海藻希瓦氏菌感染对半滑舌鳎肠道菌群结构及相关功能基因表达的影响

文章编号：2095-1191（2019）10-2300-08

Effects of infection with Shewanella algae on the microbial communities and expression of related functional genes in the intestine of Cynoglossus semilaevis

ZHANG Yan-yu1， HAN Zhuo-ran1， SUN Jing-feng1*， LYU Ai-jun1，

HU Xiu-cai1， LIU Jun-feng2

（1College of Fisheries， Tianjin Agricultural University/Tianjin Key Lab of Aqua-Ecology and Aquaculture， Tianjin 300384; 2Tianjin Yushengtang Biotechnology Co.， Ltd.， Tianjin  300404， China）

Abstract：【Objective】The purpose of this study was to investigate the changes of intestinal microbial communities and expression of related functional genes in the intestine of Cynoglossus semilaevis after artificial infection with the pathogenic Shewanella algae， and reveal the role of intestinal microbial flora and related functional genes in the process of disease occurrence and intestinal immune response in the host. 【Method】After C. semilaevis were artificially infected with the pathogenic S. algae，16S rDNA high-throughput sequencing technique was used to study the changes of intestinal microbial communities， and real-time fluorescence quantitative PCR technology was used to study the expression pattern of functional genes involved in the process of disease occurrence and immune response in the intestinal tissues. 【Result】A total of 118657 effective tags were obtained and assigned to 6732 OTUs based on a 97% sequence similarity level. The results of alpha diversity analysis showed that the indexes of Shannon and Chao1 were the highest in the control group （CG）， and the lowest in the group of 12 h post-injection（12hpi）. At phylum level，although the identified dominant intestinal bacteria were consistent between the control and infection groups， the relative abundance of the bacterial taxa varied. At genus level， the relative abundances of Elizabethkingia， Chitinophaga， Brevinema， Phenylobacterium， Pseudomonas， Lactobacillus， Marivita， andalstonia were the highest in the intestinal microbiota of CG; the relative abundances of Shewanella， Petrimonas， Proteiniphilum， and Aminobacterium in 12hpi group were more than those in other groups. Acidovorax， Bacillus， and Vibrio had the highest relative abundance in the group of 24 h post-injection （24hpi）. The expression patterns of related functional genes in the intestine of C. semilaevis were as follows：the relative expression of fructose drphosphate aldolase A gene ALDOA in 24hpi group was significantly higher than that in CG（P<0.05， the same below）; the relative expression of phospholipase b1 gene（PLB1）， heat shock protein 70 kD protein 1A gene（HSPA1A），histidine trimer nucleoside binding protein 1 gene（HINT1） and gamma glutamyltransferase 1 gene（GGT1）in 12hpi group was significantly higher than those in CG and 24hpi groups; the relative expression of trehalase gene（TREH） in 12hpi group was significantly lower than those in CG and 24hpi groups.【Conclusion】The structure of the microflora changes with the diversity of intestinal microflora of C. semilaevis decreasing after infection with S. algae. The immune function related genes（HSPA1A and HINT1） and metabolism-related enzyme genes （ALDOA， PLB1，GGT1 and TREH） are diffe-rentially expressed. It indicates that infection with S. algae results in the disturbance of intestinal microecology of C. semilaevis， and the immune function related genes and metabolism-related enzyme genes are involved in the process of immune response against bacterial infection and disease occurrence.

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