Temporal transcription profiling of sweet orange in response to PthA4-mediated Xanthomonas citri subsp. citri infection.

Yang Hu, Shuo Duan, Yunzeng Zhang, Deepak Shantharaj, Jeffrey B Jones, Nian Wang,

Phytopathology, January 18, 2016

Citrus canker, caused by Xanthomonas citri subsp. citri (Xcc), is a devastating disease of most commercial citrus varieties. In our previous study, we analyzed the transcriptional response of Valencia sweet orange to Xcc wild type and pthA4 mutant infection at 48 hours post inoculation (hpi). Using microarray analysis, two PthA4 targets, CsLOB1 and CsSWEET1, were identified. We have shown that PthA4 binds to the EBE (Effector Binding Element) of CsLOB1 and activates gene expression of this susceptibility gene. However, how PthA4 modulates host genes at different stages of infection remains to be determined. In this study, we compared the transcriptional profiles between citrus leaf tissue inoculated with Xcc306 and those inoculated with a pthA4-deletion mutant strain Xcc306∆pthA4 at 6, 48 and 120 hpi. At both 48 and 120 hpi, the PthA4-mediated infection significantly up-regulated expression of a variety of genes involved in cell wall degradation and modification, DNA packaging, G-protein, protein synthesis, sucrose metabolism and cell division functions, while the down-regulated genes were mainly enriched in photosynthesis, transport, secondary metabolism, cytochrome P450 and various plant defense associated mechanisms. To validate microarray results, gene expression of 26 genes representing genes associated with cell wall-associated, immunity system, and carbohydrate metabolism was confirmed using quantitative reverse transcription-PCR. Expression patterns of these genes at 48 and 120 hpi were consistent with the microarray results. We also identified putative Effector Binding Elements for PthA4 (EBEPthA4) in the promoter regions of multiple genes up-regulated by PthA4, to which PthA4 might bind directly to control their gene expression. Our study provided a dynamic picture of citrus genes regulated by PthA4 during the Xcc infection of citrus leaves at different stages. This study will be useful in further understanding the virulence mechanism of Xcc and identifying potential targets of PthA4.

Pubmed Link: 26780431