CCDR_1D_replicate_3
Oryza sativa
Protocols: Samples were all taken 50 days after germination. Leaf samples ~ 2 cm from three biological replicates were collected from the control untreated (0-day) and the inoculation sites after 1, 3 and 5 days after treatment and placed immediately in liquid nitrogen until transfer to -80C freezer for storage. Mist chambers assays were performed in a greenhouse located on the Louisiana State University (LSU) campus in Baton Rouge, LA. The temperature inside the chamber ranging from minimum 27 degrees celsius in the night to maximum 37 degrees celsius in the day. Natural daylight was used with day length of approximately 11 hours 30 minutes and the humidity was maintained 80-90%. Inoculation occured 50 days after germination with a PDA medium disc (0.8 cm diameter) containing Rhizoctonia solani (LR172) mycelia placed at the base of the stem and other disc placed between leaf blade and leaf sheath in the primary tiller of each plant. Total RNA from frozen seedling shoot sample was extracted as per manufacturer's protocol using RNA Plant reagent (Invitrogen Inc., USA), RNeasy kits (Qiagen Inc., USA), and treated with RNase-free DNase (Life Technologies Inc., USA). True-Seq kit (Illumina Inc.) was used to prepare RNA-seq libraries, according to the manufacture's protocol