Extended Data Fig. 6: Phylogenetic analysis of genes encoding C4 enzymes and their non-C4 isoforms.

Phylogenetic and expression (UMAP plots) analysis of NADP-ME (a), NADP-MDH (b), PPDK (c), PEPC (d) and CA (e) were presented. Seita indicates Setaria italica, Aa indicates Arundinella anomala, Sobic indicates Sorghum bicolor, Zm indicates Zea mays, Os indicates Oryza sativa, and AT indicates Arabidopsis thaliana. Genes in the light red clade encode proteins in the C4 pathway. Tree scale: 0.1. Colour scales in the UMAP plots indicate gene expression levels in individual cells. f, Expression profiles of C4-related homologs identified in A. anomala. Gene pairs were classified into five categories according to subgenome expression patterns across five tissues (aerial, ear, leaf, sheath, stem): A_dominant, B_dominant, balanced, A_only, and B_only. Heatmap colors represent normalized TPM values, with grey indicating gene loss. Red-labeled genes denote C4 core functional homologs.