Abstract Convergent trait evolution is a recurrent phenomenon in all domains of the tree of life. While some convergent traits are caused by simple sequence changes, many are associated with extensive changes to the sequence and regulation of large cohorts of genes. It is unknown how organisms traverse this expansive genotype space to assemble such complex convergent phenotypes. C 4 photosynthesis is a paradigm of large-scale phenotypic convergence. Conceptual and mathematical models propose that C 4 photosynthesis evolved from ancestral C 3 photosynthesis through sequential adaptive changes. These adaptive changes could have been rapidly assembled if modifications to the activity and abundance of enzymes of the C 4 cycle was neutral in C 3 plants. This neutrality would enable populations of C 3 plants to maintain genotypes with expression levels of C 4 enzymes analogous to those in C 4 species and thus enable rapid assembly of a functional C 4 cycle from naturally occurring genotypes given shared environmental selection. Here we show that there is substantial natural variation in expression of genes encoding C 4 cycle enzymes between natural accessions of the C 3 plant Arabidopsis thaliana . We further show through targeted transgenic experiments in the C 3 crop Oryza sativa , that high expression of the majority of C 4 cycle enzymes in rice is neutral with respect to growth, development, biomass and photosynthesis. Thus, substantial variation in the abundance and activity of C 4 cycle enzymes is permissible within the limits of operation of C 3 photosynthesis and the emergence of component parts of this complex convergent trait can be facilitated by neutral variation.

A role for neutral variation in the evolution of C4photosynthesis