Responses of Different Cannabis sativa L. Populations to Plant Density and Various Fertilization Treatments

Abstract

The increasing demand for raw materials in the paper and fermentation industries has intensified pressure on forest resources, highlighting the urgent need for sustainable alternatives. Hemp (Cannabis sativa L.), a fast-growing, non-woody fiber crop with additional medicinal and nutritional benefits, has emerged as a promising candidate. This two-year field study, conducted using a split-split plot design with three replications, evaluated the effects of four planting densities (3.6, 5.6, 9.5, and 20 plants m-2) and four nitrogen levels (0,150, 300, and 450 kg ha-1) on three hemp genotypes: Futura 75 (Germany), Isfahan, and Yazd, in the Alborz region of Iran. Significant effects were observed for all main factors and their interactions on key morphological and yield-related traits. The highest dry matter yield (22.559 kg ha-1) was obtained from the Isfahan genotype under 300 kg ha-1 nitrogen and a plant density of 20 plants m-2. The Futura 75 genotype recorded the greatest stem diameter (5.31 cm) and number of lateral stems (70 per plant) under low density (3.6 or 9.5 plants m-2) and high nitrogen input (450 kg ha-1). Leaf dry weight peaked at 103.66 g plant-1 in the Isfahan genotype with high nitrogen and low density. Yield components were generally higher in the first growing season, suggesting a notable influence of favorable environmental conditions. These findings emphasize the importance of optimizing planting density and nitrogen application according to genotype and environmental conditions. The Isfahan genotype, combined with high plant density and 300 kg ha-1nitrogen, is particularly recommended for enhancing biomass yield in semi-arid regions.

Keywords

• Cannabis sativa L
• Nitrogen fertilizer
• Plant density
• Genotype

References

1. Ahsan, S., Injamum-Ul-Hoque, M., Shaffique, S., Ayoobi, A., Rahman, M., Rahman, M. M., & Choi, H. (2024). Illuminating Cannabis sativa L.: The power of light in enhancing C. sativa growth and secondary metabolite production. Plants, 13(19), 2774. https://doi.org/10.3390/plants13192774
2. Anderson, S., Pearson, B., Kjelgren, R., & Brym, Z. (2021). Response of essential oil hemp (Cannabis sativa L.) growth, biomass, and cannabinoid profiles to varying fertigation rates. PLoS ONE, 16(6), e0252985. https://doi.org/10.1371/journal.pone.0252985
3. Andre, C. M., Hausman, J. F., & Guerriero, G. (2016). Cannabis sativa: The plant of the thousand and one molecules. Frontiers in Plant Science, 7, 19. https://doi.org/10.3389/fpls.2016.00019
4. Arian, A., & Vlosky, R. (2023). An overview of the wood products sector in Iran: 2007–2022. Journal of Forest Business Research, 2(2), 40. https://doi.org/10.62320/jfbr.v2i2.40
5. Burgel, L., Hartung, J., Pflugfelder, A., & Graeff-Hönninger, S. (2020). Impact of growth stage and biomass fractions on cannabinoid content and yield of different hemp (Cannabis sativa L.) genotypes. Agronomy, 10(3), 372. https://doi.org/10.3390/agronomy10030372
6. Calabrese, A., Campanale, M., & Iurino, F. (2023). Nitrogen supply of crops and type of fertilization and their impact on the soil: Case study of Apulia Region. Journal of Biomedical Research & Environmental Sciences, 4(6), 652–657. https://doi.org/10.37871/jbres1855
7. Campiglia, E., Gobbi, L., Marucci, A., Rapa, M., Ruggieri, R., & Vinci, G. (2020). Hemp seed production: Environmental impacts of Cannabis sativa L. agronomic practices by life cycle assessment (LCA) and carbon footprint methodologies. Sustainability, 12(16), 6570. https://doi.org/10.3390/su12166570
8. Campiglia, E., Radicetti, E., & Mancinelli, R. (2017). Plant density and nitrogen fertilization affect agronomic performance of industrial hemp (Cannabis sativa L.) in Mediterranean environment. Industrial Crops and Products, 100, 246–254. https://doi.org/10.1016/j.indcrop.2017.02.022

9. Charitos, I. A., Gagliano-Candela, R., Santacroce, L., & Bottalico, L. (2020). The cannabis spread throughout the continents and its therapeutic use in history. Endocrine, Metabolic & Immune Disorders - Drug Targets, 20(4), 473–481. https://doi.org/10.2174/1871530320666 200520095900
10. Danziger, N., & Bernstein, N. (2022). Too dense or not too dense: Higher planting density reduces cannabinoid uniformity but increases yield/area in drug-type medical cannabis. Frontiers in Plant Science, 13, 713481. https://doi.org/10.3389/fpls.2022.713481
11. Demirel, ANŞ., & Demirel, S. (2024). Thyme sediments for energy storage systems and its benefit evaluations for agricultural economics. ISPEC Journal of Agricultural Sciences, 8(1), 116-126, 2024. https:// doi.org/10.5281/zenodo.10815486
12. Deng, G., Du, G., Yang, Y., Bao, Y., & Liu, F. (2019). Planting density and fertilization evidently influence the fiber yield of hemp (Cannabis sativa L.). Agronomy, 9(7), 368. https://doi.org/10.3390/agronomy9070368
13. Fordjour, E., Manful, C. F., Sey, A. A., Javed, R., Pham, T. H., Thomas, R. H., & Cheema, M. (2023). Cannabis: A multifaceted plant with endless potentials. Frontiers in Pharmacology, 14, 1200269. https://doi.org/10.3389/fphar.2023.1200269
14. Gommers, C. M. M., Buti, S., Tarkowská, D., Pěnčík, A., Banda, J., Arricastres, V., & Pierik, R. (2018). Organ‐specific phytohormone synthesis in two Geranium species with antithetical responses to far‐red light enrichment. Plant Direct, 2(9), e00066. https://doi.org/10.1002/pld3.66
15. Herppich, W., Gusovius, H., Flemming, I., & Drastig, K. (2020). Effects of drought and heat on photosynthetic performance, water use and yield of two selected fiber hemp cultivars at a poor-soil site in Brandenburg (Germany). Agronomy, 10(9), 1361. https://doi.org/10.3390/agronomy10091361
16. Hitsuma, G., Himmapan, W., Yagihashi, T., Miyamoto, K., & Vacharangkura, T. (2021). Effects of tree density and size symmetry of competition on diameter growth in the early stages of growth in planted teak (Tectona grandis) trees in northern Thailand. Journal of Forest Research, 26(3), 195–204. https://doi.org/10.1080/13416979.2021.1902598
17. Hogewoning, S., Trouwborst, G., Meinen, E., & Van Ieperen, W. (2012). Finding the optimal growth-light spectrum for greenhouse crops. Acta Horticulturae, 956, 229–236. https://doi.org/10.17660/ActaHortic.2012.956.41
18. Hua, S., Dal-Bianco, M., & Chen, Z. (2022). Editorial: Crop yield and quality response to the interaction between environment and genetic factors. Frontiers in Genetics, 13, 823279. https://doi.org/10.3389/fgene.2022.823279
19. Ju, Z., Liu, K., Zhao, G., Ma, X., & Jia, Z. (2022). Nitrogen fertilizer and sowing density affect flag leaf photosynthetic characteristics, grain yield, and yield components of oat in a semiarid region of Northwest China. Agronomy, 12(9), 2108. https://doi.org/10.3390/agronomy12092108
20. Karaca Öner, E., Arslanoğlu, Ş. F., & İskender, R. (2023). Determination of the effects of different nitrogen and phosphorus applications on leaf area of Echinacea (Echinacea purpurea L.). ISPEC Journal of Agricultural Sciences, 7(1), 53–59. https://doi.org/10.5281/zenodo.7717862
21. Lopez, G., Ahmadi, S. H., Amelung, W., Athmann, M., Ewert, F., Gaiser, T., Gocke, M., Kautz, T., Postma, J., Rachmilevitch, S., Schaaf, G., Schnepf, A., Stoschus, A., Watt, M., Yu, P., & Seidel, S. (2023). Nutrient deficiency effects on root architecture and root-to-shoot ratio in arable crops. Frontiers in Plant Science, 13, 1067498. https://doi.org/10.3389/fpls.2022.1067498
22. Mahakosee, S., Jogloy, S., Vorasoot, N., Theerakulpisut, P., Toomsan, B., Holbrook, C., Kvien, C., & Banterng, P. (2022). Light interception and radiation use efficiency of three cassava genotypes with different plant types and seasonal variations. Agronomy, 12(11), 2888. https://doi.org/10.3390/agronomy12112888
23. Malik, M., Praus, L., Kuklina, A., Velechovský, J., Kosmáková Janatová, A., Klouček, P., Mládek, V., & Tlustoš, P. (2025). Cannabis yield and cannabinoid profile affected by plant nutrition and planting density. Industrial Crops and Products, 223, 120293. https://doi.org/10.1016/j.indcrop.2024.120293
24. Malinowska, K., Myśków, B., Czyczyło-Mysza, I., & Góralska, M. (2018). Study on changes and relationships of physiological and morphological parameters of rye subjected to soil drought stress. Acta Agrophysica, 25(1), 71–84. https://doi.org/10.31545/AAGR/93584
25. McPartland, J. M. (2018). Cannabis systematics at the levels of family, genus, and species. Cannabis and Cannabinoid Research, 3(1), 203–212. https://doi.org/10.1089/can.2018.0039
26. Mostafaei Dehnavi, M., Damerum, A., Taheri, S., Ebadi, A., Panahi, S., Hodgin, G., Brandley, B., Salami, S. A., & Taylor, G. (2024). Population genomics of a natural Cannabis sativa L. collection from Iran identifies novel genetic loci for flowering time, morphology, sex and chemotyping. BMC Plant Biology, 25, 1–17. https://doi.org/10.1186/s12870-025-06045-4
27. Mustafa, A., Athar, F., Khan, I., Chattha, M. U., Nawaz, M., Shah, A., Mahmood, A., Batool, M., Aslam, M. T., Jaremko, M., Abdelsalam, N., Ghareeb, R. Y., & Hassan, M. (2022). Improving crop productivity and nitrogen use efficiency using sulfur and zinc-coated urea: A review. Frontiers in Plant Science, 13, 942384. https://doi.org/10.3389/fpls.2022.942384
28. Oh, S., & Koh, S. (2019). Fruit development and quality of hot pepper (Capsicum annuum L.) under various temperature regimes. Korean Journal of Horticultural Science and Technology, 37(3), 320–330. https://doi.org/10.7235/hort.20190032
29. Olsen, S. R., Cole, C. G., Watanabe, F. S., & Dean, L. A. (1954). Estimation of available phosphorus in soils by extraction with sodium bicarbonate (Circular No. 939). United States Department of Agriculture.
30. Oral, E. Tunçtürk, R. Tunçtürk, M. Nohutçu, L., & Şelem, E. (2024). Effect of mycorrhiza application son some biochemical properties of soybean (Glycine max L.) Grown under salt stress conditions. ISPEC Journal of Agricultural Sciences, 8(3), 551-559, 2024. https://doi.org/10.5281/zenodo.12576435
31. Padmadevi, K., Jegadeeswari, V., Vijayalatha, K., Mohanalakshmi, M., Kalaivani, J., & Sidhdharth, G. (2024). Assessment of vegetative and flower characters in cocoa (Theobroma cacao) under different planting densities. Journal of Scientific Research and Reports, 30(6), 1–10. https://doi.org/10.9734/jsrr/2024/v30i62091
32. Shi, L. (2024). The impact of different planting densities on the growth and ecological benefits of sandland Caragana. Journal of Environmental and Building Engineering.
33. Sipahi, H. (2024). Genome-wide characterization of expansin gene family in Cannabis sativa L. ISPEC Journal of Agricultural Sciences, 8(1), 255–265. https://doi.org/10.5281/zenodo.10864859
34. Smith, R. M., Kaur, G., Orlowski, J., Singh, G., Chastain, D., Irby, T., Krutz, L., Falconer, L. L., & Cook, D. (2019). Narrow-row production system for soybeans in Mississippi Delta. Crop, Forage & Turfgrass Management, 5(1). https://doi.org/10.2134/cftm2019.02.0015
35. Soratto, R. P., Catuchi, T. A., Souza, E. F., & Garcia, J. L. N. (2017). Plant density and nitrogen fertilization on common bean nutrition and yield. Revista Ceres, 64(3), 315–323. https://doi.org/10.1590/1983-21252017v30n315rc
36. Sun, S., Huang, Z., Liu, H., Xu, J., Zheng, X., Xue, J., & Li, S. (2023). Response of grain yield to planting density and maize hybrid selection in high latitude China—A multisource data analysis. Agronomy, 13(5), 1333. https://doi.org/10.3390/agronomy13051333
37. Tang, K., Tang, Y. J., & Blankenship, R. E. (2011). Carbon metabolic pathways in phototrophic bacteria and their broader evolutionary implications. Frontiers in Microbiology, 2, 165. https://doi.org/10.3389/fmicb.2011.00165
38. Vera, C., Malhi, S., Phelps, S., May, W., & Johnson, E. N. (2010). N, P, and S fertilization effects on industrial hemp in Saskatchewan. Canadian Journal of Plant Science, 90(1), 29–37. https://doi.org/10.4141/CJPS09101
39. Wang, X., Hu, X., Lin, C., Liu, Q., Li, Y., Du, D., Mkapa, D., Zhang, W., Huang, X., & Yi, K. (2024). Agave schidigera transcriptome reveals stress-responsive phenylalanine ammonia-lyase genes in Agave. Agronomy, 14(11), 2520. https://doi.org/10.3390/agronomy14112520
40. Xu, Q., Liu, H., Li, M., & Li, P. (2023). Interactive effect of irrigation volume and planting density on growth and salt uptake in field-grown drip-irrigated Suaeda salsa (L.) Pall. Plants, 12(6), 1383. https://doi.org/10.3390/plants12061383
41. Yang, Y., Zha, W., Tang, K., Deng, G., Du, G., & Liu, F. (2021). Effect of nitrogen supply on growth and nitrogen utilization in hemp (Cannabis sativa L.). Agronomy, 11(11), 2310. https://doi.org/10.3390/agronomy11112310
42. Yasmin, A., Zeb, A., Khalil, A. W., Paracha, Gholam M.U.D., & Badshah Khattak, A. (2008). Effect of processing on anti-nutritional factors of red kidney bean (Phaseolus vulgaris) grains. Food Bioprocess Technol, 1, 415–419. https://doi.org/ 10.1007/s11947-008-0125-3
43. Zhao, Y., Huang, Y., Li, S., Chu, X., & Ye, Y. (2020). Improving the growth, lodging and yield of different density-resistance maize by optimising planting density and nitrogen fertilisation. Plant Soil and Environment, 66(10), 499–506. https://doi.org/10.17221/178/2020-PSE