养殖刀鲚与生长环境菌群PCR-DGGE指纹图谱及多样性分析

为了分析养殖刀鲚体内与生长环境菌群结构,利用PCR-DGGE技术,对养殖刀鲚鳃、胃、肠壁及肠内容物和养殖水体菌群结构进行了初步分析。PCR-DGGE指纹图谱分离显示,42条清晰条带,其中养殖水体(27)、鳃(9)、胃(13)、肠道壁(19)、肠道内容物(18)的香农指数分别为3.037、1.883、2.193、2.825、2.683;养殖水体与刀鲚鳃、胃、肠道壁及肠道内容物分别具有6、9、11、8共有带。UPGMA聚类分析显示,样品3个重复相似度都在95%以上,差异不明显;不同样品之间,养殖刀鲚鳃和胃聚为一支,具有较高的相似度(76%),同时与养殖水体相似度达29%;养殖刀鲚肠道壁和肠道内容物聚为一支,相似度为38%。回收测定所有显示条带,主要包含变形菌、放线菌、拟杆菌、柔膜菌、蓝藻细菌、厚壁菌、梭杆菌及少量未定义菌种。研究表明,PCR-DGGE技术能区分养殖刀鲚主要部位及水体微生物的结构差异和多样性,澄清养殖刀鲚及生长水体微生物区系,可为定植益生菌的开发提供参考。

Fingertprinting and diversity analysis of the predominant bacterial community in Coilia nasus from ponds and its environment

The preliminary analysis of the bacterial community structure in the water environment, gill , stomach and intestine of Coilia nasus from fishponds, including intestinal wall and intestinal contents based on 16S rDNA PCR-DGGE figure using a culture independent approach. The DGGE fingerprint showed respectively 27, 9, 13, 19, 18 obvious different Signal strength bands in the samples of water environment, gill, stomach, intestinal wall and intestinal contents of Coilia nasus , meanwhile, the Shannon Diversity index of which respectively were 3.037, 1.883, 2.193, 2.825, 2.683, these might reflect the existence of rich bacteria species. The clustering analysis to the DGGE fingerprint displayed that No significant differences between groups of Repeating sample, the similarity of which above 95%; The sample of gill and stomach of Coilia nasus get together for a branch, the bacteria structure similarity of which was 77% and that in the intestinal wall and intestinal contents of Coilia nasus was 38%. 42 DGGE bands all were successfully re-amplified, cloned, sequenced and aligned with BLAST, including Proteobacteria, Actinobacteria, Firmicutes, Tenericutes, Bacteroidetes Cyanobacteria, Fusobacteria and some unclassified bacteria. These results implied that DGGE technique could be used to analyze the bacterial genetic diversity and clarify the predominant bacterial community of Coilia nasus from fishponds and its growing environment.