The correlation of evolutionary rate with pathway position in plant terpenoid biosynthesis

Genes are expected to face stronger selective constraint and to evolve more slowly if they encode enzymes upstream as opposed to downstream in metabolic pathways, because upstream genes are more pleiotropic, being required for a wider range of end products. However, few clear examples of this trend in evolutionary rate variation exist. We examined whether genes involved in plant terpenoid biosynthesis exhibit such a pattern, using data for 40 genes from five fully sequenced angiosperms, Oryza, Vitis, Arabidopsis, Populus, and Ricinus. Our results show that d(N)/d(S) does in fact correlate with pathway position along pathways converting glucose to the terpenoid phytohormones abscissic acid, gibberellic acid (GA), and brassinosteroids. Upstream versus downstream rate variation is particularly strong in the GA pathway. In contrast, we found no or little apparent variation in d(N)/d(S) with gene copy number. We also introduce a new measure of pathway position, the Pathway Pleiotropy Index (PPI), which counts groups of enzymes between pathway branch points. We found that this measure is superior to pathway position in explaining variation in d(N)/d(S) along each pathway. Although at least 8 of the 40 genes showed evidence of positive selection, correlations of d(N)/d(S) with PPI remain significant when these genes are removed. Therefore, our results are consistent with the prediction that selective constraint is progressively relaxed along metabolic pathways.