High throughput sequencing of messenger RNA from Citrus cultivars infected by X. citri

Citrus canker, caused by the bacterium Xanthomonas citri subsp. citri (X. citri), is one of the most devastating diseases of citrus plants, and responsible for citriculture damage around the world. The identification of genes that are induced or repressed during the infection process by X. citri, through the technique of RNA-Seq, is an important tool for the understanding of genetic and molecular mechanisms that confer different levels of susceptibility to plant citrus against Xac pathogen.

To detect genetic modifications underlying Citrus Canker disease, leaves of these 8 citrus cultivars, with different levels of susceptibility to the disease, were inoculated with a suspension of X. citri isolate (treatment) or sterile water (control). At 24, 48 and 72 hours after inoculation leaves were collected and immediately stored in liquid nitrogen. mRNA was isolated for cDNA synthesis, and in vitro cloning (clustering) was performed in a flow cell using cBOT (Illumina, Ca, USA). Samples were then multiplexed and sequenced in HiScanSQ high throughput sequencing machine (Illumina, Ca, USA)(Peron et al., 2014).

Peron et al. Identification of Genes Involved in the Resistance of Tangerine Ponkan (Citrus reticulata) to Citrus Canker. XLIII Annual Meeting of SBBq Foz do Iguaçu, PR, Brazil, May 17 to 20 , 2014.