Callus induction and bioactive phenolic compounds production from Byrsonima verbascifolia (L.) DC. (Malpighiaceae)

Ana Hortência Fonsêca Castro, Karina de Queiroz Braga, Fernanda Marinez de Sousa, Mairon César Coimbra, Rafael César Russo Chagas


This study developed a methodology for callus induction in leaf segments of B. verbascifolia and evaluated the bioactive phenolic compounds production. Leaf explants were cultured in MS medium with 30 g L-1 sucrose, solidified with 7 g L- 1 agar supplemented with 2,4-D (0; 4.52; 9.05; 18.10 μM) and BAP (0; 4.44; 8.88; 17.75 μM ) in the presence and absence of light. Forty-five days after inoculation we assessed the percentage of callus induction, color, consistency, fresh and dry matter, total phenols, flavonoids, tannins contents, and chromatographic profile by HPLC-DAD method. Callus induction occurred only in medium with growth regulators. Maximal induction (100%) was found in medium containing 2,4-D combined with BAP in the presence and absence of light. We obtained friable and compact callus in yellow, green, and red. Culture media containing 4.52 μM 2,4-D + 4.44 μM BAP induced 100% of friable callus with higher fresh and dry weight in the absence of light. The callus produced higher amounts of total phenols and flavonoids than the initial explant. Total tannins were detected only in callus induced in media containing 17.75 μM BAP and 4.52 μM 2,4-D + 17.75 μM BAP, and were not found in the initial explant. The bioactive phenolic compounds detected are derived from benzoic, p-coumaric, cinnamic, gallic acids, and catechins.


Cerrado; Medicinal plant; In vitro culture; Secondary metabolites

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ABAD-GARCÍA, B. et al. A general analytical strategy for the characterization of phenolic compounds in fruit juices by high-performance liquid chromatography with diode array detection coupled to electrospray ionization and triple quadrupole mass spectrometry. Journal of Chromatography A, v. 1216, n. 28, p. 5398-5415, 2009.

ALMEIDA, S. P. de et al. Cerrado: espécies vegetais úteis. Planaltina: EMBRAPA-CPAC, 1998. 464 p.

AN, S. M.; KOH, J. S.; BOO, Y. C. p-coumaric acid not only inhibits human tyrosinase activity in vitro but also melanogenesis in cells exposed to UVB. Phytotherapy Research, v. 24, n. 8, p. 1175-1180, 2010.

ARUNYANART, S.; CHAITRAYAGUN, M. Induction of somatic embryogenesis in lotus (Nelumbo nucifera Geartn.). Scientia Horticulturae, v. 105, n. 3, p. 411-420, 2005.

ASSOCIATION OF OFFICIAL ANALYTICAL CHEMISTS. Official methods of the association of official analytical chemists. 11. ed. Washington: AOAC International, 1995. 1015 p.

CASTRO, A. H. F. et al. Calogênese e teores de fenóis e taninos totais em barbatimão [Stryphnodendron adstringens (Mart.) Coville]. Ciência e Agrotecnologia, v. 33, n. 2, p. 385-390, 2009.

CECÍLIO, A. B. et al. Screening of Brazilian medicinal plants for antiviral activity against rotavirus. Journal of Ethnopharmacology, v. 141, n. 3, p. 975-981, 2012.

CHERDSHEWASART, W.; SUBTANG, S.; DAGKAB, W. Major isoflavonoid contents of the phytoestrogen rich-herb Pueraria mirifica in comparison to Pueraria lobata. Journal of Pharmaceutical and Biomedical Analysis, v. 43, n. 2, p. 428-434, 2007.

COENEN, C.; LOMAX, T. L. Auxin-cytokinin interactions in higher plants: old problems and new tools. Trends in Plant Science, v. 2, n. 9, p. 351-356, 1997.

DONG, C.; WANG, W. Headspace solid-phase microextraction applied to the simultaneous determination of sorbic and benzoic acids in beverages. Analytica Chimica Acta, v. 562, n. 1, p. 23-29, 2006.

FERREIRA, D. F. Sisvar: a computer statistical analysis system. Ciência e Agrotecnologia, v. 35, n. 6, p. 1039-1042, 2011.

FUKUDA, N. et al. Directional blue light irradiation triggers epidermal cell elongation of abaxial side resulting in inhibition of leaf epinasty in geranium under red light condition. Scientia Horticulturae, v. 115, n. 2, p. 176-182, 2008.

GONÇALVES, C. A. et al. Gestational exposure to Byrsonima verbascifolia: teratogenicity, mutagenicity and immunomodulation evaluation in female Swiss mice. Journal of Ethnopharmacology, v. 150, n. 3, p. 843-850, 2013.

GUILHON-SIMPLICIO, F.; PEREIRA, M. de M. Aspectos químicos e farmacológicos de Byrsonima (Malpighiaceae). Química Nova, v. 34, n. 6, p. 1032-1041, 2011.

HAGERMAN, A. E. Radial diffusion method for determining tannin in plant extract. Journal of Chemical Ecology, v. 13, n. 3, p. 437-449, 1987.

KAKANI, A.; PENG, Z. ARR5 and ARR6 mediate tissue specific cross-talk between auxin and cytokinin in Arabidopsis. American Journal of Plant Sciences, v. 2, n. 4, p. 549-553, 2011.

KAUR, M. et al. Gallic acid, an active constituent of grape seed extract, exhibits anti-proliferative, pro-apoptotic and anti-tumorigenic effects against prostate carcinoma xenograft growth in nude mice. Pharmaceutical Research, v. 26, n. 9, p. 2133-2140, 2009.

KIM, S. H. et al. Gallic acid inhibits histamine release and pro-inflammatory cytokine production in mast cells. Toxicological Sciences, v. 91, n. 1, p. 123-131, 2006.

LOREDO-CARRILLO, S. E. et al. Establishment of callus from Pyrostegia venusta (Ker Gawl.) Miers and effect of abiotic stress on flavonoids and sterols accumulation. Journal of Plant Biochemistry and Biotechnology, v. 22, n. 3, p. 312-318, 2013.

MÄÄTTÄ, K. R.; KAMAL-ELDIN, A.; TÖRRÖNEN, A. R. High-performance liquid chromatography (HPLC) analysis of phenolic compounds in berries with diode array and electrospray ionization mass spectrometric (MS) detection: ribes species. Journal of Agricultural and Food Chemistry, v. 51, n. 23, p. 6736-6744, 2003.

MANEECHAI, S. et al. Flavonoid and stilbenoid production in callus cultures of Artocarpus lakoocha. Phytochemistry, v. 81, p. 42-49, 2012.

MEYER, J. E.; PE´PIN, M. F.; SMITH, M. A. L. Anthocyanin production from Vaccinium pahalae: limitations of the physical microenvironment. Journal of Biotechnology, v. 93, n. 1, p. 45-57, 2002.

MURASHIGE, T.; SKOOG, F. A revised medium for rapid growth and biossays with tabacco tissue cultures. Physiologia Plantarum, v. 15, n. 3, p. 473-497, 1962.

OKSMAN-CALDENTEY, K.; INZÉ, D. Plant cell factories in the post-genomic era: new ways to produce designer secondary metabolites. Trends in Plant Science, v. 9, n. 9, p. 433-40, 2004.

PALACIO, L. et al. Phenolic compound production in relation to differentiation in cell and tissue cultures of Larrea divaricata (Cav.). Plant Science, v. 193-194, p. 1-7, 2012.

SAHIN, G.; VERMA, S. K.; GUREL, E. Calcium and magnesium elimination enhances accumulation of cardenolides in callus cultures of endemic Digitalis species of Turkey. Plant Physiology and Biochemistry, v. 73, p. 139-143, 2013.

SAKAKIBARA, H. et al. Simultaneous determination of all polyphenols in vegetables, fruits, and teas. Journal of Agricultural and Food Chemistry, v. 51, n. 3, p. 571-581, 2003.

SILVA, L. C. et al. Cell viability of Byrsonima intermedia A. Juss Calli. Journal of Agricultural Science and Technology, v. 2, n. 8, p. 713-720, 2012.

SZOPA, A.; EKIERT, H. Production of biologically active phenolic acids in Aronia melanocarpa (Michx.) Elliott in vitro cultures cultivated on different variants of the Murashige and Skoog medium. Plant Growth Regulators, v. 72, n. 1, p. 51-58, 2014.

WOISKY, R. G.; SALATINO, A. Analysis of propolis: some parameters and procedures for chemical quality control. Journal of Apicultural Research, v. 37, n. 2, p. 99-105, 1998.

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