Gene co-expression reveals the modularity and integration of C4 and CAM in Portulaca

• Published in: Plant physiology (Bethesda) Vol. 189; no. 2; pp. 735 - 753
• Main Authors: Gilman, Ian S, Moreno-Villena, Jose J, Lewis, Zachary R, Goolsby, Eric W, Edwards, Erika J
• Format: Journal Article
• Language: English
• Published: United States Oxford University Press 01.06.2022
• Subjects: Crassulacean Acid Metabolism - genetics; Droughts; Phosphoenolpyruvate Carboxylase - genetics; Phosphoenolpyruvate Carboxylase - metabolism; Photosynthesis - genetics; Portulaca - metabolism
• ISSN: 0032-0889; 1532-2548
• DOI: 10.1093/plphys/kiac116

C4 photosynthesis and Crassulacean acid metabolism (CAM) have been considered as largely independent adaptations despite sharing key biochemical modules. Portulaca is a geographically widespread clade of over 100 annual and perennial angiosperm species that primarily use C4 but facultatively exhibit CAM when drought stressed, a photosynthetic system known as C4 + CAM. It has been hypothesized that C4 + CAM is rare because of pleiotropic constraints, but these have not been deeply explored. We generated a chromosome-level genome assembly of Portulaca amilis and sampled mRNA from P. amilis and Portulaca oleracea during CAM induction. Gene co-expression network analyses identified C4 and CAM gene modules shared and unique to both Portulaca species. A conserved CAM module linked phosphoenolpyruvate carboxylase to starch turnover during the day-night transition and was enriched in circadian clock regulatory motifs in the P. amilis genome. Preservation of this co-expression module regardless of water status suggests that Portulaca constitutively operate a weak CAM cycle that is transcriptionally and posttranscriptionally upregulated during drought. C4 and CAM mostly used mutually exclusive genes for primary carbon fixation, and it is likely that nocturnal CAM malate stores are shuttled into diurnal C4 decarboxylation pathways, but we found evidence that metabolite cycling may occur at low levels. C4 likely evolved in Portulaca through co-option of redundant genes and integration of the diurnal portion of CAM. Thus, the ancestral CAM system did not strongly constrain C4 evolution because photosynthetic gene networks are not co-regulated for both daytime and nighttime functions.