Abstract
<jats:p>C4 and CAM photosynthesis evolved repeatedly across angiosperms, yet the four palm species examined here - representing three of five Arecaceae subfamilies (~2,600 species total) - all lack both. We hypothesized that this reflects the absence of PEPC1, a phosphoenolpyruvate carboxylase isoform required for C4 carbon fixation initiation and present in commelinids (Poaceae + Bromeliaceae) but whose distribution across monocots is unknown. We surveyed six core C4 enzyme families by HMMER profiling across nine plant genomes - four palms (Cocos nucifera, Elaeis guineensis, Phoenix dactylifera, Nypa fruticans), two grasses, one bromeliad, one basal monocot, and one fern. We constructed maximum-likelihood PEPC phylogenies and performed codon-based branch-site likelihood ratio tests using PAML. All six enzyme families were present in palms. PEPC1 was detected in all commelinids (rice, maize, pineapple) but absent from all four palm species and outgroups. No palm PEPC sequence fell within the PPC-1 clade. Molecular dating indicates the Arecaceae-commelinid divergence (~120 Mya) predates PEPC1 origin (~105 Mya). Branch-site tests on the maize C4-PEPC1 branch were marginally significant (2Dl = 3.03, p ~ 0.082). The absence of PEPC1 represents an ancestral molecular barrier that likely precluded C4 evolution in palms. This identifies a lineage-specific gene duplication as a constraint on photosynthetic pathway evolution in a major tropical plant lineage.</jats:p>