AbstractCandidatus Liberibacter asiaticus (CLas) has been associated with Huanglongbing, a lethal vector-borne disease affecting citrus crops worldwide. While comparative genomics has provided preliminary insights into the metabolic capabilities of this uncultured microorganism, a comprehensive functional characterization is currently lacking. Here, we reconstructed and manually curated genome-scale metabolic models for the six CLas strains A4, FL17, gxpsy, Ishi-1, psy62, and YCPsy, in addition to a model of the closest related culturable microorganism, L. crescens BT-1. Predictions about nutrient requirements and changes in growth phenotypes of CLas were confirmed using in vitro hairy root-based assays, while the L. crescens BT-1 model was validated using cultivation assays. Host-dependent metabolic phenotypes were revealed using expression data obtained from CLas-infected citrus trees and from the CLas-harboring psyllid Diaphorina citri Kuwayama. These results identified conserved and unique metabolic traits, as well as strain-specific interactions between CLas and its hosts, laying the foundation for the development of model-driven Huanglongbing management strategies.
Globally, citrus is threatened by huanglongbing (HLB), and the lack of effective control measures is a major concern of farmers, markets, and consumers. There is compelling evidence that plant health is a function of the activities of the plant's associated microbiome. Using
Liberibacter crescens
, a culturable surrogate for the unculturable HLB-associated bacterium “
Candidatus
Liberibacter asiaticus,” we tested the hypothesis that members of the citrus microbiome produce potential anti-“
Ca
. Liberibacter asiaticus” natural products with potential anti-“
Ca
. Liberibacter asiaticus” activity. A subset of isolates obtained from the microbiome inhibited
L. crescens
growth in an agar diffusion inhibition assay. Further fractionation experiments linked the inhibitory activity of the fungus
Cladosporium cladosporioides
to the fungus-produced natural products cladosporols A, C, and D, demonstrating dose-dependent antagonism to
L. crescens
.