Análise do transcriptoma de Lippia alba (Mill.) N.E.Br. (Verbenaceae) por RNAseq visando a identificação de enzimas terpeno sintases

Abstract

Lippia alba, popularly known as erva-cidreira, is a widely distributed specie in Americas and it is found throughout Brazil. This specie has important using in popular medicine for cramp-ing, indigestion, nausea, diarrhea, dysentery, respiratory diseases, liver disorders treatment and infectious diseases such as syphilis and gonorrhea. The leaves of L. alba, which are pre-pared by infusion or decoction and orally ingested, producing an essential oil rich in terpene compounds. These compounds are of pharmacological and industrial interest, due to their use in fragrance preparation. Interestingly, the composition of essential oils change according to different abiotic factors and genetic variations such as ploidy level. In this context, the aims of this work were to characterize the transcriptome of leaves of L. alba (linalool chemotype) and to search putative enzymes sequences involved in production of secondary metabolites. The transcriptome was sequenced by Miseq platform (Illumina) running pair-end libraries 300 bp. The sequencing resulted in 47,498,310 reads (23,749,155 reads for each end sequenced) of 35-308 bp, comprising 12,148,327,567 nucleotides (-12 Gb). The de novo assembly of tran-scripts was processed by Trinity software and generated 193,532 transcripts, in 128,209 uni-genes, with N50 equal to 1,187 bp. 86,122 ORFs (Open Read Frame) were obtained and sub-mitted to BlastP algorithm, finding 75,533 sequences included in NR (Non-Redundant) pro-tein database. Approximately 78.4% of these sequences were functionally annotated using Blast2Go pipeline. Analysis of annotated sequences revealed putative enzymes for synthesis of terpenoids such as geraniol and linalool/nerolidol. For assembly and annotation validation, qPCR assay were realized by amplification of 13 endogenous control genes and 4 terpene synthases genes. The results found here corroborate transcriptome studies in non-model or-ganisms using high-performance sequencing technologies.