Response of Culture Media and Auxin on Growth and Glucosinolate Accumulation in the Hairy Root Cultures of Rocket (Eruca sativa)

Main Article Content

Sang Un Park
Nam Su Kim
Sun Ju Bong
Sook Young Lee*

Abstract

Rocket (Eruca sativa) is a domesticated plant species that is commonly eaten in salads and known to provide health benefits because of the high levels of glucosinolates, flavonols, and other compounds. Hairy root cultures (HRCs) are effective biotechnological tools for biosynthesis of secondary metabolites under various growing conditions. HRCs of rocket were treated with growth media of half-strength and full-strength Murashige-Skoog (MS) media, Gamborg’s B5 medium, and Schenk and Hildebrand (SH) medium and auxins to evaluate the growth response and the accretion of glucosinolate. The growth pattern of the hairy roots varied extensively under the different media and auxin treatments; the highest and the lowest fresh weights were recorded in HRCs grown under full-strength SH and half-strength MS media, respectively. Treatment with NAA at 1.0 mg/l produced the highest hairy root fresh weight, followed by the treatments with IAA 0.1 mg/l and IBA 1.0 mg/l. The MS media induced the highest glucosinolate accumulation, followed by B5; all media enhanced the production of glucosinolates but auxins treatments (exception for glucoerucin) did not positively enhance the production of glucosinolates. Total glucosinolate levels were increased 1.7, 1.68, 1.33, and 1.26 fold in full-strength MS, half-strength MS, half-strength B5, and B5, respectively. These findings indicated that hairy root production and glucosinolate accumulation did not follow the same trend. Although SH media and NAA 1.0 slightly enhanced hairy root production, full-strength MS media induced higher amounts of glucosinolate, and auxin treatments did not increase the accumulation of glucosinolate. We therefore propose that MS media, regardless of additional treatment, provides a valuable alternative approach for the mass production of hairy root cultures and glucosinolates in rocket.


 


Keywords: glucosinolates; growth media; auxin; hairy root; rocket salad


*Corresponding author: Tel.: +82-10-8610-6739; Fax: +82-61-555-1260


                                       E-mail: seedbank@chosun.ac.kr

Downloads

Download data is not yet available.

Article Details

Section
Research Articles

References

[1] Pasini, F., Verardo, V., Cerretani, L., Caboni, M.F. and D’Antuono, L.F., 2011. Rocket salad (Diplotaxis and Eruca spp.) sensory analysis and relation with glucosinolate and phenolic content. Journal of the Science of Food and Agriculture, 91(15), 2858-2864.
[2] Martinez-Sanchez, A., Marin, A., Llorach, R., Ferreres, F. and Gil, M.I., 2006. Controlled atmosphere preserves quality and phytonutrients in wild rocket (Diplotaxis tenuifolia). Postharvest Biology and Technology, 40(1), 26-33.
[3] Bozokalfa, K.M., Esiyok, D. and Yagmur, B., 2011. Use of multivariate analysis in mineral accumulation of rocket (Eruca sativa) accessions. Genetika-Belgrade, 43(3), 437-448.
[4] Sahoo, R.K., Kumar, M., Sukla, L.B. and Subudhi, E., 2017. Bioprospecting hot spring metagenome: lipase for the production of biodiesel. Environmental Science and Pollution Research, 24(4), 3802-3809.
[5] Bennett, R.N., Rosa, E.A.S., Mellon, F.A. and Kroon, P.A., 2006. Ontogenic profiling of glucosinolates, flavonoids, and other secondary metabolites in Eruca sativa (salad rocket), Diplotaxis erucoides (wall rocket), Diplotaxis tenuifolia (wild rocket), and Bunias orientalis (Turkish rocket). Journal of Agricultural and Food Chemistry, 54(11), 4005-4015.
[6] Azarenko, O., Jordan, M.A. and Wilson, L., 2014. Erucin, the major isothiocyanate in arugula (Eruca sativa), inhibits proliferation of MCF7 tumor cells by suppressing microtubule dynamics. PloS One, 9(6), e100599, https://doi.org/10.137/journal.pone.0100599
[7] Michael, H.N., Shafik, R.E. and Rasmy, G.E., 2011. Studies on the chemical constituents of fresh leaf of Eruca sativa extract and its biological activity as anticancer agent in vitro. Journal of Medicinal Plants Research, 5, 1184-1191.
[8] Alam, M.S., Kaur, G., Jabbar, Z., Javed, K. and Athar, M., 2007. Eruca sativa seeds possess antioxidant activity and exert a protective effect on mercuric chloride induced renal toxicity. Food and Chemical Toxicology, 45(6), 910-920.
[9] Koubaa, M., Driss, D., Bouaziz, F., Ghorbel, R.E. and Chaabouni, S.E., 2015. Antioxidant and antimicrobial activities of solvent extract obtained from rocket (Eruca sativa L.) flowers. Free Radicals and Antioxidants, 5(1), 29-34.
[10] Khoobchandani, M., Ojeswi, B.K., Ganesh, N., Srivastava, M.M., Gabbanini, S., Matera, R., Iori, R. and Valgimigli, L., 2010. Antimicrobial properties and analytical profile of traditional Eruca sativa seed oil: Comparison with various aerial and root plant extracts. Food Chemistry, 120(1), 217-224.
[11] Yehuda, H., Khatib, S., Sussan, I., Musa, R., Vaya, J. and Tamir, S., 2009. Potential skin antiinflammatory effects of 4‐methylthiobutylisothiocyanate (MTBI) isolated from rocket (Eruca sativa) seeds. Biofactors, 35(3), 295-305.
[12] Clarke, D.B., 2010. Glucosinolates, structures and analysis in food. Analytical Methods, 2(4), 310-325.
[13] Jørgensen, M.E., Nour-Eldin, H.H. and Halkier, B.A., 2015. Transport of defense compounds from source to sink: lessons learned from glucosinolates. Trends Plant Science, 20(8), 508-514.
[14] Holst, B. and Williamson, G., 2004. A critical review of the bioavailability of glucosinolates and related compounds. Natural Product Reports, 21(3), 425-447.
[15] D’Antuono, L.F., Elementi, S. and Neri, R., 2009. Exploring new potential health-promoting vegetables: glucosinolates and sensory attributes of rocket salads and related Diplotaxis and Eruca species. Journal of the Science of Food and Agriculture, 89(4), 713-722.
[16] Higdon, J.V., Delage, B., Williams, D.E. and Dashwood, R.H., 2007. Cruciferous vegetables and human cancer risk: epidemiologic evidence and mechanistic basis. Pharmacological Research, 55(3), 224-236.
[17] Herr, I. and Büchler, M.W., 2010. Dietary constituents of broccoli and other cruciferous vegetables: implications for prevention and therapy of cancer. Cancer Treatment Reviews, 36(5), 377-383.
[18] Krzyzanowska, J., Czubacka, A. and Oleszek, W., 2010. Dietary phytochemicals and human health. Advances in Experimental Medicine and Biology, 698, 74-98.
[19] Podsedek, A., 2007. Natural antioxidants and antioxidant capacity of Brassica vegetables: A review. LWT-Food Science and Technology, 40(1), 1-11.
[20] Casagrande, S.S., Wang, Y., Anderson, C. and Gary, T.L., 2007. Have Americans increased their fruit and vegetable intake? The trends between 1988 and 2002. American Journal of Preventive Medicine, 32(4), 257-263.
[21] Woodward, A.W. and Bartel, B., 2005. Auxin: regulation, action, and interaction. Annals of Botany, 95(5), 707-735.
[22] George, E.F., Hall, M.A. and De Klerk, G.-J., 2008. The components of plant tissue culture media I: macro-and micro-nutrients. In: E.F. George, M.A. Hall and G.-J. De Klerk, eds. Plant Propagation by Tissue Culture: Volume 1. The Background. 3rd ed. Dordrecht: Springer, pp. 65-113.
[23] Saad, A.I.M. and Elshahed, A.M., 2012. Plant tissue culture media. In: A. Leva and L.M.R. Rinaldi, eds. Recent Advances in Plant In Vitro Culture. Winchester: In Tech, 29-40.
[24] Washida, D., Shimomura, K., Nakajima, Y., Takido, M. and Kitanaka, S., 1998. Ginsenosides in hairy roots of a Panax hybrid. Phytochemistry, 49(8), 2331-2335.
[25] Dhakulkar, S., Ganapathi, T.R. Bhargava, S. and Bapat, V.A., 2005. Induction of hairy roots in Gmelina arborea Roxb. and production of verbascoside in hairy roots. Plant Science, 169(5), 812-818.
[26] Kumar, G.B.S., Ganapathi, T.R., Srinivas, L., Revathi, C.J. and Bapat, V.A., 2006. Expression of hepatitis B surface antigen in potato hairy roots, Plant Science, 170(5), 918-925.
[27] Kim, Y.S., Li, X., Park, W.T., Uddin, M.R., Park, N.I., Kim, Y.B., Lee, M.Y. and Park, S.U., 2012. Influence of media and auxins on growth and falvone production in hairy root cultures of baikal skullcap, Scutellaria baicalensis. Plant Omics Journal, 5(1), 24-27.
[28] Georgiev, M.I., Agostini, E., Ludwig-Müller, J. and Xu, J., 2012. Genetically transformed roots: from plant disease to biotechnological resource. Trends in Biotechnology, 30(10), 528-537.
[29] Cuong, D.M., Park, S.U., Park, C.H., Kim, N.S., Bong, S.J. and Lee S.Y., 2019. Comparative analysis of glucosinolate production in hairy roots of green and red kale (Brassica oleracea var. acephala). Preparative Biochemistry and Biotechnology, 49(8), 775-782.
[30] Cuong, D.M., Park, C.H., Bong, S.J., Kim, N.S., Kim, J.K. and Park, S.U., 2019. Enhancement of glucosinolate production in watercress (Nasturtium officinale) hairy roots by overexpressing cabbage transcription factors. Journal of Agricultural and Food Chemistry, 67(17), 4860-4867.
[31] Cuong, D.M., Kim, J.K., Bong, S.J., Baek, S.A., Jeon, J., Park, J.S. and Park, S.U., 2018. Comparative analysis of glucosinolates and metabolite profiling of green and red mustard (Brassica juncea) hairy roots. 3 Biotech, 8, 382, https://doi.org/10.1007/s13205-018-1393-x
[32] Bong, S.J., Uddin, M.R., Kim, S.-J., Park, J.S. and Park, S.U., 2015. Influence of auxins and wounding on glucosinolate biosynthesis in hairy root cultures of Chinese cabbage (Brassica rapa ssp. pekinensis). Biosciences Biotechnology Research Asia, 12(2), 1041-1046.
[33] Kim, S.-J., Park, W.T., Uddin, M.R., Kim, Y.B., Nam, S.-Y., Jho, K.H. and Park, S.U., 2013a. Glucosinolate biosynthesis in hairy root cultures of broccoli (Brassica oleracea var. italica). Natural Product Communications, 8(2), 217-220.
[34] Xue, S.-H., Luo, X.-J., Wu, Z.-H., Zhang, H.-L. and Wang, X.-Y., 2008. Cold storage and cryopreservation of hairy root cultures of medicinal plant Eruca sativa Mill., Astragalusmembranaceus and Gentianamacrophylla Pall. Plant Cell, Tissue and Organ Culture, 92(3), 251-260.
[35] Kastell, A., Schreiner, M., Knorr, D., Ulrichs, C. and Mewis, I., 2018. Influence of nutrient supply and elicitors on glucosinolate production in E. sativa hairy root cultures. Plant Cell, Tissue and Organ Culture, 132(3), 561-572.
[36] Murashige, T. and Skoog, F., 1962. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum, 15(3), 473-497.
[37] Schenk, R.U. and Hildebrandt, A.C., 1972. Medium and techniques for induction and growth of monocotyledonous and dicotyledonous plant cell cultures. Canadian Journal of Botany, 50(1), 199-204.
[38] Gamborg, O. L., Miller, R. and Ojima, K., 1968. Nutrient requirements of suspension cultures of soybean root cells. Experimental Cell Research, 50(1), 151-158.
[39] International Organization of Standardization, 1992. Rapeseed - Determination of Glucosinolates Content- Part 1, Method Using High-Performance Liquid Chromatography. (ISO 9167-1:1992). Geneva: International Organization of Standardization.
[40] Kim, S.J., Kawaharada, C., Jin, S., Hashimoto, M., Ishii, G. and Yamauchi, H., 2007. Structural elucidation of 4-(cystein-S-yl) butyl glucosinolate from the leaves of Eruca sativa. Bioscience, Biotechnology, and Biochemistry, 71(1), 114-121.
[41] Murthy, H.N., Lee, E.-J. and Paek, K.-Y., 2014. Production of secondary metabolites from cell and organ cultures: strategies and approaches for biomass improvement and metabolite accumulation. Plant Cell, Tissue and Organ Culture, 118(1), 1-16.
[42] Park, C.H, Kim, N.S., Yeo, H.J., Bong, S.J., Park, J.S., Park, N.I. and Park, S.U., 2019. Effects of culture medium on growth and glucosinolate accumulation in the hairy root cultures of watercress (Nasturtium officinale). Research Journal of Biotechnology, 14(2), 61-66.
[43] Sivakumar, G., Yu, K.W., Hahn, E.J. and Paek, K.Y., 2005. Optimization of organic nutrients for ginseng hairy roots production in large-scale bioreactors. Current Science, 89(4), 641-649.
[44] Nagella, P., Chung, I.-M. and Murthy, H.N., 2011. In vitro production of gymnemic acid from cell suspension cultures of Gymnema sylvestre R. Br. Engineering in Life Sciences, 11(5), 537-540.
[45] Cheruvathur, M.K. and Thomas, T.D., 2014. Effect of plant growth regulators and elicitors on rhinacanthin accumulation in hairy root cultures of Rhinacanthus nasutus (L.) Kurz. Plant Cell, Tissue and Organ Culture, 118(1), 169-177.
[46] Mantell, S.H. and Smith, H., 1983. Cultural factors that influence secondary metabolite accumulations in plant cell and tissue cultures. In: S.H. Mantell and H. Smith, eds. Plant Biotechnology. Cambridge: Cambridge University Press, pp. 75-108.
[47] Sahai, O. and Shuler, M., 1984. Environmental parameters influencing phenolics production by batch cultures of Nicotiana tabacum. Biotechnology and Bioengineering, 26(2), 111-120.
[48] Sauerwein, M., Yamazaki, T. and Shimomura, K., 1991. Hernandulcin in hairy root cultures of Lippia dulcis. Plant Cell Reports, 9(10), 579-581.
[49] Uddin, M.R., Park, K.W., Kim, Y.K., Park, S.U. and Pyon, J.Y., 2010. Enhancing sorgoleone levels in grain sorghum root exudates. Journal of Chemical Ecology, 36(8), 914-922.
[50] Uddin, M.R., Park, W.T., Kim, Y.K., Pyon, J.Y. and Park, S.-U., 2011. Effects of auxins on sorgoleone accumulation and genes for sorgoleone biosynthesis in sorghum roots. Journal of Agricultural and Food Chemistry, 59(24), 12948-12953.
[51] Kim, H.H, Kwon, D.Y., Bae, H., Kim, S.J., Kim, Y.B., Uddin, M.R. and Park, S.U., 2013. Influence of auxins on glucosinolate biosynthesis in hairy root cultures of broccoli (Brassica oleracea var. italica). Asian Journal of Chemistry, 25(11), 6099-6101.
[52] Lee, S.Y., Bong, S.J., Kim, J.K. and Park, S.U., 2016. Glucosinolate biosynthesis as influenced by growth media and auxin in hairy root cultures of kale (Brassica oleracea var. acephala). Emirates Journal of Food and Agriculture, 28(4), 277-282.
[53] Brown, A.F., Yousef, G.G., Jeffery, E.H., Klein, B.P., Wallig, M.A., Kushad, M.M. and Juvik, J.A., 2002. Glucosinolate profiles in broccoli: Variation in levels and implications in breeding for cancer chemoprotection. Journal of the American Society for Horticultural Science, 127(5), 807-813.
[54] Vallejo, F., Tomas-Barberán, F.A., Gonzalez Benavente-García, A. and García-Viguera, C., 2003. Total and individual glucosinolate contents in inflorescences of eight broccoli cultivars grown under various climatic and fertilization conditions. Journal of the Science of Food and Agriculture, 83(4), 307-313.
[55] Kumar, S. and Andy, A., 2012. Health promoting bioactive phytochemicals from Brassica. International Food Research Journal, 19(1), 141-152.
[56] Bálványos, I., Kursinszki, L. and Szõke, E., 2001. The effect of plant growth regulators on biomass formation and lobeline production of Lobelia inflata L. hairy root culture. Plant Growth Regulator, 34(3), 339-345.
[57] Washida, D., Shimomura, K., Takido, M. and Kitanaka, S., 2004. Auxins affected ginsenoside production and growth of hairy roots in Panax hybrid. Biological and Pharmaceutical Bulletin, 27(5), 657-660.