基于16S rRNA基因V3-V4区高通量测序分析东北自然发酵酸菜中细菌群落结构

Analysis of the Bacterial Community Structure in Naturally Fermented Cabbage of Northeast China by High-throughput Sequencing

摘要:

为探究东北自然发酵酸菜中细菌群落结构，本试验采用454 FLX+平台对东北地区传统自然发酵的16份酸菜汁样品中细菌16S rRNA基因的V3-V4区进行测序。通过454焦磷酸测序，共得到302327条有效序列。计算Chao 1指数、Shannon指数和Simpson指数，对样品菌群的Alpha多样性进行评价，发现松原和大庆样品的细菌群落具有较高的丰富度。Beta多样性分析发现黑龙江、吉林和辽宁省份样品中存在交叠细菌物种，但含量极低。Heatmap图显示各省份的优势菌属各不相同。样品群落结构分析表明大庆、黑河和松原样品中乳杆菌含量超过50%，黑河样品中乳杆菌含量高达69.6%。KEGG统计结果表明高纬度地区存在着一定数量的耐寒细菌。此外，本试验首次在发酵食品中发现污蝇解壳杆菌属细菌（wohlfahrtiimonas），显示出高通量测序技术相对传统纯培养技术的优势。

Abstract:

In order to reveal the microbial community structure in naturally fermented cabbage from Northeast China, the 454 FLX+pyrosequencing platform was used to carry out sequencing of V3-V4 regions of the 16S rRNA gene in the microbiota of the cabbage. A total of 302327 valid sequences were analyzed by 454 pyrosequencing in 16 samples. The Chao1 index, Shannon index, and Simpson index were used to evaluate the alpha diversity of the samples, and the results showed that the microbiota in samples from Songyuan and Daqing had a relatively high abundance. Beta diversity analysis found common bacterial species among the samples from Heilongjiang Province, Jilin Province, and Liaoning Province, although their content was extremely low. In addition, heatmap analysis revealed that dominant bacterial genera were significantly different among the samples from these three provinces. Analysis of the microbial community structure indicated that the content of Lactobacillus strains in the samples from Daqing, Heihe, and Songyuan was greater than 50%; the content in the sample from Heihe reached up to 69.6%. Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses showed that there were a certain number of cold-resistant bacteria in the high latitude areas. Furthermore, Wohlfahrtiimonas spp. were reported for the first time in the fermented food, which showed the advantages of high-throughput sequencing compared with the conventional culture technique.