RNA-seq analysis of gene expression changes in cuticles during the larval-pupal metamorphosis of Plutella xylostella

Abstract

The diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae) is a holometabolous insect that its cuticles must undergo the significant changes during the larval-pupal metamorphosis development. To elucidate these changes at molecular levels, RNA-seq analysis of cuticles from LLS (later fourth instar larval stage), PPS (prepupal stage) and PS (pupal stage) were performed in P. xylostella. In this paper, a total of 17,710 transcripts were obtained in the larval–pupal transition of P. xylostella, and out of which 2293 (881 up-regulated and 1412 down-regulated) and 2989 transcripts (2062 up-regulated and 927 down-regulated) were identified to be differentially expressed between LLS and PPS, as well as PPS and PS, respectively. The further GO and KEGG analysis of differentially expressed genes (DEGs) revealed that the ‘structural constituent of cuticle’, ‘chitin metabolic process’, ‘chitin binding’, ‘tyrosine metabolism’ and ‘insect hormone biosynthesis’ pathways were significantly enriched, indicating these pathways might be involved in the process of larval pupation in P. xylostella. Then, we found some genes that encoded cuticular proteins, chitinolytic enzymes, chitin synthesis enzymes, and cuticle tanning proteins changed their expression levels remarkably, indicating these genes might play important roles in the restruction (degradation and biosynthesis) of insect cuticles during the larval metamorphosis. Additionally, the significant changes in the mRNA levels of 20-hydroxyecdysone (20E) and juvenile hormone (JH) related genes suggested their crucial roles in regulating cuticle remodeling during the larval metamorphosis of P. xylostella. In conclusion, the present study provide us the comprehensive gene expression profiles to explore the molecular mechanisms of cuticle metamorphosis in P. xylostella, which laid a molecular basis to study roles of specific pathways and genes in insect development.