RNA-Seq of Plasmopara viticola 221: 24hpi Vitis vinifera cv. Muscat Ottonel detached leaves
Seedlings of Vitis vinifera cv. Muscat Ottonel were grown on stone wool in a greenhouse at 22-19 C (day-night) and a photoperiod of 16 h of light. Detached leaves were surface-sterilized with bleach,followed by three washes in sterile water. Leaves were inoculated all through their surface with 10 ml-drops of a suspension of P. viticola containing 50 000 sporangia/ml, kept in Petri dishes onwet filter paper and incubated in a growing chamber at 21C and a photoperiod of 16/8 h. 24 hours post-inoculation (hpi), successful infection was confirmed by microscopic observation of pathogen structures. Leaves were then snap-frozen with liquid nitrogen and conserved at -80C.Total RNA was extracted using a classical cetyltrimethyl-ammonium-bromide (CTAB)/phenol-chloroform extraction method followed by LiCl precipitation. Removal of residual genomic DNA was achieved by DNase treatment of RNAs using the Ambion Turbo-DNAfree kit. RNA quality was verified using an Agilent Bioanalyzer 2100 LabChip.mRNA were selected using poly-T beads and libraries were prepared at the GeT-PlaGe facility (Toulouse, France) according to Illumina protocols using the Illumina TruSeq Stranded mRNA sample prep kit.