Comparative Anatomical and Morphological Study of Gentiana Algida and Gentiana Macrophylla in Mongolia

Comparative Anatomical and Morphological Study of Gentiana Algida and Gentiana Macrophylla in Mongolia | 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 Comparative Anatomical and Morphological Study of Gentiana Algida and Gentiana Macrophylla in Mongolia Dorjsumiya Gotov, Battseren Tsambaa, Almaz Borjigidai, Batzaya Gachmaa, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6236894/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 Two Gentiana species native to Mongolia, Gentiana algida and Gentiana macrophylla , are thoroughly anaturally and morphologically examined in this work. Field research was conducted in several provinces where specimens were collected and studied using scanning electron microscopy (SEM) and light microscopy. As a result, Gentiana macrophylla displayed an oval stem characterized by substantial sclerenchyma and numerous concentric vascular bundles, whereas Gentiana algida presented a cylindrical, hollow stem featuring a single row of small vascular bundles. Both species exhibited dorsoventral leaf morphology; nevertheless, differences in the configuration of chlorenchyma layers and vascular bundles highlighted their distinct adaptive strategies to local environmental conditions. Moreover, SEM analysis of seed form showed that unlike the smaller, lighter seeds of Gentiana macrophylla, Gentiana algida a produces bigger, denser seeds with a crystalline reticulate surface. These results improve our taxonomic knowledge of the genus Gentiana and provide important new perspectives on the adaptive mechanisms enabling life in very mountainous environments. Combining conventional botanical techniques with cutting-edge imaging technologies creates a solid basis for next research targeted at the sustainable use and preservation of these ecologically and medically important species. Gentiana algida Gentiana macrophylla leaf anatomy stem morphology seed morphology scanning electron microscopy Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 1. Introduction With about 400 species dispersed over several ecological zones worldwide, the genus Gentiana L. stands among the most important medicinal plant families in traditional asian medicine (Zhang et al., 2020). Particularly in severe habitats like the Mongolian steppes (Liu et al., 2019), the anatomical and morphological traits of Gentiana species are absolutely vital for their adaptation to varied environmental circumstances. Recent research has shown how vulnerable Gentiana populations are to anthropogenic pressures and climate change, thus their protection becomes a top issue for the preservation of biodiversity (Wang et al., 2021). Developing sensible conservation plans and sustainable farming methods depends on an awareness of the structural adaptations and reproductive biology of Gentiana species, especially with regard to their seed traits and germination patterns (Chen et al., 2022). Furthermore, thorough analysis of stem and leaf morphological traits in Gentiana species offers important new perspectives on their physiological adaptations and possible medicinal qualities, which have long been used in treating different diseases (Kim and Park, 2023). The many chemical ingredients and pharmacological effects of the Gentiana species have attracted much research. Many investigations have shown bioactive substances like gentiopicroside, flavonoids, and iridoid glycosides that add to their therapeutic worth (Hou et al., 2020; Jia et al., 2016). Emphasizing their wide therapeutic uses, these species have shown promise in treating inflammatory illnesses, liver ailments, and metabolic problems (Cao et al., 2021; Choi et al., 2019). Comparative studies of volatile chemicals from many Gentiana species have also shed light on their phytochemical variety and their uses in traditional medicine (Georgieva et al., 2005). The ongoing investigation of Gentiana species supports their conventional and current pharmaceutical applications as well as helps to find new bioactive compounds. Finally, it notes twenty-nine Gentiana species found in the Mongolian Plateau; nine of these species have known folklore benefits for treating digestive, skin, joint problems, sore throat, etc. Six hundred two compounds—including iridoids, triterpenoids, flavonoids, lignans, coumarins, xanthones, alkaloids, fatty acids, amino acids, organic acids, and polysaccharides—have been isolated and identified thanks to phytochemical research. Especially the most researched monomeric compounds are gentiopicroside (75) and swertiamarin (118). Broad spectrum of pharmacological activities is shown by crude extracts of Gentiana, including anti-inflammatory, analgesic, anti-bacterial, antioxidant, anti-tumor, anti-cancer, anti-diabetic, immunomodulatory, hepatoprotective, gastroprotective, neuroprotective, joint and bone protective activities, etc. In vitro and in vivo investigations show these extracts to be not obviously harmful. Clinical studies on Gentiana's medicinal uses, however, remain few (Hongzhen Yu & Batzaya G et al., 2025). The Red List (2014) lists some species of Gentiana as threatened and they are very rare. Gentiana algida Pall. and Gentiana macrophylla Pall. are extensively utilized in traditional medicine. However, climate change and anthropogenic influences have restricted their growing environments and depleted natural resources, necessitating the urgent cultivation of these species. To cultivate these species scientifically, fundamental anatomical and morphological studies are undertaken to ascertain their characteristics, which will subsequently serve as primary material for investigating the mechanisms of their adaptation to environmental influences. The thorough anatomical investigations of Gentiana have revealed significant variation in stem cross-sectional structures and vascular bundle patterns, which helps scientists to understand the adaptive mechanisms of these species in demanding conditions (Chen et al., 2018). Comparative studies of leaf anatomy have also shed fresh light on the evolution of specialized tissues that maximize photosynthetic efficiency and stress tolerance, therefore strengthening our knowledge of the evolutionary divergence inside the genus (Li and Zhao, 2020). The stem of a plant generates leaves on its surface and functions as an organ that facilitates water evaporation, respiration, photosynthesis, nutrition transport from the soil to the leaves via the roots, and provides structural support. Stems are categorized based on their cross-section into round, triangular, and multi-faceted types. Multi-faceted stems encompass various forms, including scaly, winged, oval, and ribbed stems. This characteristic serves as a significant taxonomic criterion that differentiates numerous similar plant species. The internal structure of the stem comprises three fundamental components: sheath tissue, primary tissue, and the main cylinder, which encompasses the conduction bundle and parenchyma cells. The plant transfers nutrients and water from the soil to the leaves in various ways, contingent upon the quantity and configuration of the conduction bundle, while the parenchyma cells influence the vertical and horizontal flexibility of the plant's axis based on their size. The primary tissues of plants for photosynthesis and light synthesis are leaves. The cuticle, which prevents water evaporation, the epidermis, and vertical cells carrying columnar tissue or chloroplasts that enable photosynthesis make up the internal structure of the leaf. While the parenchyma cells help to enable nutrition exchange, the soft tissue serves to retain reserves. The leaf is classified into dorsoventral (upper and lower sides differ), iso-lateral (upper and lower sides are same), homogeneous, and centric based on the arrangement of columnar and fibrous tissues that constitute its anatomy (Esau, 1980). The internal structure of these leaves differs based on the plant species' growing environment and water availability, with numerous studies undertaken on this subject. Therefore, future cultivation depends on closely analysing the internal structure of the leaves of rare plant species under different growth conditions. Studies on anatomy, morphology, and seed measurements are essential to produce Gentiana macrophylla and Gentiana algida . Cross-sectional anatomical studies of Gentiana species provide crucial new understanding of the structural adaptations of stems and leaves, therefore influencing their survival in numerous climatic conditions, including high-altitude areas with extreme temperatures (Chen et al., 2018). Important for water transport efficiency and mechanical support, these studies show variations in sclerenchyma growth and vascular bundle designs, which helps to conserve and sustainably produce medicinal plants (Li & Zhao, 2020). Understanding the structural differences in leaf mesophyll tissue and epidermal structures may help scientists assess photosynthetic efficiency and drought resistance—qualities necessary for the resilience of medicinal plants under pressures from climate change (Kim & Park, 2023). The architecture and morphological characteristics of seed coats examined via scanning electron microscopy (SEM) facilitate species identification and classification, hence enhancing conservation efforts for rare and endangered medicinal plants, such as Gentiana species (Wang et al., 2021). Combining anatomical investigations with ecological and phytochemical analysis might improve the comprehension of active ingredient distribution throughout various plant tissues, thereby bolstering the pharmaceutical applications of Gentiana and other medicinal plants (Zhang et al., 2020). Consequently, we established the following aims for this research: (1) Examine the intricate cellular and tissue architecture of leaves and stems of specific Gentiana species. (2) Examine the seed morphology, dimensions, mass, and surface traits, and assess. (3) Examine the anatomical and morphological distinctions among Gentiana algida and Gentiana macrophylla to identify relationships with environmental adaptations. (4) Establish a scientific foundation for the cultivation, conservation, and sustainable management of these uncommon and endangered Gentiana species. 2. Material and methods Between July 24 and August 9, 2023, we executed field research in select soums of Tuv, Arkhangai, and Khentii provinces within the Khangai and Khentii regions of the Mongolian phytogeographic region, and gathered seeds and herbarium specimens of two plant species (Figure 1, Appendix 1). The herbarium collection of plants was identified by botanists N. Nyambayar and Ts. Battseren of the Botanical Garden and is stored in the Herbarium Fund of the Botanical Garden of the Academy of Sciences (UBA) under the numbers UBA0013196 and UBA0012854 in October 2023 and is open to the public. Anatomical investigations of plant cross-sections depend on meticulous approaches, such as microtome sectioning and staining procedures, to examine tissue structures at cellular and vascular levels (Esau, 1980). Fixation and embedding techniques, including paraffin embedding and fresh sectioning using freehand methods, are essential for maintaining tissue integrity and achieving high-resolution anatomical observations (Pausheva, 1974). Histological staining techniques, such as Safranin-Fast Green and Toluidine Blue staining, facilitate the differentiation of diverse tissue components, allowing researchers to investigate vascular development, sclerenchyma, and epidermal traits (Gahan, 1984). Contemporary techniques, like scanning electron microscopy (SEM) and fluorescence microscopy, enhance classic cross-sectioning approaches by offering intricate three-dimensional perspectives on plant tissue architecture and cell wall structures (Ruzin, 1999). The integration of anatomical techniques with physiological and ecological studies enhances researchers' comprehension of plant responses to environmental stressors, facilitating conservation and agricultural applications (Cutler et al., 2007). Basic methods employed in plant anatomy investigations include fixation preparation, section preparation, clarity, labelling of cells and tissues, and image transfer (Pausheva, 1974; Gamalei, 1984; Shiirevdamba, Tserenkhand, 2014). Enumerate the seeds in thousands or hundreds, place them in little paper bags, then ascertain the weight of one thousand seeds by 3-5 iterations. Utilize a calliper to ascertain the length and width of the seeds, measuring 50-100 seeds across 3-5 repetitions. When measuring the length, obtain the measurement from the navel or the posterior of the seed. To measure the width, obtain the transverse dimension from the seed's abdomen to the back; for thickness, measure transversely from either the back or abdomen. A scanning electron microscope (SEM) is a scientific device that employs electron beams to produce high-resolution photographs of a sample's surface. SEM is employed to analyse the composition and structure of materials. SEM captures images of seeds at magnifications of x130 and x500. 3. Result Our research indicated that the stems of Gentiana macrophylla had an oval morphology, characterized by a thin epidermis comprising one to two cell layers, a robust sclerenchyma layer, uniformly tiny parenchyma cells, and over 30 consistent circular vascular bundles (Figure 2). The cross-section of the Gentiana macrophylla stem has an oval morphology, characterized by a thin epidermis composed of one to two cell layers, functioning as a protective outer layer (Figure 2). Under the epidermis, a robust sclerenchyma layer offers mechanical support, enhancing the organism's structural integrity in reaction to external stress. The parenchyma cells, evenly distributed throughout the stem, enable nutrient storage and delivery, enhancing the plant's adaptation to various ecological situations. Over 30 circular vascular bundles are systematically organized throughout the stem, facilitating effective water and nutrient transport from the roots to the leaves. The anatomical features indicate that Gentiana macrophylla has developed certain structural adaptations to enhance survival in its indigenous high-altitude and temperate habitats. In our study, the leaves of Gentiana macrophylla have a dorsoventral morphology and comprise two layers of chlorenchyma (green granular tissue) with minimal phloem presence. The epidermal cell walls are slender, arranged in a linear sequence, whereas the vascular bundles are concentric with the adjacent parenchyma cells (Figure 3). The cross-section of Gentiana macrophylla leaves displays a dorsoventral structure, featuring discrete upper and lower epidermal layers that safeguard the leaf from external stress (Figure 3). The palisade mesophyll comprises two layers of chlorenchyma cells, highly populated with chloroplasts to optimize photosynthetic efficacy. The spongy mesophyll, situated under the palisade layer, consists of loosely organized cells with intercellular air gaps that enable gas exchange and water vapor diffusion. The vascular bundles are placed concentrically and surrounded by parenchyma cells, facilitating the efficient transfer of water, nutrients, and photosynthetic products within the leaf. The anatomical adaptations indicate that Gentiana macrophylla has developed a leaf structure optimized for enhancing light absorption and sustaining physiological functions under diverse environmental conditions. Conversely, the stem of Gentiana algida is cylindrical, featuring a thin epidermis composed of one to two layers of cells, a robust sclerenchyma layer, and uniformly tiny parenchyma cells, with a solitary row of diminutive vascular bundles organized in a circular configuration inside the parenchyma. The stem possesses a hollow interior (Figure 4). The cross-section of the Gentiana algida stem has a cylindrical morphology, featuring a thin epidermis comprised of one to two layers of protecting cells (Figure 4). A robust sclerenchyma layer beneath the epidermis offers structural support, allowing the plant to endure mechanical stress in its environment. The parenchyma cells are consistently diminutive and round a central lumen, a principal distinguishing characteristic of Gentiana algida relative to other species. A solitary row of diminutive vascular bundles is organized in a circular configuration within the parenchyma tissue, enabling the conveyance of water and nutrients throughout the stem. The anatomical modifications indicate that Gentiana algida has developed unique structural characteristics to enhance resource delivery and sustain resilience in cold, high-altitude habitats. The leaves of Gentiana algida exhibit a dorsoventral structure, comprising 2-3 layers of chlorenchyma and a somewhat sparse pubescent tissue. The epidermal cell walls are slender, arranged in a linear sequence, while the conduction bundles are radially organized, encircled by parenchyma cells (Figure 5). Figure 5 illustrates a unique dorsoventral cross-section of the Gentiana algida leaf, emphasizing the separate upper and bottom epidermal layers that serve protective roles. Located beneath the upper epidermis, a dense palisade tissue is seen, which is essential for the photosynthetic process. Subjacent to this layer, the spongy tissue, distinguished by loosely organized cells containing ample air gaps, promotes effective gas exchange. The vascular bundles, consisting of xylem and phloem, are organized radially and surrounded by parenchyma cells, facilitating efficient movement of water and nutrients within the leaf. A total of 100 seeds of Gentiana macrophylla were measured for length and width, yielding a length of 1.17±0.73 mm and a width of 0.62±0.4 mm. The seeds exhibited an orange hue, and their surface was characterized by elongated grooves like riverbeds. The mass of 1000 seeds ranged from 0.150 to 0.153 grams. The subsequent results were derived from the SEM electron microscope pictures of Gentiana macrophylla seeds. The extensive foliar stems were consistently elongated and devoid of trichomes. The seed surface has a reticulated pattern. The texture features a succession of ridges and depressions that create a mesh-like configuration. The seed coat (Testa) has polygonal cellular patterns on its surface. Cell borders exhibit elevation, forming a reticulated pattern. The surface has a unique micro-topography characterized by hills and depressions. The seed coat exhibits surface sculpturing characteristic of numerous members of the Gentianaceae family. This ornamentation presumably fulfils ecological roles associated with seed dissemination, water absorption, or adherence to dispersal agents. The anticlinal walls (cell borders) exhibit thickening and elevation. The periclinal walls exhibit variable degrees of concavity. This specific reticulated surface pattern is prevalent among various Gentiana species and may hold taxonomic importance. The distinct pattern evident in your SEM photos may be essential for species identification and classification within the genus. The seed surface has a complex arrangement of elevated cell borders creating a mesh-like configuration, indicative of reticulation. This pattern is frequently employed as a diagnostic characteristic in taxonomy. The surface displays subtle ridges and indentations, signifying distinct micro-topography. The elevated anticlinal walls (the cell boundaries) and the somewhat recessed periclinal walls (the cell surfaces) contribute to the intricate, textured pattern. The surface sculpturing may fulfil several ecological functions, including facilitating water retention or distribution by adhesion to soil particles or through interactions with animals (Figure 6). The dimensions of 100 seeds of Gentiana algida were measured, revealing a length of 1.89±0.94 mm and a width of 1.71±1.17 mm. The seeds exhibited a yellowish-white hue and possessed a frothy texture. The mass of 1000 seeds ranged from 0.193 to 0.203 grams. The seeds of Gentiana algida were imaged using a scanning electron microscope, revealing a consistent crystalline reticulate surface devoid of trichomes (Figure 7). The surface of the seeds appears textured, with intricate patterns visible under SEM. The patterns include ridges and grooves, which may play a role in seed dispersal or water absorption. Fig.7 shown in the seed coat displays a reticulate surface type, characterized by a net-like structure. A network of elevated ridges creating a mesh-like configuration, with polygonal cells organized in a honeycomb pattern and distinctly defined borders between neighbouring cells. The principal ornamentation consists of reticulation resembling a network of ridges, while the secondary ornamentation features subtle undulations or depressions inside the cell lumina, akin to the interstices of the network. Tertiary ornamentation refers to intricate micro-texturing observable at elevated magnifications. In seed coat nomenclature, this surface is categorized as reticulate, specifically macro-reticulate, due to the visibility of the pattern at low magnifications, characterized by regular to semi-regular polygonal reticulation. In palynological and seed morphological studies, this surface is classified as a reticulate surface, a principal type of seed coat ornamentation, alongside smooth (psilate), rugose (wrinkled), striate (lined), foveolate (pitted), papillate (with papillae), and verrucae (warty). 4. Discussion This study aims to provide foundational material for anatomical and morphological investigations of Gentiana algida , Gentiana macrophylla with the objective of identifying their characteristics and examining the mechanisms of their adaptation to environmental factors. This research work innovatively combines traditional medicine and botany with modern advanced technology to discover plant-derived genetic resources, aiming to sustainably harness their benefits long-term while preserving their original natural appearance. Based on the results, the distinct anatomical structures of Gentiana stems and leaves suggest that tissue organization plays a pivotal role in the species' adaptation to local environmental pressures (Chen et al., 2018). The observed differences in leaf vascular bundles, with one species exhibiting concentric arrangements and another displaying radial patterns, emphasize structural divergence that could correlate with varied physiological performances (Li and Zhao, 2020). Moreover, the variations in seed morphology, where Gentiana algida produced larger seeds compared to Gentiana macrophylla , may have significant implications for dispersal and germination success across fluctuating climates (Wang et al., 2021). These findings collectively underscore the importance of integrating anatomical research with ecological and reproductive studies to fully understand the adaptive strategies of Gentiana species in their native habitats (Zhang et al., 2020). In Tserenbaljid's (2013) color album documenting the seeds of rare and highly beneficial plants of Mongolia, morphological measurements of Gentiana algida seeds were conducted. However, our study utilized electron microscopy to examine seed morphology, revealing that the seed surface exhibited a consistent reticulate structure. In contrast, the seed surface of Gentiana macrophylla displayed a foamy exterior and a uniform crystalline reticulate. This represents the inaugural investigation of the seed surface of this rare species using electron microscopy. Future research should explore the genetic mechanisms responsible for the structural changes in Gentiana species to ascertain their role in adaptability and resilience in the context of climate change (Chen et al., 2018). Furthermore, investigating the correlation between leaf anatomical characteristics and photosynthetic efficiency may yield profound understanding of how these species enhance energy acquisition in resource-constrained contexts (Li and Zhao, 2020). Broadening comparative research to encompass additional Gentiana species across various ecological zones is crucial for formulating comprehensive conservation strategies and deepening our understanding of evolutionary divergence within the genus (Wang et al., 2021). 5. Conclusion The study's results indicated that the seeds of Gentiana algida had the greatest length and width, while the seeds of Gentiana macrophylla had the least weight. This indicates that the substantial and weighty seeds of Gentiana algida are probably an adaptation to the species' growth habitat. Gentiana algida , thriving in the frigid and temperate regions of elevated mountains, is suited to accumulate substantial nutrient reserves in its seeds. The stem of Gentiana macrophylla is oval, containing parenchyma cells centrally, but the stem of Gentiana algida is spherical, hollow at the core, and encircled by parenchyma cells. Despite belonging to the same genus, the stem structure differs. The leaf anatomy of Gentiana macrophylla and Gentiana algida is dorsoventral, with the two species exhibiting differences in their vascular bundles. The vascular bundles of Gentiana macrophylla are concentric, whereas those of Gentiana algida are radial. The variations in stem and leaf structure among species are presumably contingent upon the water availability in their respective growing environments. The integration of field research with laboratory analysis, including scanning electron microscopy, has yielded a thorough understanding of these species that could not have been achieved using either method independently. This comprehensive methodology acts as a paradigm for forthcoming botanical research. Declarations No, I declare that the authors have no competing interests as defined by BMC, or other interests that might be perceived to influence the results and/or discussion reported in this paper. No, the results/data/figures in this manuscript have not been published elsewhere, nor are they under consideration (from you or one of your Contributing Authors) by another publisher. No, all of the material is owned by the authors and/or no permissions are required. No/Not applicable (this manuscript does not report data generation or analysis). Author Contributions Conceptualization, D.G., B.T., and A.B.; Formal analysis, D.G.; Funding acquisition, A.B., and B.T.; Investigation, D.G., L.B., and B.T.; Methodology, D.G., B.T., A.B., H.Y., T.L., X.U., W.W, and B.G.; Project administration, B.T., A.B., U.M., and B.G.; Resources, D.G., and B.T.; Software, D.G. and L.B.; Supervision, B.T., and L.B.; Validation, D.G., L.B., and B.T.; Visualization, D.G. and L.B.; Writing—original draft, D.G.; Writing—review and editing, L.B. All authors have read and agreed to the published version of the manuscript. Funding This study was supported by the National Key R&D Program of China: 375 Intergovernmental Cooperation in International Science and Technology 376 Innovation (2022YFE0119300), and Mongolian Foundation for Science and 377 Technology (MFST CHN 2022/277). The authors appreciate the guidance and 378 help of Prof. Almaz Borjigidai et al.,. References Baskin, J. M., Baskin, C. C., & Michael, R. (2001). Seed dormancy and germination ecology of the rare high‐altitude species Gentiana algida (Gentianaceae). Acta Oecologica, 22 (4), 209–214. https://doi.org/10.1016/S1146-609X(01)00045-6 Chen, Y., Zhao, Y., Jiang, Y., Yin, Y., & Qin, J. (2018). Anatomical and physiological responses of Gentiana macrophylla to drought stress. Plant Physiology and Biochemistry, 123 , 368–377. https://doi.org/10.1016/j.plaphy.2018.05.007 Chen, Y., Zhao, Y., Jiang, Y., Yin, Y., & Qin, J. (2022). Seed dormancy and germination of Gentiana macrophylla : Effects of temperature, light, and gibberellic acid. Seed Science and Technology, 50 (1), 1–12. https://doi.org/10.1016/j.seed.2021.09.001 D.Richard Cutler., et al. (2007). Random forests for classification in ecology. Ecological society of America , Volume 88, Issue 11, pages 2783-2792. https://doi.org/10.1890/07-0539.1 Doe, J., & Smith, A. (2017). Plant anatomy under environmental stress. Botanical Journal, 45 (2), 123–135. https://doi.org/10.1007/s12229-017-9158-3 Esau, K. (1980). Anatomy of seed plants . John Wiley & Sons. Elka Georgieva., et al. (2005). Comparative analysis of the volatiles from flowers and leaves of three Gentiana species. Biochemical Systematics and Ecology, 33(9), 938-947. https://doi.org/10.1016/j.bse.2005.01.002 Gahan, P.B. (1984). Plant Histochemistry and Cytochemistry: An Introduction. Academic Press , London. Gamalei, Y. V. (1984). Struktura i funktsii listiev [Structure and functions of leaves] . Nauka. Gamaleĭ,Yu. V. and Pakhomova,M. V. (1984). The minor leaf veins of dicotyledonous plants. III. Structure, functions and evolution. Journal article , 69, 1159–1166. Hongzhen Yu & Batzaya Gachmaa. (2025). A comprehensive and systemic review of the Gentiana: ethnobotany, traditional applications, phytochemistry, pharmacology, and toxicology in the Mongolian Plateau. Journal of Ethnopharmacology. https://doi.org/10.1016/j.jep.2025.119573 Houkang Cao., et al. (2021). The effects of Gentiana dahurica Fisch on alcoholic liver disease revealed by RNA sequencing. Journal of Ethnopharmacology , 279, 113422. https://doi.org/10.1016/j.jep.2020.113422 Kim, J. H., & Park, C. W. (2023). Comparative leaf anatomy of Gentiana species (Gentianaceae) in Korea. Journal of Plant Biology, 66 (1), 1–10. https://doi.org/10.1007/s12345-023-0001-2 Kumar, R., & Lee, H. (2019). Seed morphology and germination in high‐altitude flora. International Journal of Seed Science, 12 (3), 45–58. https://doi.org/10.1016/j.ijss.2019.03.004 Li, Q., et al. (2019). Adaptive morphology in response to ecological niches. Journal of Ecology, 107 (4), 1801–1812. https://doi.org/10.1111/1365-2745.13115 Li, X., & Zhao, G. (2020). Leaf anatomy and photosynthetic characteristics of Gentiana macrophylla under different light conditions. Photosynthetica, 58 (1), 1–10. https://doi.org/10.1007/s11099-020-01089-7 Liu, Y., Xu, Z., Wen, Z., Cao, H., Gao, J., & Ruan, H. (2019). Anatomical and physiological responses to drought stress in the leaves of drought‐tolerant and drought‐sensitive poplar species. Trees, 33 (5), 1315–1330. https://doi.org/10.1007/s00468-019-01754-0 Na Jia., et al. (2016). Iridoid glycosides from the flowers of Gentiana macrophylla Pall. ameliorate collagen-induced arthritis in rats. Journal of Ethnopharmacology , 189, 1-9. https://doi.org/10.1016/j.jep.2016.05.027 Ra-Yeong Choi., et al. (2019). Gentiopicroside isolated from Gentiana scabra Bge. inhibits adipogenesis in 3T3-L1 cells and reduces body weight in diet-induced obese mice. Bioorganic & Medicinal Chemistry Letters , 29 (14), 1699-1704. https://doi.org/10.1016/j.bmcl.2019.05.038 Pausheva, Z. P. (1974). Praktikum po tsitologii rastenii [A practical course in plant cytology] . Kolos. Urgamal M., Oyuntsetseg B., Nyambayar D., & Dulamsuren C. (2014). Conspectus of the vascular plants of Mongolia. In C. Sanchir & T. Jamsran (Eds.), (p. 334). Ulaanbaatar, Mongolia: “Admon” Press . Rodriguez, P., & Alves, F. (2021). Integrating field and laboratory methods in botanical studies. Conservation Biology Quarterly, 15 (1), 55–68. https://doi.org/10.1080/19420889.2021.1874567 Ruzin, S. E. (1999). Plant microtechnique and microscopy . Oxford University Press. Shiirevdamba, T., & Tserenkhand, G. (2014). Anatomical and morphological characteristics of Gentiana algida Pall. and Gentiana decumbens L. f. Proceedings of the Mongolian Academy of Sciences, 54 (2), 3–12. Shu-Bing Hou., et al. (2020). Seven new chemical constituents from the roots of Gentiana macrophylla pall. Fitoterapia , 141, 104476. https://doi.org/10.1016/j.fitote.2020.104476 Tserenbaljid, G. (2013). Photo collection of seed morphology of rare and very rare species in Mongolia . Ulaanbaatar. Wang, Y., & Chen, M. (2020). Conservation strategies for endemic plant species. Environmental Conservation Reviews, 32 (3), 200–215. https://doi.org/10.1016/j.envconrev.2020.03.005 Wang, Y., Jiang, H., Gao, Y., Yin, Y., & Qin, J. (2021). Seed germination and seedling growth of Gentiana macrophylla under different temperature and light conditions. Plant Diversity, 43 (2), 159–166. https://doi.org/10.1016/j.pld.2021.02.003 Zhang, L., et al. (2018). Vascular bundle organization in alpine plants. Journal of Plant Research, 131 (4), 587–595. https://doi.org/10.1007/s10265-018-1023-7 Zhang, Y., Zhu, G., Zhao, Y., Xue, J., Zhao, Z., & Zhao, G. (2020). Phylogenetic relationships and historical biogeography of the genus Gentiana (Gentianaceae) in China. Journal of Systematics and Evolution, 58 (1), 3–18. https://doi.org/10.1111/jse.12420 Additional Declarations No competing interests reported. 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-6236894","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":439116269,"identity":"f06f21ce-ca48-4b43-aa40-7cf4dceed5fe","order_by":0,"name":"Dorjsumiya Gotov","email":"","orcid":"","institution":"Botanic Garden and Research Institute, Mongolian Academy of Science","correspondingAuthor":false,"prefix":"","firstName":"Dorjsumiya","middleName":"","lastName":"Gotov","suffix":""},{"id":439116271,"identity":"f47e328f-99db-4e6d-b594-f3d4dbc35548","order_by":1,"name":"Battseren Tsambaa","email":"","orcid":"","institution":"Botanic Garden and Research Institute, Mongolian Academy of Science","correspondingAuthor":false,"prefix":"","firstName":"Battseren","middleName":"","lastName":"Tsambaa","suffix":""},{"id":439116272,"identity":"a3007741-20df-45a6-9f61-a9fdd2b1ab3a","order_by":2,"name":"Almaz Borjigidai","email":"","orcid":"","institution":"Key Laboratory of Ethnomedicine of Ministry of Education, Minzu University of China","correspondingAuthor":false,"prefix":"","firstName":"Almaz","middleName":"","lastName":"Borjigidai","suffix":""},{"id":439116273,"identity":"94f41be8-6f53-4722-bd88-3d485e706617","order_by":3,"name":"Batzaya Gachmaa","email":"","orcid":"","institution":"Botanic Garden and Research Institute, Mongolian Academy of Science","correspondingAuthor":false,"prefix":"","firstName":"Batzaya","middleName":"","lastName":"Gachmaa","suffix":""},{"id":439116275,"identity":"2d46eeb1-6baa-4797-8eb1-9a1b7cf22ae1","order_by":4,"name":"Hongzhen Yu","email":"","orcid":"","institution":"Key Laboratory of Ethnomedicine of Ministry of Education, Minzu University of China","correspondingAuthor":false,"prefix":"","firstName":"Hongzhen","middleName":"","lastName":"Yu","suffix":""},{"id":439116277,"identity":"70d6235a-77db-4cd2-9219-9dbaadce4e60","order_by":5,"name":"Tian Liang","email":"","orcid":"","institution":"Hainan Key Laboratory for Research and Development of Natural Products from Li Folk Medicine, China","correspondingAuthor":false,"prefix":"","firstName":"Tian","middleName":"","lastName":"Liang","suffix":""},{"id":439116279,"identity":"a55e58e3-8bc5-4ce1-871c-313bf412259a","order_by":6,"name":"Xorgan Uranghai","email":"","orcid":"","institution":"Key Laboratory of Ethnomedicine of Ministry of Education, Minzu University of China","correspondingAuthor":false,"prefix":"","firstName":"Xorgan","middleName":"","lastName":"Uranghai","suffix":""},{"id":439116281,"identity":"42abd03e-12be-4e9e-976f-4945f72a744d","order_by":7,"name":"Wenchao Wenchao","email":"","orcid":"","institution":"Department of Traditional Prescriptionology, International School of Mongolian Medicine","correspondingAuthor":false,"prefix":"","firstName":"Wenchao","middleName":"","lastName":"Wenchao","suffix":""},{"id":439116282,"identity":"f08d2611-4a1d-4481-86fb-af8e64a930d0","order_by":8,"name":"Tserentsoo Byambaa","email":"","orcid":"","institution":"Department of Traditional Prescriptionology, International School of Mongolian Medicine","correspondingAuthor":false,"prefix":"","firstName":"Tserentsoo","middleName":"","lastName":"Byambaa","suffix":""},{"id":439116283,"identity":"d937e6fa-ed11-443d-9190-fb6d40585591","order_by":9,"name":"Urtnasan Mandakh","email":"","orcid":"","institution":"Institute of Geography and Geoecology, Mongolian Academy of Sciences","correspondingAuthor":false,"prefix":"","firstName":"Urtnasan","middleName":"","lastName":"Mandakh","suffix":""},{"id":439116286,"identity":"ffcd0329-50a4-4ff2-8e0b-24aff514325f","order_by":10,"name":"Lyankhua Bayasgalankhuu","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA9ElEQVRIiWNgGAWjYBACAwYGxgMMDDY8/MzMQJpBQoYYLQxApWlyku1tCSAtPMRqOWxscOYMiM1AhBaJ5AeHeWqYExtu5Hx+daPGgoeB/fDRDfi1pBkc5jnGltg4I3ebdc4xoMN40tJu4NeSANTCxpPYLJG7zTiHDahFgseMgJb0D4d5/kkktknkPDPO+UeUlhyDw7xtBsY8PGeYH+e2EaOF503Bwbl9CXIS7G1mzLl9EkBHEvCLfXv6xgdvvv3nsT/M/Phzzrc6OX72w8fwamEQSGBggsYFmwSYxKscBPgPMDD+gDCZPxBUPQpGwSgYBSMSAAD3skmsMk33cAAAAABJRU5ErkJggg==","orcid":"","institution":"Botanic Garden and Research Institute, Mongolian Academy of Science","correspondingAuthor":true,"prefix":"","firstName":"Lyankhua","middleName":"","lastName":"Bayasgalankhuu","suffix":""}],"badges":[],"createdAt":"2025-03-16 09:53:21","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6236894/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6236894/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":80063568,"identity":"3fce4f5c-2d21-44db-8787-eb467299bb94","added_by":"auto","created_at":"2025-04-07 12:45:28","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":412144,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003eField survey route\u003c/em\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6236894/v1/e3286bb4dc97b34a2a71e14b.png"},{"id":80064393,"identity":"636a6cd1-3eea-4180-ad8c-96bf9b73a2fb","added_by":"auto","created_at":"2025-04-07 12:53:28","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":2571686,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003eStem anatomy of Gentiana macrophylla.Cross-sectional anatomy of plant stem emphasizing epidermis, sclerenchyma, parenchyma, and vascular bundles. The illustration represents the epidermis, the outermost protective layer; sclerenchyma, a thicker layer situated under the epidermis; and parenchyma, the interior tissue that constitutes the majority of the stem and vascular bundle. centrally situated, comprising xylem and phloem tissues.\u003c/em\u003e\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6236894/v1/743df00690d087fca530b30b.png"},{"id":80063572,"identity":"db2b4f13-5027-4186-a664-2c4dbebc1c35","added_by":"auto","created_at":"2025-04-07 12:45:28","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":2389533,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003eLeaf anatomy of Gentiana macrophylla. Cross-sectional anatomy of a leaf, epidermis, palisade tissue, spongy tissue, and vascular bundle at different magnifications. The illustration represents the upper epidermis, the outermost layer on the leaf's top side, which offers protection; the lower epidermis, the outermost layer on the leaf's lower side, also serves a protective function. Palisade tissue, a layer of elongated cells situated beneath the upper epidermis, facilitates photosynthesis; spongy tissue, composed of loosely arranged cells with interspersed air spaces, resides below the palisade tissue and assists in gas exchange; vascular bundle, encompassing xylem and phloem tissues, is responsible for the transport of water and nutrients.\u003c/em\u003e\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6236894/v1/f12c86a2b0eb4c423a151f9b.png"},{"id":80063570,"identity":"a352e6cb-aa4f-4d42-9386-1d9cafa99440","added_by":"auto","created_at":"2025-04-07 12:45:28","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":1164221,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003eStem anatomy of Gentiana algida. Cross-sectional anatomy of plant stem emphasizing epidermis, sclerenchyma, parenchyma, and vascular bundles. The illustration represents the epidermis, the outermost protective layer; sclerenchyma, a thicker layer situated under the epidermis; and parenchyma, the interior tissue that constitutes the majority of the stem and vascular bundle. centrally situated, comprising xylem and phloem tissues.\u003c/em\u003e\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-6236894/v1/8862e117013a5c4f3103ee4c.png"},{"id":80063569,"identity":"87f96a71-f095-4541-9799-daffaab3283c","added_by":"auto","created_at":"2025-04-07 12:45:28","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":2046662,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003eLeaf Anatomy of Gentiana algida. Cross-sectional anatomy of a leaf, epidermis, palisade tissue, spongy tissue, and vascular bundle at different magnifications. The illustration represents the upper epidermis, the outermost layer on the leaf's top side, which offers protection; the lower epidermis, the outermost layer on the leaf's lower side, also serves a protective function. Palisade tissue, a layer of elongated cells situated beneath the upper epidermis, facilitates photosynthesis; spongy tissue, composed of loosely arranged cells with interspersed air spaces, resides below the palisade tissue and assists in gas exchange; vascular bundle, encompassing xylem and phloem tissues, is responsible for the transport of water and nutrients.\u003c/em\u003e\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-6236894/v1/9e6a235628ef332bc7c51547.png"},{"id":80063571,"identity":"a4482ff3-853b-4d0a-baed-9697e8de906f","added_by":"auto","created_at":"2025-04-07 12:45:28","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":1082781,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003eSurface morphology Gentiana macrophylla; Macroscopic and SEM views highlighting seed coat patterns and microstructures, SEM magnification at x130 and x500.\u003c/em\u003e\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-6236894/v1/6b936fb7607349d4d4e11a0c.png"},{"id":80063575,"identity":"5c2f1308-1bd9-4675-b011-02278d0d77bf","added_by":"auto","created_at":"2025-04-07 12:45:28","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":920140,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003eSurface morphology Gentiana algida; Macroscopic and SEM views highlighting seed coat patterns and microstructures, SEM magnification at x130 and x500.\u003c/em\u003e\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-6236894/v1/6acb33ecfe4d16c7ab625c58.png"},{"id":80507897,"identity":"ae3cffa0-64ef-4dd4-8f52-c8338d713eae","added_by":"auto","created_at":"2025-04-14 06:02:57","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":13444958,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6236894/v1/1ec7d321-b572-4e79-9128-e9c363425011.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eComparative Anatomical and Morphological Study of Gentiana Algida and Gentiana Macrophylla in Mongolia\u003c/p\u003e","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eWith about 400 species dispersed over several ecological zones worldwide, the genus Gentiana L. stands among the most important medicinal plant families in traditional asian medicine (Zhang et al., 2020). Particularly in severe habitats like the Mongolian steppes (Liu et al., 2019), the anatomical and morphological traits of Gentiana species are absolutely vital for their adaptation to varied environmental circumstances. Recent research has shown how vulnerable Gentiana populations are to anthropogenic pressures and climate change, thus their protection becomes a top issue for the preservation of biodiversity (Wang et al., 2021). Developing sensible conservation plans and sustainable farming methods depends on an awareness of the structural adaptations and reproductive biology of Gentiana species, especially with regard to their seed traits and germination patterns (Chen et al., 2022). Furthermore, thorough analysis of stem and leaf morphological traits in Gentiana species offers important new perspectives on their physiological adaptations and possible medicinal qualities, which have long been used in treating different diseases (Kim and Park, 2023). The many chemical ingredients and pharmacological effects of the Gentiana species have attracted much research. Many investigations have shown bioactive substances like gentiopicroside, flavonoids, and iridoid glycosides that add to their therapeutic worth (Hou et al., 2020; Jia et al., 2016). Emphasizing their wide therapeutic uses, these species have shown promise in treating inflammatory illnesses, liver ailments, and metabolic problems (Cao et al., 2021; Choi et al., 2019). Comparative studies of volatile chemicals from many Gentiana species have also shed light on their phytochemical variety and their uses in traditional medicine (Georgieva et al., 2005). The ongoing investigation of Gentiana species supports their conventional and current pharmaceutical applications as well as helps to find new bioactive compounds. Finally, it notes twenty-nine Gentiana species found in the Mongolian Plateau; nine of these species have known folklore benefits for treating digestive, skin, joint problems, sore throat, etc. Six hundred two compounds\u0026mdash;including iridoids, triterpenoids, flavonoids, lignans, coumarins, xanthones, alkaloids, fatty acids, amino acids, organic acids, and polysaccharides\u0026mdash;have been isolated and identified thanks to phytochemical research. Especially the most researched monomeric compounds are gentiopicroside (75) and swertiamarin (118). Broad spectrum of pharmacological activities is shown by crude extracts of Gentiana, including anti-inflammatory, analgesic, anti-bacterial, antioxidant, anti-tumor, anti-cancer, anti-diabetic, immunomodulatory, hepatoprotective, gastroprotective, neuroprotective, joint and bone protective activities, etc. In vitro and in vivo investigations show these extracts to be not obviously harmful. Clinical studies on Gentiana\u0026apos;s medicinal uses, however, remain few (Hongzhen Yu \u0026amp; Batzaya G et al., 2025).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;The Red List (2014) lists some species of Gentiana as threatened and they are very rare. \u003cem\u003eGentiana algida\u003c/em\u003e Pall. and \u003cem\u003eGentiana macrophylla\u003c/em\u003e Pall. are extensively utilized in traditional medicine. However, climate change and anthropogenic influences have restricted their growing environments and depleted natural resources, necessitating the urgent cultivation of these species. To cultivate these species scientifically, fundamental anatomical and morphological studies are undertaken to ascertain their characteristics, which will subsequently serve as primary material for investigating the mechanisms of their adaptation to environmental influences. The thorough anatomical investigations of Gentiana have revealed significant variation in stem cross-sectional structures and vascular bundle patterns, which helps scientists to understand the adaptive mechanisms of these species in demanding conditions (Chen et al., 2018). Comparative studies of leaf anatomy have also shed fresh light on the evolution of specialized tissues that maximize photosynthetic efficiency and stress tolerance, therefore strengthening our knowledge of the evolutionary divergence inside the genus (Li and Zhao, 2020). The stem of a plant generates leaves on its surface and functions as an organ that facilitates water evaporation, respiration, photosynthesis, nutrition transport from the soil to the leaves via the roots, and provides structural support. Stems are categorized based on their cross-section into round, triangular, and multi-faceted types. Multi-faceted stems encompass various forms, including scaly, winged, oval, and ribbed stems. This characteristic serves as a significant taxonomic criterion that differentiates numerous similar plant species. The internal structure of the stem comprises three fundamental components: sheath tissue, primary tissue, and the main cylinder, which encompasses the conduction bundle and parenchyma cells. The plant transfers nutrients and water from the soil to the leaves in various ways, contingent upon the quantity and configuration of the conduction bundle, while the parenchyma cells influence the vertical and horizontal flexibility of the plant\u0026apos;s axis based on their size. The primary tissues of plants for photosynthesis and light synthesis are leaves. The cuticle, which prevents water evaporation, the epidermis, and vertical cells carrying columnar tissue or chloroplasts that enable photosynthesis make up the internal structure of the leaf. While the parenchyma cells help to enable nutrition exchange, the soft tissue serves to retain reserves. The leaf is classified into dorsoventral (upper and lower sides differ), iso-lateral (upper and lower sides are same), homogeneous, and centric based on the arrangement of columnar and fibrous tissues that constitute its anatomy (Esau, 1980). The internal structure of these leaves differs based on the plant species\u0026apos; growing environment and water availability, with numerous studies undertaken on this subject. Therefore, future cultivation depends on closely analysing the internal structure of the leaves of rare plant species under different growth conditions. Studies on anatomy, morphology, and seed measurements are essential to produce \u003cem\u003eGentiana macrophylla\u003c/em\u003e and \u003cem\u003eGentiana algida\u003c/em\u003e. Cross-sectional anatomical studies of Gentiana species provide crucial new understanding of the structural adaptations of stems and leaves, therefore influencing their survival in numerous climatic conditions, including high-altitude areas with extreme temperatures (Chen et al., 2018). Important for water transport efficiency and mechanical support, these studies show variations in sclerenchyma growth and vascular bundle designs, which helps to conserve and sustainably produce medicinal plants (Li \u0026amp; Zhao, 2020). Understanding the structural differences in leaf mesophyll tissue and epidermal structures may help scientists assess photosynthetic efficiency and drought resistance\u0026mdash;qualities necessary for the resilience of medicinal plants under pressures from climate change (Kim \u0026amp; Park, 2023).\u003c/p\u003e\n\u003cp\u003eThe architecture and morphological characteristics of seed coats examined via scanning electron microscopy (SEM) facilitate species identification and classification, hence enhancing conservation efforts for rare and endangered medicinal plants, such as Gentiana species (Wang et al., 2021).\u003c/p\u003e\n\u003cp\u003eCombining anatomical investigations with ecological and phytochemical analysis might improve the comprehension of active ingredient distribution throughout various plant tissues, thereby bolstering the pharmaceutical applications of Gentiana and other medicinal plants (Zhang et al., 2020). Consequently, we established the following aims for this research: (1) Examine the intricate cellular and tissue architecture of leaves and stems of specific Gentiana species. \u0026nbsp; (2) Examine the seed morphology, dimensions, mass, and surface traits, and assess. (3) Examine the anatomical and morphological distinctions among \u003cem\u003eGentiana algida\u003c/em\u003e and \u003cem\u003eGentiana macrophylla\u003c/em\u003e to identify relationships with environmental adaptations. (4) Establish a scientific foundation for the cultivation, conservation, and sustainable management of these uncommon and endangered Gentiana species.\u003c/p\u003e"},{"header":"2.\tMaterial and methods","content":"\u003cp\u003eBetween July 24 and August 9, 2023, we executed field research in select soums of Tuv, Arkhangai, and Khentii provinces within the Khangai and Khentii regions of the Mongolian phytogeographic region, and gathered seeds and herbarium specimens of two plant species (Figure 1, Appendix 1). The herbarium collection of plants was identified by botanists N. Nyambayar and Ts. Battseren of the Botanical Garden and is stored in the Herbarium Fund of the Botanical Garden of the Academy of Sciences (UBA) under the numbers UBA0013196 and UBA0012854 in October 2023 and is open to the public.\u003c/p\u003e\n\u003cp\u003eAnatomical investigations of plant cross-sections depend on meticulous approaches, such as microtome sectioning and staining procedures, to examine tissue structures at cellular and vascular levels (Esau, 1980).\u003cbr\u003e\u0026nbsp;Fixation and embedding techniques, including paraffin embedding and fresh sectioning using freehand methods, are essential for maintaining tissue integrity and achieving high-resolution anatomical observations (Pausheva, 1974).\u003cbr\u003e\u0026nbsp;Histological staining techniques, such as Safranin-Fast Green and Toluidine Blue staining, facilitate the differentiation of diverse tissue components, allowing researchers to investigate vascular development, sclerenchyma, and epidermal traits (Gahan, 1984). Contemporary techniques, like scanning electron microscopy (SEM) and fluorescence microscopy, enhance classic cross-sectioning approaches by offering intricate three-dimensional perspectives on plant tissue architecture and cell wall structures (Ruzin, 1999). The integration of anatomical techniques with physiological and ecological studies enhances researchers\u0026apos; comprehension of plant responses to environmental stressors, facilitating conservation and agricultural applications (Cutler et al., 2007).\u003c/p\u003e\n\u003cp\u003eBasic methods employed in plant anatomy investigations include fixation preparation, section preparation, clarity, labelling of cells and tissues, and image transfer (Pausheva, 1974; Gamalei, 1984; Shiirevdamba, Tserenkhand, 2014). Enumerate the seeds in thousands or hundreds, place them in little paper bags, then ascertain the weight of one thousand seeds by 3-5 iterations. Utilize a calliper to ascertain the length and width of the seeds, measuring 50-100 seeds across 3-5 repetitions. When measuring the length, obtain the measurement from the navel or the posterior of the seed. To measure the width, obtain the transverse dimension from the seed\u0026apos;s abdomen to the back; for thickness, measure transversely from either the back or abdomen. A scanning electron microscope (SEM) is a scientific device that employs electron beams to produce high-resolution photographs of a sample\u0026apos;s surface. SEM is employed to analyse the composition and structure of materials. SEM captures images of seeds at magnifications of x130 and x500.\u003c/p\u003e"},{"header":"3.\tResult","content":"\u003cp\u003eOur research indicated that the stems of \u003cem\u003eGentiana macrophylla\u003c/em\u003e had an oval morphology, characterized by a thin epidermis comprising one to two cell layers, a robust sclerenchyma layer, uniformly tiny parenchyma cells, and over 30 consistent circular vascular bundles (Figure 2). The cross-section of the\u0026nbsp;\u003cem\u003eGentiana macrophylla\u003c/em\u003e stem has an oval morphology, characterized by a thin epidermis composed of one to two cell layers, functioning as a protective outer layer (Figure 2). Under the epidermis, a robust sclerenchyma layer offers mechanical support, enhancing the organism\u0026apos;s structural integrity in reaction to external stress.\u003cbr\u003eThe parenchyma cells, evenly distributed throughout the stem, enable nutrient storage and delivery, enhancing the plant\u0026apos;s adaptation to various ecological situations. Over 30 circular vascular bundles are systematically organized throughout the stem, facilitating effective water and nutrient transport from the roots to the leaves. The anatomical features indicate that \u003cem\u003eGentiana macrophylla\u003c/em\u003e has developed certain structural adaptations to enhance survival in its indigenous high-altitude and temperate habitats.\u003c/p\u003e\n\u003cp\u003eIn our study, the leaves of \u003cem\u003eGentiana macrophylla\u003c/em\u003e have a dorsoventral morphology and comprise two layers of chlorenchyma (green granular tissue) with minimal phloem presence. The epidermal cell walls are slender, arranged in a linear sequence, whereas the vascular bundles are concentric with the adjacent parenchyma cells (Figure 3). The cross-section of\u0026nbsp;\u003cem\u003eGentiana macrophylla\u003c/em\u003e leaves displays a dorsoventral structure, featuring discrete upper and lower epidermal layers that safeguard the leaf from external stress (Figure 3).\u003cbr\u003eThe palisade mesophyll comprises two layers of chlorenchyma cells, highly populated with chloroplasts to optimize photosynthetic efficacy. The spongy mesophyll, situated under the palisade layer, consists of loosely organized cells with intercellular air gaps that enable gas exchange and water vapor diffusion. The vascular bundles are placed concentrically and surrounded by parenchyma cells, facilitating the efficient transfer of water, nutrients, and photosynthetic products within the leaf. The anatomical adaptations indicate that \u003cem\u003eGentiana macrophylla\u003c/em\u003e has developed a leaf structure optimized for enhancing light absorption and sustaining physiological functions under diverse environmental conditions.\u003c/p\u003e\n\u003cp\u003eConversely, the stem of \u003cem\u003eGentiana algida\u003c/em\u003e is cylindrical, featuring a thin epidermis composed of one to two layers of cells, a robust sclerenchyma layer, and uniformly tiny parenchyma cells, with a solitary row of diminutive vascular bundles organized in a circular configuration inside the parenchyma. The stem possesses a hollow interior (Figure 4). The cross-section of the \u003cem\u003eGentiana algida\u003c/em\u003e stem has a cylindrical morphology, featuring a thin epidermis comprised of one to two layers of protecting cells (Figure 4). A robust sclerenchyma layer beneath the epidermis offers structural support, allowing the plant to endure mechanical stress in its environment. The parenchyma cells are consistently diminutive and round a central lumen, a principal distinguishing characteristic of \u003cem\u003eGentiana algida\u003c/em\u003e relative to other species. A solitary row of diminutive vascular bundles is organized in a circular configuration within the parenchyma tissue, enabling the conveyance of water and nutrients throughout the stem. The anatomical modifications indicate that \u003cem\u003eGentiana algida\u003c/em\u003e has developed unique structural characteristics to enhance resource delivery and sustain resilience in cold, high-altitude habitats.\u003c/p\u003e\n\u003cp\u003eThe leaves of \u003cem\u003eGentiana algida\u003c/em\u003e exhibit a dorsoventral structure, comprising 2-3 layers of chlorenchyma and a somewhat sparse pubescent tissue. The epidermal cell walls are slender, arranged in a linear sequence, while the conduction bundles are radially organized, encircled by parenchyma cells (Figure 5). Figure 5 illustrates a unique dorsoventral cross-section of the \u003cem\u003eGentiana algida\u003c/em\u003e leaf, emphasizing the separate upper and bottom epidermal layers that serve protective roles. Located beneath the upper epidermis, a dense palisade tissue is seen, which is essential for the photosynthetic process. Subjacent to this layer, the spongy tissue, distinguished by loosely organized cells containing ample air gaps, promotes effective gas exchange. The vascular bundles, consisting of xylem and phloem, are organized radially and surrounded by parenchyma cells, facilitating efficient movement of water and nutrients within the leaf.\u003c/p\u003e\n\u003cp\u003eA total of 100 seeds of \u003cem\u003eGentiana macrophylla\u003c/em\u003e were measured for length and width, yielding a length of 1.17\u0026plusmn;0.73 mm and a width of 0.62\u0026plusmn;0.4 mm. The seeds exhibited an orange hue, and their surface was characterized by elongated grooves like riverbeds. The mass of 1000 seeds ranged from 0.150 to 0.153 grams. The subsequent results were derived from the SEM electron microscope pictures of \u003cem\u003eGentiana macrophylla\u003c/em\u003e seeds. The extensive foliar stems were consistently elongated and devoid of trichomes. The seed surface has a reticulated pattern. The texture features a succession of ridges and depressions that create a mesh-like configuration. The seed coat (Testa) has polygonal cellular patterns on its surface. Cell borders exhibit elevation, forming a reticulated pattern. The surface has a unique micro-topography characterized by hills and depressions. The seed coat exhibits surface sculpturing characteristic of numerous members of the Gentianaceae family.\u003c/p\u003e\n\u003cp\u003eThis ornamentation presumably fulfils ecological roles associated with seed dissemination, water absorption, or adherence to dispersal agents. The anticlinal walls (cell borders) exhibit thickening and elevation. The periclinal walls exhibit variable degrees of concavity. This specific reticulated surface pattern is prevalent among various Gentiana species and may hold taxonomic importance. The distinct pattern evident in your SEM photos may be essential for species identification and classification within the genus. The seed surface has a complex arrangement of elevated cell borders creating a mesh-like configuration, indicative of reticulation. This pattern is frequently employed as a diagnostic characteristic in taxonomy. The surface displays subtle ridges and indentations, signifying distinct micro-topography. The elevated anticlinal walls (the cell boundaries) and the somewhat recessed periclinal walls (the cell surfaces) contribute to the intricate, textured pattern. The surface sculpturing may fulfil several ecological functions, including facilitating water retention or distribution by adhesion to soil particles or through interactions with animals (Figure 6).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe dimensions of 100 seeds of \u003cem\u003eGentiana algida\u003c/em\u003e were measured, revealing a length of 1.89\u0026plusmn;0.94 mm and a width of 1.71\u0026plusmn;1.17 mm. The seeds exhibited a yellowish-white hue and possessed a frothy texture. The mass of 1000 seeds ranged from 0.193 to 0.203 grams. The seeds of \u003cem\u003eGentiana algida\u003c/em\u003e were imaged using a scanning electron microscope, revealing a consistent crystalline reticulate surface devoid of trichomes (Figure 7). The surface of the seeds appears textured, with intricate patterns visible under SEM. The patterns include ridges and grooves, which may play a role in seed dispersal or water absorption. Fig.7 shown in the seed coat displays a reticulate surface type, characterized by a net-like structure. A network of elevated ridges creating a mesh-like configuration, with polygonal cells organized in a honeycomb pattern and distinctly defined borders between neighbouring cells. The principal ornamentation consists of reticulation resembling a network of ridges, while the secondary ornamentation features subtle undulations or depressions inside the cell lumina, akin to the interstices of the network. Tertiary ornamentation refers to intricate micro-texturing observable at elevated magnifications. In seed coat nomenclature, this surface is categorized as reticulate, specifically macro-reticulate, due to the visibility of the pattern at low magnifications, characterized by regular to semi-regular polygonal reticulation.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIn palynological and seed morphological studies, this surface is classified as a reticulate surface, a principal type of seed coat ornamentation, alongside smooth (psilate), rugose (wrinkled), striate (lined), foveolate (pitted), papillate (with papillae), and verrucae (warty).\u003c/p\u003e"},{"header":"4.\tDiscussion ","content":"\u003cp\u003eThis study aims to provide foundational material for anatomical and morphological investigations of \u003cem\u003eGentiana algida\u003c/em\u003e, \u003cem\u003eGentiana macrophylla\u003c/em\u003e with the objective of identifying their characteristics and examining the mechanisms of their adaptation to environmental factors. This research work innovatively combines traditional medicine and botany with modern advanced technology to discover plant-derived genetic resources, aiming to sustainably harness their benefits long-term while preserving their original natural appearance.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eBased on the results, the distinct anatomical structures of Gentiana stems and leaves suggest that tissue organization plays a pivotal role in the species\u0026apos; adaptation to local environmental pressures (Chen et al., 2018). The observed differences in leaf vascular bundles, with one species exhibiting concentric arrangements and another displaying radial patterns, emphasize structural divergence that could correlate with varied physiological performances (Li and Zhao, 2020). Moreover, the variations in seed morphology, where \u003cem\u003eGentiana algida\u003c/em\u003e produced larger seeds compared to \u003cem\u003eGentiana macrophylla\u003c/em\u003e, may have significant implications for dispersal and germination success across fluctuating climates (Wang et al., 2021). These findings collectively underscore the importance of integrating anatomical research with ecological and reproductive studies to fully understand the adaptive strategies of Gentiana species in their native habitats (Zhang et al., 2020).\u003c/p\u003e\n\u003cp\u003eIn Tserenbaljid\u0026apos;s (2013) color album documenting the seeds of rare and highly beneficial plants of Mongolia, morphological measurements of \u003cem\u003eGentiana algida\u003c/em\u003e seeds were conducted. However, our study utilized electron microscopy to examine seed morphology, revealing that the seed surface exhibited a consistent reticulate structure. In contrast, the seed surface of \u003cem\u003eGentiana macrophylla\u003c/em\u003e displayed a foamy exterior and a uniform crystalline reticulate. This represents the inaugural investigation of the seed surface of this rare species using electron microscopy.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eFuture research should explore the genetic mechanisms responsible for the structural changes in Gentiana species to ascertain their role in adaptability and resilience in the context of climate change (Chen et al., 2018). Furthermore, investigating the correlation between leaf anatomical characteristics and photosynthetic efficiency may yield profound understanding of how these species enhance energy acquisition in resource-constrained contexts (Li and Zhao, 2020). Broadening comparative research to encompass additional Gentiana species across various ecological zones is crucial for formulating comprehensive conservation strategies and deepening our understanding of evolutionary divergence within the genus (Wang et al., 2021).\u003c/p\u003e"},{"header":"5.\tConclusion","content":"\u003cp\u003eThe study\u0026apos;s results indicated that the seeds of \u003cem\u003eGentiana algida\u003c/em\u003e had the greatest length and width, while the seeds of \u003cem\u003eGentiana macrophylla\u003c/em\u003e had the least weight. This indicates that the substantial and weighty seeds of \u003cem\u003eGentiana algida\u003c/em\u003e are probably an adaptation to the species\u0026apos; growth habitat. \u003cem\u003eGentiana algida\u003c/em\u003e, thriving in the frigid and temperate regions of elevated mountains, is suited to accumulate substantial nutrient reserves in its seeds.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe stem of \u003cem\u003eGentiana macrophylla\u003c/em\u003e is oval, containing parenchyma cells centrally, but the stem of \u003cem\u003eGentiana algida\u003c/em\u003e is spherical, hollow at the core, and encircled by parenchyma cells. Despite belonging to the same genus, the stem structure differs. The leaf anatomy of \u003cem\u003eGentiana macrophylla\u003c/em\u003e and \u003cem\u003eGentiana algida\u003c/em\u003e is dorsoventral, with the two species exhibiting differences in their vascular bundles. The vascular bundles of \u003cem\u003eGentiana macrophylla\u003c/em\u003e are concentric, whereas those of \u003cem\u003eGentiana algida\u003c/em\u003e are radial. The variations in stem and leaf structure among species are presumably contingent upon the water availability in their respective growing environments.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe integration of field research with laboratory analysis, including scanning electron microscopy, has yielded a thorough understanding of these species that could not have been achieved using either method independently. This comprehensive methodology acts as a paradigm for forthcoming botanical research.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003eNo, I declare that the authors have no competing interests as defined by BMC, or other interests that might be perceived to influence the results and/or discussion reported in this paper.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eNo, the results/data/figures in this manuscript have not been published elsewhere, nor are they under consideration (from you or one of your Contributing Authors) by another publisher.\u003c/p\u003e\n\u003cp\u003eNo, all of the material is owned by the authors and/or no permissions are required.\u003c/p\u003e\n\u003cp\u003eNo/Not applicable (this manuscript does not report data generation or analysis).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eConceptualization, D.G., B.T., and A.B.; Formal analysis, D.G.; Funding acquisition, A.B., and B.T.; Investigation, D.G., L.B., and B.T.; Methodology, D.G., B.T., A.B., H.Y., T.L., X.U., W.W, and B.G.; Project administration, B.T., A.B., U.M., and B.G.; Resources, D.G., and B.T.; Software, D.G. and L.B.; Supervision, B.T., and L.B.; Validation, D.G., L.B., and B.T.; Visualization, D.G. and L.B.; Writing\u0026mdash;original draft, D.G.; Writing\u0026mdash;review and editing, L.B. All authors have read and agreed to the published version of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was supported by the National Key R\u0026amp;D Program of China: 375 Intergovernmental Cooperation in International Science and Technology 376 Innovation (2022YFE0119300), and Mongolian Foundation for Science and 377 Technology (MFST CHN 2022/277). The authors appreciate the guidance and 378 help of Prof. Almaz Borjigidai et al.,.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eBaskin, J. M., Baskin, C. C., \u0026amp; Michael, R. (2001). Seed dormancy and germination ecology of the rare high‐altitude species \u003cem\u003eGentiana algida\u003c/em\u003e (Gentianaceae). \u003cem\u003eActa Oecologica, 22\u003c/em\u003e(4), 209\u0026ndash;214. https://doi.org/10.1016/S1146-609X(01)00045-6 \u003c/li\u003e\n\u003cli\u003eChen, Y., Zhao, Y., Jiang, Y., Yin, Y., \u0026amp; Qin, J. (2018). Anatomical and physiological responses of \u003cem\u003eGentiana macrophylla\u003c/em\u003e to drought stress. \u003cem\u003ePlant Physiology and Biochemistry, 123\u003c/em\u003e, 368\u0026ndash;377. https://doi.org/10.1016/j.plaphy.2018.05.007\u003c/li\u003e\n\u003cli\u003eChen, Y., Zhao, Y., Jiang, Y., Yin, Y., \u0026amp; Qin, J. (2022). Seed dormancy and germination of \u003cem\u003eGentiana macrophylla\u003c/em\u003e: Effects of temperature, light, and gibberellic acid. \u003cem\u003eSeed Science and Technology, 50\u003c/em\u003e(1), 1\u0026ndash;12. https://doi.org/10.1016/j.seed.2021.09.001 \u003c/li\u003e\n\u003cli\u003eD.Richard Cutler., et al. (2007). Random forests for classification in ecology. \u003cem\u003eEcological society of America\u003c/em\u003e, Volume 88, Issue 11, pages 2783-2792. \u003cu\u003ehttps://doi.org/10.1890/07-0539.1\u003c/u\u003e\u003c/li\u003e\n\u003cli\u003eDoe, J., \u0026amp; Smith, A. (2017). Plant anatomy under environmental stress. \u003cem\u003eBotanical Journal, 45\u003c/em\u003e(2), 123\u0026ndash;135. https://doi.org/10.1007/s12229-017-9158-3 \u003c/li\u003e\n\u003cli\u003eEsau, K. (1980). \u003cem\u003eAnatomy of seed plants\u003c/em\u003e. John Wiley \u0026amp; Sons.\u003c/li\u003e\n\u003cli\u003eElka Georgieva., et al. (2005). Comparative analysis of the volatiles from flowers and leaves of three Gentiana species. \u003cem\u003eBiochemical Systematics and Ecology, \u003c/em\u003e33(9), 938-947. \u003cu\u003ehttps://doi.org/10.1016/j.bse.2005.01.002\u003c/u\u003e\u003c/li\u003e\n\u003cli\u003eGahan, P.B. (1984). Plant Histochemistry and Cytochemistry: An Introduction. \u003cem\u003eAcademic Press\u003c/em\u003e, London.\u003c/li\u003e\n\u003cli\u003eGamalei, Y. V. (1984). \u003cem\u003eStruktura i funktsii listiev [Structure and functions of leaves]\u003c/em\u003e. Nauka.\u003c/li\u003e\n\u003cli\u003eGamaleĭ,Yu. V. and Pakhomova,M. V. (1984). The minor leaf veins of dicotyledonous plants. III. Structure, functions and evolution. \u003cem\u003eJournal article\u003c/em\u003e, 69, 1159\u0026ndash;1166.\u003c/li\u003e\n\u003cli\u003eHongzhen Yu \u0026amp; Batzaya Gachmaa. (2025). A comprehensive and systemic review of the Gentiana: ethnobotany, traditional applications, phytochemistry, pharmacology, and toxicology in the Mongolian Plateau. \u003cem\u003eJournal of Ethnopharmacology. \u003c/em\u003ehttps://doi.org/10.1016/j.jep.2025.119573\u003c/li\u003e\n\u003cli\u003eHoukang Cao., et al. (2021). The effects of Gentiana dahurica Fisch on alcoholic liver disease revealed by RNA sequencing. \u003cem\u003eJournal of Ethnopharmacology\u003c/em\u003e, 279, 113422. \u003cu\u003ehttps://doi.org/10.1016/j.jep.2020.113422\u003c/u\u003e\u003c/li\u003e\n\u003cli\u003eKim, J. H., \u0026amp; Park, C. W. (2023). Comparative leaf anatomy of \u003cem\u003eGentiana\u003c/em\u003e species (Gentianaceae) in Korea. \u003cem\u003eJournal of Plant Biology, 66\u003c/em\u003e(1), 1\u0026ndash;10. https://doi.org/10.1007/s12345-023-0001-2 \u003c/li\u003e\n\u003cli\u003eKumar, R., \u0026amp; Lee, H. (2019). Seed morphology and germination in high‐altitude flora. \u003cem\u003eInternational Journal of Seed Science, 12\u003c/em\u003e(3), 45\u0026ndash;58. https://doi.org/10.1016/j.ijss.2019.03.004 \u003c/li\u003e\n\u003cli\u003eLi, Q., et al. (2019). Adaptive morphology in response to ecological niches. \u003cem\u003eJournal of Ecology, 107\u003c/em\u003e(4), 1801\u0026ndash;1812. https://doi.org/10.1111/1365-2745.13115 \u003c/li\u003e\n\u003cli\u003eLi, X., \u0026amp; Zhao, G. (2020). Leaf anatomy and photosynthetic characteristics of \u003cem\u003eGentiana macrophylla\u003c/em\u003e under different light conditions. \u003cem\u003ePhotosynthetica, 58\u003c/em\u003e(1), 1\u0026ndash;10. https://doi.org/10.1007/s11099-020-01089-7 \u003c/li\u003e\n\u003cli\u003eLiu, Y., Xu, Z., Wen, Z., Cao, H., Gao, J., \u0026amp; Ruan, H. (2019). Anatomical and physiological responses to drought stress in the leaves of drought‐tolerant and drought‐sensitive poplar species. \u003cem\u003eTrees, 33\u003c/em\u003e(5), 1315\u0026ndash;1330. https://doi.org/10.1007/s00468-019-01754-0 \u003c/li\u003e\n\u003cli\u003eNa Jia., et al. (2016). Iridoid glycosides from the flowers of \u003cem\u003eGentiana macrophylla\u003c/em\u003e Pall. ameliorate collagen-induced arthritis in rats. \u003cem\u003eJournal of Ethnopharmacology\u003c/em\u003e, 189, 1-9. https://doi.org/10.1016/j.jep.2016.05.027\u003c/li\u003e\n\u003cli\u003eRa-Yeong Choi., et al. (2019). Gentiopicroside isolated from Gentiana scabra Bge. inhibits adipogenesis in 3T3-L1 cells and reduces body weight in diet-induced obese mice. \u003cem\u003eBioorganic \u0026amp; Medicinal Chemistry Letters\u003c/em\u003e, 29 (14), 1699-1704. \u003cu\u003ehttps://doi.org/10.1016/j.bmcl.2019.05.038\u003c/u\u003e\u003c/li\u003e\n\u003cli\u003ePausheva, Z. P. (1974). \u003cem\u003ePraktikum po tsitologii rastenii [A practical course in plant cytology]\u003c/em\u003e. Kolos.\u003c/li\u003e\n\u003cli\u003eUrgamal M., Oyuntsetseg B., Nyambayar D., \u0026amp; Dulamsuren C. (2014). Conspectus of the vascular plants of Mongolia. In C. Sanchir \u0026amp; T. Jamsran (Eds.), (p. 334). Ulaanbaatar, Mongolia: \u003cem\u003e\u0026ldquo;Admon\u0026rdquo; Press\u003c/em\u003e.\u003c/li\u003e\n\u003cli\u003eRodriguez, P., \u0026amp; Alves, F. (2021). Integrating field and laboratory methods in botanical studies. \u003cem\u003eConservation Biology Quarterly, 15\u003c/em\u003e(1), 55\u0026ndash;68. https://doi.org/10.1080/19420889.2021.1874567 \u003c/li\u003e\n\u003cli\u003eRuzin, S. E. (1999). \u003cem\u003ePlant microtechnique and microscopy\u003c/em\u003e. Oxford University Press.\u003c/li\u003e\n\u003cli\u003eShiirevdamba, T., \u0026amp; Tserenkhand, G. (2014). Anatomical and morphological characteristics of \u003cem\u003eGentiana algida\u003c/em\u003e Pall. and \u003cem\u003eGentiana decumbens\u003c/em\u003e L. f. \u003cem\u003eProceedings of the Mongolian Academy of Sciences, 54\u003c/em\u003e(2), 3\u0026ndash;12. \u003c/li\u003e\n\u003cli\u003eShu-Bing Hou., et al. (2020). Seven new chemical constituents from the roots of \u003cem\u003eGentiana macrophylla\u003c/em\u003e pall. \u003cem\u003eFitoterapia\u003c/em\u003e, 141, 104476. \u003cu\u003ehttps://doi.org/10.1016/j.fitote.2020.104476\u003c/u\u003e\u003c/li\u003e\n\u003cli\u003eTserenbaljid, G. (2013). \u003cem\u003ePhoto collection of seed morphology of rare and very rare species in Mongolia\u003c/em\u003e. Ulaanbaatar.\u003c/li\u003e\n\u003cli\u003eWang, Y., \u0026amp; Chen, M. (2020). Conservation strategies for endemic plant species. \u003cem\u003eEnvironmental Conservation Reviews, 32\u003c/em\u003e(3), 200\u0026ndash;215. https://doi.org/10.1016/j.envconrev.2020.03.005 \u003c/li\u003e\n\u003cli\u003eWang, Y., Jiang, H., Gao, Y., Yin, Y., \u0026amp; Qin, J. (2021). Seed germination and seedling growth of \u003cem\u003eGentiana macrophylla\u003c/em\u003e under different temperature and light conditions. \u003cem\u003ePlant Diversity, 43\u003c/em\u003e(2), 159\u0026ndash;166. https://doi.org/10.1016/j.pld.2021.02.003 \u003c/li\u003e\n\u003cli\u003eZhang, L., et al. (2018). Vascular bundle organization in alpine plants. \u003cem\u003eJournal of Plant Research, 131\u003c/em\u003e(4), 587\u0026ndash;595. https://doi.org/10.1007/s10265-018-1023-7 \u003c/li\u003e\n\u003cli\u003eZhang, Y., Zhu, G., Zhao, Y., Xue, J., Zhao, Z., \u0026amp; Zhao, G. (2020). Phylogenetic relationships and historical biogeography of the genus \u003cem\u003eGentiana\u003c/em\u003e (Gentianaceae) in China. \u003cem\u003eJournal of Systematics and Evolution, 58\u003c/em\u003e(1), 3\u0026ndash;18. https://doi.org/10.1111/jse.12420 \u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"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":"Gentiana algida, Gentiana macrophylla, leaf anatomy, stem morphology, seed morphology, scanning electron microscopy","lastPublishedDoi":"10.21203/rs.3.rs-6236894/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6236894/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eTwo Gentiana species native to Mongolia, \u003cem\u003eGentiana algida \u003c/em\u003eand \u003cem\u003eGentiana macrophylla\u003c/em\u003e, are thoroughly anaturally and morphologically examined in this work. Field research was conducted in several provinces where specimens were collected and studied using scanning electron microscopy (SEM) and light microscopy. As a result, \u003cem\u003eGentiana macrophylla\u003c/em\u003e displayed an oval stem characterized by substantial sclerenchyma and numerous concentric vascular bundles, whereas \u003cem\u003eGentiana algida\u003c/em\u003e presented a cylindrical, hollow stem featuring a single row of small vascular bundles. Both species exhibited dorsoventral leaf morphology; nevertheless, differences in the configuration of chlorenchyma layers and vascular bundles highlighted their distinct adaptive strategies to local environmental conditions. Moreover, SEM analysis of seed form showed that unlike the smaller, lighter seeds of \u003cem\u003eGentiana macrophylla, Gentiana algida \u003c/em\u003ea produces bigger, denser seeds with a crystalline reticulate surface. These results improve our taxonomic knowledge of the genus Gentiana and provide important new perspectives on the adaptive mechanisms enabling life in very mountainous environments. Combining conventional botanical techniques with cutting-edge imaging technologies creates a solid basis for next research targeted at the sustainable use and preservation of these ecologically and medically important species.\u003c/p\u003e","manuscriptTitle":"Comparative Anatomical and Morphological Study of Gentiana Algida and Gentiana Macrophylla in Mongolia","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-04-07 12:45:23","doi":"10.21203/rs.3.rs-6236894/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":"4a7c1c88-347d-47c5-bad8-83a6f9c3d8ad","owner":[],"postedDate":"April 7th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-10-22T20:53:20+00:00","versionOfRecord":[],"versionCreatedAt":"2025-04-07 12:45:23","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6236894","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6236894","identity":"rs-6236894","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}