Whole-Genome Evolutionary Analyses of Non-Endosymbiotic Organelle-Targeting Nuclear Genes Reveal Their Genetic Evolution in 12 Representative Poaceae Species

Chloroplasts and mitochondria, descendants of ancient prokaryotes via endosymbiosis, occupy a pivotal position in plant growth and development due to their intricate connections with the nuclear genome. Genes encoded by the nuclear genome but relocated to or being functional within these organelles are commonly referred as organelle-targeting nuclear genes (ONGs). These genes are essential for maintaining cytonuclear coordination, thereby determining the stability of the life cycle. While molecular function and cytonuclear coordination of some endosymbiosis-derived ONGs (E-ONGs) have been extensively studied, the evolutionary history and characteristics from a more widespread range of non-endosymbiosis-derived ONGs (NE-ONGs) remain largely enigmatic. In this study, we focused on 12 representative species within the Poaceae family to systematically identify NE-ONGs and investigated their evolutionary history and functional significance on a phylogenetic timescale. Upon aligning these 12 species' evolutionary histories, we observed the following phenomena: (i) an exploration of NE-ONGs between the BOP and PACMAD clades unveiled dynamic compositions, potentially influencing their photosynthetic divergence; (ii) the majority of the abundant species-specific NE-ONGs exist in a single-copy status, and functional enrichment analysis further underscored their specialized roles, which could be crucial for species adaptation; and (iii) comparative analyses between plasmid- and mitochondria-related NE-ONGs (pNE-ONGs and mNE-ONGs) revealed a prevalence of pNE-ONGs, indicating tighter control for chloroplast function in Poaceae. In summary, this study offers novel insights into the cytonuclear co-evolutionary dynamics in Poaceae speciation and draws attention to crop improvement by using NE-ONGs.