MS/MS proteomics from: <em>Citrus sinensis</em> leaves in response to Diaphorina citri infestation and Huanglongbing disease
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Huanglongbing (HLB) is a fatal citrus disease currently threatening all commercially relevant citrus varieties worldwide. In the United States, the putative causative agent, Candidatus Liberibacter asiaticus (CLas), is transmitted by the vector Diaphorina citri, commonly known as the Asian citrus psyllid (ACP). Progress towards understanding CLas and its role in HLB has been hindered by the inability to culture the bacterium and the inability to confidently detect it in diseased trees during the asymptomatic stage. To identify early changes in citrus metabolism in response to inoculation of CLas using its natural psyllid vector, leaves from Madam Vinous sweet orange (Citrus sinensis (L.) Osbeck) trees were exposed to CLas-positive ACP or CLas-negative ACP and longitudinally analyzed using transcriptomics (RNA sequencing), proteomics (liquid chromatography-tandem mass spectrometry), and metabolomics (proton nuclear magnetic resonance). At 4 weeks post-exposure (wpe) to psyllids, the initial HLB plant response was primarily to the ACP and, to a lesser extent, the presence or absence of CLas. Additionally, analysis of 4, 8, 12, and 16 wpe identified 17 genes and one protein as consistently differentially expressed between leaves exposed to CLas-positive ACP versus CLas-negative ACP.. The results of this study inform identification of early detection molecular targets and contribute to the broader understanding of vector-transmitted plant pathogen interactions. There are two datasets related to this research MS/MS Proteomics: 10.25338/B83H1Z 1H NMR based metabolomics: 10.25338/B83P9Q Methods Whole Citrus sinensis leaves were collected throughout this longitudinal analysis. Citrus leaf proteins were extracted and precipitated, followed by peptide sample preparation, gel analysis, reduction/alkylation/trypsin digestion, C18 column cleanup, and mass spectrometry data acquisition. Mass spectrometry was performed on a Fusion Exploris (Thermo Fisher Scientific) mass spectrometer with a Thermo Easy-nLC HPLC with autosampler. The resulting .raw files are provided in this submission.



