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Peptide conjugated morpholinos for management of the huanglongbing pathosystem

Sandoval‐Mojica, Andrés F, Altman, Sidney, Hunter, Wayne B, Pelz‐Stelinski, Kirsten S
Pest management science 2020 v.76 no.9 pp. 3217-3224
Candidatus Liberibacter asiaticus, Citrus, Diaphorina citri, Psyllidae, Wolbachia, bacteria, disease transmission, essential genes, fruit diseases, fruit trees, gene expression, greening disease, imagos, insect vectors, pathogens, peptides, pest management, symbionts, viability
BACKGROUND: ‘Candidatus Liberibacter asiaticus’ (CLas) is the causal agent of the devastating citrus disease Huanglongbing (HLB) and is transmitted by the insect vector Diaphorina citri (Hemiptera: Liviidae). A potential approach for treating CLas infection is the use of synthetic nucleic acid‐like oligomers to silence bacterial gene expression. Peptide conjugated morpholinos (PPMOs) targeting essential genes in CLas and the psyllid vector's endosymbiotic bacteria, Wolbachia (‐Diaphorina, wDi), were evaluated using in vitro and in vivo assays. RESULTS: Expression of the wDi gyrA gene was significantly reduced following incubation of wDi cells with PPMOs. In addition, the viability of isolated wDi cells was greatly reduced when treated with PPMOs as compared to untreated cells. Feeding D. citri adults with a complementary PPMO (CLgyrA‐14) showed significantly reduced (70% lower) expression of the CLas gyrA gene. CLas relative density was significantly lower in the psyllids fed with CLgyrA‐14, when compared to untreated insects. Psyllids that were treated with CLgyrA‐14 were less successful in transmitting the pathogen into uninfected plants, compared to untreated insects. CONCLUSION: The expression of essential genes in the D. citri symbiont, wDi and the HLB pathogen were suppressed in response to PPMO treatments. This study demonstrates the potential of PPMOs as a novel strategy for management of bacterial pathogens of fruit trees, such as HLB. © 2020 Society of Chemical Industry