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为研究不同养殖时期刺参Apostichopus japonicus肠道内的细菌菌群结构,采用PCR-DGGE技术对DGGE凝胶中的主要条带进行切割、回收、克隆、测序。结果表明:3、4、5、6、9、10月,大连瓦房店海区养殖刺参肠道内细菌16S r DNA-V3可变区序列经PCR扩增及DGGE电泳后分别获得49、49、54、53、52、51个条带;3月与9月的菌群结构相似度最高,为67.9%,4月与10月次之,为63.8%;经条带序列分析得到43个主要条带对应的核酸序列,11个与未培养菌相似,32个与归属为7个细菌类群的细菌序列相似,包括α-变形菌纲、γ-变形菌纲、δ-变形菌纲、黄杆菌纲、蓝藻纲、绿弯菌门、疣微菌门;α-变形菌纲、γ-变形菌纲、黄杆菌纲是不同养殖时期刺参肠道内的主要细菌群落,其中γ-变形菌纲所占比例最高,达25.00%~35.71%,除3月和9月外,黄杆菌纲的相对含量均高于α-变形菌纲。研究表明,春秋两季刺参肠道内菌群结构呈现类似的演替变化规律,α-变形菌纲所占比例下降,而γ-变形菌纲与黄杆菌纲所占比例上升。
In order to study the bacterial flora in the intestinal tract of Apostichopus japonicus in different culture periods, the major bands in DGGE gel were cut, recovered, cloned and sequenced by PCR-DGGE. The results showed that the bacterial 16S r DNA-V3 variable region sequences of the cultured sea cucumbers in the Wafangdian area of Dalian were obtained by PCR amplification and DGGE electrophoresis after 49, 49, 54,53,52,51 bands; the similarity of the flora in March and September was the highest, 67.9%, followed by 63.8% in April and October; 43 bands With corresponding nucleic acid sequences, 11 were similar to uncultured bacteria, and 32 were similar to the bacterial sequences belonging to 7 bacterial groups, including α-Proteobacteria, γ-Proteobacteria, δ-Proteobacteria, , Cyanobacteria, Green Beans, and Verrucae; α-Proteobacteria, γ-Proteobacteria and Flavobacterium are the main bacterial communities in the colon of the sea cucumbers in different culture periods, among which γ-Proteobacteria accounted for The highest proportion, up to 25.00% ~ 35.71%, in addition to March and September, the relative content of the flavobacteria are higher than the alpha-Proteobacteria. The results showed that the structure of S. glabra in the spring and autumn showed similar succession, the proportion of α-Proteobacteria decreased, while the proportion of γ-Proteobacteria and the species of Flavobacterium increased.