mRNA extracted from whole plant infected with Acidovorax avenae
Acidovorax avenae subsp. avenae obtained from the Culture Collection of the Instituto Biológico (IBSBF) was grown in NA medium (beef extract 3 g/L; Peptone 5 g/L; NaCl 5 g/L) at 28ºC. In vitro-grown sugarcane plantlets were maintained at 28°C with an irradiance of 60 µmol photons m-2 s-1 and 12 h photoperiod. After the development of a root system, vigorous and pathogen-free plants were divided in two halves with a scalpel. One half was inoculated by immersing the root system for 5 minute in a suspension of A. avenae in distilled water (106 CFU mL-1), and then washed twice with distilled water in order to eliminate superficial bacteria. The other half was used as control, immersing the root system in distilled water by 5 minute and then washed twice with distilled water. Inoculated and control plants were transferred to MS medium and kept there for seven days. After this period, whole plants were harvested and examined for bacterial colonization by plate counting. Total RNA was isolated from whole plants using Trizol (Invitrogen, CA, USA) as described by the manufacturer. The mRNA-seq library was constructed following the TruSeq RNA Sample Prep Kit.