Additional file 9: Figure S5. of Transcription analysis of neonicotinoid resistance in Mediterranean (MED) populations of B. tabaci reveal novel cytochrome P450s, but no nAChR mutations associated with the phenotype

Heatmap spearman correlation matrix of significant DE (LogFC > 1 FDR < 0.05) unigenes based on their TMM-FPKM and GO enrichment analysis. The relative expression levels of each gene (row) in the susceptible and resistant samples (column) are shown. Up- and down-regulated genes are represented respectively by red and yellow. Clustering analysis was performed using Euclidean distance and dward algorithm with optimal leaf order (unigenes) or Spearman’s rank correlation (12 samples) using mean-centering transformation. The cluster was cut in three different parts using “cutree” and the unigenes of each sub cluster were recovered for GO enrichment analysis with Blast2GO for GO enrichment analysis using all DE as reference. Clustering of the samples grouped the neonicotinoid resistant samples together. Based on the transcription level of the 3,745 DE unigenes, three sub-clusters showing marked differences in normalized FPKM were identified. Assigning known transcripts to molecular functions revealed differences in the GO enrichment in the three clusters, displayed in the figure. The sub-clusters A1 and A2 group unigenes with lower expression profile in the susceptible strain and differ in expression profiles between the resistant strains. The sub-cluster A1 includes transcripts highly over-expressed in GR4-AS and GR4-AP and the more frequent GO terms included transferase activity (24 %), ATP binding (23 %) and heme binding (20 %). The sub-cluster A2 displayed the more visible and more abundant differences between the GR9-IS and the other resistant strains and revealed an enrichment in terms related to cytochrome P450s such as heme binding, iron ion binding and oxidoreductase activity (60 % of all overrepresented GO terms). Terms related to hydrolase activity were also enriched in sub-cluster A2. The cluster B represents unigenes mostly up regulated in the susceptible strain. The cluster B is the most diversified with 16 GO terms enriched, the majority of which were related with signal transduction. (PDF 261 kb