★ Identify 95 new immunity-related genes including most members of the Toll, IMD, JAK-STAT, and MAPK-JNK-p38 pathways. ★ Detect minor changes in mRNA levels of the pathway members and dramatic elevations of transcripts for effector proteins. ★ Reveal the attacin gene family expansion using the RNA-Seq data. ★ Support immunogenome analysis with a deep, quantitative transcriptomic study.
As a biochemical model, Manduca sexta has substantially contributed to our knowledge on insect innate immunity. The RNA-Seq approach was implemented in three studies to examine tissue immunotranscriptomes of this species. With the latest and largest focusing on highly regulated process- and tissue-specific genes, we further analyzed the same set of data using BLAST2GO to explore functional aspects of the larval fat body (F) and hemocyte (H) transcriptomes with (I) or without (C) immune challenge. Using immunity-related sequences from other insects, we found 383 homologous contigs and compared them with those discovered based on relative abundance changes. The major overlap of the two lists validated our previous research designed for gene discovery and transcript profiling in organisms lacking sequenced genomes. By concatenating the contigs, we established a repertoire of 232 immunity-related genes encoding proteins for pathogen recognition (16%), signal transduction (53%), microbe killing (13%) and others (18%). We examined their transcript levels along with attribute classifications and detected prominent differences in nine of the 30 level 2 gene ontology (GO) categories. The increase in extracellular proteins (155%) was consistent with the highly induced synthesis of defense molecules (e.g., antimicrobial peptides) in fat body after the immune challenge. We identified most members of the putative Toll, IMD, MAPK-JNK-p38 and JAK-STAT pathways and small changes in their mRNA levels. Together, these findings set the stage for on-going analysis of the M. sexta immunogenome.