Plant Science 229 (2014) 238–246 Contents lists available at ScienceDirect Plant Science j ourna l ho me pa g e: www.elsevier.com/locate/plantsci De novo assembly of Eugenia uniflora L. transcriptome and identification of genes from the terpenoid biosynthesis pathway Frank Guzman a,b , Franceli Rodrigues Kulcheski a,b , Andreia Carina Turchetto-Zolet a , Rogerio Margis a,b,c,∗ a PPGGBM, Departamento de Genética, Universidade Federal do Rio Grande do Sul - UFRGS, Porto Alegre, RS, Brazil b PPGBCM, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul - UFRGS, Porto Alegre, RS, Brazil c Departamento de Biofisica, Universidade Federal do Rio Grande do Sul - UFRGS, Porto Alegre, RS, Brazil a r t i c l e i n f o Article history: Received 14 August 2014 Received in revised form 7 October 2014 Accepted 10 October 2014 Available online 22 October 2014 Keywords: Eugenia uniflora Myrtaceae Transcriptome Terpene synthase Oxidosqualene cyclase Secondary metabolism a b s t r a c t Pitanga (Eugenia uniflora L.) is a member of the Myrtaceae family and is of particular interest due to its medicinal properties that are attributed to specialized metabolites with known biological activities. Among these molecules, terpenoids are the most abundant in essential oils that are found in the leaves and represent compounds with potential pharmacological benefits. The terpene diversity observed in Myrtaceae is determined by the activity of different members of the terpene synthase and oxidosqualene cyclase families. Therefore, the aim of this study was to perform a de novo assembly of transcripts from E. uniflora leaves and to annotation to identify the genes potentially involved in the terpenoid biosynthesis pathway and terpene diversity. In total, 72,742 unigenes with a mean length of 1048 bp were identified. Of these, 43,631 and 36,289 were annotated with the NCBI non-redundant protein and Swiss-Prot databases, respectively. The gene ontology categorized the sequences into 53 functional groups. A metabolic path- way analysis with KEGG revealed 8,625 unigenes assigned to 141 metabolic pathways and 40 unigenes predicted to be associated with the biosynthesis of terpenoids. Furthermore, we identified four putative full-length terpene synthase genes involved in sesquiterpenes and monoterpenes biosynthesis, and three putative full-length oxidosqualene cyclase genes involved in the triterpenes biosynthesis. The expression of these genes was validated in different E. uniflora tissues. © 2014 Elsevier Ireland Ltd. All rights reserved. 1. Introduction Eugenia uniflora L., commonly known as Brazilian cherry, pitanga or nangapiri, belongs to the Myrtaceae family [1]. E. uniflora L. is a shrubby tree with edible cherry-like fruits, is native to South Amer- ica and occurs in different vegetation types and ecosystems, with high adaptability to different soil and climate conditions [2]. Its leaves have been used in folk medicine for the treatment of different diseases due to the presence of different compounds in its essen- tial oils [3]. For this reason, E. uniflora has been the focus of several Abbreviations: EC, enzyme commission number; GO, gene ontology; MCMC, Markov chain Monte Carlo; ORF, open reading frame; TPS, terpene synthase; OSC, oxidosqualene cyclase; wAIC, Akaike’s information criterion. ∗ Corresponding author at: Centre of Biotechnology and PPGBCM, Laboratory of Genomes and Plant Population, building 43431, Federal University of Rio Grande do Sul - UFRGS, P.O. Box 15005, CEP 91501-970, Porto Alegre, RS, Brazil. Tel.: +55 51 33087766; fax: +55 5133087309. E-mail addresses: rogerio.margis@gmail.com, rogerio.margis@ufrgs.br (R. Margis). phytochemical studies in recent years. These studies reported some pharmacological properties that were reviewed by Lim [4] and that encompass antioxidant, antimicrobial, antihyperglycemic, antihypertriglycerimedic, hypotensive, vasorelaxant, antiviral, antinociceptive, hypothermic, central nervous system-related, diuretic, anti-inflammatory, antidiarrheal, muscle contractile, try- panocidal, antibiotic potentiating and toxicity activities. Among the main compounds identified in the essential oils of E. uniflora leaves, terpenes are the most abundant. These compounds are one of the most important in the plant kingdom because they form such a large class of plant specialized metabolites and play a number of roles in the interaction between a plant and its envi- ronment [5]. The more abundant terpenes identified in E. uniflora correspond to different types of sesquiterpenes (C15) and monoter- penes (C10) [6–10]. The amount of terpene found in the essential oils of E. uniflora specimens from different regions varies depend- ing on soil composition, sample collection season, altitude and the method used to extract the essential oils [11]. The terpene diversity reported in plant species is a conse- quence of the enzymatic activity of terpene synthases (TPS) and http://dx.doi.org/10.1016/j.plantsci.2014.10.003 0168-9452/© 2014 Elsevier Ireland Ltd. All rights reserved.