Deciphering the common bean resistance against the anthracnose disease: Transcriptome and identification of genes responsive to the compatible and incompatible interaction Phaseolus vulgaris/Colletotricuhm lindemuthianum
Transcriptomic and gene expression studies of the phytopathogenic system Phaseolus vulgaris vs Colletotrichum lindemuthianum that have been carried out in the Laboratory of Cell and Molecular Biology of CENA (USP) have revealed tissue-specific gene groups with a role in the common bean defense mechanism against the anthracnose pathogen. However, the pattern of decisive transcripts for the localized biological responses (cell-specific) has not yet been described for this system, and may be achieved through the combination of laser microdissection microscopy and RNA-seq techniques. As hypotheses of this study are: 1) there are differences in gene expression among cells from leaf tissues during compatible (Diamante Negro vs. race 81) and incompatible (IAC-UNA vs. race 81) interaction of common bean and the anthracnose pathogen (host-pathogen contact); 2) there are differences in gene expression among cells adjacent to the site of contact host-pathogen, considering time of interaction; 3) there is reproducibility of the model to be defined in others genotypes resistant to the disease. Thus, target-cells from leaf-tissues infected with the race 81 of the C. lindemuthianum will be excised by laser microdissection microscopy (Leica), from which total RNA will be isolated and processed for sequencing in the MiSeq Personal Sequencer equipment (Illumina). Sequence analyses and comparisons among transcriptomic data will be carried out using bioinformatic programs as Galaxy, Blast2GO and MapMan. The results will be validated by quantitative Reverse Trancriptase-PCR (RT-qPCR). Finally, the validation of the genes as involved in the plant defense will be determined by testing them in others common bean genotypes resistant to anthracnose.
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