Résumé
The objective of this study was to obtain an overview of the active genes in Caturra variety arabica coffee leaves when subjected to various abiotic stresses and to analyze these genes identified in silico using the RTqPCR technique. The plant material consisted of in vitro arabica coffee plants exposed for 3 hours under different stresses: drought - low relative humidity 9%, cold 5 °C, heat 40 °C, high light intensity 200 μM m-2 s-1 and exogenous application of 10 μM abscisic acid. After the stress period, the coffee leaves were collected for RNA extraction. Preparation of the mRNA library was done through Illumina's TruSeq Stranded mRNA Sample Preparation Kit. The sequencing was performed on the HiSeq 2500 platform (Illumina) using the SBS (Synthesis by Sequencing) technique by MGX - Montpellier Genomix. Differential expression analysis was performed from the raw reads counts using the DESeq2 statistical package. The in silico result presented a total of 17,399 differentially expressed genes among all the stresses tested, being 2.217, 812, 4,263, 8.008 and 2.099 for the exogenous ABA stress, cold, drought, heat and oxidative stress, respectively. Using these data, the most expressive candidate genes were selected under the stress conditions tested and their differential expression profiles were confirmed by RT-qPCR experiments. Through in silico experiments of this study it was possible to identify several candidate genes responding to the different stresses applied to coffee plants, which may help in understanding the genetic determinism of tolerance to abiotic stresses in coffee trees. The identification and characterization of these genes make it possible to carry out new studies of differential expression analysis between field and greenhouse plants, and the prospecting of natural variability using different genetic materials, which allow the identification and validation of specific polymorphisms and alleles for the development of molecular markers associated with tolerance to various abiotic stresses.