Transcriptome of Sitophilus zeamais: whole adults challenged by Gram+ or Gram- bacteria
RNA-seq libraries were constructed from a pool of both nave weevils and weevils challenged with Gram-positive or Gram-negative bacteria. Briefly, septic injuries were made on adults challenged with 10^5 bacteria of the genus Micrococcus luteus (strain CCM169, Gram-positive) or Dickeya dadantii (strain A470, Gram-negative). A total of 69 nl of bacterial suspension were injected into the hemolymph using a Nanoject II (Drummond Scientific, Broomall, PA, USA). Five individuals were randomly sampled at 6h and 12h post-injection for each bacterial challenge leading to a total of 20 adults. Total RNA from these 20 adults was extracted with Trizol reagent (Invitrogen, ThermoFisher Scientific, Waltham, MA, USA) following the manufacturers instructions. RNA was incubated with 1 U/g of RQ1 RNase-free DNase (Promega, Madison, WI, USA) for 30 min, at 37 C then purified with the NucleoSpin RNA Clean-up kit (Macherey-Nagel, Dren, Germany). Total RNA concentration and quality were checked using the Qubit Fluorometer (ThermoFisher Scientific) and the Agilent 2200 TapeStation (Agilent Technologies, Santa Clara, CA, USA). Twenty nave adults were randomly selected and total RNA was extracted using the same procedure. The pool was prepared by mixing 1 g of RNA from nave weevils with 1 g of RNA from weevils challenged with bacteria. RNA-seq libraries were then prepared using the SENSE mRNA-seq library preparation kit (Lexogen, GmbH, Vienna, Austria) following the manufacturers instructions and sequenced on an Illumina NextSeq500 platform (2x150bp, Illumina, San Diego, CA, USA) at the sequencing platform of the IGFL (Institut de Gnomique Fonctionnelle de Lyon, Ecole Normale Suprieure de Lyon, France).