Expression rewiring and methylation of non-coding RNAs involved in rhizome phenotypic variations of lotus ecotypes

Non-coding RNAs (ncRNAs), including miRNAs, lncRNAs, and circRNAs, emerge as crucial components for gene regulation. Nelumbo nucifera (lotus), a horticulturally important plant, differentiates into a temperate ecotype of enlarged rhizomes and a tropical ecotype of thin rhizomes. Nevertheless, whether and how ncRNAs can be rewired in expression and differentially methylated contributing to adaptive divergence of this storage organ in lotus ecotypes is unclear. Herein, we study the expression behaviors and DNA methylation patterns of ncRNAs in temperate and tropical lotus rhizomes. By whole transcriptome sequencing, we found both mRNAs and lncRNAs have divergent expression patterns between ecotypes, whereas miRNAs and circRNAs tended to be accession-specific or noisier in expression. The differentially expressed ncRNAs are involved in phenotypic differentiation of lotus rhizome between ecotypes, as the genes that interacted with them in the competing endogenous RNA network are enriched in functions including carbohydrate metabolism and plant hormone signaling, being critical to rhizome enlargement. Intriguingly, ncRNA-targeted genes are less prone to show positive selection or differential expression during ecotypic divergence due to constraints from ncRNA-mRNA interactions. The methylation levels of ncRNAs generally tend to be higher in temperate lotus than in tropical lotus, and differential methylation of lncRNAs also tends to have expression changes. Overall, our study of ncRNAs and their targets highlights the role of ncRNAs in rhizome growth variation between lotus ecotypes through expression rewiring and methylation modification.