Melatonin Is Involved in Citrus Response to the Pathogen Huanglongbing via Modulation of Phytohormonal Biosynthesis

The auxin-like metabolite melatonin regulates the production of citrus phytohormones upon infection with the huanglongbing pathogen and infestation with its vector. Huanglongbing (HLB) is a devastating citrus disease worldwide that is putatively caused by Candidatus Liberibacter asiaticus and transmitted by Diaphorina citri. Melatonin is a ubiquitously distributed auxin-like metabolite found in both prokaryotes and eukaryotes. In this study, we used integrative metabolomic and transcriptomic approaches to investigate the potential role of melatonin in citrus response against HLB and to understand the relationships between melatonin and the stress-associated phytohormones at molecular and metabolic levels. Melatonin was detected in the leaves of Valencia sweet orange (Citrus sinensis) after derivatization with N-methyl-N-trimethylsilyltrifluoroacetamide using a targeted gas chromatography-mass spectrometry running in selective ion monitoring mode-based method. Ca. L. asiaticus infection and D. citri infestation significantly increased endogenous melatonin levels in Valencia sweet orange leaves and upregulated the expression of its biosynthetic genes (CsTDC, CsT5H, CsSNAT, CsASMT, and CsCOMT). However, infection with Ca. L. asiaticus had a greater effect than did infestation with D. citri. Melatonin induction was positively correlated with salicylic acid content, but not that of trans-jasmonic acid. Moreover, melatonin supplementation enhanced the endogenous contents of the stress-associated phytohormones (salicylates, auxins, trans-jasmonic acid, and abscisic acid) and the transcript levels of their biosynthetic genes. Furthermore, melatonin supplementation diminished the Ca. L. asiaticus titer within the infected leaves, which suggests that melatonin might play an antibacterial role against this bacterium and gram-negative bacteria in general. These findings provide a better understanding of the melatonin-mediated defensive response against HLB via modulation of multiple hormonal pathways. Understanding the role of melatonin in citrus defense to HLB may provide a novel therapeutic strategy to mitigate the disease.