Enhancing sustainable cultivation: The role of plant growth promoting fungi in optimizing Cucumis sativus L. growth

Enhancing sustainable cultivation: The role of plant growth promoting fungi in optimizing Cucumis sativus L. growth | 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 Enhancing sustainable cultivation: The role of plant growth promoting fungi in optimizing Cucumis sativus L. growth Rukhsana Qadir, Abdul Hamid Wani, Mohd Yaqub Bhat, Bilal Ahmad Dar, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6156187/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 9 You are reading this latest preprint version Abstract Rhizosphere and root associated fungi, commonly known as plant growth promoting fungi (PGPF), facilitate the growth and productivity of Cucumis sativus L. through the formation of mutualistic relationships with the plant roots. These fungi enhance nutrient uptake, promote root development and bolster stress tolerance via the secretion of growth regulating hormones. Despite numerous PGP fungal species have been isolated from the rhizosphere of C. sativus , their specific roles in modulating various growth parameters of the C. sativus remain insufficiently defined. In the present study, effect of seven PGP fungal species were evaluated such as Penicillium chrysogenum , Paecilomyces variotii , Talaromyces purpureogenus , Paecilomyces brunneolus , Aspergillus flavus , Trichoderma viride , and Trichoderma atroviride on the growth parameters of C. sativus . Our results revealed that all PGP fungal species positively influenced growth parameters such as, root length, shoot length, fresh and dry biomass, leaf area, specific leaf area, moisture content, flower number, leaf area ratio, leaf weight ratio and chlorophyll content, albeit with varying degrees of efficiency. Notably, Trichoderma atroviride exhibited a more pronounced impact on plant growth as compared to other PGP fungal species and the control. Our results suggest that these PGP fungal species may contribute to sustainable cultivation practices by enhancing crop yield and reducing the dependency on chemical fertilizers and pesticides. Cucumis sativus (PGPF) Growth parameters Figures Figure 1 Figure 2 1. Introduction The growing global population and the corresponding rise in demand for agricultural production have resulted in the extensive use of pesticides and chemical fertilizers worldwide (Penuelas et al., 2023 ). The use of chemical fertilizers enhances crop yields however, substantial amounts of these chemicals accumulate in the soil, posing potential health risks and disrupting the soil's dynamics (Indira et al., 2022 ; Vanshree et al., 2022 ; Zhou et al., 2024 ). Moreover, chemical fertilizers are expensive, contributing significantly to the overall cost of agricultural production (Alexander et al., 2023 ). Rhizosphere and root associated fungi, commonly referred as plant growth promoting fungi (PGPF), confer numerous benefits to plants. These fungi play a pivotal role in biogeochemical and nutrient cycles, facilitating the utilization of decomposing organic matter through mineral solubilization (Thepbandit and Athinuwat, 2024 ). Modern agricultural practices emphasize minimizing the reliance on chemical fertilizers and promoting the application of plant growth promoting fungi (PGPF) as alternative strategies to enhance sustainable farming methods (Chaudhary et al., 2022 ). PGPF have been utilized as biofertilizers and as supplementary agents to chemical fertilizers in the cultivation of field vegetables and forage crops (Kumar et al., 2022 ). PGPF, are vital constituents of the rhizospheric soil and generally represent a larger proportion of the soil biomass compared to bacteria (Malviya et al., 2011 ; Adedayo and Babalola, 2023 ). PGPF, also produce bioactive compounds, including plant hormones (gibberellins, auxins, cytokinins, and siderophores) and enhance plant resilience to abiotic stresses (Naimuzzaman et al., 2024 ). Furthermore, rhizospheric fungi can suppress plant pathogens through various antagonistic mechanisms, including the secretion of hydrolytic enzymes, mycoparasitism, competition for saprophytic colonization, and the induction of systemic resistance in host plants (Tyśkiewicz et al., 2022 ). Previous study has also demonstrated that soil treated with (PGPF), species significantly enhances plant growth, phosphorus uptake, and yield in various important crops, such as brinjal (Li et al., 2021 ), wheat (Mohamed et al., 2022 ), maize (Ain et al., 2024 ). Several other studies were carried on PGPF genera, including Gliocladium , Penicillium , Phoma , Phytophthora , Rhizoctonia , Talaromyces , and Trichoderma , as effective in promoting growth and development in crops like orange (Omomowo et al., 2020 ), apple (Kuzin et al., 2020 ), pear (Cantabella et al., 2020 ), tomato, (Attia et al., 2022 ) and carrot (Pandit et al., 2022 ). Additionally, these fungi contribute to enhancing the innate immunity of plants and stimulating the production of essential secondary metabolites. Cucumis sativus L. commonly known as cucumber, is one of the most globally consumed vegetable. Its popularity is due to its nutritional benefits, adaptability to diverse climatic conditions, and versatility in culinary applications, making it a valuable crop in the agricultural market. Producing high quality C. sativus significantly boosts market sales and enhances farmer income, making it a valuable crop choice for sustainable agriculture. In this study, a greenhouse experiment was carried out to elucidate the roles of various PGP fungal species isolated from the rhizosphere of C. sativus . To evaluate the effects of various PGP fungal species on growth parameters of test plant, seven isolated PGP fungal species such as Penicillium chrysogenum , Paecilomyces variotii , Talaromyces purpureogenus , Paecilomyces brunneolus , Aspergillus flavus , Trichoderma viride and Trichoderma atroviride were evaluated for their efficacy on growth parameter of test plant such as root length, shoot length, fresh mass, dry mass, leaf area, specific leaf area, moisture content, number of flowers, leaf area ratio, leaf weight ratio and chlorophyll content were evaluated. This avenue of inquiry aims to elucidate the role PGP fungal species within the dynamic interface between plant roots and soil and also helps in developing innovative strategies to improve the resilience and productivity of agricultural systems in the face of increasing challenges. 2. Materials and methods For the preparation fungal inoculum, seven different fungal species Penicillium chrysogenum , Paecilomyces variotii , Talaromyces purpureogenus , Paecilomyces brunneolus , Aspergillus flavus , Trichoderma viride and Trichoderma atroviride were cultured on PDA for 5 days at 28°C. After this incubation period, the actively growing fungal plates were flooded with sterile water separately and gentle agitation was employed to liberate the spores from the mycelia. The liberated spores were then carefully separated from the mycelial debris by passing them through a sieve and were then collected in a beaker. The spore concentration was then adjusted by dilution with sterile water to a final concentration of 10 4 to 10 6 spores/ml. The concentration of fungal spores was selected based on an OD reading of 0.1 of the suspension at 600 nm using a Spectrophotometer. To carry out the Pot house experiment, soil was collected from Kashmir University Botanical Garden and passed through 2mm sieve to remove debris and combined with sand in 3:1 ratio. The soil was packed in sterile polythene bags and autoclaved at 15lb pressure for 20 to 30 min. To ensure the absence of any living microorganisms or contaminants that could interfere with the experiment, the potting mixture was autoclaved thrice. Following the first autoclave cycle, the material was autoclaved again the following day. This allowed any remaining heat-resistant spores within the material to potentially germinate overnight and subsequently be eradicated. The third autoclave cycle was performed on the third day to ensure thorough eradication of any potential pathogens, including bacteria, fungi, or weed seeds. C. sativus seeds were sterilized in (2% NaOCl for 10 minutes followed by streptomycin (100 µg/ml) + cycloheximide (100 µg/ml) for 12 hrs and placed on sterile wet filter paper in 150 mm diameter Petri dishes at 28˚C and 95% humidity. After sterilization, the inoculated seeds were sown into a seedling tray containing sterilized soil at 26˚C and 60% relative humidity. Furthermore, the C. sativus seedlings at the three-leaf and heart stage were transplanted from the tray to 15cm clay pots (5 replicates for each treatment in a completely randomized block design) filled with triple autoclaved sand and soil in a ratio of 3:1. The pots were placed in a green house, the pots were irrigated with double distilled water every alternate day and the fungal spore suspension (10 ml in each pot) were applied individually at intervals of 7 days. Plants were grown in the green house for 3 months during which various observations like root length, shoot lengths, fresh mass, dry mass, specific leaf area etc were recorded. 3. Statistical analysis Data were analysed as a one-way ANOVA using the IBM SPSS Statistics 25 software package (IBM, USA). Significant differences between treatments were identified using Duncan’s new multiple range test at the p < 0.05 level of significance. 4. Results The effect of seven PGP fungal species were evaluated such as Penicillium chrysogenum , Paecilomyces variotii , Talaromyces purpureogenus , Paecilomyces brunneolus , Aspergillus flavus , Trichoderma viride , and Trichoderma atroviride on the growth parameters of C. sativus . Results revealed (Fig. 1; Table 1) that all the PGP fungal species considerably improved growth parameters such as root length, shoot length, fresh and dry biomass, leaf area, specific leaf area, moisture content, flower number, leaf area ratio, leaf weight ratio and chlorophyll content of C. sativus . Root length was highest (12.1 cm) under Trichoderma atroviride and lowest (3.36 cm) under control. ANOVA and post-hoc analysis revealed that the increase in root length under the Trichoderma atroviride treatment is significantly higher compared to other treatments. Shoot length varied from a highest of 58.66 cm under Trichoderma atroviride and lowest of 33 cm under control treatment. Like root and shoot length, shoot fresh mass was also highest (18.22 g/plant) under Trichoderma atroviride and lowest (8.78 g/plant) under the control treatment. Shoot dry mass also differed under different treatments and was significantly higher (4.09 g/plant) under Trichoderma atroviride and was lowest (1.33 g/plant) under control treatment. The number of flowers ranged from 10.66 under Trichoderma atroviride to 2.33 under control. ANOVA and post-hoc analysis revealed that treatments such as Trichoderma atroviride , Trichoderma viride , Aspergillus flavus , Paecilomyces brunneolus , Talaromyces purpureogenus , Paecilomyces variotii , Penicillium chrysogenum and control were significantly different from one another. Furthermore, the moisture content percent was highest in control (85.05%) and lowest in Trichoderma atroviride (77.92%). ANOVA and post-hoc analysis revealed that treatments such as Trichoderma atroviride , Trichoderma viride , Aspergillus flavus , Paecilomyces brunneolus , Talaromyces purpureogenus , Paecilomyces variotii , Penicillium chrysogenum and control were not significantly different from one another. Leaf area was highest in Trichoderma atroviride (112.35 cm 2 ) followed by the other fungi and least was observed in control (49.3cm 2 ). Specific leaf area was highest in Paecilomyces variotii (166.42 cm 2 /g) and lowest in Trichoderma atroviride (82.42 cm 2 /g). Leaf area ratio was highest in Talaromyces purpureogenus (34.56 cm 2 /g) and lowest in Trichoderma atroviride (21.33 cm 2 /g). There were no significant differences between Paecilomyces variotii and control. Leaf area ratio was highest in Talaromyces purpureogenus (34.56 cm 2 /g) and lowest in Trichoderma atroviride (21.33 cm 2 /g). Leaf weight ratio was highest in Talaromyces purpureogenus and Trichoderma atroviride (0.25 each). Additionally, chlorophyll content was highest in Trichoderma atroviride (7.95 mg/g) and lowest in control (1.95 mg/g). The effects of different treatments Penicillium chrysogenum, Paecilomyces variotii, Talaromyces purpureogenus , Paecilomyces brunneolus , Aspergillus flavus , Trichoderma viride and Trichoderma atroviride on root length, shoot length, fresh mass, dry mass, leaf area, number of flowers and chlorophyll content revealed significant differences in these growth parameters across the treatments, as compared to control, suggesting that the rhizospheric fungi have a discernible impact on plant growth compared to the control group. Each parameter was measured in five replicates (n = 5), and the results are presented as mean values with their respective standard deviations (SD). 5. Discussion The study presents a comprehensive evaluation of the effects of plant growth promoting fungi (PGPF) on the growth parameters of C. sativus . Previous studies have also demonstrated that specific fungal species can stimulate key growth parameters, including seed germination and other developmental stages. Trichoderma species, which are well-documented for their biostimulators effects, likely due to their ability to produce phytohormones and solubilize essential nutrients (Asghar et al., 2024 ) have also been shown to elicit systemic resistance in plants, aiding them in coping with abiotic and biotic stresses (Gupta and Bar, 2020 ). The use of these fungal inoculants provides insight into reducing chemical inputs while maintaining high crop yield. Chemical fertilizers and pesticides, while effective, are increasingly scrutinized due to their impact on human and environmental health, as well as the development of pathogen resistance (Indira et al., 2022 ; Vanshree et al., 2022 ). In the context of plant growth enhancement, different fungal treatments were used to evaluate how these fungal species, which include Penicillium chrysogenum , Paecilomyces variotii , Talaromyces purpureogenus , Paecilomyces brunneolus , Aspergillus flavus , Trichoderma viride , and Trichoderma atroviride , impact various growth parameters of C. sativus . The results highlight that C. sativus seedlings treated with diverse rhizospheric plant growth promoting fungal species, particularly Trichoderma atroviride , significantly improves growth parameters like root and shoot length, fresh and dry weight, and leaf chlorophyll content. For instance, Trichoderma atroviride -treated plants exhibited the highest root length (12.1 cm) and shoot length (58.66 cm), indicating a robust root system and enhanced vegetative growth. Interestingly, treatments also affected moisture retention, with control showing the highest moisture content, contrasting with Trichoderma atroviride , which exhibited the lowest. This variation in moisture content could reflect the different capacities of fungal species to enhance water use efficiency and dry mass accumulation or adapt to fluctuating water availability in the soil matrix. Moreover, Paecilomyces variotii , and Talaromyces purpureogenus , which showed significant improvements in specific leaf area and leaf area ratio, could be contributing to an optimized plant architecture that maximizes light capture, a feature particularly advantageous in competitive plant environments. This investigation aligns with current research trends that emphasize sustainable agricultural practices and the reduced use of chemical fertilizers and pesticides. Given the context of global food security and the associated challenges posed by environmental stressors on crop productivity, this study offers a promising approach to addressing the yield demands of C. sativus which is widely cultivated and consumed in world. 6. Conclusion This study highlights that plant growth promoting fungi (PGPF), can serve as eco-friendly bio stimulants, enhancing crop resilience and productivity while minimizing dependence on pesticides and chemical fertilizers. The study also demonstrates that PGP fungal species significantly improved growth parameters, including root and shoot length, biomass accumulation and chlorophyll content which indicates strengthened plant structure and photosynthetic efficiency. The observed bio-enhancement suggests that such fungi could play a vital role in advancing food security and eco-friendly crop management practices. Declarations ACKNOWLEDGEMENTS We are highly thankful to head Department of Botany and Plant Pathology, Mycology and Microbiology Laboratory, University of Kashmir for providing feasible facilities to carry out this work. AUTHORS CONTRIBUTIONS All authors listed in this submission have made significant contributions to the conceptualization and design of the study, as well as the analysis and interpretation of the data. Each author has actively participated in drafting the manuscript, critically revising it for key intellectual content, and approving the final version for publication Data availability statement No datasets were generated or analysed during the current study CONFLICT OF INTEREST The authors declared that there is no conflict of interest. Funding Nil Ethics, Consent to participate, Consent to publish Not applicable Clinical trial Not applicable References Adedayo, A. A., & Babalola, O. O. (2023). Fungi that promote plant growth in the rhizosphere boost crop growth. Journal of Fungi , 9 (2), 239. https://doi.org/10.3390/jof9020239 Ain, Q. U., Hussain, H. A., Zhang, Q., Maqbool, F., Ahmad, M., Mateen, A., ... & Imran, A. (2024). Coordinated influence of Funneliformis mosseae and different plant growth-promoting bacteria on growth, root functional traits, and nutrient acquisition by maize. Mycorrhiza , 1-12. DOI:10.1007/s00572-024-01165-5 Alexander, P., Arneth, A., Henry, R., Maire, J., Rabin, S., & Rounsevell, M. D. (2023). High energy and fertilizer prices are more damaging than food export curtailment from Ukraine and Russia for food prices, health and the environment. Nature Food , 4 (1), 84-95. DOI: 10.1038/s43016-022-00659-9 Asghar, W., Craven, K. D., Kataoka, R., Mahmood, A., Asghar, N., Raza, T., & Iftikhar, F. (2024). The application of Trichoderma spp., an old but new useful fungus, in sustainable soil health intensification: A comprehensive strategy for addressing challenges. Plant Stress , 100455. https://doi.org/10.1016/j.stress.2024.100455 Attia, M. S., Abdelaziz, A. M., Al-Askar, A. A., Arishi, A. A., Abdelhakim, A. M., & Hashem, A. H. (2022). Plant growth-promoting fungi as biocontrol tool against Fusarium wilt disease of tomato plant. Journal of Fungi , 8 (8), 775. https://doi.org/10.3390/jof8080775 Cantabella, D., Dolcet-Sanjuan, R., Casanovas, M., Solsona, C., Torres, R., & Teixidó, N. (2020). Inoculation of in vitro cultures with rhizosphere microorganisms improve plant development and acclimatization during immature embryo rescue in nectarine and pear breeding programs. Scientia Horticulturae , 273 , 109643. DOI:10.1016/j.scienta.2020.109643 Chaudhary, P., Singh, S., Chaudhary, A., Sharma, A., & Kumar, G. (2022). Overview of biofertilizers in crop production and stress management for sustainable agriculture. Frontiers in Plant Science , 13 , 930340. https://doi.org/10.3389/fpls.2022.930340 Gupta, R., & Bar, M. (2020). Plant immunity, priming, and systemic resistance as mechanisms for Trichoderma spp. biocontrol. Trichoderma : Host Pathogen Interactions and Applications, 81-110. DOI:10.1007/978-981-15-3321-1_5 Indira D. P., Manjula M., Bhavani R.V. (2022) Agrochemicals, Environment, and Human Health; Annual Review of Environment and Resources, 47, 399-421. https://doi.org/10.1146/annurev-environ-120920-111015 Malviya, J. M., Kiran Singh, K. S., & Vaibhavi Joshi, V. J. (2011). Effect of phosphate solubilizing fungi on growth and nutrient uptake of ground nut (Arachis hypogaea) plants. Adv Biores, 2,110-113. Kumar, S., Sindhu, S. S., & Kumar, R. (2022). Biofertilizers: An ecofriendly technology for nutrient recycling and environmental sustainability. Current Research in Microbial Sciences , 3 , 100094. https://doi.org/10.1016/j.crmicr.2021.100094 Kuzin, A., Solovchenko, A., Stepantsova, L., & Pugachev, G. (2020). Soil fertility management in apple orchard with microbial biofertilizers. In E3S Web of Conferences (Vol. 222, p. 03020). EDP Sciences. DOI:10.1051/e3sconf/202022203020 Li, X., Li, D., Yan, J., Zhang, Y., Wang, H., Zhang, J., ... & Li, B. (2021). Effect of plant-growth-promoting fungi on eggplant ( Solanum melongena L.) in new reclamation land. Agriculture , 11 (11), 1036. https://doi.org/10.3390/agriculture11111036 Mohamed, A. H., Abd el-Megeed, F. H., Hassanein, N. M., Youseif, S. H., Farag, P. F., Saleh, S. A., ... & Abdel-Azeem, A. M. (2022). Native rhizospheric and endophytic fungi as sustainable sources of plant growth promoting traits to improve wheat growth under low nitrogen input. Journal of Fungi , 8 (2), 94. https://doi.org/10.3390/jof8020094 Naimuzzaman, M., Rahman, F., Alvi, A. T., Yeasmin, L., Mittra, P. K., & Roy, S. K. (2024). Plant growth–promoting fungi in plants: Insights from stress tolerance mechanism. In Beneficial Microbes for Sustainable Agriculture Under Stress Conditions ,469-511. Academic Press. https://doi.org/10.1016/b978-0-443-13193-6.00023-3 Omomowo, I. O., Adedayo, A. A., & Omomowo, O. I. (2020). Biocontrol potential of rhizospheric fungi from moringa oleifera, their phytochemicals and secondary metabolite assessment against spoilage fungi of sweet orange ( citrus sinensis ). Asian Journal of Applied Sciences , 8 (1). https://doi.org/10.24203/ajas.v8i1.6047 Pandit, M. A., Kumar, J., Gulati, S., Bhandari, N., Mehta, P., Katyal, R., ... & Kaur, J. (2022). Major biological control strategies for plant pathogens. Pathogens , 11 (2), 273. https://doi.org/10.3390/pathogens11020273 Penuelas, J., Coello, F., & Sardans, J. (2023). A better use of fertilizers is needed for global food security and environmental sustainability. Agriculture & Food Security , 12 (1), 1-9. https://doi.org/10.1186/s40066-023-00409-5 · Sun, X., Liu, F., Jiang, W., Zhang, P., Zhao, Z., Liu, X., ... & Sun, Q. (2023). Talaromyces purpurogenus Isolated from Rhizosphere Soil of Maize Has Efficient Organic Phosphate-Mineralizing and Plant Growth-Promoting Abilities. Sustainability , 15(7), 5961. DOI:10.3390/su15075961 Thepbandit, W., & Athinuwat, D. (2024). Rhizosphere microorganisms supply availability of soil nutrients and induce plant defense. Microorganisms , 12 (3), 558. https://doi.org/10.3390/microorganisms12030558 Tyśkiewicz, R., Nowak, A., Ozimek, E., & Jaroszuk-ściseł, J. (2022). Trichoderma : The current status of its application in agriculture for the biocontrol of fungal phytopathogens and stimulation of plant growth. International Journal of Molecular Sciences , 23 (4), 2329. https://doi.org/10.3390/ijms23042329 Vanshree, C. R., Singhal, M., Sexena, M., Sankhla, M. S., Parihar, K., Jadhav, E. B., Yadav, C. S. (2022). Microbes as biocontrol agent: From crop protection till food security. In Relationship Between Microbes and the Environment for Sustainable Ecosystem Services. 1: 215-237. https://doi.org/10.1016/B978-0-323-89938-3.00011-6 Zhou, W., Li, M., & Achal, V. (2024). A comprehensive review on environmental and human health impacts of chemical pesticide usage. Emerging Contaminants , 100410. https://doi.org/10.1016/j.emcon.2024.100410 Table 1 Table 1 is available in the Supplementary Files section. Additional Declarations No competing interests reported. Supplementary Files Table1.docx Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 21 Apr, 2025 Reviews received at journal 17 Apr, 2025 Reviews received at journal 10 Apr, 2025 Reviewers agreed at journal 08 Apr, 2025 Reviewers agreed at journal 04 Apr, 2025 Reviewers invited by journal 02 Apr, 2025 Editor assigned by journal 31 Mar, 2025 Submission checks completed at journal 29 Mar, 2025 First submitted to journal 29 Mar, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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-6156187","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":445560380,"identity":"da6080b5-3270-44d9-950f-b77f205a2451","order_by":0,"name":"Rukhsana Qadir","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABFUlEQVRIie3Pv0rEMBzA8ZRCuvTu1pQTfIWUQm6p56v0KMTlhrp1cAgIdZFz7b1F5F6gR7BdArdmcDiXTg51kQ6H2voHQVp6o0O+0y8hH5IAoNP9yyD8mXAGsD+ftOO+WR1JIho6DBgsOJYAUAmDZwNkllL6VF89no4LcS9ibJpecTvbB7HvAUs88A5yomjh2XnpriWNthLDMZHSZYGkBNiUqg6C0EUyBbB9zxJvGbZNopYuWyTCB8gmfcSp38Q53z23BBmbdJDQHI0SseDq8xZscPRFSC+xSzodrUS4VmXUkCBEMr9Mm794sO8vFiVO/SrOVrtw88IO7/PJzTWvqth37yyRd5Hfsr8bsOPQANHpdDrddx+wBW4JQfwQMwAAAABJRU5ErkJggg==","orcid":"","institution":"University of Kashmir","correspondingAuthor":true,"prefix":"","firstName":"Rukhsana","middleName":"","lastName":"Qadir","suffix":""},{"id":445560381,"identity":"fa6cd090-58f9-4d4f-b1e6-70c4b683cd72","order_by":1,"name":"Abdul Hamid Wani","email":"","orcid":"","institution":"University of Kashmir","correspondingAuthor":false,"prefix":"","firstName":"Abdul","middleName":"Hamid","lastName":"Wani","suffix":""},{"id":445560382,"identity":"0d934aa8-d3fa-4a21-a2e6-96399011d85f","order_by":2,"name":"Mohd Yaqub Bhat","email":"","orcid":"","institution":"University of Kashmir","correspondingAuthor":false,"prefix":"","firstName":"Mohd","middleName":"Yaqub","lastName":"Bhat","suffix":""},{"id":445560383,"identity":"f794ec47-fe1e-48b0-8332-180b60b9f8d2","order_by":3,"name":"Bilal Ahmad Dar","email":"","orcid":"","institution":"University of Kashmir","correspondingAuthor":false,"prefix":"","firstName":"Bilal","middleName":"Ahmad","lastName":"Dar","suffix":""},{"id":445560384,"identity":"658526ef-f8ad-452c-9899-326092b60579","order_by":4,"name":"Humeera Yousuf","email":"","orcid":"","institution":"University of Kashmir","correspondingAuthor":false,"prefix":"","firstName":"Humeera","middleName":"","lastName":"Yousuf","suffix":""}],"badges":[],"createdAt":"2025-03-04 17:08:19","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6156187/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6156187/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":81103612,"identity":"73eac6f3-bdf1-4838-aa37-969636a446ed","added_by":"auto","created_at":"2025-04-22 09:11:37","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":534188,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of plant growth promoting fungi on growth parameters of\u003cem\u003e C. sativus\u003c/em\u003e. Letters represent the significant differences p \u0026lt; 0.05\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6156187/v1/e3863d5fd31d08d2df020397.png"},{"id":81103930,"identity":"99cf711a-0712-49b0-9806-030e68a88103","added_by":"auto","created_at":"2025-04-22 09:19:37","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":288124,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of plant growth promoting fungal species on growth parameters of \u003cem\u003eC. sativus \u003c/em\u003eWhere T0: Control, T1:\u003cem\u003ePenicillium chrysogenum, \u003c/em\u003eT2: \u003cem\u003ePaecilomyces variotii,\u003c/em\u003e T3: \u003cem\u003eTalaromyces purpureogenus\u003c/em\u003e, T4: \u003cem\u003ePaecilomyces brunneolus\u003c/em\u003e, T5: \u003cem\u003eAspergillus flavus\u003c/em\u003e, T6: \u003cem\u003eTrichoderma viride\u003c/em\u003e T7:\u003cem\u003e Trichoderma atroviride\u003c/em\u003e\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-6156187/v1/9daaeb671e526875085150a8.png"},{"id":81105545,"identity":"20d0181f-9838-48b7-a6b0-bb3c0e995106","added_by":"auto","created_at":"2025-04-22 09:35:38","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1295922,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6156187/v1/b606e320-5904-4186-b388-70081d3069b3.pdf"},{"id":81103624,"identity":"a42c5f70-4679-4dde-aed5-00fa7ecaf238","added_by":"auto","created_at":"2025-04-22 09:11:39","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":675173,"visible":true,"origin":"","legend":"","description":"","filename":"Table1.docx","url":"https://assets-eu.researchsquare.com/files/rs-6156187/v1/6dd6ad2b56ab8110023b1704.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Enhancing sustainable cultivation: The role of plant growth promoting fungi in optimizing Cucumis sativus L. growth","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eThe growing global population and the corresponding rise in demand for agricultural production have resulted in the extensive use of pesticides and chemical fertilizers worldwide (Penuelas et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). The use of chemical fertilizers enhances crop yields however, substantial amounts of these chemicals accumulate in the soil, posing potential health risks and disrupting the soil's dynamics (Indira et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Vanshree et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Zhou et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Moreover, chemical fertilizers are expensive, contributing significantly to the overall cost of agricultural production (Alexander et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Rhizosphere and root associated fungi, commonly referred as plant growth promoting fungi (PGPF), confer numerous benefits to plants. These fungi play a pivotal role in biogeochemical and nutrient cycles, facilitating the utilization of decomposing organic matter through mineral solubilization (Thepbandit and Athinuwat, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Modern agricultural practices emphasize minimizing the reliance on chemical fertilizers and promoting the application of plant growth promoting fungi (PGPF) as alternative strategies to enhance sustainable farming methods (Chaudhary et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). PGPF have been utilized as biofertilizers and as supplementary agents to chemical fertilizers in the cultivation of field vegetables and forage crops (Kumar et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). PGPF, are vital constituents of the rhizospheric soil and generally represent a larger proportion of the soil biomass compared to bacteria (Malviya et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Adedayo and Babalola, \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003ePGPF, also produce bioactive compounds, including plant hormones (gibberellins, auxins, cytokinins, and siderophores) and enhance plant resilience to abiotic stresses (Naimuzzaman et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Furthermore, rhizospheric fungi can suppress plant pathogens through various antagonistic mechanisms, including the secretion of hydrolytic enzymes, mycoparasitism, competition for saprophytic colonization, and the induction of systemic resistance in host plants (Tyśkiewicz et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Previous study has also demonstrated that soil treated with (PGPF), species significantly enhances plant growth, phosphorus uptake, and yield in various important crops, such as brinjal (Li et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), wheat (Mohamed et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), maize (Ain et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Several other studies were carried on PGPF genera, including \u003cem\u003eGliocladium\u003c/em\u003e, \u003cem\u003ePenicillium\u003c/em\u003e, \u003cem\u003ePhoma\u003c/em\u003e, \u003cem\u003ePhytophthora\u003c/em\u003e, \u003cem\u003eRhizoctonia\u003c/em\u003e, \u003cem\u003eTalaromyces\u003c/em\u003e, and \u003cem\u003eTrichoderma\u003c/em\u003e, as effective in promoting growth and development in crops like orange (Omomowo et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), apple (Kuzin et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), pear (Cantabella et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), tomato, (Attia et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) and carrot (Pandit et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Additionally, these fungi contribute to enhancing the innate immunity of plants and stimulating the production of essential secondary metabolites.\u003c/p\u003e \u003cp\u003e \u003cem\u003eCucumis sativus\u003c/em\u003e L. commonly known as cucumber, is one of the most globally consumed vegetable. Its popularity is due to its nutritional benefits, adaptability to diverse climatic conditions, and versatility in culinary applications, making it a valuable crop in the agricultural market. Producing high quality \u003cem\u003eC. sativus\u003c/em\u003e significantly boosts market sales and enhances farmer income, making it a valuable crop choice for sustainable agriculture. In this study, a greenhouse experiment was carried out to elucidate the roles of various PGP fungal species isolated from the rhizosphere of \u003cem\u003eC. sativus\u003c/em\u003e. To evaluate the effects of various PGP fungal species on growth parameters of test plant, seven isolated PGP fungal species such as \u003cem\u003ePenicillium chrysogenum\u003c/em\u003e, \u003cem\u003ePaecilomyces variotii\u003c/em\u003e, \u003cem\u003eTalaromyces purpureogenus\u003c/em\u003e, \u003cem\u003ePaecilomyces brunneolus\u003c/em\u003e, \u003cem\u003eAspergillus flavus\u003c/em\u003e, \u003cem\u003eTrichoderma viride\u003c/em\u003e and \u003cem\u003eTrichoderma atroviride\u003c/em\u003e were evaluated for their efficacy on growth parameter of test plant such as root length, shoot length, fresh mass, dry mass, leaf area, specific leaf area, moisture content, number of flowers, leaf area ratio, leaf weight ratio and chlorophyll content were evaluated. This avenue of inquiry aims to elucidate the role PGP fungal species within the dynamic interface between plant roots and soil and also helps in developing innovative strategies to improve the resilience and productivity of agricultural systems in the face of increasing challenges.\u003c/p\u003e"},{"header":"2. Materials and methods","content":"\u003cp\u003eFor the preparation fungal inoculum, seven different fungal species \u003cem\u003ePenicillium chrysogenum\u003c/em\u003e, \u003cem\u003ePaecilomyces variotii\u003c/em\u003e, \u003cem\u003eTalaromyces purpureogenus\u003c/em\u003e, \u003cem\u003ePaecilomyces brunneolus\u003c/em\u003e, \u003cem\u003eAspergillus flavus\u003c/em\u003e, \u003cem\u003eTrichoderma viride\u003c/em\u003e and \u003cem\u003eTrichoderma atroviride\u003c/em\u003e were cultured on PDA for 5 days at 28\u0026deg;C. After this incubation period, the actively growing fungal plates were flooded with sterile water separately and gentle agitation was employed to liberate the spores from the mycelia. The liberated spores were then carefully separated from the mycelial debris by passing them through a sieve and were then collected in a beaker. The spore concentration was then adjusted by dilution with sterile water to a final concentration of 10\u003csup\u003e4\u003c/sup\u003e to 10\u003csup\u003e6\u003c/sup\u003e spores/ml. The concentration of fungal spores was selected based on an OD reading of 0.1 of the suspension at 600 nm using a Spectrophotometer. To carry out the Pot house experiment, soil was collected from Kashmir University Botanical Garden and passed through 2mm sieve to remove debris and combined with sand in 3:1 ratio. The soil was packed in sterile polythene bags and autoclaved at 15lb pressure for 20 to 30 min. To ensure the absence of any living microorganisms or contaminants that could interfere with the experiment, the potting mixture was autoclaved thrice. Following the first autoclave cycle, the material was autoclaved again the following day. This allowed any remaining heat-resistant spores within the material to potentially germinate overnight and subsequently be eradicated. The third autoclave cycle was performed on the third day to ensure thorough eradication of any potential pathogens, including bacteria, fungi, or weed seeds. \u003cem\u003eC. sativus\u003c/em\u003e seeds were sterilized in (2% NaOCl for 10 minutes followed by streptomycin (100 \u0026micro;g/ml)\u0026thinsp;+\u0026thinsp;cycloheximide (100 \u0026micro;g/ml) for 12 hrs and placed on sterile wet filter paper in 150 mm diameter Petri dishes at 28˚C and 95% humidity. After sterilization, the inoculated seeds were sown into a seedling tray containing sterilized soil at 26˚C and 60% relative humidity. Furthermore, the \u003cem\u003eC. sativus\u003c/em\u003e seedlings at the three-leaf and heart stage were transplanted from the tray to 15cm clay pots (5 replicates for each treatment in a completely randomized block design) filled with triple autoclaved sand and soil in a ratio of 3:1. The pots were placed in a green house, the pots were irrigated with double distilled water every alternate day and the fungal spore suspension (10 ml in each pot) were applied individually at intervals of 7 days. Plants were grown in the green house for 3 months during which various observations like root length, shoot lengths, fresh mass, dry mass, specific leaf area etc were recorded.\u003c/p\u003e"},{"header":"3. Statistical analysis","content":"\u003cp\u003eData were analysed as a one-way ANOVA using the IBM SPSS Statistics 25 software package (IBM, USA). Significant differences between treatments were identified using Duncan\u0026rsquo;s new multiple range test at the p\u0026thinsp;\u0026lt;\u0026thinsp;0.05 level of significance.\u003c/p\u003e"},{"header":"4. Results","content":"\u003cp\u003eThe effect of seven PGP fungal species were evaluated such as \u003cem\u003ePenicillium chrysogenum\u003c/em\u003e, \u003cem\u003ePaecilomyces variotii\u003c/em\u003e, \u003cem\u003eTalaromyces purpureogenus\u003c/em\u003e, \u003cem\u003ePaecilomyces brunneolus\u003c/em\u003e, \u003cem\u003eAspergillus flavus\u003c/em\u003e, \u003cem\u003eTrichoderma viride\u003c/em\u003e, and \u003cem\u003eTrichoderma atroviride\u003c/em\u003e on the growth parameters of \u003cem\u003eC. sativus\u003c/em\u003e. Results revealed (Fig.\u0026nbsp;1; Table\u0026nbsp;1) that all the PGP fungal species considerably improved growth parameters such as root length, shoot length, fresh and dry biomass, leaf area, specific leaf area, moisture content, flower number, leaf area ratio, leaf weight ratio and chlorophyll content of \u003cem\u003eC. sativus\u003c/em\u003e. Root length was highest (12.1 cm) under \u003cem\u003eTrichoderma atroviride\u003c/em\u003e and lowest (3.36 cm) under control. ANOVA and post-hoc analysis revealed that the increase in root length under the \u003cem\u003eTrichoderma atroviride\u003c/em\u003e treatment is significantly higher compared to other treatments. Shoot length varied from a highest of 58.66 cm under \u003cem\u003eTrichoderma atroviride\u003c/em\u003e and lowest of 33 cm under control treatment. Like root and shoot length, shoot fresh mass was also highest (18.22 g/plant) under \u003cem\u003eTrichoderma atroviride\u003c/em\u003e and lowest (8.78 g/plant) under the control treatment. Shoot dry mass also differed under different treatments and was significantly higher (4.09 g/plant) under \u003cem\u003eTrichoderma atroviride\u003c/em\u003e and was lowest (1.33 g/plant) under control treatment. The number of flowers ranged from 10.66 under \u003cem\u003eTrichoderma atroviride\u003c/em\u003e to 2.33 under control. ANOVA and post-hoc analysis revealed that treatments such as \u003cem\u003eTrichoderma atroviride\u003c/em\u003e, \u003cem\u003eTrichoderma viride\u003c/em\u003e, \u003cem\u003eAspergillus flavus\u003c/em\u003e, \u003cem\u003ePaecilomyces brunneolus\u003c/em\u003e, \u003cem\u003eTalaromyces purpureogenus\u003c/em\u003e, \u003cem\u003ePaecilomyces variotii\u003c/em\u003e, \u003cem\u003ePenicillium chrysogenum\u003c/em\u003e and control were significantly different from one another. Furthermore, the moisture content percent was highest in control (85.05%) and lowest in \u003cem\u003eTrichoderma atroviride\u003c/em\u003e (77.92%). ANOVA and post-hoc analysis revealed that treatments such as \u003cem\u003eTrichoderma atroviride\u003c/em\u003e, \u003cem\u003eTrichoderma viride\u003c/em\u003e, \u003cem\u003eAspergillus flavus\u003c/em\u003e, \u003cem\u003ePaecilomyces brunneolus\u003c/em\u003e, \u003cem\u003eTalaromyces purpureogenus\u003c/em\u003e, \u003cem\u003ePaecilomyces variotii\u003c/em\u003e, \u003cem\u003ePenicillium chrysogenum\u003c/em\u003e and control were not significantly different from one another. Leaf area was highest in \u003cem\u003eTrichoderma atroviride\u003c/em\u003e (112.35 cm\u003csup\u003e2\u003c/sup\u003e) followed by the other fungi and least was observed in control (49.3cm\u003csup\u003e2\u003c/sup\u003e). Specific leaf area was highest in \u003cem\u003ePaecilomyces variotii\u003c/em\u003e (166.42 cm\u003csup\u003e2\u003c/sup\u003e/g) and lowest in \u003cem\u003eTrichoderma atroviride\u003c/em\u003e (82.42 cm\u003csup\u003e2\u003c/sup\u003e/g). Leaf area ratio was highest in \u003cem\u003eTalaromyces purpureogenus\u003c/em\u003e (34.56 cm\u003csup\u003e2\u003c/sup\u003e/g) and lowest in \u003cem\u003eTrichoderma atroviride\u003c/em\u003e (21.33 cm\u003csup\u003e2\u003c/sup\u003e/g). There were no significant differences between \u003cem\u003ePaecilomyces variotii\u003c/em\u003e and control. Leaf area ratio was highest in \u003cem\u003eTalaromyces purpureogenus\u003c/em\u003e (34.56 cm\u003csup\u003e2\u003c/sup\u003e/g) and lowest in \u003cem\u003eTrichoderma atroviride\u003c/em\u003e (21.33 cm\u003csup\u003e2\u003c/sup\u003e/g). Leaf weight ratio was highest in \u003cem\u003eTalaromyces purpureogenus\u003c/em\u003e and \u003cem\u003eTrichoderma atroviride\u003c/em\u003e (0.25 each). Additionally, chlorophyll content was highest in \u003cem\u003eTrichoderma atroviride\u003c/em\u003e (7.95 mg/g) and lowest in control (1.95 mg/g). The effects of different treatments \u003cem\u003ePenicillium chrysogenum, Paecilomyces variotii, Talaromyces purpureogenus\u003c/em\u003e, \u003cem\u003ePaecilomyces brunneolus\u003c/em\u003e, \u003cem\u003eAspergillus flavus\u003c/em\u003e, \u003cem\u003eTrichoderma viride\u003c/em\u003e and \u003cem\u003eTrichoderma atroviride\u003c/em\u003e on root length, shoot length, fresh mass, dry mass, leaf area, number of flowers and chlorophyll content revealed significant differences in these growth parameters across the treatments, as compared to control, suggesting that the rhizospheric fungi have a discernible impact on plant growth compared to the control group. Each parameter was measured in five replicates (n\u0026thinsp;=\u0026thinsp;5), and the results are presented as mean values with their respective standard deviations (SD).\u003c/p\u003e"},{"header":"5. Discussion","content":"\u003cp\u003eThe study presents a comprehensive evaluation of the effects of plant growth promoting fungi (PGPF) on the growth parameters of \u003cem\u003eC. sativus\u003c/em\u003e. Previous studies have also demonstrated that specific fungal species can stimulate key growth parameters, including seed germination and other developmental stages. \u003cem\u003eTrichoderma\u003c/em\u003e species, which are well-documented for their biostimulators effects, likely due to their ability to produce phytohormones and solubilize essential nutrients (Asghar et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2024\u003c/span\u003e) have also been shown to elicit systemic resistance in plants, aiding them in coping with abiotic and biotic stresses (Gupta and Bar, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). The use of these fungal inoculants provides insight into reducing chemical inputs while maintaining high crop yield. Chemical fertilizers and pesticides, while effective, are increasingly scrutinized due to their impact on human and environmental health, as well as the development of pathogen resistance (Indira et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Vanshree et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn the context of plant growth enhancement, different fungal treatments were used to evaluate how these fungal species, which include \u003cem\u003ePenicillium chrysogenum\u003c/em\u003e, \u003cem\u003ePaecilomyces variotii\u003c/em\u003e, \u003cem\u003eTalaromyces purpureogenus\u003c/em\u003e, \u003cem\u003ePaecilomyces brunneolus\u003c/em\u003e, \u003cem\u003eAspergillus flavus\u003c/em\u003e, \u003cem\u003eTrichoderma viride\u003c/em\u003e, and \u003cem\u003eTrichoderma atroviride\u003c/em\u003e, impact various growth parameters of \u003cem\u003eC. sativus\u003c/em\u003e. The results highlight that C. \u003cem\u003esativus\u003c/em\u003e seedlings treated with diverse rhizospheric plant growth promoting fungal species, particularly \u003cem\u003eTrichoderma atroviride\u003c/em\u003e, significantly improves growth parameters like root and shoot length, fresh and dry weight, and leaf chlorophyll content. For instance, \u003cem\u003eTrichoderma atroviride\u003c/em\u003e -treated plants exhibited the highest root length (12.1 cm) and shoot length (58.66 cm), indicating a robust root system and enhanced vegetative growth. Interestingly, treatments also affected moisture retention, with control showing the highest moisture content, contrasting with \u003cem\u003eTrichoderma atroviride\u003c/em\u003e, which exhibited the lowest. This variation in moisture content could reflect the different capacities of fungal species to enhance water use efficiency and dry mass accumulation or adapt to fluctuating water availability in the soil matrix. Moreover, \u003cem\u003ePaecilomyces variotii\u003c/em\u003e, and \u003cem\u003eTalaromyces purpureogenus\u003c/em\u003e, which showed significant improvements in specific leaf area and leaf area ratio, could be contributing to an optimized plant architecture that maximizes light capture, a feature particularly advantageous in competitive plant environments. This investigation aligns with current research trends that emphasize sustainable agricultural practices and the reduced use of chemical fertilizers and pesticides. Given the context of global food security and the associated challenges posed by environmental stressors on crop productivity, this study offers a promising approach to addressing the yield demands of \u003cem\u003eC. sativus\u003c/em\u003e which is widely cultivated and consumed in world.\u003c/p\u003e"},{"header":"6. Conclusion","content":"\u003cp\u003eThis study highlights that plant growth promoting fungi (PGPF), can serve as eco-friendly bio stimulants, enhancing crop resilience and productivity while minimizing dependence on pesticides and chemical fertilizers. The study also demonstrates that PGP fungal species significantly improved growth parameters, including root and shoot length, biomass accumulation and chlorophyll content which indicates strengthened plant structure and photosynthetic efficiency. The observed bio-enhancement suggests that such fungi could play a vital role in advancing food security and eco-friendly crop management practices.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eACKNOWLEDGEMENTS\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe are highly thankful to head Department of Botany and Plant Pathology, Mycology and Microbiology Laboratory, University of Kashmir for providing feasible facilities to carry out this work.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAUTHORS CONTRIBUTIONS\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors listed in this submission have made significant contributions to the conceptualization and design of the study, as well as the analysis and interpretation of the data. Each author has actively participated in drafting the manuscript, critically revising it for key intellectual content, and approving the final version for publication\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNo datasets were generated or analysed during the current study\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCONFLICT OF INTEREST\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declared that there is no conflict of interest.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNil\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics, Consent to participate, Consent to publish\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eClinical trial\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;Not applicable\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAdedayo, A. A., \u0026amp; Babalola, O. O. (2023). Fungi that promote plant growth in the rhizosphere boost crop growth. \u003cem\u003eJournal of Fungi\u003c/em\u003e, \u003cem\u003e9\u003c/em\u003e(2), 239. https://doi.org/10.3390/jof9020239\u003c/li\u003e\n\u003cli\u003eAin, Q. U., Hussain, H. A., Zhang, Q., Maqbool, F., Ahmad, M., Mateen, A., ... \u0026amp; Imran, A. (2024). Coordinated influence of \u003cem\u003eFunneliformis mosseae\u003c/em\u003e and different plant growth-promoting bacteria on growth, root functional traits, and nutrient acquisition by maize. \u003cem\u003eMycorrhiza\u003c/em\u003e, 1-12. DOI:10.1007/s00572-024-01165-5\u003c/li\u003e\n\u003cli\u003eAlexander, P., Arneth, A., Henry, R., Maire, J., Rabin, S., \u0026amp; Rounsevell, M. D. (2023). High energy and fertilizer prices are more damaging than food export curtailment from Ukraine and Russia for food prices, health and the environment. \u003cem\u003eNature Food\u003c/em\u003e, \u003cem\u003e4\u003c/em\u003e(1), 84-95. DOI: 10.1038/s43016-022-00659-9\u003c/li\u003e\n\u003cli\u003eAsghar, W., Craven, K. D., Kataoka, R., Mahmood, A., Asghar, N., Raza, T., \u0026amp; Iftikhar, F. (2024). The application of \u003cem\u003eTrichoderma\u003c/em\u003e spp., an old but new useful fungus, in sustainable soil health intensification: A comprehensive strategy for addressing challenges. \u003cem\u003ePlant Stress\u003c/em\u003e, 100455. https://doi.org/10.1016/j.stress.2024.100455\u003c/li\u003e\n\u003cli\u003eAttia, M. S., Abdelaziz, A. M., Al-Askar, A. A., Arishi, A. A., Abdelhakim, A. M., \u0026amp; Hashem, A. H. (2022). Plant growth-promoting fungi as biocontrol tool against \u003cem\u003eFusarium \u003c/em\u003ewilt disease of tomato plant. \u003cem\u003eJournal of Fungi\u003c/em\u003e, \u003cem\u003e8\u003c/em\u003e(8), 775. https://doi.org/10.3390/jof8080775\u003c/li\u003e\n\u003cli\u003eCantabella, D., Dolcet-Sanjuan, R., Casanovas, M., Solsona, C., Torres, R., \u0026amp; Teixid\u0026oacute;, N. (2020). Inoculation of in vitro cultures with rhizosphere microorganisms improve plant development and acclimatization during immature embryo rescue in nectarine and pear breeding programs. \u003cem\u003eScientia Horticulturae\u003c/em\u003e, \u003cem\u003e273\u003c/em\u003e, 109643. DOI:10.1016/j.scienta.2020.109643\u003c/li\u003e\n\u003cli\u003eChaudhary, P., Singh, S., Chaudhary, A., Sharma, A., \u0026amp; Kumar, G. (2022). Overview of biofertilizers in crop production and stress management for sustainable agriculture. \u003cem\u003eFrontiers in Plant Science\u003c/em\u003e, \u003cem\u003e13\u003c/em\u003e, 930340. https://doi.org/10.3389/fpls.2022.930340\u003c/li\u003e\n\u003cli\u003eGupta, R., \u0026amp; Bar, M. (2020). Plant immunity, priming, and systemic resistance as mechanisms for \u003cem\u003eTrichoderma\u003c/em\u003e spp. biocontrol. \u003cem\u003eTrichoderma\u003c/em\u003e: Host Pathogen Interactions and Applications, 81-110. DOI:10.1007/978-981-15-3321-1_5\u003c/li\u003e\n\u003cli\u003eIndira D. P., Manjula M., Bhavani R.V. (2022) Agrochemicals, Environment, and Human Health; \u003cem\u003eAnnual Review of Environment and Resources,\u003c/em\u003e 47, 399-421. https://doi.org/10.1146/annurev-environ-120920-111015\u003c/li\u003e\n\u003cli\u003eMalviya, J. M., Kiran Singh, K. S., \u0026amp; Vaibhavi Joshi, V. J. (2011). Effect of phosphate solubilizing fungi on growth and nutrient uptake of ground nut (Arachis hypogaea) plants. \u003cem\u003eAdv Biores,\u003c/em\u003e2,110-113.\u003c/li\u003e\n\u003cli\u003eKumar, S., Sindhu, S. S., \u0026amp; Kumar, R. (2022). Biofertilizers: An ecofriendly technology for nutrient recycling and environmental sustainability. \u003cem\u003eCurrent Research in Microbial Sciences\u003c/em\u003e, \u003cem\u003e3\u003c/em\u003e, 100094. https://doi.org/10.1016/j.crmicr.2021.100094\u003c/li\u003e\n\u003cli\u003eKuzin, A., Solovchenko, A., Stepantsova, L., \u0026amp; Pugachev, G. (2020). Soil fertility management in apple orchard with microbial biofertilizers. In \u003cem\u003eE3S Web of Conferences\u003c/em\u003e (Vol. 222, p. 03020). EDP Sciences. DOI:10.1051/e3sconf/202022203020\u003c/li\u003e\n\u003cli\u003eLi, X., Li, D., Yan, J., Zhang, Y., Wang, H., Zhang, J., ... \u0026amp; Li, B. (2021). Effect of plant-growth-promoting fungi on eggplant (\u003cem\u003eSolanum melongena\u003c/em\u003e L.) in new reclamation land. \u003cem\u003eAgriculture\u003c/em\u003e, \u003cem\u003e11\u003c/em\u003e(11), 1036. https://doi.org/10.3390/agriculture11111036\u003c/li\u003e\n\u003cli\u003eMohamed, A. H., Abd el-Megeed, F. H., Hassanein, N. M., Youseif, S. H., Farag, P. F., Saleh, S. A., ... \u0026amp; Abdel-Azeem, A. M. (2022). Native rhizospheric and endophytic fungi as sustainable sources of plant growth promoting traits to improve wheat growth under low nitrogen input. \u003cem\u003eJournal of Fungi\u003c/em\u003e, \u003cem\u003e8\u003c/em\u003e(2), 94. https://doi.org/10.3390/jof8020094\u003c/li\u003e\n\u003cli\u003eNaimuzzaman, M., Rahman, F., Alvi, A. T., Yeasmin, L., Mittra, P. K., \u0026amp; Roy, S. K. (2024). Plant growth\u0026ndash;promoting fungi in plants: Insights from stress tolerance mechanism. In \u003cem\u003eBeneficial Microbes for Sustainable Agriculture Under Stress Conditions\u003c/em\u003e,469-511. Academic Press. https://doi.org/10.1016/b978-0-443-13193-6.00023-3\u003c/li\u003e\n\u003cli\u003eOmomowo, I. O., Adedayo, A. A., \u0026amp; Omomowo, O. I. (2020). Biocontrol potential of rhizospheric fungi from moringa oleifera, their phytochemicals and secondary metabolite assessment against spoilage fungi of sweet orange (\u003cem\u003ecitrus sinensis\u003c/em\u003e). \u003cem\u003eAsian Journal of Applied Sciences\u003c/em\u003e, \u003cem\u003e8\u003c/em\u003e(1). https://doi.org/10.24203/ajas.v8i1.6047\u003c/li\u003e\n\u003cli\u003ePandit, M. A., Kumar, J., Gulati, S., Bhandari, N., Mehta, P., Katyal, R., ... \u0026amp; Kaur, J. (2022). Major biological control strategies for plant pathogens. \u003cem\u003ePathogens\u003c/em\u003e, \u003cem\u003e11\u003c/em\u003e(2), 273. https://doi.org/10.3390/pathogens11020273\u003c/li\u003e\n\u003cli\u003ePenuelas, J., Coello, F., \u0026amp; Sardans, J. (2023). A better use of fertilizers is needed for global food security and environmental sustainability. \u003cem\u003eAgriculture \u0026amp; Food Security\u003c/em\u003e, \u003cem\u003e12\u003c/em\u003e(1), 1-9. https://doi.org/10.1186/s40066-023-00409-5 \u0026middot;\u003c/li\u003e\n\u003cli\u003eSun, X., Liu, F., Jiang, W., Zhang, P., Zhao, Z., Liu, X., ... \u0026amp; Sun, Q. (2023). \u003cem\u003eTalaromyces purpurogenus\u003c/em\u003e Isolated from Rhizosphere Soil of Maize Has Efficient Organic Phosphate-Mineralizing and Plant Growth-Promoting Abilities. \u003cem\u003eSustainability\u003c/em\u003e, 15(7), 5961. DOI:10.3390/su15075961\u003c/li\u003e\n\u003cli\u003eThepbandit, W., \u0026amp; Athinuwat, D. (2024). Rhizosphere microorganisms supply availability of soil nutrients and induce plant defense. \u003cem\u003eMicroorganisms\u003c/em\u003e, \u003cem\u003e12\u003c/em\u003e(3), 558. https://doi.org/10.3390/microorganisms12030558\u003c/li\u003e\n\u003cli\u003eTyśkiewicz, R., Nowak, A., Ozimek, E., \u0026amp; Jaroszuk-ściseł, J. (2022). \u003cem\u003eTrichoderma\u003c/em\u003e: The current status of its application in agriculture for the biocontrol of fungal phytopathogens and stimulation of plant growth. \u003cem\u003eInternational Journal of Molecular Sciences\u003c/em\u003e, \u003cem\u003e23\u003c/em\u003e(4), 2329. https://doi.org/10.3390/ijms23042329\u003c/li\u003e\n\u003cli\u003eVanshree, C. R., Singhal, M., Sexena, M., Sankhla, M. S., Parihar, K., Jadhav, E. B., Yadav, C. S. (2022). Microbes as biocontrol agent: From crop protection till food security. In \u003cem\u003eRelationship Between Microbes and the Environment for Sustainable Ecosystem Services.\u003c/em\u003e 1: 215-237. https://doi.org/10.1016/B978-0-323-89938-3.00011-6\u003c/li\u003e\n\u003cli\u003eZhou, W., Li, M., \u0026amp; Achal, V. (2024). A comprehensive review on environmental and human health impacts of chemical pesticide usage. \u003cem\u003eEmerging Contaminants\u003c/em\u003e, 100410. https://doi.org/10.1016/j.emcon.2024.100410\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Table 1","content":"\u003cp\u003eTable 1 is available in the Supplementary Files section.\u003c/p\u003e\n"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"discover-plants","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"Learn more about [Discover Plants](https://link.springer.com/journal/44372)","snPcode":"44372","submissionUrl":"https://submission.springernature.com/new-submission/44372/3","title":"Discover Plants","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Discover Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Cucumis sativus, (PGPF), Growth parameters","lastPublishedDoi":"10.21203/rs.3.rs-6156187/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6156187/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eRhizosphere and root associated fungi, commonly known as plant growth promoting fungi (PGPF), facilitate the growth and productivity of \u003cem\u003eCucumis sativus\u003c/em\u003e L. through the formation of mutualistic relationships with the plant roots. These fungi enhance nutrient uptake, promote root development and bolster stress tolerance via the secretion of growth regulating hormones. Despite numerous PGP fungal species have been isolated from the rhizosphere of \u003cem\u003eC. sativus\u003c/em\u003e, their specific roles in modulating various growth parameters of the \u003cem\u003eC. sativus\u003c/em\u003e remain insufficiently defined. In the present study, effect of seven PGP fungal species were evaluated such as \u003cem\u003ePenicillium chrysogenum\u003c/em\u003e, \u003cem\u003ePaecilomyces variotii\u003c/em\u003e, \u003cem\u003eTalaromyces purpureogenus\u003c/em\u003e, \u003cem\u003ePaecilomyces brunneolus\u003c/em\u003e, \u003cem\u003eAspergillus flavus\u003c/em\u003e, \u003cem\u003eTrichoderma viride\u003c/em\u003e, and \u003cem\u003eTrichoderma atroviride\u003c/em\u003e on the growth parameters of \u003cem\u003eC. sativus\u003c/em\u003e. Our results revealed that all PGP fungal species positively influenced growth parameters such as, root length, shoot length, fresh and dry biomass, leaf area, specific leaf area, moisture content, flower number, leaf area ratio, leaf weight ratio and chlorophyll content, albeit with varying degrees of efficiency. Notably, \u003cem\u003eTrichoderma atroviride\u003c/em\u003e exhibited a more pronounced impact on plant growth as compared to other PGP fungal species and the control. Our results suggest that these PGP fungal species may contribute to sustainable cultivation practices by enhancing crop yield and reducing the dependency on chemical fertilizers and pesticides.\u003c/p\u003e","manuscriptTitle":"Enhancing sustainable cultivation: The role of plant growth promoting fungi in optimizing Cucumis sativus L. growth","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-04-22 09:11:33","doi":"10.21203/rs.3.rs-6156187/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-04-21T05:51:59+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-04-17T17:56:30+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-04-10T17:51:54+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"48836036843298671884400717974572912624","date":"2025-04-08T09:54:57+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"120117361510880671994930287374490061240","date":"2025-04-04T08:27:22+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-04-02T06:31:26+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-03-31T07:49:09+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-03-29T10:29:47+00:00","index":"","fulltext":""},{"type":"submitted","content":"Discover Plants","date":"2025-03-29T10:28:38+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"discover-plants","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"Learn more about [Discover Plants](https://link.springer.com/journal/44372)","snPcode":"44372","submissionUrl":"https://submission.springernature.com/new-submission/44372/3","title":"Discover Plants","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Discover Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"0f75f5b3-ae82-43ba-a4ba-3e275840b86b","owner":[],"postedDate":"April 22nd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2025-06-06T19:08:23+00:00","versionOfRecord":[],"versionCreatedAt":"2025-04-22 09:11:33","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6156187","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6156187","identity":"rs-6156187","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}