Biocatalysis and Agricultural Biotechnology 22 (2019) 101408 Available online 30 October 2019 1878-8181/© 2019 Elsevier Ltd. All rights reserved. Stimulation of 6-benzylaminopurine and meta-topolin-induced in vitro shoot organogenesis and production of favonoids of Amburana cearensis (Allem~ ao) A.C. Smith) J� essica Nascimento Costa Vasconcelos a , Alone Lima Brito a , Amanda Lima Pinheiro a , Dinah Ise Jimenez Gonçalves e Costa Pinto a , Jackson Roberto Guedes da Silva Almeida b , Taliane Leila Soares a, * , Jos� e Raniere Ferreira de Santana a a Universidade Estadual de Feira de Santana, Av. Transnordestina, s/n - Novo Horizonte, Feira de Santana, BA, 44036-900, Brazil b Universidade Federal do Vale do S~ ao Francisco, Av. Jos� e de S� a Maniçoba, s/n, Petrolina, PE, 56304-917, Brazil A R T I C L E INFO Keywords: Cumaru In vitro culture Growth regulators BAP Mtop Secondary metabolites ABSTRACT Amburana cearensis (cumarú) is a tree species native to the semi-arid region of northeastern Brazil. Extracts obtained from its bark, seeds and leaves are widely used in folk medicine. The growing demand for phytotherapic preparations from this species makes it necessary to develop techniques for commercial cultivation to supply inputs to the pharmaceutical industry. In this respect, in vitro culture is a viable alternative to meet this demand. The objective of this study was to evaluate the effect of two cytokinins, 6-benzylaminopurine (BAP) and meta- topolin (Mtop), on the direct organogenesis and to establish the phytochemical profle of A. cearensis. In vitro multiplication was tested from leaf and cotyledon segments, inoculated in woody plant medium (WPM) sup- plemented with varying concentrations of BAP (0.00, 2.22, 4.44, 6.66 or 8.88 μM) or Mtop (0.0, 2.0, 4.0, 6.0, 8.0 or 10.0 μM). Phytochemical screening and high-performance liquid chromatography were carried out to establish the phytochemical profle of extracts from the aerial part of cumarú, obtained from plants resulting from con- ventional conditions in greenhouse, clonal shoots and from in vitro culture. The results allowed establishing a new protocol for in vitro regeneration of A. cearensis using Mtop. The extracts these plants grown in vitro con- tained a higher quantity of favonoids than those grown under conventional conditions. The results obtained from this study provide valuable new information to support future works, so that the medicinal properties and the secondary metabolites produced by this plant can be further investigated in beneft to human health in sustainable form. 1. Introduction Amburana cearensis (Allem~ ao) A. C. Smith is a species of the family Fabaceae, native to the Caatinga (shrubland) biome (Pereira et al., 2017), found in the Brazilian semi-arid region (Araruna et al., 2013). Extracts from this leguminous tree, popularly known as cumaru, are often used in traditional medicine, especially to treat respiratory dis- eases like colds, bronchitis and asthma (Araruna et al., 2013). These preparations have anti-infammatory, antimicrobial, anticoagulant, vasodilatory, antispasmodic and antithrombotic activities, due to the presence of secondary metabolites such as coumarins, favonoids and phenolic glycosides (Canuto and Silveira, 2010; Pereira et al., 2017). Besides medicinal uses, A. cearensis is widely extracted for its wood. This extractivisim, along with environmental degradation and habitat loss, has drastically reduced the population of the species. The produc- tion of plants via direct organogenesis of shoots utilizing the normal ontogenetic pathway for development of branches from axillary meri- stems is one of the most important in vitro propagation methods (Gahan and George, 2008). This tissue culture technique allows the production of uniform plantlets with high quality, in turn able to produce Abbreviations: BAP, benzylaminopurine; HPLC, high-performance liquid chromatography; Mtop, meta-topolin; WPM, woody plant medium; UV, ultraviolet. * Corresponding author. E-mail addresses: jessicancvasconcelos@yahoo.com.br (J.N.C. Vasconcelos), lima_brito@yahoo.com.br (A.L. Brito), amanda_pinheeiro@hotmail.com (A.L. Pinheiro), dinahagro@hotmail.com (D.I.J.G.C. Pinto), jackson.guedes@univasf.edu.br (J.R.G.S. Almeida), talialeila@gmail.com (T.L. Soares), jose.raniere@ gmail.com (J.R.F. Santana). Contents lists available at ScienceDirect Biocatalysis and Agricultural Biotechnology journal homepage: http://www.elsevier.com/locate/bab https://doi.org/10.1016/j.bcab.2019.101408 Received 14 August 2019; Received in revised form 21 October 2019; Accepted 28 October 2019