Bioinformatics analysis and investigation of the effects of different fertilizer treatments on the expression profile of Cannabinoid Synthase gene in Cannabis sativa

Yıl 2025, Cilt: 1 Sayı: 2, 79 - 88, 31.10.2025

Alireza Mirzaei , Jamshid Fooladi

Öz

Cannabis sativa is one of the most important plants producing cannabidiol, and it has a wide application in medicine. This study was conducted to investigate the effects of nitrogen, phosphorus, potassium, iron, manganese, and magnesium treatments on the expression profile of Cannabinoid Synthase gene in Cannabis sativa clones. The results of this study showed that fertilization has a direct effect on expression of Cannabinoid Synthase gene. In addition, potassium treatment stimulated the expression of Cannabinoid Synthase gene more than the other treatments studied. The results of the bioinformatics analysis showed that clones 4 and 5 were in the same category and were 55% similar to each other. In addition, the three clones studied in Cannabis sativa were in the same category compared to other related genes in other species. Also, protein blast analysis of Cannabinoid Synthase gene in different species showed that Morus alba, Arabidopsis thaliana, Phleum pretense, and Cynodon dactylon species have more structural similarity than the corresponding Cannabinoid Synthase gene in Glycine max. On the other hand, the study of the expression profile of Cannabinoid Synthase gene in Arabidopsis thaliana showed that the expression level of this gene was constant in the leaf and that fewer expression changes were seen in the Cannabinoid Synthase gene compared to other organs. In general, this research provided an accurate estimate of the expression analysis of the Cannabinoid Synthase gene in Cannabis sativa and other reference species, which provides researchers with a useful perspective on the extraction of cannabidiol from Cannabis sativa.

Anahtar Kelimeler

Cannabinoid , gene expression , Phylogenetic analysis , fertilization , Cannabis sativa

Biyoinformatik analiz ve farklı gübre uygulamalarının Cannabis sativa'da Kannabinoid Sentaz geninin ekspresyon profili üzerindeki etkilerinin araştırılması

Öz

Cannabis sativa, kannabidiol üreten en önemli bitkilerden biri olup tıpta geniş bir uygulamaya sahiptir. Bu çalışma, azot, fosfor, potasyum, demir, manganez ve magnezyum uygulamalarının Cannabis sativa klonlarında Kannabinoid Sentaz geninin ifade profilindeki değişikliklere olan etkilerini araştırmak için yürütülmüştür. Bu çalışmanın sonuçları, döllenmenin kontrol durumuna kıyasla Kannabinoid Sentaz geninin ifadesini artırmada doğrudan bir etkiye sahip olduğunu göstermiştir. Bu sonuçlara göre Kannabinoid Sentaz geninin yapraktaki ve klon 4'teki ifade düzeyinin diğer ilgili klonlardan daha yüksek olduğunu belirlenmiştir. Ek olarak, potasyum uygulaması Kannabinoid Sentaz geninin ifadesini çalışılan diğer uygulamalardan daha fazla uyarmıştır. Bu çalışmanın biyoenformatik analiz sonuçları, 4 ve 5 numaralı klonların aynı kategoride olduğunu ve birbirlerine %55 oranında benzediğini göstermektedir. Ayrıca, Cannabis sativa'da çalışılan üç klon, diğer türlerdeki diğer ilgili genlerle karşılaştırıldığında aynı kategoride olduğu görülmektedir. Farklı türlerde Kannabinoid Sentaz geninin protein blast analizi, Morus alba, Arabidopsis thaliana, Phleum pretense ve Cynodon dactylon türlerinin Glycine max'taki karşılık gelen Kannabinoid Sentaz geninden daha fazla yapısal benzerliğe sahip olduğunu göstermektdir. Öte yandan, Arabidopsis thaliana'da Kannabinoid Sentaz geninin ifade profilinin incelenmesi, bu genin ifade düzeyinin yaprakta sabit olduğunu ve diğer organlarla karşılaştırıldığında Kannabinoid Sentaz geninde daha az ifade değişikliğinin görüldüğünü göstermektdir. Genel olarak bu araştırma, Cannabis sativa ve diğer referans türlerinde Kanabinoid Sentaz geninin ifade analizinin doğru bir tahminini sunarak araştırmacılara Cannabis sativa'dan kannabidiolün çıkarılması konusunda yararlı bir bakış açısı sağladığı sonucuna varılmıştır.

Anahtar Kelimeler

Kanabinoid , gen ifadesi , filogenetik analiz , döllenme , Cannabis sativa

Kaynakça

• Adamek, K. (2023). Genome Sequencing Reveals Intra-Plant and Clonal Genetic Diversity in Cannabis (Doctoral dissertation) University of Guelph.
• Aliferis, K. A., Bernard-Perron, D. (2020). Cannabinomics: Application of metabolomics in cannabis (Cannabis sativa L.) research and development. Frontiers in Plant Science, 11, 554.
• Bruni, N., Della Pepa, C., Oliaro-Bosso, S., Pessione, E., Gastaldi, D., Dosio, F. (2018). Cannabinoid deliverysystems for pain and inflammation treatment. Molecules, 23(10), 2478.
• Chaachouay, N., Qureshi, R., Benkhnigue, O., Azeroual, A., Zidane, L. Marijuana (Cannabis sativa L. Cannabaceae). In Comprehensive Guide to Hallucinogenic Plants ,CRC Press. pp.57-76.
• Chandra, S., Lata, H., Khan, I. A., ElSohly, M. A. (2017). Cannabis sativa L.: Botany and horticulture. In Cannabis sativa l.-botany and biotechnology, pp. 79-100. Cham: Springer International Publishing.
• Conrad, C. (1997). Hemp for health: The medicinal and nutritional uses of Cannabis sativa. Inner Traditions/Bear & Co.
• Dey, M., Bera, S., Tyagi, P., Pal, L. (2025). Mechanisms and strategic prospects of cannabinoids use: Potential applications in antimicrobial food packaging—A review. Comprehensive Reviews in Food Science and Food Safety, 24(1), e70113.
• Duggan, P. J. (2021). The chemistry of cannabis and cannabinoids. Australian Journal of Chemistry, 74(6), 369-387.
• Folina, A., Roussis, I., Kouneli, V., Kakabouki, I., Karidogianni, S., Bilalis, D. (2019). Opportunities for cultivation of medical Cannabis (Cannabis sativa L.) in Greece. Scientific Papers. Series A. Agronomy, 62(1).
• Gonçalves, J., Rosado, T., Soares, S., Simão, A. Y., Caramelo, D., Luís, Â., Duarte, A. P. (2019). Cannabis and its secondary metabolites: Their use as therapeutic drugs, toxicological aspects, and analytical determination. Medicines, 6(1), 31.
• Hesami, M., Pepe, M., Alizadeh, M., Rakei, A., Baiton, A., Jones, A. M. P. (2020). Recent advances in cannabis biotechnology. Industrial Crops and Products, 158, 113026.
• Iftikhar, A., Zafar, U., Ahmed, W., Shabbir, M. A., Sameen, A., Sahar, A., Aadil, R. M. (2021). Applications of Cannabis sativa L. in food and its therapeutic potential: From a prohibited drug to a nutritional supplement. Molecules, 26(24), 7699.
• Kim, S. (2024). Genetic and Environmental Factors Shaping Cannabis Phenotypes A Study on Temperature Effects and Genetic Regulation of Anthocyanin Accumulation in Cannabis sativa (Doctoral dissertation).
• Kopustinskiene, D. M., Masteikova, R., Lazauskas, R., Bernatoniene, J. (2022). Cannabis sativa L. bioactive compounds and their protective role in oxidative stress and inflammation. Antioxidants, 11(4), 660.
• Krüger, M., van Eeden, T., Beswa, D. (2022). Cannabis sativa cannabinoids as functional ingredients in snack foods—Historical and developmental aspects. Plants, 11(23), 3330.
• Lata, H., Chandra, S., Techen, N., Khan, I. A., ElSohly, M. A. (2009). Molecular analysis of genetic stability of micropropagated plants of Cannabis sativa L. using ISSR markers. Planta Medica, 75(04), P-6.
• Li, H., Liu, Y., Tian, D., Tian, L., Ju, X., Qi, L., Liang, C. (2020). Overview of cannabidiol (CBD) and its analogues: Structures, biological activities, and neuroprotective mechanisms in epilepsy and Alzheimer’s disease. European journal of medicinal chemistry, 192, 112163.
• Malabadi, R. B., Chalannavar, R. K., Divakar, M. S., Swathi, B., Komalakshi, K. V., Kamble, A. A., Munhoz, R. (2025). Industrial Cannabis sativa (Fiber or Hemp): 3D printing-hempcrete-a sustainable building material. World Journal of Advanced Engineering Technology and Sciences, 14(02), 253-282.
• Mykhailenko, O., Jalil, B., McGaw, L. J., Echeverría, J., Takubessi, M., Heinrich, M. (2025). Climate change and the sustainable use of medicinal plants: a call for “new” research strategies. Frontiers in Pharmacology, 15, 1496792.
• Nafis, A., Kasrati, A., Jamali, C. A., Mezrioui, N., Setzer, W., Abbad, A., Hassani, L. (2019). Antioxidant activity and evidence for synergism of Cannabis sativa (L.) essential oil with antimicrobial standards. Industrial Crops and Products, 137, 396-400.
• Naz, S., Ahmed, W., Ramadan, M. F. (2023). Cannabis sativa leaf essential oil fractions and bioactive compounds: chemistry, functionality and health-enhancing traits. Journal of Food Measurement and Characterization, 17(5), 4575-4593.
• Ogwu, M. C., Izah, S. C., Joshua, M. T. (2025). Ecological and environmental determinants of phytochemical variability in forest trees. Phytochemistry Reviews, 1-29.
• Onofri, C., Mandolino, G. (2017). Genomics and molecular markers in Cannabis sativa L. Cannabis sativa L.-botany and biotechnology, 319-342.
• Pagano, C., Navarra, G., Coppola, L., Avilia, G., Bifulco, M., Laezza, C. (2022). Cannabinoids: therapeutic use in clinical practice. International journal of molecular sciences, 23(6), 3344.
• Pathak, S., Nadar, R., Deruiter, J., Ramesh, S., Ramapuram, J. B., Govidarajulu, M., Dhanasekaran, M. (2022). Cannabis as a unique and valuable nutraceutical formulation for the current and future global wellbeing. In Cannabis/marijuana for healthcare (pp. 271-299). Singapore: Springer Nature Singapore.
• Pellati, F., Borgonetti, V., Brighenti, V., Biagi, M., Benvenuti, S., Corsi, L. (2018). Cannabis sativa L. and nonpsychoactive cannabinoids: their chemistry and role against oxidative stress, inflammation, and cancer. BioMed research international, 2018(1), 1691428.
• Rehman, M., Fahad, S., Du, G., Cheng, X., Yang, Y., Tang, K., Deng, G. (2021). Evaluation of hemp (Cannabis sativa L.) as an industrial crop: a review. Environmental Science and Pollution Research, 28(38), 52832-52843.
• Sirangelo, T. M., Ludlow, R. A., Spadafora, N. D. (2022). Multi-omics approaches to study molecular mechanisms in Cannabis sativa. Plants, 11(16), 2182.
• Soydan, S., Arda, B., Altuntaş, İ., Erbaş, O. (2021). Beneficial Effects of Cannabis Sativa Extract on Oxidative Stress. Journal of Experimental and Basic Medical Sciences, 2(3), 336-342.
• Tănase Apetroaei, V., Pricop, E. M., Istrati, D. I., Vizireanu, C. (2024). Hemp Seeds (Cannabis sativa L.) as a Valuable Source of Natural Ingredients for Functional Foods—A Review. Molecules, 29(9), 2097.
• Trancoso, I., de Souza, G. A., dos Santos, P. R., dos Santos, K. D., de Miranda, R. M. D. S. N., da Silva, A. L. P. M., Campostrini, E. (2022). Cannabis sativa L.: Crop management and abiotic factors that affect phytocannabinoid production. Agronomy, 12(7), 1492.
• Turek, A. (2025). A Review of Chemical Variability and Metal Contamination of Herbaceous Plants in Terms of Health Safety—A Case Study of Tanacetum vulgare L. Applied Sciences, 15(2), 911.
• Vanhaelen, M., Lejoly, J., Hanocq, M., Molle, L. (2017). Climatic and geographical aspects of medicinal plant constituents. In The Medicinal Plant Industry (pp. 59-76). Routledge.
• Xu, J., Bai, M., Song, H., Yang, L., Zhu, D., Liu, H. (2022). Hemp (Cannabis sativa subsp. sativa) Chemical composition and the application of hempseeds in food formulations. Plant Foods for Human Nutrition, 77(4), 504-513.