基于16S rRNA基因分析双斑东方鲀肠道微生物多样性

Microbial Diversity in Intestinal Tract of Wild and Cultured Puffer Takifugu bimaculatus Based on 16S rRNA Gene Sequence

为探究野生和养殖双斑东方鲀肠道微生物的群落结构,对厦门同安海域野生和养殖双斑东方鲀肠道内容物进行16S rRNA基因的V3~V4区测序,通过Illumina Miseq PE300高通量测序,对6份样品进行分析,共得到优质序列99500条,相似度为97%的聚类分析显示,野生双斑东方鲀运算分类单元数目为524,其中独有的运算分类单元数目为68,养殖双斑东方鲀运算分类单元数目为576,其中独有的运算分类单元数目为120。通过计算Chao1、observed-species、PD_whole_tree、香农指数进行Alpha多样性分析,结果显示,养殖双斑东方鲀细菌多样性和丰富度高于野生双斑东方鲀。门水平分析表明,变形菌门为野生双斑东方鲀和养殖双斑东方鲀的优势菌。产河鲀毒素相关的希瓦氏菌属、纤毛菌属、发光杆菌属、假交替单胞菌属在野生双斑东方鲀肠道微生物中占有一定比例,而在养殖双斑东方鲀中未发现。所获结果揭示了野生和养殖双斑东方鲀肠道微生物中优势菌群的组成,为探究河鲀毒素的产毒机制提供科学证据。

The V3—V4 regions of 16S rRNA were sequenced in the intestinal contents of wild puffer Takifugu bimaculatus collected from Tong′an sea area,Xiamen,China and cultured puffer by Through Illumina Miseq PE300 high throughput to explore the community structure of intestinal microorganisms.A total of 99500 superior sequences were obtained from six samples and the cluster analysis with 97%similarity showed that the number of OTU was 524 with unique OTU of 68 in wild puffer and 576 in cultured puffer with unique OTU of 120.Alpha diversity analysis was carried out from the calculation results of 4 indices(Chao1,observed-species,PD-whole-tree and Shannon).The results showed that there were higher diversity and richness of bacteria in cultured puffer than those in wild puffer.From the data in the phylum level Proteobacteria were of the dominant bacteria in wild and cultured puffer.Shewanella,Propionigenium,Photobacterium and Pseudoalteromonas producing tetrodotoxin were found in the wild puffer,and not found in the cultured puffer.The findings provided scientific evidence for exploring the toxicity-producing mechanism of tetrodotoxin,as the composition of dominant microflora was revealed in intestinal microorganisms of wild and cultured puffer.