Assessment of Soil Seed Bank Dynamics and Regeneration Potential Across Three Different Forest Types in the Himalayan Foothills

Assessment of Soil Seed Bank Dynamics and Regeneration Potential Across Three Different Forest Types in the Himalayan Foothills | 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 Assessment of Soil Seed Bank Dynamics and Regeneration Potential Across Three Different Forest Types in the Himalayan Foothills Pema Choki Lepcha, Prabhakar Manori, Manish Kumar, Vikaspal Singh, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7984952/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract This study examines the soil seed bank potential in three forest types in Dehradun district: Sal, Pine, and Mixed Deodar. Soil samples were collected and monitored for seedling emergence to assess their roles in natural regeneration and biodiversity conservation. Species composition, density, diversity indices, and inter-forest similarities were analyzed using statistical and ordination techniques. A total of 371 individuals from 15 species across 10 families were observed, with herbs being the dominant growth form (66.67%) and Asteraceae as the most represented family (20%). Seed density varied among forest types, with Pine forest having the highest germination potential and maximum seed density of 1291.67 seeds/m² for Sorghum halepense . Deodar forest had an intermediate density dominated by Cassia tora (638.89 seeds/m²), while Sal forest had the lowest density but supported the highest species richness (10 species) and diversity indices (Shannon = 2.07, Simpson = 0.85). Jaccard’s similarity coefficient indicated greater similarity between Sal and Pine forests (0.583), with Deodar showing the least overlap with Sal (0.286). NMDS ordination confirmed distinct community assemblages among the three forest types, with characteristic indicator species identified for each. These results demonstrate significant variation in soil seed bank composition and regenerative capacity across forest types. Pine forests have dense but less diverse seed banks, mainly dominated by grasses, while Sal forests have balanced and diverse assemblages, supporting broad regeneration potential. Deodar forests exhibit lower diversity due to dense canopy and thick litter layers. The presence of invasive species like Parthenium hysterophorus in all sites highlights ecological risks to native regeneration. Biodiversity Conservation Regeneration Soil Seed Bank Species Diversity Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction Based on the Global Forest Watch report (2024), Uttarakhand experienced a loss of 30 hectares of humid primary forest, accounting for 0.13% of its total tree cover loss during the same period. The total area of humid primary forest in Uttarakhand decreased by 0.32% in this timeframe, indicating a concerning rate of forest depletion in the region. Soil Seed Bank Analysis is a valuable tool for comprehending future forest trends and informing conservation strategies. The soil seed bank serves as a reservoir of viable seeds within the soil, playing a crucial role in plant population dynamics, biodiversity preservation, and ecosystem resilience. It is essential for natural regeneration, species succession, and ecological restoration efforts (Thompson and Grime, 1979 ; Baskin and Baskin, 2014 ). Understanding the composition and dynamics of soil seed banks is particularly vital in forest ecosystems, where vegetation recovery post-disturbances relies heavily on the existing seed population. Roberts ( 1981 ) defines the soil seed bank as the collection of viable seeds present in the soil. Baker ( 1989 ) further elaborates that this reservoir consists of seeds that have not yet germinated but have the potential to replace adult plants lost due to natural causes, diseases, disturbances, or consumption by animals, including humans. All viable seeds within the soil or mixed with soil debris contribute to the soil seed bank (Simpson et al., 1989 ). Large quantities of seeds can remain dormant in the soil for extended periods, and can sprout when conditions are favorable (Warr et al. 1993 ). Soil seed banks play a crucial role in understanding vegetation history, as the composition of vegetation in terms of plant species is influenced by seed production, dispersal, seed longevity, and soil depth (V. Anju et al. 2022 ). Seed banks are key determinants of future vegetation, particularly following a disturbance (Warr et al. 1993 ). Dehradun, situated in the Doon Valley at the foothills of the western Himalayas, is known for its remarkable ecological diversity. It is located at the meeting point of the Shivalik range, the outer Himalayas, and the Indo-Gangetic plains, creating diverse climatic and soil conditions that support a variety of forest types (Champion and Seth, 1968 ). The region encompasses moist deciduous forests dominated by sal ( Shorea robusta ), pine ( Pinus roxburghii ) forests, mixed broadleaf forests, and riverine forests, among others. These diverse forest ecosystems exhibit differences in microclimatic conditions, human impact, disturbance patterns, and vegetation structure, all of which impact the composition and abundance of their soil seed banks. While regeneration studies have been conducted by various researchers, there is limited information available on the diversity of soil seed banks in different forest types in Uttarakhand. The present study aims to address this gap by investigating the soil seed bank diversity in Sal, Pine, and mixed Deodar-dominated forest stands in the Himalayan foothills. Methodology Study Area This study took place in Dehradun district, situated between the Himalayas to the north and the Shivalik Hills to the south, with geographical coordinates of 30.3165o N latitude and 78.0322o E longitude. The major forest types in the area include Sal forests at lower elevations, Pine forests at mid elevations, and Oak and Rhododendron at higher elevations. Site I: Shorea robusta (Sal) Forest: Located at latitude 30.37752 N and longitude 77.931215 E, soil samples were collected from the Sal Forest in Manduwala, part of the Dehradun Forest Division. Site II: Pinus roxburghii (Chir Pine) Forest: Coordinates at latitude 30.453615 N and longitude 77.944039 E. Soil samples were collected from the Pine Forest in Koti, within the Chakrata Forest Division. Site III: Mixed Cedrus deodara (Deodar) Forest: Coordinates at latitude 30.4449540 N and longitude 78.076609 E. This study site is located in Mussoorie, which is under the jurisdiction of the Mussoorie Forest Division. Soil Sampling In this study, 1 m × 1 m quadrats were randomly placed across the study site to evaluate species distribution and abundance. A distance of 50 meters was maintained between consecutive quadrats to ensure spatial independence and reduce sampling bias. A total of 10 quadrats were established at each study site to provide a representative sample for statistical analysis. Two soil samples were collected from each quadrat—one from a depth of 0–5 cm and the other from 5–10 cm. Therefore, 20 soil samples were collected from each site, resulting in a total of 60 samples across all three sites. In the laboratory, the two depth samples from each plot were combined and thoroughly mixed before being air-dried for 48 hours. Any visible debris, such as stones, twigs, and other extraneous material, was carefully removed during the drying process. After drying and cleaning, the soil samples were placed in labelled vegetation trays and kept in a controlled setting without existing vegetation to prevent external interference with natural seed germination. No external soil enrichments, such as fertilizers or farmyard manure (FYM), were added to observe the natural regeneration potential of the seed bank. Irrigation was carried out manually, initially every alternate day, and later daily as the summer season progressed and evaporation rates increased to maintain adequate soil moisture. Data Collection Seedling emergence was regularly monitored, and the number of germinations from each tray was recorded. Seedlings were uprooted by hand as they reached a stage where species identification was possible, to minimize disturbance and allow other seeds to continue germinating. Species were identified based on morphological traits such as size, shape, color, and surface texture of leaves and stems. The weekly recording of the number of individuals of each species was conducted over an eight-week period. Data Analysis Soil seed bank germination was compared among different forest types using the Kruskal–Wallis test (Kruskal & Wallis, 1952), followed by Dunn’s post-hoc pairwise comparisons (Dunn’s, 1964). Community structure was evaluated using diversity indices (species richness, Shannon, Simpson, and evenness), and compositional differences were visualized with non-metric multidimensional scaling (NMDS) based on Bray–Curtis dissimilarity. All statistical analyses were performed using R software (Version 4.5.0) with appropriate packages for non-parametric testing, diversity metrics, and ordination. Seed density was calculated as the number of seedlings (viable seeds) per unit area (expressed as seeds/m²) based on the total number of seedlings that emerged from each soil sample. Result Soil seed bank density and morphology of plants A total of 371 individuals emerged from the soil seed banks of three different forest types, consisting of a total of 15 species, namely Ageratum houstonianum, Sorghum halepense, Dioscorea bulbifera, Commelina diffusa, Parthenium hysterophorus, Cassia tora, Alysicarpus ovalifolius, Commelina benghalensis, Scindapsus officinalis, Solanum xanthocarpum, Artemisia vulgaris, Lindenbergia indica, Oxalis exilis, Cedrus deodara , and one unidentified species representing 10 families. The Asteraceae family was the most dominant, with 3 species (20%), followed by Fabaceae and Commelinaceae with 2 species each (13.33%). The rest of the families had only one species each (6.67%). Herbs were the dominant growth form, comprising 66.67% (10 species), followed by climbers at 13.33% (2 species). Dicots accounted for 53.33% (8 species), monocots for 33.33% (5 species), and Gymnosperms for 6.67% (1 species). The highest seed density recorded was 1291.67 seeds/m 2 for Sorghum halepense in Pine forest, followed by Cassia tora with 638.89 seeds/m 2 in Deodar forest. The lowest density recorded was 13.89 seeds/m 2 for Scindapsus officinalis , Solanum xanthocarpum (in Sal forest), and Cedrus deodara in Deodar forest. In the Sal forest, the highest seed density recorded was 166.67 seeds/m 2 for Ageratum houstonianum . Kruskal-Wallis with Dunn Post-hoc The Kruskal-Wallis test showed a significant variation in soil seed bank germination among the three forest types, and Dunn’s post-hoc analysis clarified the pairwise contrasts. The median and IQR values with significance are presented in Table 1 . The data was highly skewed, leading to the use of Median with IQR. The results showed that Pine forest had the highest germination potential, forming a distinct significance group ("b"), while Deodar and Sal forests clustered together in group "a," indicating no statistical difference between them. This suggests that the soil seed bank under Pine stands supports significantly greater germination potential compared to Deodar and Sal forests, which maintain relatively lower and similar seed bank germination levels. Table 1 Median and IQR with significance Forest Median_IQR Significance 1 Deodar 0 (0–1) a 2 Pine 1 (0–4) b 3 Sal 0 (0–0) a The violin plot (Fig. 2 ) indicates that Pine forest soils produced the highest number of germinants per tray, with some replicates exceeding 20 individuals, placing it in a distinct significance group. The wider distribution and higher upper tail in the Pine category suggest a much richer and more responsive soil seed bank compared to Deodar and Sal stands, which show relatively sparse and similar germination densities. Diversity Indices The diversity metrics indicate significant differences in soil seed bank composition among the three forest types. Sal forest has the highest species richness (10 taxa), Shannon diversity (2.07), Simpson index (0.85), and evenness (0.90), indicating a well-balanced community. Pine forest has slightly lower richness (9) and moderate diversity (Shannon 1.62; Simpson 0.74), with a slightly skewed community. Deodar forest has the lowest richness (8), Shannon (1.35), and Simpson (0.67), with reduced evenness (0.65), indicating a less diverse and uneven seed bank dominated by a few species. Overall, Sal forest maintains the most diverse and balanced soil seed bank, Pine forest is intermediate, and Deodar forest has the least diverse and even assemblage (Fig. 3 ). Similarity among soil seed banks The Jaccard’s similarity coefficient values in Table 2 show that Sal and Pine forests have the highest similarity in soil seed bank, followed by Pine and Deodar forests. The lowest similarity was found between Sal and Deodar forests. Table 2 Jaccard’s Similarity Coefficient of soil seed banks Sal Pine Deodar Sal 1 0.583 0.286 Pine 0.583 1 0.417 Deodar 0.286 0.417 1 Non-metric Multidimensional Scaling (NMDS) The NMDS ordination (Bray–Curtis) (Fig. 4 ) shows clear compositional differences in the soil seed bank among Deodar, Pine, and Sal forests, with a low stress indicating a good fit of the ordination. Points representing the three forest types are spatially separated, indicating distinct community assemblages within each forest. Deodar plots (red circles) cluster towards positive NMDS1 values, closely associated with species vectors such as Lindernia crustacea and Lindernia ciliata . Pine plots (green triangles) are located in the lower central portion of the ordination, showing stronger alignment with species like Artemisia vulgaris and Sorghum halepense , suggesting these taxa are characteristic of Pine forest seed banks. Sal plots (blue squares) group on the negative NMDS1 axis, with associations to Commelina diffusa , Alysicarpus ovalifolius , and Parthenium hysterophorus . Species vectors extending outward indicate the strength and direction of each taxon’s contribution to community structure, with longer vectors (e.g., Artemisia vulgaris, Lindernia crustacea ) having a greater influence on ordination patterns. Overall, the ordination confirms that forest type strongly influences seed bank composition, with Pine, Sal, and Deodar supporting floristically distinct assemblages and characteristic indicator species. Plate 1- Soil sample collection and seed germination Discussion This study identified 15 plant species from 10 families, with herbs being the dominant growth form (66.67%), followed by climbers (13.33%). The Asteraceae family was the most dominant (20%), followed by Fabaceae (13.33%). The highest seed density recorded was 1291.67 seeds/m2, while the lowest was 13.89 seeds/m2. Bekele et al. ( 2022 ) found 56 species from 27 families, with herbs (78.6%) and the Asteraceae family (20%) being the most dominant, consistent with the current study. Birhanu et al. ( 2022 ) observed 2133 species from 27 families, with Asteraceae as the most diverse family, followed by Solanaceae. They also found herbs to be the dominant growth form, with a seedling density of 4637 seeds/m 2 . Phartyal et al. ( 2023 ) studied the Tungnath Alpine Meadow and identified 13 species (10 dicots, 3 monocots) with an average seed density of 2141 seeds/m 2 at a soil depth of 10 cm. In the present study, 15 species (1 unidentified) were observed, including 8 dicots, 5 monocots, and 1 Gymnosperm. Mittal et al. ( 2021 ) conducted a comparative study in Kumaon Central Himalaya Forests, noting the highest emergence of grasses in Pine forests and the lowest in Sal forest soils, while forbs had maximum emergence in Deodar-Oak forest soils. The greater species richness observed in the Sal Forest (10) may be due to its more open canopy structure, allowing more light to penetrate and supporting a wider variety of understory species. Baral and Ghimire ( 2020 ) also found higher seedling density in semi-open Shorea robusta forests compared to closed-canopy stands in the Terai Sal forest in Western Nepal. In contrast, the Pine Forest, while dominated by fewer species, had a significantly higher density of Sorghum halepense , suggesting potential monoculture formation within the soil seed bank. Several studies have shown a positive impact of forest fires on maintaining plant biodiversity in Pine forests (Kuuluvainen and Rouvinen, 2000 ; Gorshkov and Stavrova, 2002 ; Marozas et al. 2007 ; Konsam et al. 2020 ). The Mixed Deodar Forest had lower diversity and evenness, likely due to its dense canopy cover and thick litter layer, typical of coniferous forests. Previous research suggests that soil seed banks in coniferous forests tend to have lower species richness and density compared to deciduous forests (Granström, 1988; Leckie et al. 2000 ). Interestingly, the dominant species in each forest type consistently had higher Simpson’s and Shannon-Wiener diversity indices, indicating an uneven species distribution with a few species dominating. This trend was also observed in studies by Grime 1988, Dougall & Dodd 1997 , and You et al. 2025 , showing that species dominance in subtropical forest seed banks can lead to reduced community evenness and diversity. The current study revealed variations in seed density and diversity indices in the seed soil banks of different forests. Similar findings were reported by Zoghloul 2008, Heydari et al. 2013 , Madawala et al. 2016 , Tessema et al . 2017, Shiferaw et al. 2018 , Mmusi et al. 2021 , Durate et al . 2022, and Zhu et al. 2023 in their respective studies on various forest ecosystems. Factors such as seed size and dispersal, viability, longevity, predator activities, dormancy, light availability, and other environmental conditions influence the density and diversity of soil seed banks (Leck et al. 1989 ; Whitmore, 1991 ; Teketay and Granstrom, 1995; Teketay 1998 ; Matus et al. 2005 ; Chen et al. 2019 ; Benvenuti and Mazzoncini, 2021 ; Zhao et al. 2022 ). In the current study, the highest Jaccard's similarity coefficient was found between Sal and Pine forests (0.583), with Pine and Deodar forests showing intermediate similarity (0.417) and Sal and Deodar forests exhibiting the lowest similarity (0.286) in terms of soil seed banks. Bekele et al. ( 2022 ) reported a higher similarity in species composition between grassland and shrub land (0.52), followed by forest and shrub land (0.47), while the forest and bare land had the lowest similarity coefficient. Birhanu et al. ( 2022 ) noted a very low similarity (0.11) between seedlings emerging from the soil seed bank and the above-ground vegetation. Godefroid et al. ( 2006 ) observed a decrease in similarity between vegetation and the seed bank in managed temperate forest ecosystems, with pine to oak plantations showing a decreasing trend. The widespread presence of invasive species like Parthenium hysterophorus in all three forest types indicates a significant distribution and underscores its potential threat to the regeneration of native species. Bajwa et al. ( 2016 ) also observed similar patterns, noting the aggressive and competitive nature of Parthenium hysterophorus in various forest ecosystems, where it can outcompete native flora, disrupt ecological balance, and reduce biodiversity. The introduction of invasive plant species has a detrimental impact on biodiversity and ecosystem functioning, contributing to global ecological and economic challenges (Fourie, 2012 ; Gioria et al. 2012 ; Shiferaw et al. 2018 ). Conclusions and recommendations The study revealed significant variations in the composition of soil seed banks among Sal, Pine, and Deodar forests in the Himalayan foothills. Sal forests exhibited the highest species richness and balanced diversity, indicating strong regenerative potential. In contrast, pine forests had the highest seed density but the lowest diversity, while Deodar forests showed low diversity due to their dense canopy and litter accumulation. The presence of the invasive species Parthenium hysterophorus poses a threat to native regeneration. To address this, forest management strategies should focus on controlling invasive species, preserving soil seed banks for regeneration, and implementing site-specific restoration techniques. Incorporating seed bank research into long-term monitoring efforts can enhance forest resilience and biodiversity conservation in the area. Declarations Funding declaration- No funding was received for the present work. Consent to Publish declaration- not applicable. Ethics declaration : Generative AI tool (Chat GPT) was used for language improvement. Clinical trial number : not applicable. Competing Interest : There is no competing interest. Consent to Participate declaration : not applicable. Author Contribution Pema Choki Lepcha did filed and lab work (the collection of samples and observation of seedling grwoth) under the supervision of Prabhakar Manori. Primary manuscript written work, Statistical analysis and preparation of figures were done by Prabhakar Manori and Manish Kumar. Manuscript formatting and arrangement of text were collectively done by Viskaspal Singh Rawat, Sandhya Goswami and Anil Kumar Uniyal. All authors reviewed and gave their suggestions for finalizing the manuscript. Acknowledgement The authors would like to express their sincere gratitude to the Department of Forestry, Dolphin PG Institute of Biomedical and Natural Sciences, for providing the necessary materials and laboratory facilities required to carry out this study. The authors also acknowledge the support and cooperation of all staff members who contributed to the successful completion of this work. This research did not receive any specific grant or funding from public, commercial, or not-for-profit agencies. Data Availability The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request. References Bajwa AA, Chauhan BS, Farooq M, Adkins SW. What do we really know about alien plant invasion? A review of the invasion mechanism of one of the world's worst weeds. Planta. 2016;244:39–57. https://doi.org/10.1007/s00425-016-2510-x . Baker HG. 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Plate Plate 1 is available in the Supplementary Files section. Additional Declarations No competing interests reported. Supplementary Files Plate1.png Plate 1- Soil sample collection and seed germination Cite Share Download PDF Status: Posted Version 1 posted 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. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-7984952","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":549414066,"identity":"032e261b-41bd-4102-bf05-87cebb41a60c","order_by":0,"name":"Pema Choki Lepcha","email":"","orcid":"","institution":"Dolphin (PG) Institute of Bio Medical and Natural Science","correspondingAuthor":false,"prefix":"","firstName":"Pema","middleName":"Choki","lastName":"Lepcha","suffix":""},{"id":549414067,"identity":"e3452e1b-8015-4728-99da-affe2b77fff8","order_by":1,"name":"Prabhakar Manori","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA0UlEQVRIiWNgGAWjYPACCTk29gYgbWBBvBZjfp4DIC0SxFuTOHNGAlgvYaXmYoefPfi4xyJxw83nVzf8KJBg4G/vTsCrxXJ2mrnhjGcSxhtu55Td7AE6TOLM2Q14tRjcTjCT5jkgIQvUknaDB6jFQCKXkJb0byAtjBtunkm7+Yc4LTlgWxRnzmA/dpsoWyxn55RJzjgACuQcttsyBhI8BP1iLp2+TeLDgTpgVB5/dvPNHxs5/vZeAg5DMHnAbB68ytG0sD8gqHoUjIJRMApGJgAACuBHAONDuBcAAAAASUVORK5CYII=","orcid":"","institution":"Dolphin (PG) Institute of Bio Medical and Natural Science","correspondingAuthor":true,"prefix":"","firstName":"Prabhakar","middleName":"","lastName":"Manori","suffix":""},{"id":549414068,"identity":"d4e1a461-a327-4040-8e23-cd04ea5a7087","order_by":2,"name":"Manish Kumar","email":"","orcid":"","institution":"Dolphin (PG) Institute of Bio Medical and Natural 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1","display":"","copyAsset":false,"role":"figure","size":113948,"visible":true,"origin":"","legend":"\u003cp\u003eLocation map ofViolin plot of Kruskal-Wallis with Dunn’s Post-hoc teststudy site.\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7984952/v1/580d0a8e70e71a59b16fe271.jpeg"},{"id":96760490,"identity":"93163787-fab2-4b10-ad23-0dccfb301edf","added_by":"auto","created_at":"2025-11-25 19:00:07","extension":"jpeg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":114426,"visible":true,"origin":"","legend":"\u003cp\u003eViolin plot of Kruskal-Wallis with Dunn’s Post-hoc test\u003c/p\u003e","description":"","filename":"floatimage2.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7984952/v1/b92f77f64a7498003fae318d.jpeg"},{"id":96914552,"identity":"39807fb8-371f-48e0-8ddb-eac6e8772698","added_by":"auto","created_at":"2025-11-27 14:06:03","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":116721,"visible":true,"origin":"","legend":"\u003cp\u003eHeat Map of Diversity Indices\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-7984952/v1/721879a0c5c45904a3bc5c5a.png"},{"id":96760494,"identity":"2f945787-84e8-4c9e-977f-f6835c87e162","added_by":"auto","created_at":"2025-11-25 19:00:07","extension":"jpeg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":147639,"visible":true,"origin":"","legend":"\u003cp\u003eNMDS Ordination with Species Vector (Bray-Curtis)\u003c/p\u003e","description":"","filename":"floatimage4.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7984952/v1/e710c643df482c2b29db223d.jpeg"},{"id":97694537,"identity":"6916530e-4a25-40e2-b389-7bbb779e96f9","added_by":"auto","created_at":"2025-12-08 11:24:22","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1126121,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7984952/v1/ef6c620b-74eb-4928-b073-400a59c012ad.pdf"},{"id":96760489,"identity":"6b5a3e6f-9afe-49ec-a94c-e87690bd7618","added_by":"auto","created_at":"2025-11-25 19:00:07","extension":"png","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":1513296,"visible":true,"origin":"","legend":"\u003cp\u003ePlate 1- Soil sample collection and seed germination\u003c/p\u003e","description":"","filename":"Plate1.png","url":"https://assets-eu.researchsquare.com/files/rs-7984952/v1/5a6ba45e10a8909866b22718.png"}],"financialInterests":"No competing interests reported.","formattedTitle":"Assessment of Soil Seed Bank Dynamics and Regeneration Potential Across Three Different Forest Types in the Himalayan Foothills","fulltext":[{"header":"Introduction","content":"\u003cp\u003eBased on the Global Forest Watch report (2024), Uttarakhand experienced a loss of 30 hectares of humid primary forest, accounting for 0.13% of its total tree cover loss during the same period. The total area of humid primary forest in Uttarakhand decreased by 0.32% in this timeframe, indicating a concerning rate of forest depletion in the region. Soil Seed Bank Analysis is a valuable tool for comprehending future forest trends and informing conservation strategies.\u003c/p\u003e\u003cp\u003eThe soil seed bank serves as a reservoir of viable seeds within the soil, playing a crucial role in plant population dynamics, biodiversity preservation, and ecosystem resilience. It is essential for natural regeneration, species succession, and ecological restoration efforts (Thompson and Grime, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e1979\u003c/span\u003e; Baskin and Baskin, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Understanding the composition and dynamics of soil seed banks is particularly vital in forest ecosystems, where vegetation recovery post-disturbances relies heavily on the existing seed population.\u003c/p\u003e\u003cp\u003eRoberts (\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e1981\u003c/span\u003e) defines the soil seed bank as the collection of viable seeds present in the soil. Baker (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e1989\u003c/span\u003e) further elaborates that this reservoir consists of seeds that have not yet germinated but have the potential to replace adult plants lost due to natural causes, diseases, disturbances, or consumption by animals, including humans. All viable seeds within the soil or mixed with soil debris contribute to the soil seed bank (Simpson et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e1989\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eLarge quantities of seeds can remain dormant in the soil for extended periods, and can sprout when conditions are favorable (Warr et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e1993\u003c/span\u003e). Soil seed banks play a crucial role in understanding vegetation history, as the composition of vegetation in terms of plant species is influenced by seed production, dispersal, seed longevity, and soil depth (V. Anju et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Seed banks are key determinants of future vegetation, particularly following a disturbance (Warr et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e1993\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eDehradun, situated in the Doon Valley at the foothills of the western Himalayas, is known for its remarkable ecological diversity. It is located at the meeting point of the Shivalik range, the outer Himalayas, and the Indo-Gangetic plains, creating diverse climatic and soil conditions that support a variety of forest types (Champion and Seth, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e1968\u003c/span\u003e). The region encompasses moist deciduous forests dominated by sal (\u003cem\u003eShorea robusta\u003c/em\u003e), pine (\u003cem\u003ePinus roxburghii\u003c/em\u003e) forests, mixed broadleaf forests, and riverine forests, among others. These diverse forest ecosystems exhibit differences in microclimatic conditions, human impact, disturbance patterns, and vegetation structure, all of which impact the composition and abundance of their soil seed banks.\u003c/p\u003e\u003cp\u003eWhile regeneration studies have been conducted by various researchers, there is limited information available on the diversity of soil seed banks in different forest types in Uttarakhand. The present study aims to address this gap by investigating the soil seed bank diversity in Sal, Pine, and mixed Deodar-dominated forest stands in the Himalayan foothills.\u003c/p\u003e"},{"header":"Methodology","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003eStudy Area\u003c/h2\u003e\u003cp\u003eThis study took place in Dehradun district, situated between the Himalayas to the north and the Shivalik Hills to the south, with geographical coordinates of 30.3165o N latitude and 78.0322o E longitude. The major forest types in the area include Sal forests at lower elevations, Pine forests at mid elevations, and Oak and Rhododendron at higher elevations.\u003c/p\u003e\u003cp\u003eSite I: \u003cem\u003eShorea robusta\u003c/em\u003e (Sal) Forest: Located at latitude 30.37752 N and longitude 77.931215 E, soil samples were collected from the Sal Forest in Manduwala, part of the Dehradun Forest Division.\u003c/p\u003e\u003cp\u003eSite II: \u003cem\u003ePinus roxburghii\u003c/em\u003e (Chir Pine) Forest: Coordinates at latitude 30.453615 N and longitude 77.944039 E. Soil samples were collected from the Pine Forest in Koti, within the Chakrata Forest Division.\u003c/p\u003e\u003cp\u003eSite III: Mixed \u003cem\u003eCedrus deodara\u003c/em\u003e (Deodar) Forest: Coordinates at latitude 30.4449540 N and longitude 78.076609 E. This study site is located in Mussoorie, which is under the jurisdiction of the Mussoorie Forest Division.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eSoil Sampling\u003c/h3\u003e\n\u003cp\u003eIn this study, 1 m \u0026times; 1 m quadrats were randomly placed across the study site to evaluate species distribution and abundance. A distance of 50 meters was maintained between consecutive quadrats to ensure spatial independence and reduce sampling bias. A total of 10 quadrats were established at each study site to provide a representative sample for statistical analysis. Two soil samples were collected from each quadrat\u0026mdash;one from a depth of 0\u0026ndash;5 cm and the other from 5\u0026ndash;10 cm. Therefore, 20 soil samples were collected from each site, resulting in a total of 60 samples across all three sites. In the laboratory, the two depth samples from each plot were combined and thoroughly mixed before being air-dried for 48 hours. Any visible debris, such as stones, twigs, and other extraneous material, was carefully removed during the drying process.\u003c/p\u003e\u003cp\u003eAfter drying and cleaning, the soil samples were placed in labelled vegetation trays and kept in a controlled setting without existing vegetation to prevent external interference with natural seed germination. No external soil enrichments, such as fertilizers or farmyard manure (FYM), were added to observe the natural regeneration potential of the seed bank. Irrigation was carried out manually, initially every alternate day, and later daily as the summer season progressed and evaporation rates increased to maintain adequate soil moisture.\u003c/p\u003e\n\u003ch3\u003eData Collection\u003c/h3\u003e\n\u003cp\u003eSeedling emergence was regularly monitored, and the number of germinations from each tray was recorded. Seedlings were uprooted by hand as they reached a stage where species identification was possible, to minimize disturbance and allow other seeds to continue germinating.\u003c/p\u003e\u003cp\u003eSpecies were identified based on morphological traits such as size, shape, color, and surface texture of leaves and stems. The weekly recording of the number of individuals of each species was conducted over an eight-week period.\u003c/p\u003e\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\u003ch2\u003eData Analysis\u003c/h2\u003e\u003cp\u003eSoil seed bank germination was compared among different forest types using the Kruskal\u0026ndash;Wallis test (Kruskal \u0026amp; Wallis, 1952), followed by Dunn\u0026rsquo;s post-hoc pairwise comparisons (Dunn\u0026rsquo;s, 1964). Community structure was evaluated using diversity indices (species richness, Shannon, Simpson, and evenness), and compositional differences were visualized with non-metric multidimensional scaling (NMDS) based on Bray\u0026ndash;Curtis dissimilarity. All statistical analyses were performed using R software (Version 4.5.0) with appropriate packages for non-parametric testing, diversity metrics, and ordination. Seed density was calculated as the number of seedlings (viable seeds) per unit area (expressed as seeds/m\u0026sup2;) based on the total number of seedlings that emerged from each soil sample.\u003c/p\u003e\u003c/div\u003e"},{"header":"Result","content":"\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\u003ch2\u003eSoil seed bank density and morphology of plants\u003c/h2\u003e\u003cp\u003eA total of 371 individuals emerged from the soil seed banks of three different forest types, consisting of a total of 15 species, namely \u003cem\u003eAgeratum houstonianum, Sorghum halepense, Dioscorea bulbifera, Commelina diffusa, Parthenium hysterophorus, Cassia tora, Alysicarpus ovalifolius, Commelina benghalensis, Scindapsus officinalis, Solanum xanthocarpum, Artemisia vulgaris, Lindenbergia indica, Oxalis exilis, Cedrus deodara\u003c/em\u003e, and one unidentified species representing 10 families. The Asteraceae family was the most dominant, with 3 species (20%), followed by Fabaceae and Commelinaceae with 2 species each (13.33%). The rest of the families had only one species each (6.67%). Herbs were the dominant growth form, comprising 66.67% (10 species), followed by climbers at 13.33% (2 species). Dicots accounted for 53.33% (8 species), monocots for 33.33% (5 species), and Gymnosperms for 6.67% (1 species).\u003c/p\u003e\u003cp\u003eThe highest seed density recorded was 1291.67 seeds/m\u003csup\u003e2\u003c/sup\u003e for \u003cem\u003eSorghum halepense\u003c/em\u003e in Pine forest, followed by \u003cem\u003eCassia tora\u003c/em\u003e with 638.89 seeds/m\u003csup\u003e2\u003c/sup\u003e in Deodar forest. The lowest density recorded was 13.89 seeds/m\u003csup\u003e2\u003c/sup\u003e for \u003cem\u003eScindapsus officinalis\u003c/em\u003e, \u003cem\u003eSolanum xanthocarpum\u003c/em\u003e (in Sal forest), and \u003cem\u003eCedrus deodara\u003c/em\u003e in Deodar forest. In the Sal forest, the highest seed density recorded was 166.67 seeds/m\u003csup\u003e2\u003c/sup\u003e for \u003cem\u003eAgeratum houstonianum\u003c/em\u003e.\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eKruskal-Wallis with Dunn Post-hoc\u003c/h3\u003e\n\u003cp\u003eThe Kruskal-Wallis test showed a significant variation in soil seed bank germination among the three forest types, and Dunn\u0026rsquo;s post-hoc analysis clarified the pairwise contrasts. The median and IQR values with significance are presented in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. The data was highly skewed, leading to the use of Median with IQR. The results showed that Pine forest had the highest germination potential, forming a distinct significance group (\"b\"), while Deodar and Sal forests clustered together in group \"a,\" indicating no statistical difference between them. This suggests that the soil seed bank under Pine stands supports significantly greater germination potential compared to Deodar and Sal forests, which maintain relatively lower and similar seed bank germination levels.\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\u003eMedian and IQR with significance\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"1\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eForest Median_IQR Significance\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e1 Deodar 0 (0\u0026ndash;1) a\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e2 Pine 1 (0\u0026ndash;4) b\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e3 Sal 0 (0\u0026ndash;0) a\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eThe violin plot (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e) indicates that Pine forest soils produced the highest number of germinants per tray, with some replicates exceeding 20 individuals, placing it in a distinct significance group. The wider distribution and higher upper tail in the Pine category suggest a much richer and more responsive soil seed bank compared to Deodar and Sal stands, which show relatively sparse and similar germination densities.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\n\u003ch3\u003eDiversity Indices\u003c/h3\u003e\n\u003cp\u003eThe diversity metrics indicate significant differences in soil seed bank composition among the three forest types. Sal forest has the highest species richness (10 taxa), Shannon diversity (2.07), Simpson index (0.85), and evenness (0.90), indicating a well-balanced community. Pine forest has slightly lower richness (9) and moderate diversity (Shannon 1.62; Simpson 0.74), with a slightly skewed community. Deodar forest has the lowest richness (8), Shannon (1.35), and Simpson (0.67), with reduced evenness (0.65), indicating a less diverse and uneven seed bank dominated by a few species. Overall, Sal forest maintains the most diverse and balanced soil seed bank, Pine forest is intermediate, and Deodar forest has the least diverse and even assemblage (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\u003ch2\u003eSimilarity among soil seed banks\u003c/h2\u003e\u003cp\u003eThe Jaccard\u0026rsquo;s similarity coefficient values in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e show that Sal and Pine forests have the highest similarity in soil seed bank, followed by Pine and Deodar forests. The lowest similarity was found between Sal and Deodar forests.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eJaccard\u0026rsquo;s Similarity Coefficient of soil seed banks\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"4\"\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\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSal\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003ePine\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eDeodar\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSal\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.583\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.286\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.583\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.417\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDeodar\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.286\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.417\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\u003ch2\u003eNon-metric Multidimensional Scaling (NMDS)\u003c/h2\u003e\u003cp\u003eThe NMDS ordination (Bray\u0026ndash;Curtis) (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e) shows clear compositional differences in the soil seed bank among Deodar, Pine, and Sal forests, with a low stress indicating a good fit of the ordination. Points representing the three forest types are spatially separated, indicating distinct community assemblages within each forest. Deodar plots (red circles) cluster towards positive NMDS1 values, closely associated with species vectors such as \u003cem\u003eLindernia crustacea\u003c/em\u003e and \u003cem\u003eLindernia ciliata\u003c/em\u003e. Pine plots (green triangles) are located in the lower central portion of the ordination, showing stronger alignment with species like \u003cem\u003eArtemisia vulgaris\u003c/em\u003e and \u003cem\u003eSorghum halepense\u003c/em\u003e, suggesting these taxa are characteristic of Pine forest seed banks.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eSal plots (blue squares) group on the negative NMDS1 axis, with associations to \u003cem\u003eCommelina diffusa\u003c/em\u003e, \u003cem\u003eAlysicarpus ovalifolius\u003c/em\u003e, and \u003cem\u003eParthenium hysterophorus\u003c/em\u003e. Species vectors extending outward indicate the strength and direction of each taxon\u0026rsquo;s contribution to community structure, with longer vectors (e.g., \u003cem\u003eArtemisia vulgaris, Lindernia crustacea\u003c/em\u003e) having a greater influence on ordination patterns. Overall, the ordination confirms that forest type strongly influences seed bank composition, with Pine, Sal, and Deodar supporting floristically distinct assemblages and characteristic indicator species.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003ePlate 1- Soil sample collection and seed germination\u003c/p\u003e\u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eThis study identified 15 plant species from 10 families, with herbs being the dominant growth form (66.67%), followed by climbers (13.33%). The Asteraceae family was the most dominant (20%), followed by Fabaceae (13.33%). The highest seed density recorded was 1291.67 seeds/m2, while the lowest was 13.89 seeds/m2. Bekele et al. (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) found 56 species from 27 families, with herbs (78.6%) and the Asteraceae family (20%) being the most dominant, consistent with the current study. Birhanu et al. (\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) observed 2133 species from 27 families, with Asteraceae as the most diverse family, followed by Solanaceae. They also found herbs to be the dominant growth form, with a seedling density of 4637 seeds/m\u003csup\u003e2\u003c/sup\u003e. Phartyal et al. (\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2023\u003c/span\u003e) studied the Tungnath Alpine Meadow and identified 13 species (10 dicots, 3 monocots) with an average seed density of 2141 seeds/m\u003csup\u003e2\u003c/sup\u003e at a soil depth of 10 cm. In the present study, 15 species (1 unidentified) were observed, including 8 dicots, 5 monocots, and 1 Gymnosperm. Mittal et al. (\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) conducted a comparative study in Kumaon Central Himalaya Forests, noting the highest emergence of grasses in Pine forests and the lowest in Sal forest soils, while forbs had maximum emergence in Deodar-Oak forest soils.\u003c/p\u003e\u003cp\u003eThe greater species richness observed in the Sal Forest (10) may be due to its more open canopy structure, allowing more light to penetrate and supporting a wider variety of understory species. Baral and Ghimire (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) also found higher seedling density in semi-open \u003cem\u003eShorea robusta\u003c/em\u003e forests compared to closed-canopy stands in the Terai Sal forest in Western Nepal. In contrast, the Pine Forest, while dominated by fewer species, had a significantly higher density of \u003cem\u003eSorghum halepense\u003c/em\u003e, suggesting potential monoculture formation within the soil seed bank. Several studies have shown a positive impact of forest fires on maintaining plant biodiversity in Pine forests (Kuuluvainen and Rouvinen, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2000\u003c/span\u003e; Gorshkov and Stavrova, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Marozas et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Konsam et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe Mixed Deodar Forest had lower diversity and evenness, likely due to its dense canopy cover and thick litter layer, typical of coniferous forests. Previous research suggests that soil seed banks in coniferous forests tend to have lower species richness and density compared to deciduous forests (Granstr\u0026ouml;m, 1988; Leckie et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2000\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eInterestingly, the dominant species in each forest type consistently had higher Simpson\u0026rsquo;s and Shannon-Wiener diversity indices, indicating an uneven species distribution with a few species dominating. This trend was also observed in studies by Grime 1988, Dougall \u0026amp; Dodd \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e1997\u003c/span\u003e, and You et al. \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2025\u003c/span\u003e, showing that species dominance in subtropical forest seed banks can lead to reduced community evenness and diversity.\u003c/p\u003e\u003cp\u003eThe current study revealed variations in seed density and diversity indices in the seed soil banks of different forests. Similar findings were reported by Zoghloul 2008, Heydari et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2013\u003c/span\u003e, Madawala et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2016\u003c/span\u003e, Tessema \u003cem\u003eet al\u003c/em\u003e. 2017, Shiferaw et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2018\u003c/span\u003e, Mmusi et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2021\u003c/span\u003e, Durate \u003cem\u003eet al\u003c/em\u003e. 2022, and Zhu et al. \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2023\u003c/span\u003e in their respective studies on various forest ecosystems. Factors such as seed size and dispersal, viability, longevity, predator activities, dormancy, light availability, and other environmental conditions influence the density and diversity of soil seed banks (Leck et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e1989\u003c/span\u003e; Whitmore, \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e1991\u003c/span\u003e; Teketay and Granstrom, 1995; Teketay \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e1998\u003c/span\u003e; Matus et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Chen et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Benvenuti and Mazzoncini, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Zhao et al. \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn the current study, the highest Jaccard's similarity coefficient was found between Sal and Pine forests (0.583), with Pine and Deodar forests showing intermediate similarity (0.417) and Sal and Deodar forests exhibiting the lowest similarity (0.286) in terms of soil seed banks. Bekele et al. (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) reported a higher similarity in species composition between grassland and shrub land (0.52), followed by forest and shrub land (0.47), while the forest and bare land had the lowest similarity coefficient. Birhanu et al. (\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) noted a very low similarity (0.11) between seedlings emerging from the soil seed bank and the above-ground vegetation. Godefroid et al. (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2006\u003c/span\u003e) observed a decrease in similarity between vegetation and the seed bank in managed temperate forest ecosystems, with pine to oak plantations showing a decreasing trend.\u003c/p\u003e\u003cp\u003eThe widespread presence of invasive species like \u003cem\u003eParthenium hysterophorus\u003c/em\u003e in all three forest types indicates a significant distribution and underscores its potential threat to the regeneration of native species. Bajwa et al. (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) also observed similar patterns, noting the aggressive and competitive nature of \u003cem\u003eParthenium hysterophorus\u003c/em\u003e in various forest ecosystems, where it can outcompete native flora, disrupt ecological balance, and reduce biodiversity. The introduction of invasive plant species has a detrimental impact on biodiversity and ecosystem functioning, contributing to global ecological and economic challenges (Fourie, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Gioria et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Shiferaw et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e"},{"header":"Conclusions and recommendations","content":"\u003cp\u003eThe study revealed significant variations in the composition of soil seed banks among Sal, Pine, and Deodar forests in the Himalayan foothills. Sal forests exhibited the highest species richness and balanced diversity, indicating strong regenerative potential. In contrast, pine forests had the highest seed density but the lowest diversity, while Deodar forests showed low diversity due to their dense canopy and litter accumulation. The presence of the invasive species \u003cem\u003eParthenium hysterophorus\u003c/em\u003e poses a threat to native regeneration. To address this, forest management strategies should focus on controlling invasive species, preserving soil seed banks for regeneration, and implementing site-specific restoration techniques. Incorporating seed bank research into long-term monitoring efforts can enhance forest resilience and biodiversity conservation in the area.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eFunding declaration-\u003c/strong\u003e No funding was received for the present work.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to Publish declaration-\u003c/strong\u003e not applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics declaration\u003c/strong\u003e: Generative AI tool (Chat GPT) was used for language improvement.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eClinical trial number\u003c/strong\u003e: not applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interest\u003c/strong\u003e: There is no competing interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to Participate declaration\u003c/strong\u003e: not applicable.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003ePema Choki Lepcha did filed and lab work (the collection of samples and observation of seedling grwoth) under the supervision of Prabhakar Manori. Primary manuscript written work, Statistical analysis and preparation of figures were done by Prabhakar Manori and Manish Kumar. Manuscript formatting and arrangement of text were collectively done by Viskaspal Singh Rawat, Sandhya Goswami and Anil Kumar Uniyal. All authors reviewed and gave their suggestions for finalizing the manuscript.\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eThe authors would like to express their sincere gratitude to the Department of Forestry, Dolphin PG Institute of Biomedical and Natural Sciences, for providing the necessary materials and laboratory facilities required to carry out this study. The authors also acknowledge the support and cooperation of all staff members who contributed to the successful completion of this work. This research did not receive any specific grant or funding from public, commercial, or not-for-profit agencies.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eThe datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eBajwa AA, Chauhan BS, Farooq M, Adkins SW. What do we really know about alien plant invasion? A review of the invasion mechanism of one of the world's worst weeds. Planta. 2016;244:39\u0026ndash;57. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s00425-016-2510-x\u003c/span\u003e\u003cspan address=\"10.1007/s00425-016-2510-x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBaker HG. (1989). 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Afr J Environ Sci Technol. 2008;2(7):172\u0026ndash;84. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.5897/AJEST.9000036\u003c/span\u003e\u003cspan address=\"10.5897/AJEST.9000036\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhao YT, Wang GD, Zhao ML, Wang M, Jiang M. Direct and indirect effects of soil salinization on soil seed banks in salinizing wetlands in the Songnen Plain, China. Sci Total Environ. 2022;819:152035. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.scitotenv.2021.152035\u003c/span\u003e\u003cspan address=\"10.1016/j.scitotenv.2021.152035\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhu T, Fang Q, Jia L, Zou Y, Wang X, Qu C, Yu J, Yang J. Diversity of soil seed bank and influencing factors in the nascent wetland of the Yellow River Delta. Front Plant Sci. 2023;14:1249139. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fpls.2023.1249139\u003c/span\u003e\u003cspan address=\"10.3389/fpls.2023.1249139\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"},{"header":"Plate ","content":"\u003cp\u003ePlate 1 is available in the Supplementary Files section.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Biodiversity, Conservation, Regeneration, Soil Seed Bank, Species Diversity","lastPublishedDoi":"10.21203/rs.3.rs-7984952/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7984952/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThis study examines the soil seed bank potential in three forest types in Dehradun district: Sal, Pine, and Mixed Deodar. Soil samples were collected and monitored for seedling emergence to assess their roles in natural regeneration and biodiversity conservation. Species composition, density, diversity indices, and inter-forest similarities were analyzed using statistical and ordination techniques.\u003c/p\u003e\u003cp\u003eA total of 371 individuals from 15 species across 10 families were observed, with herbs being the dominant growth form (66.67%) and Asteraceae as the most represented family (20%). Seed density varied among forest types, with Pine forest having the highest germination potential and maximum seed density of 1291.67 seeds/m\u0026sup2; for \u003cem\u003eSorghum halepense\u003c/em\u003e. Deodar forest had an intermediate density dominated by \u003cem\u003eCassia tora\u003c/em\u003e (638.89 seeds/m\u0026sup2;), while Sal forest had the lowest density but supported the highest species richness (10 species) and diversity indices (Shannon\u0026thinsp;=\u0026thinsp;2.07, Simpson\u0026thinsp;=\u0026thinsp;0.85). Jaccard\u0026rsquo;s similarity coefficient indicated greater similarity between Sal and Pine forests (0.583), with Deodar showing the least overlap with Sal (0.286). NMDS ordination confirmed distinct community assemblages among the three forest types, with characteristic indicator species identified for each.\u003c/p\u003e\u003cp\u003eThese results demonstrate significant variation in soil seed bank composition and regenerative capacity across forest types. Pine forests have dense but less diverse seed banks, mainly dominated by grasses, while Sal forests have balanced and diverse assemblages, supporting broad regeneration potential. Deodar forests exhibit lower diversity due to dense canopy and thick litter layers. The presence of invasive species like \u003cem\u003eParthenium hysterophorus\u003c/em\u003e in all sites highlights ecological risks to native regeneration.\u003c/p\u003e","manuscriptTitle":"Assessment of Soil Seed Bank Dynamics and Regeneration Potential Across Three Different Forest Types in the Himalayan Foothills","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-25 19:00:02","doi":"10.21203/rs.3.rs-7984952/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"4fd4ba02-5c02-4055-83aa-c508a96cd738","owner":[],"postedDate":"November 25th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-12-08T11:23:51+00:00","versionOfRecord":[],"versionCreatedAt":"2025-11-25 19:00:02","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7984952","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7984952","identity":"rs-7984952","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}