Contents lists available at ScienceDirect Industrial Crops & Products journal homepage: www.elsevier.com/locate/indcrop 1 H-NMR-based metabolomic of plant cell suspension cultures of Thevetia peruviana treated with salicylic acid and methyl jasmonate Dary Mendoza a,b, ⁎ , Juan Pablo Arias a , Olmedo Cuaspud a , Nuria Esturau-Escofet c, ⁎⁎ , Circe C. Hernández-Espino c , Eduardo Rodríguez de San Miguel d , Mario Arias a a Grupo de Investigación en Biotecnología Industrial, Laboratorio de Bioconversiones, Universidad Nacional de Colombia, Calle 59A No.63-20 Bloque 19A-313, Medellín, Colombia b Grupo de Productos Naturales y Bioquímica de Macromoléculas, Facultad de Ciencias, Universidad del Atlántico, Km 7 vía a Puerto Colombia, Barranquilla, Colombia c Instituto de Química, Universidad Nacional Autónoma de México, Ciudad Universitaria, 04510 Cd., México, Mexico d Departamento de Química Analítica, Facultad de Química, Universidad Nacional Autónoma de México, Ciudad Universitaria, 04510 Cd., México, Mexico ARTICLE INFO Keywords: Metabolic profle Methyl jasmonate Plant cell culture Salicylic acid Thevetia peruviana ABSTRACT The metabolomic profle of plant cell suspension cultures of T. peruviana treated with salicylic acid (SA), methyl jasmonate (MeJA), and their combination (SA + MeJA) was determined using proton nuclear magnetic re- sonance spectroscopy ( 1 H-NMR and J-Resolved) and multivariate data analysis (MVDA). The qualitative and quantitative variations in the metabolite pool were detected by comparing control cells suspensions against cells suspensions treated with elicitors, 24–144 h after treatment. MVDA showed a clear separation between control and treatments conditions. The MeJA caused a metabolic reprogramming in cells that afected primary meta- bolism and phenolic biosynthesis at 72 h after elicitor addition. Upon MeJA elicitation, glucose, proline and glutamine content increased while sucrose content decreased. These metabolic responses could be important for obtaining energy, carbon skeletons, and equivalent redox (NADPH) necessary for de novo phenolic compounds biosynthesis. Benzyl-glucosides, chlorogenic, ferulic and phenylacetic acid were the principal phenolic com- pounds identifed upon MeJA-treatment. Combination of SA and MeJA showed that SA reverted the efect of MeJA on T. peruviana metabolism, specifcally on phenolic biosynthesis. These results could allow us to identify hot spots susceptible to stimulate phenolic compounds production of pharmaceutical interest in cell suspension cultures of T. peruviana. 1. Introduction Thevetia peruviana (Pers.) K. Schum is an ornamental shrub be- longing to the order Gentianales, Apocynaceae family. It is widely distributed in tropical and sub-tropical regions of Central and South America, Asia, and Africa where it is known for its cardiac properties (Bandara et al., 2010). Fruits and leaves of T. peruviana also store phenolic compounds with potential usage for the development of anti- infammatory (Rahman et al., 2017), anti-microbial (Hassan et al., 2011; Dabur et al., 2007), and antineoplastic drugs (Haldar et al., 2015; Ramos-Silva et al., 2017). Cell suspension cultures of T. peruviana have been established to promote the production of phytochemicals with potential pharmaceu- tical value (Arias et al., 2010). Recent studies have shown that these cultures produce high levels of total phenolic compounds (TPC) at shake fask scale (Arias et al., 2016). In addition, treatment of these cultures with two chemical elicitors of phenolic compounds biosynth- esis, salicylic acid (SA) and methyl jasmonate (MeJA), at a concentra- tion of 300 μM and 3 μM respectively, increased the accumulation of TPC. Interestingly, the treatment of cultures with the combination SA + MeJA increased the accumulation of some metabolites, while the accumulation of others decreased (Mendoza et al., 2018), suggesting that there is a complex interaction on the signaling pathways of these elicitors that afects phenolic compounds biosynthesis. Jasmonates (JA and MeJA) and SA are phytochemical hormones that play key roles in the regulation of defense signaling network against pathogen or herbivorous insect attack (Pieterse et al., 2012), promoting accumulation of secondary metabolites for plant defense such as phytoalexins (Siciliano et al., 2015; Gundlach et al., 1992; Klessig and Malamy, 1994). SA and MeJA have been applied https://doi.org/10.1016/j.indcrop.2019.04.012 Received 12 October 2018; Received in revised form 29 March 2019; Accepted 7 April 2019 ⁎ Corresponding author at: Laboratorio de Bioconversiones, Universidad Nacional de Colombia, Calle 59A No.63-20 Bloque 19A-313, Medellín, Colombia. ⁎⁎ Corresponding author. E-mail addresses: dalmendozame@unal.edu.co, darymendoza@mail.uniatlantico.edu.co (D. Mendoza), nesturau@iquimica.unam.mx (N. Esturau-Escofet). Industrial Crops & Products 135 (2019) 217–229 0926-6690/ © 2019 Elsevier B.V. All rights reserved. T