Liu, Yangyang

Abstract Citrus Huanglongbing and Citrus tristeza are two diseases that affect the citrus industry worldwide. The pathogens causing these diseases are the phloem-limited bacteria ‘Candidatus Liberibacter spp.’ (mainly Ca. L. asiaticus, CLas) and citrus tristeza virus (CTV). We recently found that both CLas and CTV could be acquired and retained by the Asian citrus psyllid Diaphorina citri. However, the mechanism through which CLas and CTV interact with the insect vectors and plant hosts has not been defined. In this study, an electrical penetration graph was used to study the feeding behavior of D. citri adults on four groups of Citrus reticulata Blanco cv. Hongjü plants: healthy, CLas-infected, CTV-infected, and CTV-CLas coinfected plants. Liquid chromatography with tandem mass spectrometry (LC–MS/MS) was applied to analyze the metabolites of the four groups of plants. The combined results are as follows: (1) The lowest number of metabolites were enriched in CTV-infected plants, which hardly influenced the feeding behavior of D. citri, suggesting that mild CTV strain (CT31) infection caused limited disorders in citrus plants compared with CLas infection; (2) Increased levels of L-arabinose and kaempferol in CTV-infected and CLas-CTV coinfected plants were suggested to contribute to increased penetration time during feeding of D. citri. CLas-infection increases the difficulty of finding appropriate feeding sites by the vector and results in xylem feeding for certain duration; (3) A significant reduction in α-linolenic acid metabolism in CLas-infected plants was found to be related to methyl jasmonate signaling, which induced resistance to D. citri and increased the duration of salivation. This effect was reversed by coinfection with CTV and was consistent with the phloem structure and carbohydrate accumulation alteration; (4) Stress response-associated 2'-hydroxygenistein and sakuranetin were highly upregulated flavonoid in CTV-CLas coinfected plants. This combinged with the anatomical alterations might interfere with D. citri feeding in the citrus phloem, as reflected by the time reduction of sap-sucking there. These findings will provide new insights into the interactions between CTV and CLas in citrus and the insect vector D. citri that transmiting these pathogens.

Diaphorina citri Kuwayama (D. citri) is an insect vector of phloem-limited ‘Candidatus Liberibacter asiatus’ (CLas), the presumed pathogen of citrus Huanglongbing (HLB). Recently, our lab has preliminarily found it acquired and transmitted Citrus tristeza virus (CTV), which was previously suggested to be vectored by species of aphids. However, the influences of one of the pathogens on the acquisition and transmission efficiency of the other pathogen remain unknown. In this study, CLas and CTV acquisition and transmission by D. citri at different development stages under field and laboratory conditions were determined. CTV could be detected from the nymphs, adults, and honeydew of D. citri but not from the eggs and exuviates of them. CLas in plants might inhibit CTV acquisition by D. citri as lower CTV–positive rates and CTV titers were detected in D. citri collected from HLB-affected trees compared to those from CLas–free trees. D. citri were more likely to obtain CTV than CLas from host plants co-infected with the two pathogens. Intriguingly, CTV in D. citri facilitated the acquisition and transmission of CLas, but CLas carried by D. citri had no significant effect on the transmission of CTV by the same vector. Molecular detection and microscopy methods confirmed the enrichment of CTV in the midgut after a 72-h acquisition access period. Collectively, these results raise essential scientific questions for further research on the molecular mechanism of pathogen transmission by D. citri and provide new ideas for the comprehensive prevention and control of HLB and CTV.

“Candidatus Liberibacter asiaticus” (CLas) is a phloem-restricted α-proteobacterium that is associated with citrus huanglongbing (HLB), which is the most destructive disease that affects all varieties of citrus. Although midrib is usually used as a material for CLas detection, we recently found that the bacterium was enriched in fruits, especially in the fruit pith. However, no study has revealed the molecular basis of these two parts in responding to CLas infection. Therefore, we performed transcriptome and UHPLC–MS-based targeted and untargeted metabolomics analyses in order to organize the essential genes and metabolites that are involved. Transcriptome and metabolome characterized 4834 differentially expressed genes (DEGs) and 383 differentially accumulated metabolites (DAMs) between the two materials, wherein 179 DEGs and 44 DAMs were affected by HLB in both of the tissues, involving the pathways of phenylpropanoid biosynthesis, phytohormone signaling transduction, starch and sucrose metabolism, and photosynthesis. Notably, we discovered that the gene expression that is related to beta-glucosidase and endoglucanase was up-regulated in fruits. In addition, defense-related gene expression and metabolite accumulation were significantly down-regulated in infected fruits. Taken together, the decreased amount of jasmonic acid, coupled with the reduced accumulation of phenylpropanoid and the increased proliferation of indole-3-acetic acid, salicylic acid, and abscisic acid, compared to leaf midribs, may contribute largely to the enrichment of CLas in fruit piths, resulting in disorders of photosynthesis and starch and sucrose metabolism.