Dual RNA-seq reveals the effect of the flgM gene of Pseudomonas plecoglossicida on the immune response of Epinephelus coioides

Abstract

Pseudomonas plecoglossicida is an important and highly pathogenic bacterium for aquaculture and causes serious losses. The expression level of flgM was found to be significantly upregulated post-infection compared with in vitro results, which was confirmed by quantitative real-time PCR. RNAi significantly reduced the expression level of flgM mRNA of P. plecoglossicida. Compared with infection with the wild-type strain, infection with the flgM-RNAi strain resulted in a delay in death and a 75% reduction in the mortality of Epinephelus coioides, followed by alleviation of the symptoms in E. coioides spleen. Moreover, compared with infection with the wild-type strain, infection with the flgM-RNAi strain of P. plecoglossicida resulted in a significant change in the transcriptome of the spleens of infected E. coioides and P. plecoglossicida. KEGG analysis for E. coioides showed that genes of 17 immune pathways were most affected by flgM-RNAi of P. plecoglossicida. Among them, the expression of mhc2, zap70, rhoh, tlr2, ca79a, hcst and cd32 in E. coioides spleen was predicted to be negatively related to flgM in P. plecoglossicida but positively related to genes involved in communication, metabolism and motility.

Keywords: Dual RNA-eq; Epinephelus coioides; Pseudomonas plecoglossicida; flgM; immune response.