The evolution, variation and expression patterns of the annexin gene family in the maize pan-genome.

Annexins (Anns) are a family of evolutionarily conserved, calcium-dependent, phospholipid-binding proteins that play critical roles in plant growth, development, and stress responses. Utilizing the pan-genome of 26 high-quality maize genomes, we identified 12 Ann genes, comprising 9 core genes (present in all 26 lines) and 3 near-core genes (present in 24-25 lines). This highlights the limitations of studying ZmAnn genes based on a single reference genome. Evaluating the Ka/Ks values of Ann genes in 26 varieties revealed that ZmAnn10 was under positive selection in certain varieties, while the remaining genes had Ka/Ks values less than 1, indicating purifying selection. Phylogenetic analysis divided ZmAnn proteins into six groups, with group VI containing only ZmAnn12. Structural variation in certain varieties altered the conserved domains, generating many atypical genes. Transcriptome analysis showed that different Ann members have distinct expression patterns in various tissues and under different abiotic and biotic stress treatments. Weighted gene co-expression network analysis of transcriptome data from various maize tissues under cold stress identified four Ann genes (ZmAnn2, ZmAnn6, ZmAnn7, ZmAnn9) involved in co-expression modules. Overall, this study utilized high-quality maize pangenomes to perform a bioinformatic analysis of ZmAnn genes, providing a foundation for further research on ZmAnn genes.