Effects of Growth Regulators on Biomass and the Production of Secondary Metabolites in Peppermint (Mentha pi-perita) Micropropagated in Vitro


The effects of plant growth regulators on peppermint (Mentha piperita) cultured in vitro were studied for the purpose of maximizing growth and essential oil production in micropropagated plants. The basal medium was experimentally supplemented with the auxin 4-indol-3-ylbutyric acid (IBA) and the cytokinin 6-benzylaminopurine (BAP) individually and in combination. Supplementation with BAP alone resulted in the highest values for root length, root dry weight, shoot length, and numbers of nodes, leaves, and ramifications. Treatment with IBA alone or with IBA + BAP resulted in a ~50% increase in shoot fresh weight. The production of secondary metabolites was affected only by the addition of cytokinin, which resulted in a ~40% increase in the total yield of essential oils (EOs). Similar trends were observed for yields of the major EO components (menthone, menthol, pulegone, and menthofuran). Our findings demonstrate that the application of growth regulators increases EO production and biomass concomitantly in an herbaceous species rich in commercially valuable terpenes.

Share and Cite:

Santoro, M. , Nievas, F. , Zygadlo, J. , Giordano, W. and Banchio, E. (2013) Effects of Growth Regulators on Biomass and the Production of Secondary Metabolites in Peppermint (Mentha pi-perita) Micropropagated in Vitro. American Journal of Plant Sciences, 4, 49-55. doi: 10.4236/ajps.2013.45A008.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] N. S. Sangwan, A. H. A. Farooqi, F. Shabih and R. S. Sangwan, “Regulation of Essential Oil Production in Plants,” Plant Growth Regulation, Vol. 34, No. 1, 2001, pp. 3-21. doi:10.1023/A:1013386921596
[2] D. Ram, M. Ram and R. Singh, “Optimization of Water and Nitrogen Application to Menthol Mint (Mentha arvensis L.) through Sugarcane Trash Mulch in a Sandy Loam Soil of Semi-Arid Subtropical Climate,” Bioresource Technology, Vol. 97, No. 7, 2006, pp. 886-893. doi:10.1016/j.biortech.2005.04.047
[3] W. Schwab, R. Davidovich-Rikanati and E. Lewinsohn, “Biosynthesis of Plant-Derived Flavor Compounds,” The Plant Journal, Vol. 54, No. 4, 2008, pp. 712-732. doi:10.1111/j.1365-313X.2008.03446.x
[4] R. Croteau, T. Kutchan and N. Lewis, “Natural Products (Secondary Metabolites),” In: B. Buchanan, W. Gruissem and R. Joneas, Eds., Biochemistry and Molecular Biology of Plants, American Society of Plant Biologists, Rockville, 2000, pp. 1250-1268.
[5] P. Harrewijn, A. M. Van Oosten and P. G. Piron, “Natural Terpenoids as Messengers. A Multidisciplinary Study of Their Production, Biological Functions and Practical Applications,” Kluwer Academic Publishers, London, 2001.
[6] F. Bourgaud, A. Gravot, S. Milesi and E. Gontier, “Production of Plant Secondary Metabolites: A Historical Perspective,” Plant Science, Vol. 161, No. 13, 2001, pp. 839- 851. doi:10.1016/S0168-9452(01)00490-3
[7] R. Verpoorte, A. Contin and J. Memelink, “Biotechnology for the Production of Plant Secondary Metabolites,” Phytochemistry Reviews, Vol. 1, No. 1, 2002, pp. 13-25. doi:10.1023/A:1015871916833
[8] M. Vanisree and H.-S. Tsay, “Plant Cell Cultures: Production of Biologically Important Secondary Metabolites from Medicinal Plants of Taiwan,” In: O. Kayser and W. Quax Eds., Medicinal Plant Biotechnology. From Basic Research to Industrial Application, Wiley-VCH Verlag GmbH & Co., KGaA, Weinheim, 2007, pp. 267-285.
[9] R. Verpoorte, R. van der Heijden and J. Memelink, “Engineering the Plant Cell Factory for Secondary Metabolite Production,” Transgenic Research, Vol. 9, No. 4-5, 2000, pp. 323-343. doi:10.1023/A:1008966404981
[10] R. Rao and G. A. Ravishankar, “Plant Cell Cultures: Chemical Factories of Secondary Metabolites,” Biotechnology Advances, Vol. 20, No. 2, 2002, pp. 101-153. doi:10.1016/S0734-9750(02)00007-1
[11] S. V. Tisserat and R. Silman, “Influence of Modified Oxygen and Carbon Dioxide Atmospheres on Mint and Thyme Plant Growth, Morphogenesis and Secondary Metabolism in Vitro,” Plant Cell Reports, Vol. 20, No. 10, 2002, pp. 912-916. doi:10.1007/s00299-001-0428-6
[12] G. W. Turner, J. Gershenzon and R. B. Croteau, “Distribution of Peltate Glandular Trichomes on Developing Leaves of Peppermint,” Plant Physiology, Vol. 124, No. 2, 2000, pp. 655-664. doi:10.1104/pp.124.2.655
[13] B. Tisserat and S. Vaughn, “Essential Oils Enhanced by Ultra-High Carbon Dioxide Levels from Lamiaceae Species Grown in Vitro and in Vivo,” Plant Cell Reports, Vol. 20, No. 4, 2001, pp. 361-368. doi:10.1007/s002990100327
[14] Y. N. Shukla and A. H. Farooqi, “Utilization of Plant Growth Regulators in Aromatic Plant Production,” Medicinal & Aromatic Plants, Vol. 12, No. 3, 1990, pp. 152- 157.
[15] T. Murashige and F. A. Skoog, “Revised Medium for Rapid Growth and Bio Assay with Tobacco Tissue Culture,” Plant Physiology, Vol. 15, No. 3, 1962, pp. 473-497. doi:10.1111/j.1399-3054.1962.tb08052.x
[16] M. Santoro, J. Zygadlo, W. Giordano and E. Banchio, “Volatile Organic Compounds from Rhizobacteria Increase Biosynthesis of Essential Oils and Growth Parameters in Peppermint (Mentha piperita),” Plant Physiology and Biochemistry, Vol. 49, No. 11, 2011, pp. 1177-1182. doi:10.1016/j.plaphy.2011.07.016
[17] M. C. Mok, “Cytokinins and Plant Development—An Overview,” In: D. W. S. Mok and M. C. Mok, Eds., Cytokinins. Chemistry, Activity, and Function, CRC Press, Boca Raton, 1994, pp. 155-166.
[18] A. A. Hafiz, S. D. Johnson and J. V. Staden, “Promoting Branching of a Potential Biofuel crop Jatropha curcas L. by Foliar Application of PGR,” Plant Growth Regulation, Vol. 58, No. 3, 2009, pp. 287-295. doi:10.1007/s10725-009-9377-9
[19] C. Liu, J. Zhu, Z. Liu, L. Li, R. Pan and L. Jin, “Exogenous Auxin Effects on Growth and Phenotype of Normal and Hairy Roots of Pueraria lobata (Wild.) Ohwi,” Plant Growth Regulation, Vol. 38, No. 1, 2002, pp. 37-43. doi:10.1023/A:1020904528045
[20] J. F. Harbage and D. P. Stimart, “Effect of pH and 1H- Indole-3-Butyric Acid (IBA) on Rooting of Apple Microcuttings,” Journal of the American Society for Horticultural Science, Vol. 121, No. 6, 1996, pp. 1049-1053.
[21] V. R. Affonso, H. R. Bizzo, C. L. Salguiero Lage and A. Sato, “Influence of Growth Regulators in Biomass Production and Volatile Profile of in Vitro Plantlets of Thymus vulgaris L.,” Journal of Agricultural and Food Chemistry, Vol. 57, No. 14, 2009, pp. 6392-6395. doi:10.1021/jf900816c
[22] P. Baskaran, B. Ncube and J. Van Staden, “In Vitro Propagation and Secondary Product Production by Merwilla plúmbea (Lindl.) Speta,” Plant Growth Regulation, Vol. 67, No. 13, 2012, pp. 235-245. doi:10.1007/s10725-012-9682-6
[23] L. Maes and A. Goossens, “Hormone-Mediated Promotion of Trichome Initiation in Plants Is Conserved but Utilizes Species- and Trichome-Specific Regulatory Mechanisms,” Plant Signaling & Behavior, Vol. 5, No. 2, 2010, pp. 205-207. doi:10.4161/psb.5.2.11214
[24] H. A. Farooqi and S. Sharma, “Effect of Growth Retardants on Growth and Essential Oil Content in Japanese mint. P1,” Growth Regulation, Vol. 7, No. 1, 1988, pp. 39-45. doi:10.1007/BF00121688
[25] D. Fraternale, L. Giamperi, D. Ricci, M. B. L. Rocchi, L. Guidi, F. Epifanio and M. C. Marcotullio, “The Effect of TRIA on Micropropagation and on Secretory System of Thymus mastichina,” Plant Cell, Tissue and Organ Culture, Vol. 74, No. 1, 2003, pp. 87-97. doi:10.1023/A:1023321024040
[26] D. A. Herms and W. J. Mattson, “The Dilemma of Plants: to Grow or Defend,” The Quarterly Review of Biology, Vol. 67, 1992, pp. 283-335. doi:10.1086/417659
[27] R. Matyssek, R. Agerer, D. Ernst, J.-C. Munch, W. Osswald, H. Pretzsch, E. Priesack, H. Schnyder and D. Treutter, “The Plant’s Capacity in Regulating Resource Demand,” Plant Biology, Vol. 7, No. 1, 2005, pp. 560- 580. doi:10.1055/s-2005-872981
[28] S. S. Mahmoud and R. B. Croteau, “Strategies for Transgenic Manipulation of Monoterpene Biosynthesis in Plants—Review,” Trends in Plant Science, Vol. 7, No. 8, 2002, pp. 366-373. doi:10.1016/S1360-1385(02)02303-8

Copyright © 2023 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.