Three replicates were analysed in both microarray and QRT-PCR experiments. Vertical bars represent standard deviations. Conclusions Sustainable control measures for bacterial blight in Africa will depend on understanding and characterizing those of the microbe’s genes involved in the rice-Xoo interaction. We therefore focused our study on analysing and characterizing Xoo MAI1 at the transcriptional level.
For this we constructed a Xoo MAI1 SSH array, performed in planta gene expression analysis and selected and validated by QRT-PCR various gene expressions to generate robust and reliable data. Although the SSH microarray may not be as sensitive as QRT-PCR for some genes, results included several candidate genes whose regulation and function will need to be elucidated to better understand the Xoo-rice interactions. Our ICG-001 study shows that the regulation of gene expression in the Xoo strain MAI1 is controlled at different time points during pathogen infection. We identified conserved mechanisms for which some were reported in other Xoo-plant interactions but not yet described for African strains. We also identified differentially regulated genes specific to the Xoo strain MAI1. Several homologues
of Xoo MAI1 differentially expressed genes were located in the vicinity of IS elements in the Xoo BAI3 genome. The role played by these IS elements in controlling neighbouring-gene selleck inhibitor expression needs to be elucidated. More data on African Xoo strains also need to be generated. Recently, the sequencing of various African Xoo and Xoc strains has been initiated at our laboratory and others. With this information, the full-length cDNA of desired genes can be easily obtained and their specific functions in pathogenicity studied, using available gene knockout technology. Functional characterization of the proteins/genes related to virulence will be of particular importance in understanding the complex RG-7388 purchase interaction between
Xoo MAI1 and rice. Our work constitutes a significant contribution towards the biology of an emerging and devastating pathogen under a specific, but insufficiently Adenosine triphosphate studied, environment in West Africa. Methods DNA microarray construction Two subtracted DNA libraries (SSH) were previously constructed in our laboratory and partially characterized . For the first library (MAI1-PXO86), the tester was the African Xoo MAI1 (race A3) and the driver the Philippine Xoo PXO86 (Phil race 2). For the second library (MAI1-BLS256), the same tester was used, with Xoc BLS256 being the driver. We randomly selected 2112 clones from MAI1-PXO86 library and 2304 from MAI1-BLS256. From the MAI1-PXO86 SSH library, we selected another 88 clones that represented a non-redundant set of sequences selected from a previous analysis of 265 sequences from that library .