General Medicine

Pathogens from the fastidious, phloem-restricted ‘Candidatus Liberibacter’ species cause the devastating Huanglongbing (HLB) disease in citrus worldwide and cause diseases on many solanaceous crops and plants in the Apiaceae family. However, little is known about the pathogenic mechanisms due to the difficulty in culturing the corresponding ‘Ca. Liberibacter’ species. Here, we report that the citrus HLB pathogen ‘Ca. L. asiaticus’ uses an active salicylate hydroxylase SahA to degrade salicylic acid (SA) and suppress plant defenses. Purified SahA protein displays strong enzymatic activity to degrade SA and its derivatives. Overexpression of SahA in transgenic tobacco plants abolishes SA accumulation and hypersensitive response (HR) induced by nonhost pathogen infection. By degrading SA, ‘Ca. L. asiaticus’ not only enhances the susceptibility of citrus plants to both nonpathogenic and pathogenic Xanthomonas citri but also attenuates the responses of citrus plants to exogenous SA. In addition, foliar spraying of 2,1,3-benzothiadiazole and 2,6-dichloroisonicotinic acid, SA functional analogs not degradable by SahA, displays comparable (and even better) effectiveness with SA in suppressing ‘Ca. L. asiaticus’ population growth and HLB disease progression in infected citrus trees under field conditions. This study demonstrates one or more pathogens suppress plant defenses by degrading SA and establish clues for developing novel SA derivatives-based management approaches to control the associated plant diseases.

Huanglongbing, a destructive disease of citrus, is caused by the fastidious bacterium ‘Candidatus Liberibacter asiaticus’ and transmitted by Asian citrus psyllid, Diaphorina citri. The impact of ‘Ca. L. asiaticus’ infection or D. citri infestation on Valencia sweet orange (Citrus sinensis) leaf metabolites was investigated using gas chromatography mass spectrometry, followed by gene expression analysis for 37 genes involved in jasmonic acid (JA), salicylic acid (SA), and proline-glutamine pathways. The total amino acid abundance increased after ‘Ca. L. asiaticus’ infection, while the total fatty acids increased dramatically after infestation with D. citri, compared with control plants. Seven amino acids (glycine, l-isoleucine, l-phenylalanine, l-proline, l-serine, l-threonine, and l-tryptophan) and five organic acids (benzoic acid, citric acid, fumaric acid, SA, and succinic acid) increased in ‘Ca. L. asiaticus’-infected plants. On the other hand, the abundance of trans-JA and its precursor α-linolenic increased in D. citri-infested plants. Surprisingly, the double attack of both D. citri infestation and ‘Ca. L. asiaticus’ infection moderated the metabolic changes in all chemical classes studied. In addition, the gene expression analysis supported these results. Based on these findings, we suggest that, although amino acids such as phenylalanine are involved in citrus defense against ‘Ca. L. asiaticus’ infection through the activation of an SA-mediated pathway, fatty acids, especially α-linolenic acid, are involved in defense against D. citri infestation via the induction of a JA-mediated pathway.

Huanglongbing (HLB) is currently the largest threat to global citrus production. We examined the effect of HLB pathogen ‘Candidatus Liberibacter asiaticus’ infection or infestation by its vector, Diaphorina citri, on ‘Valencia’ sweet orange leaf pigments using high-performance liquid chromatography, followed by gene expression analysis for 46 involved genes in carotenoid and chlorophyll biosynthesis pathways. Both ‘Ca. L. asiaticus’ and D. citri alter the total citrus leaf pigment balance with a greater impact by ‘Ca. L. asiaticus’. Although zeaxanthin was accumulated in ‘Ca. L. asiaticus’-infected leaves, chlorophyllide a was increased in D. citri-infested plants. Our findings support the idea that both ‘Ca. L. asiaticus’ and D. citri affect the citrus pigments and promote symptom development but using two different mechanisms. ‘Ca. L. asiaticus’ promotes chlorophyll degradation but accelerates the biosynthesis of carotenoid pigments, resulting in accumulation of abscisic acid and its precursor, zeaxanthin. Zeaxanthin also has a photoprotective role. By contrast, D. citri induced the degradation of most carotenoids and accelerated chlorophyll biosynthesis, leading to chlorophyllide a accumulation. Chlorophyllide a might have an antiherbivory role. Accordingly, we suggest that citrus plants try to defend themselves against ‘Ca. L. asiaticus’ or D. citri using multifaceted defense systems, based on the stressor type. These findings will help in better understanding the tritrophic interactions among plant, pathogen, and vector.