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Recent reports in model plant species have highlighted a role for DNA methylation pathways in the regulation of the somatic-to-reproductive transition in the ovule, suggesting that apomixis (asexual reproduction through seeds) likely relies on RdDM downregulation. Our aim was therefore to explore this hypothesis by characterizing genes involved in DNA methylation in the apomictic grass  Eragrostis curvula . We explored floral transcriptomes to identify homologs of three candidate genes, for which mutations in  Arabidopsis  and maize mimic apomixis ( AtAGO9/ZmAGO104 ,  AtCMT3/ZmDMT102/ZmDMT105 , and  AtDDM1/ZmCHR106 ), and compared both their spatial and temporal expression patterns during reproduction in sexual and apomictic genotypes. Quantitative expression analyses revealed contrasting expression patterns for the three genes in apomictic vs sexual plants.  In situ  hybridization corroborated these results for two candidates,  EcAGO104  and  EcDMT102 , and revealed an unexpected ectopic pattern for the  AGO  gene during germ line differentiation in apomicts. Although our data partially support previous results obtained in sexual plant models, they suggest that rather than an RdDM breakdown in the ovule, altered localization of  AtAGO9/ZmAGO104  expression is required for achieving diplospory in  E .  curvula . The differences in the RdDM machinery acquired during plant evolution might have promoted the emergence of the numerous apomictic paths observed in plants.

Temporal and spatial expression of genes involved in DNA methylation during reproductive development of sexual and apomictic Eragrostis curvula