Conservation of m5C in tRNAs in the kingdom Plantae

Here we use bisulfite conversion of RNA combined with high-throughput IIlumina sequencing (RBS-seq) to identify single-nucleotide resolution of m5C sites in transfer RNAs of all three sub-cellular transcriptomes across six diverse species that include, the single-celled algae Nannochloropsis oculata, the macro algae Caulerpa taxifolia and multi-cellular higher plants Arabidopsis thaliana, Brassica rapa, Triticum durum and Ginkgo biloba. Overall design: RNA Bisulfite sequencing was used to identify candidate m5C sites in tRNAs of diverse plant species. RNA-seq was used to examine abundance of tRNAs in Arabidopsis thaliana wild type. Arabidopsis thaliana RNA Bisulfite-Seq libraries were performed in biological triplicate. All other samples had only one biological replicate. Illumina sequencing was performed on a MiSeq platform.