Genome-wide transcriptome variation landscape in Ruta chalepensis organs revealed potential genes responsible for rutin biosynthesis

Abstract

Ruta chalepensis L., most commonly known as 'fringed rue,' is an excellent and valuable bioactive plant that produces a range of complex flavonoids, of which rutin is the major compound present in this plant of great pharmaceutical and medicinal significance. The present study is a pioneering attempt to examine the changes in the transcriptomic landscape of leaf, stem, and root tissues and correlate this with rutin quantity in each tissue in order to identify the candidate genes responsible for rutin biosynthesis and to increase genomic resources in fringed rue. Comparative transcriptome sequencing of leaves, stems and roots were performed using the NovaSeq 6000 platform. The de novo transcriptome assembly generated 254,685 transcripts representing 154,018 genes with GC content of 42.60 % and N50 of 2280 bp. Searching assembled transcripts against UniRef90 and SwissProt databases annotated 79.7 % of them as protein coding. The leaf tissues had the highest rutin content followed by stems and roots. Several differentially expressed genes and transcripts relating to rutin biosynthesis were identified in leaves comparing with roots or stems comparing with roots. All the genes known to be involved in rutin biosynthesis showed up-regulation in leaves as compared with roots. These results were confirmed by gene ontology (GO) and pathway enrichment analyses. Up-regulated genes in leaves as compared with roots enriched GO terms with relation to rutin biosynthesis e.g. action of flavonol synthase, biosynthetic mechanism of malonyl-CoA, and action of monooxygenase. Phylogenetic analysis of the rhamnosyltransferase (RT) gene showed that it was highly homologues with RT sequence from Citrus species and all were located in the same clade. This transcriptomic dataset will serve as an important public resource for future genomics and transcriptomic studies in R. chalepensis and will act as a benchmark for the identification and genetic modification of genes involved in the biosynthesis of secondary metabolites.