title: mRNA extracted from inoculated plants supplemented with nitrogen
design: In vitro-grown sugarcane plantlets from SP70-1143 cultivar were maintained at 28*C with an irradiance of 60 *mol photons m-2 s-1 and 12 h photoperiod. After the development of a root system, the plantlets were transferred to hydroponic condition. In this system the plants were grown in a modified Hoagland solution (Hoagland & Arnon, 1950) and inoculated with G. diazotrophicus (PAL5) for four days. Then, the plants were transferred to Hoagland solution plus 0.5 mM Ca2(NO3-) and 5 mM Ca2(NO3-). As a control non-inoculated plants received the same amount of sterile culture medium and were transferred to Hoagland's solution supplemented with 0.5mM Ca2(NO3-) and 5 mM Ca2(NO3-). Plant roots were collected after seven days of treatment with nitrogen and frozen in liquid nitrogen. For RNAseq libraries construction, total RNA was extracted from frozen material according to Logeman et al. (1987). The quality and quantification of RNA samples was checked via agarose gel and Agilent 2100 Bioanalyzer TM (Agilent Technologies, Aplo Alto, CA). The biological samples were used for RNAseq Illumina sequencing at FASTERIS (Switzerland).