Changes in m 6 A RNA methylation are associated with male sterility in wolfberry

Background N 6 -methyladenosine (m 6 A) modification is the most abundant type of RNA modification in eukaryotic cells, playing pivotal roles in multiple plant growth and development processes. Yet the potential role of m 6 A in conferring the trait of male sterility in plants remains unknown. Results In this study, we performed RNA-sequencing (RNA-Seq) and m 6 A-sequencing (m 6 A-Seq) of RNAs obtained from the anther tissue of two wolfberry lines: ‘Ningqi No.1’ ( LB 1) and its natural male sterile mutant ‘Ningqi No.5’ ( LB 5). Based on the newly assembled transcriptome, we established transcriptome-wide m 6 A maps for LB 1 and LB 5 at the single nucleus pollen stage. We found that the gene XLOC_021201, a homolog of m 6 A eraser-related gene ALKBH10 in Arabidopsis thaliana , was significantly differentially expressed between LB 1 and LB 5. We also identified 1642 and 563 m 6 A-modified genes with hypermethylated and hypomethylated patterns, respectively, in LB 1 compared with LB 5. We found the hypermethylated genes significantly enriched in biological processes related to energy metabolism and lipid metabolism, while hypomethylation genes were mainly linked to cell cycle process, gametophyte development, and reproductive process. Among these 2205 differentially m 6 A methylated genes, 13.74% (303 of 2205) were differentially expressed in LB 1 vis-à-vis LB 5. Conclusions This study constructs the first m 6 A transcriptome map of wolfberry and establishes an association between m 6 A and the trait of male sterility in wolfberry.