Salicylic acid as a stimulator of phenolic metabolism in hairy root cultures of Ocimum basilicum L.

  • V. V. Aliekseyshyna Educational and Scientific Center "Institute of Biology and Medicine" of Taras Shevchenko National University of Kyiv, Ukraine, 03022, Kyiv, Academician Glushkov Avenue, 2
  • O. V. Bulko Institute of Cell Biology and Genetic Engineering, NAS of Ukraine, Ukraine, 03143, Kyiv, Akademika Zabolotnogo str., 148 https://orcid.org/0000-0002-8048-0168
  • L. G. Lioshyna Institute of Cell Biology and Genetic Engineering, NAS of Ukraine, Ukraine, 03143, Kyiv, Akademika Zabolotnogo str., 148 https://orcid.org/0000-0003-1858-2785
Keywords: Ocimum basilicum, culture of transformed roots, hairy root, elicitation, salicylic acid, secondary metabolites, phenolic compounds

Abstract

Aim. To investigate the effect of salicylic acid as an elicitor on the accumulation of phenolic compounds in hairy root cultures of two Ocimum basilicum cultivars (purple and lemon). Methods. Basil seeds were introduced into in vitro culture following surface sterilization and cultivated on MS nutrient medium. Hairy root cultures were obtained using the standard Agrobacterium rhizogenes-mediated transformation of leaf explants. To stimulate secondary metabolite synthesis, cultures were elicited with salicylic acid at concentrations of 100 µM, 200 µM, and 300 µM. The total phenolic content was determined spectrophotometrically using the Folin–Denis method. Results. Hairy root cultures of both basil cultivars were successfully established. The application of salicylic acid was found to enhance phenolic compound accumulation in the obtained cultures. For the purple basil cultivar, the maximum phenolic content was observed at a concentration of 200 µM, whereas for the lemon basil cultivar, the optimum was 100 µM salicylic acid. Conclusions. Salicylic acid elicitation is an effective approach for stimulating the biosynthesis of phenolic compounds in hairy root cultures of basil. The optimal elicitor concentration is cultivar-dependent.

References

Bais H. P., Walker T. S., Schweizer H. P., Vivanco J. M. Root-specific elicitation and antimicrobial activity of rosmarinic acid in hairy root cultures of Ocimum basilicum. Plant Physiology and Biochemistry. 2002. Vol. 40, No. 11. P. 983–995. https://doi.org/10.1016/S0981-9428(02)01460-2.

Bhaskar R., Xavier L., Udayakumaran G., Kumar D., Venkatesh R., Nagella P. Biotic elicitors: a boon for the in vitro production of plant secondary metabolites. Plant Cell, Tissue and Organ Culture. 2022. Vol. 149, No. 1. P. 7–24. https://doi.org/10.1007/s11240-021-02131-1.

Chen J.-Y., Wen P.-F., Kong W.-F., Pan Q.-H., Zhan J.-C., Li J.-M., Wan S.-B., Huang W.-D. Effect of salicylic acid on phenylpropanoids and phenylalanine ammonia-lyase in harvested grape berries. Postharvest Biology and Technology. 2006. Vol. 40, No. 1. P. 64–72. https://doi.org/10.1016/j.postharvbio.2005.12.017.

Gamborg O. L., Miller R. A., Ojima K. Nutrient requirements of suspension cultures of soybean root cells. Experimental Cell Research. 1968. Vol. 50. P. 151–158.

Georgiev M. I., Pavlov A. I., Bley T. Hairy root type plant in vitro systems as sources of bioactive substances. Applied Microbiology and Biotechnology. 2007. Vol. 74, No. 6. P. 1175–1185. https://doi.org/10.1007/s00253-007-0856-5.

Kwon D. Y., Kim Y. B., Kim J. K., Park S. U. Production of rosmarinic acid and correlated gene expression in hairy root cultures of green and purple basil (Ocimum basilicum L.). Preparative Biochemistry and Biotechnology. 2021. Vol. 51, No. 1. P. 35–43. https://doi.org/10.1080/10826068.2020.1789990.

Murashige T., Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Plant Physiology. 1962. Vol. 15, No. 3. P. 473–497.

Pandey H., Pandey P., Singh S., Gupta R., Banerjee S. Production of anti-cancer triterpene (betulinic acid) from callus cultures of different Ocimum species and its elicitation. Protoplasma. 2015. Vol. 252, No. 2. P. 647–655. https://doi.org/10.1007/s00709-014-0711-3.

Pulvirenti L., Ferreri T., Strano T., Zappalà F., Infantino S., Caruso C., Morrone R., D’Antona N., Barbera A. C., Carrubba A., Napoli E. Effects of photoperiod on phytochemical profiles of four Ocimum basilicum genotypes in a hydroponic climate chamber. Frontiers in Agronomy. 2025. Vol. 7. P. 1653993. https://doi.org/10.3389/fagro.2025.1653993.

Singleton V. L., Orthofer R., Lamuela-Raventós R. M. Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent. Methods in Enzymology. 1999. Vol. 299. P. 152–178.

Srivastava S., Conlan X. A., Adholeya A. Chaill D M. Elite hairy roots of Ocimum basilicum as a new source of rosmarinic acid and antioxidants. Plant Cell, Tissue and Organ Culture. 2016. Vol. 126. P. 19–32. https://doi.org/10.1007/s11240-016-0973-x.

Srivastava S., Lal R. K., Yadav K., Pant Y., Bawitlung L., Kumar P., Mishra A., Gupta P., Pal A., Rout P. K., Chanotiya C. S. Chemical composition of phenylpropanoid-rich chemotypes of Ocimum basilicum L. and their antimicrobial activities. Industrial Crops and Products. 2022. Vol. 183. P. 114978. https://doi.org/10.1016/j.indcrop.2022.114978.

Torres R. G., Casanova L., Carvalho J., Marcondes M.C., Costa S. S., Sola-Penna M., Zancan P. Ocimum basilicum but not Ocimum gratissimum presents cytotoxic effects on human breast cancer cell line MCF-7, inducing apoptosis and triggering mTOR/Akt/p70S6K pathway. Journal of Bioenergetics and Biomembranes. 2018. Vol. 50, No. 2. P. 93–105. https://doi.org/10.1007/s10863-018-9750-3.

Vogt T. Phenylpropanoid biosynthesis. Molecular Plant. 2010. Vol. 3, No. 1. P. 2–20. https://doi.org/10.1093/mp/ssp106.

Zhakipbekov K., Turgumbayeva A., Akhelova S., Bekmuratova K., Blinova O., Utegenova G., Shertaeva K., Sadykov N., Tastambek K., Saginbazarova A., Urazgaliyev K., Tulegenova G., Zhalimova Z., Karasova Z. Antimicrobial and other pharmacological properties of Ocimum basilicum (Lamiaceae). Molecules. 2024. Vol. 29, No. 2. P. 388. https://doi.org/10.3390/molecules29020388.