Discovery of Conserved and Novel MicroRNAs in Galdieria sulphuraria

As a thermoacidophilic microalga, Galdieria sulphuraria has a unique biological function. MicroRNA (miRNA) plays an important regulating role in plant various stress responses. In this study, we identified lots of conserved and novel miRNAs in G. sulphuraria (gsu-miRNAs), and predicted their putative targets for the first time. Conserved and novel gsu-miRNAs were predicted via deep sequencing on the Illumina HiSeq 4000 platform combined with bioinformatics analysis with a series of filtration criteria. Characterization of gsu-miRNAs and their targets were searched by different bioinformatics software. Some gsu-miRNAs were validated by Northern blot and RT-PCR analysis. MiRNA target gene function was predicted via GO and KEGG analysis. The interrelationship between gsu-miRNAs and target genes was constructed via Cytoscape networks analysis. A total of 134 gsu-miRNAs belonging to 124 MIRNA families were identified. Characterization analysis and experimental validation revealed that most of them were credible. A few miRNAs showed conservatism between G. sulphuraria and 20 representative plants. 1,589 putative miRNA targets were predicted. GO analysis revealed that the genes targeted by gsu-miRNAs involved in some important physiological processes of this alga, such as the ETC, and KEGG pathway analysis revealed that RNA transport and the PPP were predicted to be the two most enriched pathways. Cytoscape networks between miRNAs and target genes indicated their various interactions. Research on gsu-miRNAs, which act as key regulators during gene expression in G. sulphuraria will open a new avenue for further developing this thermoacidophilic alga at the post-transcriptional level.