The transcriptomic responses of C4 grasses to subambient CO2 and low light are largely species specific and only refined by photosynthetic subtype

Three subtypes of C₄ photosynthesis exist (NADP-ME, NAD-ME and PEPCK), each known to be beneficial under specific environmental conditions. However, the influence of photosynthetic subtype on transcriptomic plasticity, as well as the genes underpinning this variability, remain largely unknown. Here, we comprehensively investigate the responses of six C₄ grass species, spanning all three C₄ subtypes, to two controlled environmental stresses: low light (200 µmol m⁻² sec⁻¹) and glacial CO₂ (subambient; 180 ppm). We identify a susceptibility within NADP-ME species to glacial CO₂. Notably, although glacial CO₂ phenotypes could be tied to C₄ subtype, biochemical and transcriptomic responses to glacial CO₂ were largely species specific. Nevertheless, we were able to identify subtype specific subsets of significantly differentially expressed transcripts which link resource acquisition and allocation to NADP-ME species susceptibility to glacial CO₂. Here, low light phenotypes were comparable across species with no clear subtype response, while again, transcriptomic responses to low light were largely species specific. However, numerous functional similarities were noted within the transcriptomic responses to low light, suggesting these responses are functionally relatively conserved. Additionally, PEPCK species exhibited heightened regulation of transcripts related to metabolism in response to both stresses, likely tied to their C₄ metabolic pathway. These results highlight the influence that both species and subtype can have on plant responses to abiotic stress, building on our mechanistic understanding of acclimation within C₄ grasses and highlighting avenues for future crop improvements.