Unraveling a tangled skein: Evolutionary analysis of the bacterial gibberellin biosynthetic operon

ABSTRACT Gibberellin (GA) phytohormones are ubiquitous regulators of growth and developmental processes in vascular plants. The convergent evolution of GA production by plant-associated bacteria, including both symbiotic, nitrogen-fixing rhizobia and phytopathogens, suggests that manipulation of GA signaling is a powerful mechanism for microbes to gain an advantage in these interactions. Although homologous operons encode GA biosynthetic enzymes in both rhizobia and phytopathogens, notable genetic heterogeneity and scattered operon distribution in these lineages suggests distinct functions for GA in varied plant-microbe interactions. Therefore, deciphering GA operon evolutionary history could provide crucial evidence for understanding the distinct biological roles for bacterial GA production. To further establish the genetic composition of the GA operon, two operon-associated genes that exhibit limited distribution among rhizobia were biochemically characterized, verifying their roles in GA biosynthesis. Additionally, a maximum-parsimony ancestral gene block reconstruction algorithm was employed to characterize loss, gain, and horizontal gene transfer (HGT) of GA operon genes within alphaproteobacteria rhizobia, which exhibit the most heterogeneity among GA operon-containing bacteria. Collectively, this evolutionary analysis reveals a complex history for HGT of both individual genes and the entire GA operon, and ultimately provides a basis for linking genetic content to bacterial GA functions in diverse plant-microbe interactions.