Short RNAs associated to Cauliflower mosaic virus infections in Canola and Turnip

Three plants of two species in Brassica genus, i.e. B. rapa cv. "Purple To White Globe" and B. napus cv. "SariGol", highly susceptible and tolerant hosts of Cauliflower mosaic virus (CaMV), respectively, were mechanically inoculated with CaMV strain NY8153.Phosphate buffer was used as control in mock-inoculated plants. RNAs, in three biological repeats, were extracted 20 days post-inoculation (dpi) collecting ten leaves from mock-inoculated and virus-infected plants using Trizol reagent (Invitrogen, Carlsbad, CA) according to the manufacturer’s instructions. Five µg of total RNA were resolved in 15% SDS-PAGE gel. Total RNA pattern was visualized upon Ethidium Bromide staining and sRNAs in the size range of 20-30 nt were eluted from gel. sRNAs replicates were quantified and mixed in equal molarity. Libraries were obtained using NEXTflex Small RNA-Seq Kit v3 (Bio Scientific) according to manufacturer’s instructions. Libraries were eluted from a 6% non-denaturing PAGE. The quality of libraries was checked by BioPharma QC, and then, submitted for sequencing with a HiSeq 2500 Illumina platform. Short RNA fractionation and characterization