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Candidatus  Liberibacter asiaticus ( C Las), a bacterium transmitted by the Asian citrus psyllid,  Diaphorina citri , is the causal agent of citrus greening disease, or Huanglongbng (HLB). Currently, vector population suppression with insecticides and tree removal are the most effective strategies for managing the HLB pathosystem. In this study, we assessed the bactericidal capabilities of 2′-deoxy-2′-fluoro- d -arabinonucleic acid antisense oligonucleotides (FANA ASO) both in vitro and in vivo by (1) confirming their capacity to penetrate insect cells, (2) silencing bacterial essential genes, and (3) quantifying reductions in bacterial titer and  D. citri  transmission. We confirmed that FANA ASO are able to penetrate insect cells without the use of a delivery agent. Expression of an essential gene in the  D. citri  endosymbiont,  Wolbachia  (wDi), significantly decreased by 30% following incubation with a wDi-specific FANA ASO. Viability of isolated wDi cells also decreased in response to the FANA ASO treatment. Delivery of a  C Las-specific FANA ASO to infected adult  D. citri  in feeding assays resulted in significant silencing of a  C Las essential gene.  C Las relative density and transmission were significantly lower among  D. citri  fed FANA ASO in diet compared to untreated insects. Root infusions of a  C Las-specific FANA ASO into infected  Citrus  trees significantly reduced  C Las titer during a 30-day trial. Our results suggest that FANA ASO targeting insect-transmitted plant bacteria or insect endosymbionts may be useful tool for integrated management of agricultural pathogens.

Antibacterial FANA oligonucleotides as a novel approach for managing the Huanglongbing pathosystem