Identification of 35 C-Type Lectins in the Oriental Armyworm, Mythimna separata (Walker)

SIMPLE SUMMARY: The oriental armyworm Mythimna separata is a lepidopteral agricultural pest that causes serious damage to many crops, such as maize, wheat, and sorghum. To control this pest, it is advisable to take comprehensive measures, including the use of chemical pesticides, microbial pesticides, and cultural practices. However, microbial pesticides (entomopathogens) can be eliminated by the insect immune system. C-type lectins (CTLs) are a family of pattern-recognition receptors that recognize carbohydrates and mediate immune responses. C-type lectins in the oriental armyworm have not yet been identified and characterized. In this study, a transcriptome of M. separata larvae was constructed and a total of 35 CTLs containing single or dual carbohydrate-recognition domains (CRDs) were identified from unigenes. Phylogenetic analyses, sequence alignments and structural predictions were performed. Gene expression profiles in different developmental stages, naïve larval tissues, and bacteria/fungi-challenged larvae were analyzed. Overall, our findings indicate that most dual-CRD CTLs are expressed in mid-late-stage larvae, pupae, and adults. Bacterial and fungal challenges can stimulate the expression of many CTLs in larval hemocytes, fat body, and midgut. Our data suggest the importance of CTLs in immune responses of M. separata. ABSTRACT: Insect C-type lectins (CTLs) play vital roles in modulating humoral and cellular immune responses. The oriental armyworm, Mythimna separata (Walker) (Lepidoptera: Noctuidae) is a migratory pest that causes significant economic loss in agriculture. CTLs have not yet been systematically identified in M. separata. In this study, we first constructed a transcriptome of M. separata larvae, generating a total of 45,888 unigenes with an average length of 910 bp. Unigenes were functionally annotated in six databases: NR, GO, KEGG, Pfam, eggNOG, and Swiss-Prot. Unigenes were enriched in functional pathways, such as those of signal transduction, endocrine system, cellular community, and immune system. Thirty-five unigenes encoding C-type lectins were identified, including CTL-S1~CTL-S6 (single CRD) and IML-1~IML-29 (dual CRD). Phylogenetic analyses showed dramatic lineage-specific expansions of IMLs. Sequence alignment and structural modeling identified potential ligand-interacting residues. Real-time qPCR revealed that CTL-Ss mainly express in eggs and early stage larvae, while IMLs mainly express in mid-late-stage larvae, pupae, and adults. In naïve larvae, hemocytes, fat body, and epidermis are the major tissues that express CTLs. In larvae challenged by Escherichia coli, Staphylococcus aureus, or Beauveria bassiana, the expression of different CTLs was stimulated in hemocytes, fat body and midgut. The present study will help further explore functions of M. separata CTLs.