold stratification 4° C 15 days 5.25 ef Water immersion for 17 hrs 2.38 fg Water immersion for 24 hrs 2.88 fg control 0 g Ethrel - 50 ppm (16 hr) 0 g Ethrel − 25 ppm (16 hr) 0 g It was observed that the pre-sowing treatments effectively alleviated seed dormancy in wild finger millet accessions, IE 8414 and IE 8416, with efficacy varying significantly across treatments and between genotypes. Among the treatments evaluated, chemical applications demonstrated the highest overall effectiveness. Thiourea at 0.5% concentration for 16hr consistently emerged as the most effective treatment for breaking dormancy. Following thiourea (0.5% for 16hr), the next most effective treatments were potassium nitrate (KNO₃) at 0.2%, thiourea at 1% for 16hr, and KNO₃ at 0.5%. GA₃ treatments showed moderate effectiveness, with 500 ppm for 2hrs performing slightly better than 1000 ppm for 2hr. Physical treatments were considerably less effective; cold stratification at 4°C for 7 days showed some benefit over stratification for 15 days, and water immersion for 24 hours was marginally better than immersion for 17 hours. Treatments involving ethrel at either 25 ppm for 16hr or 50 ppm for 16hr were entirely ineffective, showing results equivalent to the untreated control. Genotype-specific responses were notable. For genotype, IE 8414, the order of treatment effectiveness began with thiourea (0.5% for 16hr) as the best, followed sequentially by KNO₃ (0.2%), thiourea (1% for 16hr), GA₃ (1000 ppm for 2 hr), water immersion (24 hours), GA₃ (500 ppm for 2 hr), and cold stratification (7 days). Treatments including water immersion (17 hours), KNO₃ (0.5%), and cold stratification (15 days) showed similarly low effectiveness, while ethrel treatments and the control were the least effective. A significant genotype × treatment interaction altered this pattern for IE 8416. Here, thiourea (0.5% for 16hr) was again the most effective, followed by KNO₃ (0.2%), then KNO₃ (0.5%), thiourea (1% for 16hr), GA₃ (500 ppm for 2 hr), and GA₃ (1000 ppm for 2 hr). Cold stratification for 7 days was more beneficial than for 15 days. Interestingly, water immersion for 17 hours was slightly more effective than the control but less effective than immersion for 24 hours in this genotype. Ethrel treatments remained completely ineffective, matching the control. Crucially, most treatments induced a substantial positive response in IE 8416, enhancing germination by approximately 50% compared to the control. The persistent dormancy observed despite extended cold stratification or water immersion (hydropriming), coupled with observed impermeable seed coats and a lack of imbibition and radicle emergence, strongly indicates the presence of physical dormancy in these genotypes (Baskin and Baskin 2004 ). Alarmingly, treatments utilizing ethylene-releasing compounds (ethrel at 25 for 16 hr and 50 ppm for 16hr) and gibberellic acid (GA₃ at 500 and 1000 ppm) resulted in severe phytotoxicity. Ethrel caused near-total (100%) seed mortality, while GA₃ treatments resulted in mortality approaching 100%. Affected seeds frequently exhibited microbial rot or fungal proliferation. Microscopic examination revealed structural damage to both the seed coat and endosperm, confirming the phytotoxic effects of these compounds at the applied concentrations.Generally, the responsiveness of seed to pre-sowing treatments is genotype and species-specific (Corbineau 2024 ), but dormancy-breaking treatments are standardised considering benefits at a larger scale. Ethrel (25 ppm for 16 hr) has been shown to enhance seed quality, germination, and seedling vigour in foxtail millet ( Setaria italica L.) by effectively breaking dormancy, with treated seeds exhibiting 86.4% germination, increased seedling length and reduced electrolyte leakage (Sebastian et al., 2015). It has been reported that ethrel (25 ppm 16 hr) enhanced seed quality, germination, and seedling vigour in crops such as soybean and rice (Ishibashi et al. 2013 ; Zhang et al. 2024 ). However, higher concentration of ethylene has negative impacts on germination in peanut, (Cui et al. 2025 ). In the case of GA 3 , it has a positive impact on the alpha-amylase activity, which had a key role in the regulation of seed germination (Brasileira et al. 2002 ). The alpha-activity estimation in both genotypes may provide an understanding of their differential response towards GA3 treatment. Though there was a huge difference in germination % between the genotypes due to GA 3 treatment, no significant increase in germination % was observed when the concentration of GA 3 increased from 500 ppm for 2 hr to 1000 ppm for 2hr. In the case of thiourea, it is the establishment of the proper redox equilibrium within a cellular environment that plays a key role in stimulatory effects on germination and growth, and the development of various crops (Vikas Yadav Patade et al. 2020). The study carried out by(Gupta et al. 2011 ) has also reported the negative impact of the increase in KNO 3 concentration on germination. The mechanism involved in it needs to be elucidated at the biochemical and molecular levels. Further, treatments like cold stratification and hydro priming treatments have been ineffective; the combination of these treatments with various chemical treatments can be studied in the future and their species-specific impact needs further elucidation in terms of finger millet wilds. Conclusion From the above study, it was concluded that [email protected] % for 16 hr was most effective in alleviating dormancy of freshly harvested wild finger millet germplasm. This treatment offers an effective method for breaking seed dormancy in wild finger millet species and can be adopted by genebanks, researchers, and breeders to facilitate improved germination and promote broader utilization of these genetic resources. Identification of treatments with similar potential and a combination of chemical treatments and hydropriming or water soaking can be more cost-effective and environmentally friendly. The availability of standardized pre-sowing treatments for finger millet wild species for alleviating the impact of dormancy can improve the efficiency of conservation and distribution of finger millet wild genetic resources. Further, the distribution of these genetic resources can initiate several pre-breeding programs for trait exploration. Declarations Conflict of Interest declaration: The authors declare that they have no conflict of interest. Funding Statement: This study was undertaken as part of the CGIAR Genebank Initiative and is part of the Research Program on Accelerated Crop Improvement, ICRISAT. Author Contribution O.P. Wrote the manuscriptK.J. Prepared the figures and tables.V.M. and K.S. 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In: Aryadeep Roychoudhury, Durgesh Kumar Tripathi (eds) Protective Chemical Agents in the Amelioration of Plant Abiotic Stress: Biochemical and Molecular Perspectives Zhang K, Khan MN, Luo T, Bi J, Hu L, Luo L (2024) Seed Priming with Gibberellic Acid and Ethephon Improved Rice Germination under Drought Stress via Reducing Oxidative and Cellular Damage. J Soil Sci Plant Nutr 24:2679–2693. https://doi.org/10.1007/s42729-024-01691-3 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 11 Dec, 2025 Read the published version in Genetic Resources and Crop Evolution → Version 1 posted Editorial decision: Revision requested 14 Sep, 2025 Reviews received at journal 13 Sep, 2025 Reviewers agreed at journal 10 Sep, 2025 Reviewers invited by journal 10 Sep, 2025 Editor assigned by journal 10 Sep, 2025 Submission checks completed at journal 10 Sep, 2025 First submitted to journal 03 Sep, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7526433","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":514786628,"identity":"fdf27e88-d1c5-43cc-b912-618b7aa6a88d","order_by":0,"name":"Ovais Peerzada","email":"data:image/png;base64,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","orcid":"","institution":"Genebank, International Crops Research Institute for the Semi-Arid Tropics","correspondingAuthor":true,"prefix":"","firstName":"Ovais","middleName":"","lastName":"Peerzada","suffix":""},{"id":514786629,"identity":"98bef9ea-1932-4352-a145-82367132a22c","order_by":1,"name":"Vetriventhan Mani","email":"","orcid":"","institution":"Genebank, International Crops Research Institute for the Semi-Arid Tropics","correspondingAuthor":false,"prefix":"","firstName":"Vetriventhan","middleName":"","lastName":"Mani","suffix":""},{"id":514786630,"identity":"c4b2fa2e-6bb9-453b-81ae-10bbeec49ca3","order_by":2,"name":"Kommineni Jagadeesh","email":"","orcid":"","institution":"Professor Jayashankar Telangana State Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Kommineni","middleName":"","lastName":"Jagadeesh","suffix":""},{"id":514786631,"identity":"955b210e-4437-4a30-8499-c6ecdb46e4c1","order_by":3,"name":"Kuldeep Singh","email":"","orcid":"","institution":"Genebank, International Crops Research Institute for the Semi-Arid Tropics","correspondingAuthor":false,"prefix":"","firstName":"Kuldeep","middleName":"","lastName":"Singh","suffix":""}],"badges":[],"createdAt":"2025-09-03 11:23:26","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7526433/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7526433/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s10722-025-02655-x","type":"published","date":"2025-12-11T15:58:17+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":91528660,"identity":"6c5ebd02-4e4a-435b-868a-c7f5138a0265","added_by":"auto","created_at":"2025-09-17 11:31:33","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":293092,"visible":true,"origin":"","legend":"\u003cp\u003eBar graph representing treatment means of germination of finger millet wild genotypes % under various pre-sowing seed treatment\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eNote: \u003c/strong\u003eT1- Control, T2\u003cstrong\u003e- \u003c/strong\u003eCold stratification 4° C 7 days, T3- Cold stratification 4° C 15 days , T4- Water immersion for 17 hrs, T5- Water immersion for 24 hrs, T6- KNO₃ - 0.2% (16 hr), T7- KNO₃ - 0.5% (16 hr), T8- Ethrel - 25 ppm (16 hr), T9- Ethrel - 50 ppm (16 hr), T10- Thiourea - 0.5 % (16 hr), T11- Thiourea - 1 % (16 hr), T12- GA₃ - 500ppm (2 hr), T13- GA₃- 1000ppm (2 hr)\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7526433/v1/9f8dc898a3dbbf75f5f6f604.jpeg"},{"id":91528666,"identity":"b68be6f2-2acd-43c0-b7f7-47a0793fb5a0","added_by":"auto","created_at":"2025-09-17 11:31:33","extension":"jpeg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":875651,"visible":true,"origin":"","legend":"\u003cp\u003eGermination test of Finger millet wild germplasm (IE 8416) using\u003cstrong\u003e \u003c/strong\u003ethe \"Top of Paper\" method under Thiourea treatment (0.5% for 16 hr) (A) and control (B)\u003c/p\u003e","description":"","filename":"floatimage2.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7526433/v1/f41f5339822337790f50ea51.jpeg"},{"id":91528662,"identity":"5c2828eb-bbfe-4088-8a00-96f67981dc16","added_by":"auto","created_at":"2025-09-17 11:31:33","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":66554,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eGenotype x Treatment means of germination % under various pre-sowing seed treatments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eNote: \u003c/strong\u003eT1- control, T2\u003cstrong\u003e- \u003c/strong\u003eCold stratification 4° C 7 days, T3- Cold stratification 4° C 15 days , T4- Water immersion for 17 hrs, T5- Water immersion for 24 hrs, T6- KNO₃ - 0.2%, T7- KNO₃ - 0.5%, T8- Ethrel - 25 ppm (16 hrs), T9- Ethrel - 50 ppm (16 hrs), T10- Thiourea - 0.5 % (16 hrs), T11- Thiourea - 1 % (16 hrs), T12- GA₃ - 500ppm (2 hr), T13- GA₃- 1000ppm (2 hr)\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-7526433/v1/398a44ac22fad9a247f9ff8b.png"},{"id":98244299,"identity":"c3dc1629-357e-48a8-b4e9-81c6410342c9","added_by":"auto","created_at":"2025-12-15 16:13:58","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2010156,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7526433/v1/0fa1c186-2b98-4342-b75b-75b0f704af93.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Optimization of pre-sowing treatments to break seed dormancy in wild finger millet (Eleusine africana L) for enhanced conservation and utilization of genetic resources","fulltext":[{"header":"Introduction","content":"\u003cp\u003eFinger millet is one of the most important millets among dry land crops with an allotetraploid nature (2n\u0026thinsp;=\u0026thinsp;4x\u0026thinsp;=\u0026thinsp;36) and C4 photosynthetic machinery. Its cultivation has been spread across several countries i.e., India, Ethiopia, Nepal, Uganda, Tanzania, Kenya, Zimbabwe, Zambia, Malawi, Eritrea, Mozambique, Rwanda, which accounts for around 20% of global millet area and 26% of global millet production (Gebreyohannes et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). India is one of the largest global producers of finger millet, with its cultivation spread across 1.162\u0026nbsp;million ha and with production of 1.692\u0026nbsp;million tons and productivity of 1456 Kg/ha (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.indiastat.com/\u003c/span\u003e\u003cspan address=\"https://www.indiastat.com/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). In addition, it possesses a better nutritional profile with superiority in several nutrients (Backiyalakshmi et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). In the current scenario of the intensified impact of climate change, genetic resources, specifically the crop wild species, hold the key to addressing the spiraling challenges. Crop wild relatives are the reservoir of several desirable alleles for various traits(Dida et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) and play a crucial role in breeding for traits such as drought tolerance, pest resistance, and enhanced nutritional profiles (Hilu and Dewet \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e1976\u003c/span\u003e). However, seed dormancy has been identified as one of the major hurdles among the wild species and needs to be addressed.\u003c/p\u003e\u003cp\u003eSeed dormancy is the inability of the seed to germinate, though the embryo is intact and viable. While dormancy enhances survival in natural ecosystems, it poses a significant challenge in ex-situ conservation, where rapid and uniform germination is essential for conservation, regeneration, and breeding programs. It has been a major hurdle for the freshly harvested seeds for their immediate use, which has enhanced the need for standardization of several pre-sowing seed treatments for overcoming it. Seed dormancy is a complex trait, affected by multiple genes and environmental factors (Graeber et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Lu et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) and an important component of plant adaptation (Donohue et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Huang et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). It is regulated by the antagonistic hormones, abscisic acid (ABA) and gibberellic acid (GA) (Finkelstein et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Shu et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2016\u003c/span\u003ee et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Moreover, seed dormancy is also considered an undesirable trait by farmers, affecting crop establishment. So, the dormancy-breaking pre-treatments have been found most useful in most of the crops, i.e., rice, wheat, maize, sorghum, pearl millet, chickpea, pigeon pea and many other crops (Burton \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e1969\u003c/span\u003e; Sahu et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e1993\u003c/span\u003e; Matus-C\u0026aacute;diz and Hucl \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Shanmugavalli et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Khadraji and Qaddoury \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Lamichaney et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Compared to cultivated, the strong dormancy mechanisms present in wild genotypes often result in poor germination under standard conditions, complicating their conservation and utilization (Long et al., 2015). The study conducted by (Kannababu et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2025\u003c/span\u003e) reported several genes regulating seed vigour related traits in finger millet. The impact of seed dormancy breaking treatments on the expression of genes regulating seed vigour and related traits can elucidate further mechanisms involved.\u003c/p\u003e\u003cp\u003eCurrently, ICRISAT genebank holds a global finger millet collection of 7536 accessions collected from 25 countries including 206 wild germplasm accessions (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://genebank.icrisat.org/\u003c/span\u003e\u003cspan address=\"https://genebank.icrisat.org/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). Seed viability testing, typically conducted through germination tests, is a critical component of routine genebank operations. To ensure that germination results accurately reflect true seed viability, it is essential to apply an effective dormancy-breaking treatment, but limited information regarding the effective pre-sowing seed treatments and standardized protocol to overcome seed dormancy in wild species of finger millet is limiting their efficient conservation as well as exploration of their potential in crop improvement programs. Hence, the current study was initiated to unveil the impact of various pre-seed treatments to develop a more efficient treatment for routine use in viability testing for conservation and also to enhance the utility of crop wild relatives of finger millet.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003ePlant material\u003c/h2\u003e\u003cp\u003eThe current study was carried out during the year 2023-24 at ICRISAT, Genebank, Patancheru, Hyderabad. The seeds belonging to the two wild finger millet accessions (IE 8414 and IE 8416) were sown in the glass house at ICRISAT in the month of October 2023. The seeds were grown in 12-inch pots in a mixture of red soil, sand and vermicompost in the ratio of 3:2:1. At the time of sowing, 2 g DAP was added to the mixture, and 2 g of urea was applied after 25 days of sowing. After thinning, only five seedlings were maintained in each pot, and the seeds from both accessions were harvested at maturity. The seeds were carefully harvested, threshed, and after harvesting, a sample of seeds from the accessions was taken in grip seal bags to determine the initial seed moisture content, which was 16.2% (Oven dry method). Seeds were dried in the sun (max. 30\u003csup\u003e\u0026deg;\u003c/sup\u003eC, min. 22\u003csup\u003e\u0026deg;\u003c/sup\u003eC) to bring the moisture content down to 12.6% and then transferred to the genebank drying room maintained at 15\u0026deg;C and 15% RH for further drying. The seeds were dried to a uniform moisture content (7.1%) before sampling for germination testing. The seeds were taken out from the drying room and kept at room temperature (25\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u003csup\u003e0\u003c/sup\u003eC) for saturation before being immersed in different chemical solutions.\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eExperimental design and treatments\u003c/h3\u003e\n\u003cp\u003eThe current experiment was carried out in a factorial completely randomized design with genotypes and treatments as two different factors and four replications. The combinations of genotype \u0026times; treatments were applied randomly across replications.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eDifferent kinds of pre-sowing seed treatments used for breaking seed dormancy\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"3\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eS. No\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eTreatment\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eConcentration/Condition\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eControl\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eNormal Incubation (25\u003csup\u003e0\u003c/sup\u003eC for 10 days)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCold stratification\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e4\u0026deg;C for 7 days\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCold stratification\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e4\u0026deg;C for 15 days\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eHydropriming\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSoaking in distilled water for 17 hr\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eHydropriming\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSoaking in distilled water for 24 hr\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eChemical treatment\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003ePotassium nitrate (KNO₃) at 0.2% (16 hr)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eChemical treatment\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003ePotassium nitrate (KNO₃) at 0.5% (16 hr)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eChemical treatment\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eEthrel 25 ppm (16 hr)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eChemical treatment\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eEthrel 50 ppm (16 hr)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eChemical treatment\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eThiourea 0.5% (16 hr)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eChemical treatment\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eThiourea 1% (16 hr)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e12\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eChemical treatment\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eGibberellic acid (GA₃) at 500 ppm (2 hr)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eChemical treatment\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eGibberellic acid (GA₃) at 1000 ppm (2 hr)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\n\u003ch3\u003eSeed Viability and dormancy evaluation\u003c/h3\u003e\n\u003cp\u003eBefore applying dormancy-breaking treatments, the seed viability of both the wild accessions was assessed using the tetrazolium (TZ) test following ISTA guidelines (ISTA \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). For each accession, 100 seeds were preconditioned by soaking in distilled water at room temperature (23\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026deg;C) for 16 hours. The seeds were then longitudinally and carefully dissected under a magnifying lens to expose the embryo. Dissected seeds were immersed in a 1% tetrazolium chloride solution and incubated in the dark at 25\u0026deg;C for two hours. Following staining, seeds were rinsed twice with distilled water. Seeds were carefully examined under a Tagarno magnifying microscope for evaluation. Seeds were considered viable if the embryo was uniformly stained red. The viability percentage for each accession was calculated as the proportion of seeds with completely stained embryos relative to the total number of seeds tested.\u003c/p\u003e\u003cp\u003eFor dormancy evaluation, uniform and intact seeds were selected from the available seed, and a total of 400 seeds were randomly selected from them and divided into four replications of 100 seeds each. The seeds were then subjected to different kinds of pre-sowing seed treatments, including cold stratification and hydropriming with exposure for different number of days and variable periods, respectively. Moreover, the treatments include exposure to chemicals like potassium nitrate (KNO₃), thiourea, and also hormones like ethrel and gibberellic acid at different concentrations and periods \u003cb\u003e(\u003c/b\u003eTable\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e\u003cb\u003e).\u003c/b\u003e Following the treatments, after the soaking period was over, the seeds were taken out and washed thoroughly with distilled water before placing them on petri dishes for germination. Germination testing was performed at the seed laboratory of ICRISAT genebank, following the International Seed Testing Association rules (ISTA \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) using the \"Top of Paper\" method, where seeds were evenly spaced on moistened germination sheets in petri dishes. The dishes were covered and incubated in a growth chamber under controlled conditions (25\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C temperature and 85\u0026ndash;90% relative humidity). Germination was recorded on the tenth day by counting normal seedlings, and the germination percentage was calculated as\u003c/p\u003e\u003cp\u003eGermination percentage (%) = (Number of germinated seeds / Total seeds) * 100\u003c/p\u003e\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\u003ch2\u003eStatistical analysis\u003c/h2\u003e\u003cp\u003eGermination percentage data were arcsine square root transformed before statistical analysis to stabilize the variance. The data were analyzed based on the linear model in which genotype, treatment, genotype \u0026times; treatment, and replication are considered fixed factors using R software (version 4.2.2) and the packages emmeans and multcompView. The interaction (Genotype \u0026times; Treatment) and main effect mean (treatment and genotype) were calculated. Further, post-hoc analysis was performed using the Duncan multiple range test (DMRT) (α\u0026thinsp;=\u0026thinsp;0.05).\u003c/p\u003e\u003c/div\u003e"},{"header":"Results and Discussion","content":"\u003cp\u003eEmbryos of seeds from both wild finger millet accessions exhibited a uniform red coloration upon staining with 1% tetrazolium chloride (TZ) solution, indicating that all tested seeds were viable. The viability percentage recorded was 100% for the accession IE 8416, while it was 99% for the accession IE 8414. The complete and consistent staining of the embryo tissues suggests that cellular respiration was active, confirming the physiological integrity and viability of the seeds across both accessions. The differential impact of various dormancy-breaking treatments imposed on wild accessions of finger millet was evident \u003cstrong\u003e(\u003c/strong\u003eTable \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e\u003cstrong\u003e).\u003c/strong\u003e Highly significant variation in germination rates among different treatments (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) and between accessions (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001), with a significant genotype \u0026times; treatment interaction effect (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001), was observed. The control groups of both \u003cem\u003eE. africana\u003c/em\u003e accessions (IE 8414 and IE 8416) demonstrated absolute germination failure (0% \u0026plusmn; 0.00), confirming the persistence of primary dormancy under standard conditions (25\u003csup\u003e0\u003c/sup\u003e C for 10 days) \u003cstrong\u003e(\u003c/strong\u003eTable \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e\u003cstrong\u003e).\u003c/strong\u003e In addition, a significant difference in germination (%) among the two wild genotypes has also been observed.\u003c/p\u003e\n\u003cp\u003e\u003c/p\u003e\n\u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eAnalysis of variance (ANOVA) for the germination percentage of \u003cem\u003eEleusine africana\u003c/em\u003e genotypes under various treatments\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSource of Variation\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eDF\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSum Sq\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMean Sq\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eF-Value\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePr(\u0026gt;\u0026thinsp;F)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAccession\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5835\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5835\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e558.27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;2e-16***\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTreatment\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e47798\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3983\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e381.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;2e-16***\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAccession* treatment\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6138\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e512\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e48.94\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;2e-16***\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eResiduals\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e815\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTotal\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e103\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e60,586\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\"\u003eNote: Significance codes: *** indicates P\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\n \u003ch2\u003eGenotype*Treatment means\u003c/h2\u003e\n \u003cp\u003eAmong the various pre-sowing treatments \u003cstrong\u003e(\u003c/strong\u003eTable \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e\u003cstrong\u003e)\u003c/strong\u003e, similar to the control, no germination was observed in the case of ethrel under both concentrations (50 ppm and 25 ppm) for a period of 16 hr in both accessions. Though cold stratification of 4\u0026deg;C for 7 days and 15 days has induced germination in both accessions, the germination percentage was lower than in other treatments, and differential response was observed in the two accessions. Moreover, water immersion treatments (17 and 24 hr) have not been very effective and resulted in poor germination (ranged between 2.38\u0026ndash;2.88%) in the case of both accessions. GA\u003csub\u003e3\u003c/sub\u003ehas a significant positive impact on germination, but it was limited only to one of the two varieties i.e., effective in IE 8416 with germination around 50% under both the concentrations i.e., 500 ppm (2 hr) and 1000 ppm (2 hr). No significant difference in germination was observed in different concentrations of gibberellic acid i.e., 500 ppm for 2 hr and 1000 ppm for 2 hr. Similar to GA\u003csub\u003e3\u003c/sub\u003e, KNO\u003csub\u003e3\u003c/sub\u003e treatment for a period of 16 hr also had a differential impact on germination in two accessions, with a greater positive impact on IE 8416. In addition, it also varied based on concentration (0.2% and 0.5%), which was not observed in the case of GA\u003csub\u003e3\u003c/sub\u003e. The increase in the concentration of KNO₃ resulted in a significant decrease in germination and it was much more drastic in the case of IE 8414 (50.25\u0026ndash;0.25%). Apart from all the treatments, thiourea is the chemical treatment that has shown the highest germination percentage (\u0026gt;\u0026thinsp;90%) in both the accessions under 0.5% for 16 hr while further increase in concentration (1% for 16 hrs) resulted in reduction in germination (~\u0026thinsp;30%) .\u003c/p\u003e\n \u003cdiv align=\"left\" class=\"colspec\"\u003e\u003cbr\u003e\u003c/div\u003e\n \u003ctable id=\"Tab3\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eGrouping of genotypes*treatment means of germination% based on Duncan multiple range test\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003eDuncan\u0026apos;s Multiple Range Test _ Treatments\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eAccession\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTreatment\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMean\u0026thinsp;\u0026plusmn;\u0026thinsp;SEM\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003egroups\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8416\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCold stratification 4\u0026deg; C 15 days\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e10.25\u0026thinsp;\u0026plusmn;\u0026thinsp;1.49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003egh\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8414\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCold stratification 4\u0026deg; C 15 days\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ei\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8416\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCold stratification 4\u0026deg; C (7 days)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e13.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.65\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eg\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8414\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCold stratification 4\u0026deg; C (7 days)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ei\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8414\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eControl (25\u003csup\u003e0\u003c/sup\u003e for 10 days)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ei\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8416\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eControl (25\u003csup\u003e0\u003c/sup\u003e for 10 days)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ei\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8414\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eEthrel - 50 ppm(16 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ei\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8416\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eEthrel - 50 ppm (16 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ei\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8414\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eEthrel \u0026minus;\u0026thinsp;25 ppm (16 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ei\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8416\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eEthrel \u0026minus;\u0026thinsp;25 ppm (16 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ei\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8416\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGA₃ \u0026minus;\u0026thinsp;500 ppm (2 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e51.00\u0026thinsp;\u0026plusmn;\u0026thinsp;4.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ecd\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8414\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGA₃ \u0026minus;\u0026thinsp;500 ppm (2 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.75\u0026thinsp;\u0026plusmn;\u0026thinsp;0.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ei\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8416\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGA₃- 1000 ppm (2 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e46.25\u0026thinsp;\u0026plusmn;\u0026thinsp;3.90\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ed\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8414\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGA₃- 1000 ppm (2 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.00\u0026thinsp;\u0026plusmn;\u0026thinsp;1.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ei\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8416\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eKNO₃ \u0026minus;\u0026thinsp;0.2% (16 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e70.75\u0026thinsp;\u0026plusmn;\u0026thinsp;2.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eb\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8414\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eKNO₃ \u0026minus;\u0026thinsp;0.2% (16 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e50.75\u0026thinsp;\u0026plusmn;\u0026thinsp;0.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ecd\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8416\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eKNO₃ \u0026minus;\u0026thinsp;0.5% (16 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e53.75\u0026thinsp;\u0026plusmn;\u0026thinsp;4.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ec\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8414\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eKNO₃ \u0026minus;\u0026thinsp;0.5% (16 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ei\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8416\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eThiourea \u0026minus;\u0026thinsp;0.5%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e95.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ea\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8414\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eThiourea \u0026minus;\u0026thinsp;0.5%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e91.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ea\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8416\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eThiourea \u0026minus;\u0026thinsp;1%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e36.25\u0026thinsp;\u0026plusmn;\u0026thinsp;2.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ee\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8414\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eThiourea \u0026minus;\u0026thinsp;1%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e27.50\u0026thinsp;\u0026plusmn;\u0026thinsp;2.84\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ef\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8416\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWater immersion (17 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4.50\u0026thinsp;\u0026plusmn;\u0026thinsp;1.19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehi\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8414\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWater immersion (17 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ei\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8416\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWater immersion (24 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.75\u0026thinsp;\u0026plusmn;\u0026thinsp;0.75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehi\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIE 8414\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWater immersion (24 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.00\u0026thinsp;\u0026plusmn;\u0026thinsp;1.68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ei\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003c/p\u003e\n \u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003c/div\u003e\n\u003ch3\u003eTreatment means\u003c/h3\u003e\n\u003cp\u003eThe means of germination % for various treatments \u003cstrong\u003e(\u003c/strong\u003eTable \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e\u003cstrong\u003e)\u003c/strong\u003e and grouping based on significant differences \u003cstrong\u003e(\u003c/strong\u003eFig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e\u003cstrong\u003e)\u003c/strong\u003e can provide an overview of the impact of various treatments on finger millet wilds. No significant difference was observed among the treatments, control, ethrel (25 ppm for 16 hr and 50 ppm for 16 hr) and water immersion (17 hrs and 24 hrs) and these are not effective in breaking seed dormancy. The cold stratification treatment of 4\u0026deg;C for 7 days and 15 days treatments are also similar and had minimal impact on enhancing germination. No significant increase in germination percentage when the concentration of GA\u003csub\u003e3\u003c/sub\u003e was increased from 500 ppm to 1000 ppm (24.63% and 25.88%). In the case of KNO\u003csub\u003e3\u003c/sub\u003e for a period of 16 hr, a lower concentration of 0.2% shown a greater positive impact than a higher concentration (0. 5%) with germination of 60.75% and 27% respectively. Finally, thiourea (0.5%) for a period of 16 hr is the chemical treatment that has been most effective and enhanced germination levels of \u0026gt;\u0026thinsp;90%, but a drastic reduction in was witnessed when its concentration was increased to 1%.\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n \u003cdiv align=\"left\" class=\"colspec\"\u003e\u003cbr\u003e\u003c/div\u003e\n \u003ctable id=\"Tab4\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eTreatment grouping based on Germination % using Duncan multiple range test (DMRT)\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTreatment\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMean\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eGroup\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eThiourea \u0026minus;\u0026thinsp;0.5% (16 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e93.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ea\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eKnO₃ \u0026minus;\u0026thinsp;0.2% (16 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e60.75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eb\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eThiourea \u0026minus;\u0026thinsp;1% (16 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e31.88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ec\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGA₃- 1000ppm (2 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e24.63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ed\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGA₃ \u0026minus;\u0026thinsp;500ppm (2 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e25.88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ed\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eKnO₃ \u0026minus;\u0026thinsp;0.5% (16 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e27.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ed\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCold stratification 4\u0026deg; C 7 days\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ee\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCold stratification 4\u0026deg; C 15 days\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eef\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWater immersion for 17 hrs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003efg\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWater immersion for 24 hrs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003efg\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003econtrol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eg\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eEthrel - 50 ppm (16 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eg\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eEthrel \u0026minus;\u0026thinsp;25 ppm (16 hr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eg\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003eIt was observed that the pre-sowing treatments effectively alleviated seed dormancy in wild finger millet accessions, IE 8414 and IE 8416, with efficacy varying significantly across treatments and between genotypes. Among the treatments evaluated, chemical applications demonstrated the highest overall effectiveness. Thiourea at 0.5% concentration for 16hr consistently emerged as the most effective treatment for breaking dormancy. Following thiourea (0.5% for 16hr), the next most effective treatments were potassium nitrate (KNO₃) at 0.2%, thiourea at 1% for 16hr, and KNO₃ at 0.5%. GA₃ treatments showed moderate effectiveness, with 500 ppm for 2hrs performing slightly better than 1000 ppm for 2hr. Physical treatments were considerably less effective; cold stratification at 4\u0026deg;C for 7 days showed some benefit over stratification for 15 days, and water immersion for 24 hours was marginally better than immersion for 17 hours. Treatments involving ethrel at either 25 ppm for 16hr or 50 ppm for 16hr were entirely ineffective, showing results equivalent to the untreated control.\u003c/p\u003e\n\u003cp\u003eGenotype-specific responses were notable. For genotype, IE 8414, the order of treatment effectiveness began with thiourea (0.5% for 16hr) as the best, followed sequentially by KNO₃ (0.2%), thiourea (1% for 16hr), GA₃ (1000 ppm for 2 hr), water immersion (24 hours), GA₃ (500 ppm for 2 hr), and cold stratification (7 days). Treatments including water immersion (17 hours), KNO₃ (0.5%), and cold stratification (15 days) showed similarly low effectiveness, while ethrel treatments and the control were the least effective. A significant genotype \u0026times; treatment interaction altered this pattern for IE 8416. Here, thiourea (0.5% for 16hr) was again the most effective, followed by KNO₃ (0.2%), then KNO₃ (0.5%), thiourea (1% for 16hr), GA₃ (500 ppm for 2 hr), and GA₃ (1000 ppm for 2 hr). Cold stratification for 7 days was more beneficial than for 15 days. Interestingly, water immersion for 17 hours was slightly more effective than the control but less effective than immersion for 24 hours in this genotype. Ethrel treatments remained completely ineffective, matching the control. Crucially, most treatments induced a substantial positive response in IE 8416, enhancing germination by approximately 50% compared to the control.\u003c/p\u003e\n\u003cp\u003eThe persistent dormancy observed despite extended cold stratification or water immersion (hydropriming), coupled with observed impermeable seed coats and a lack of imbibition and radicle emergence, strongly indicates the presence of physical dormancy in these genotypes (Baskin and Baskin \u003cspan class=\"CitationRef\"\u003e2004\u003c/span\u003e). Alarmingly, treatments utilizing ethylene-releasing compounds (ethrel at 25 for 16 hr and 50 ppm for 16hr) and gibberellic acid (GA₃ at 500 and 1000 ppm) resulted in severe phytotoxicity. Ethrel caused near-total (100%) seed mortality, while GA₃ treatments resulted in mortality approaching 100%. Affected seeds frequently exhibited microbial rot or fungal proliferation. Microscopic examination revealed structural damage to both the seed coat and endosperm, confirming the phytotoxic effects of these compounds at the applied concentrations.Generally, the responsiveness of seed to pre-sowing treatments is genotype and species-specific (Corbineau \u003cspan class=\"CitationRef\"\u003e2024\u003c/span\u003e), but dormancy-breaking treatments are standardised considering benefits at a larger scale. Ethrel (25 ppm for 16 hr) has been shown to enhance seed quality, germination, and seedling vigour in foxtail millet (\u003cem\u003eSetaria italica\u003c/em\u003e L.) by effectively breaking dormancy, with treated seeds exhibiting 86.4% germination, increased seedling length and reduced electrolyte leakage (Sebastian et al., 2015). It has been reported that ethrel (25 ppm 16 hr) enhanced seed quality, germination, and seedling vigour in crops such as soybean and rice (Ishibashi et al. \u003cspan class=\"CitationRef\"\u003e2013\u003c/span\u003e; Zhang et al. \u003cspan class=\"CitationRef\"\u003e2024\u003c/span\u003e). However, higher concentration of ethylene has negative impacts on germination in peanut, (Cui et al. \u003cspan class=\"CitationRef\"\u003e2025\u003c/span\u003e). In the case of GA\u003csub\u003e3\u003c/sub\u003e, it has a positive impact on the alpha-amylase activity, which had a key role in the regulation of seed germination (Brasileira et al. \u003cspan class=\"CitationRef\"\u003e2002\u003c/span\u003e). The alpha-activity estimation in both genotypes may provide an understanding of their differential response towards GA3 treatment. Though there was a huge difference in germination % between the genotypes due to GA\u003csub\u003e3\u003c/sub\u003e treatment, no significant increase in germination % was observed when the concentration of GA\u003csub\u003e3\u003c/sub\u003e increased from 500 ppm for 2 hr to 1000 ppm for 2hr.\u003c/p\u003e\n\u003cp\u003eIn the case of thiourea, it is the establishment of the proper redox equilibrium within a cellular environment that plays a key role in stimulatory effects on germination and growth, and the development of various crops (Vikas Yadav Patade et al. 2020). The study carried out by(Gupta et al. \u003cspan class=\"CitationRef\"\u003e2011\u003c/span\u003e) has also reported the negative impact of the increase in KNO\u003csub\u003e3\u003c/sub\u003e concentration on germination. The mechanism involved in it needs to be elucidated at the biochemical and molecular levels. Further, treatments like cold stratification and hydro priming treatments have been ineffective; the combination of these treatments with various chemical treatments can be studied in the future and their species-specific impact needs further elucidation in terms of finger millet wilds.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eFrom the above study, it was concluded that \u003cem\[email protected]%\u003c/em\u003e for 16 hr was most effective in alleviating dormancy of freshly harvested wild finger millet germplasm. This treatment offers an effective method for breaking seed dormancy in wild finger millet species and can be adopted by genebanks, researchers, and breeders to facilitate improved germination and promote broader utilization of these genetic resources. Identification of treatments with similar potential and a combination of chemical treatments and hydropriming or water soaking can be more cost-effective and environmentally friendly. The availability of standardized pre-sowing treatments for finger millet wild species for alleviating the impact of dormancy can improve the efficiency of conservation and distribution of finger millet wild genetic resources. Further, the distribution of these genetic resources can initiate several pre-breeding programs for trait exploration.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eConflict of Interest\u0026nbsp;declaration:\u003c/strong\u003e The authors declare that they have no conflict of interest.\u003c/p\u003e\n\u003ch2\u003eFunding Statement:\u003c/h2\u003e\n\u003cp\u003eThis study was undertaken as part of the CGIAR Genebank Initiative and is part of the Research Program on Accelerated Crop Improvement, ICRISAT.\u003c/p\u003e\n\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\n\u003cp\u003eO.P. Wrote the manuscriptK.J. Prepared the figures and tables.V.M. and K.S. 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In: Aryadeep Roychoudhury, Durgesh Kumar Tripathi (eds) Protective Chemical Agents in the Amelioration of Plant Abiotic Stress: Biochemical and Molecular Perspectives\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhang K, Khan MN, Luo T, Bi J, Hu L, Luo L (2024) Seed Priming with Gibberellic Acid and Ethephon Improved Rice Germination under Drought Stress via Reducing Oxidative and Cellular Damage. J Soil Sci Plant Nutr 24:2679\u0026ndash;2693. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s42729-024-01691-3\u003c/span\u003e\u003cspan address=\"10.1007/s42729-024-01691-3\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"genetic-resources-and-crop-evolution","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"gres","sideBox":"Learn more about [Genetic Resources and Crop Evolution](https://www.springer.com/journal/10722)","snPcode":"10722","submissionUrl":"https://submission.nature.com/new-submission/10722/3","title":"Genetic Resources and Crop Evolution","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Wild relatives, genetic conservation, climate resilience, dormancy, and germination","lastPublishedDoi":"10.21203/rs.3.rs-7526433/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7526433/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eCrop wild relatives of finger millet, a part of its genetic resources, are key to exploring the genetic potential of the crop for several climate-resilient and nutritional traits. Seed dormancy in wild \u003cem\u003especies\u003c/em\u003e accessions poses a significant challenge to the conservation, evaluation, distribution, and their utilization in pre-breeding activities. In the present study, we tested 12 treatments to break dormancy and enhance germination in two wild accessions (IE 8414 and IE 8416). Germination of these two accessions under control conditions was 0%. Among the treatments tested, Thiourea at 0.5% for 16 hrs was the most effective treatment, achieving germination rates of 91.00% in IE 8414 and 95.25% in IE 8416. Potassium nitrate (KNO₃) at 0.2% for 16 hrs also showed significant efficacy, with 50.75% germination in IE 8414 and 70.75% in IE 8416. Gibberellic acid (GA₃) at 500 ppm for 2 hrs induced 51.00% germination in IE 8416, while\u0026thinsp;\u0026lt;\u0026thinsp;1% in IE 8414. Chemical treatments have been more effective, and while other treatments such as ethrel, cold stratification, and water immersion, were largely ineffective, with germination rates remaining below 5%. These findings highlight the potential of Thiourea and KNO₃ as effective tools for breaking dormancy in wild \u003cem\u003eEleusine\u003c/em\u003e accessions.\u003c/p\u003e","manuscriptTitle":"Optimization of pre-sowing treatments to break seed dormancy in wild finger millet (Eleusine africana L) for enhanced conservation and utilization of genetic resources","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-17 11:31:28","doi":"10.21203/rs.3.rs-7526433/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-09-14T10:01:21+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-09-13T17:07:09+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"11583557937966355042731256556087517858","date":"2025-09-10T09:21:20+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-09-10T08:41:37+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-09-10T08:37:51+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-09-10T08:36:44+00:00","index":"","fulltext":""},{"type":"submitted","content":"Genetic Resources and Crop Evolution","date":"2025-09-03T11:18:10+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"genetic-resources-and-crop-evolution","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"gres","sideBox":"Learn more about [Genetic Resources and Crop Evolution](https://www.springer.com/journal/10722)","snPcode":"10722","submissionUrl":"https://submission.nature.com/new-submission/10722/3","title":"Genetic Resources and Crop Evolution","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"f98ad139-a1d8-4584-9a19-662620c60630","owner":[],"postedDate":"September 17th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-12-15T16:07:21+00:00","versionOfRecord":{"articleIdentity":"rs-7526433","link":"https://doi.org/10.1007/s10722-025-02655-x","journal":{"identity":"genetic-resources-and-crop-evolution","isVorOnly":false,"title":"Genetic Resources and Crop Evolution"},"publishedOn":"2025-12-11 15:58:17","publishedOnDateReadable":"December 11th, 2025"},"versionCreatedAt":"2025-09-17 11:31:28","video":"","vorDoi":"10.1007/s10722-025-02655-x","vorDoiUrl":"https://doi.org/10.1007/s10722-025-02655-x","workflowStages":[]},"version":"v1","identity":"rs-7526433","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7526433","identity":"rs-7526433","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}