东北传统发酵酸菜中微生物群落多样性及时空分布规律研究

As a traditional fermented food in northeast of China，sour pickled cabbages is usually naturally fermented with abundant microbiota. However, so far it is still not very clear about the distribution and diversity of microbiota involved in naturally fermented sour pickled cabbages. In the present study, samples of naturally fermented sour pickled cabbages with different area or fermentation periods by traditional method were collected from village families in the northeast of China，which are chosen as a model to investigate the overview of community structure and population dynamics of fermented food using pyrosequencing tagged by 16S rRNA gene and ITS amplicons. Based on the analysis of sequence screening, blast, DNA distance calculation and the abundance of-based coverage, etc. by software and procedures including BLASTN, PHYLIP 3.68, DOTUR program, MEGAN 5.0 and so on, finely the diversity and change rules of microbiota involved in naturally fermented sour pickled cabbages can be revealed through the phylogenetic trees. In conclusion,the technology adopt in the project is very rapid and accurate without cloning or electrophoresis, the data are more objective and roundly，it has important significance for further studies，excavation and protection of microbial resource in northeast of China.

酸菜是我国东北地区传统自然发酵食品，其中蕴藏着丰富的微生物资源。然而，其中的微生物群落结构及变化规律至今仍未得到全面准确的认识。 本项目以采自我国东北三省不同区域和不同发酵阶段的传统自然发酵酸菜为试材，从宏基因组学角度出发，主要利用较为先进的焦磷酸测序技术，对样品中的微生物DNA进行大规模测序，再利用BLASTN、PHYLIP 3.68、DOTUR program、MEGAN 5.0等软件和程序，进行序列筛选、比对、DNA distance等计算和参数分析，最终构建各样品中微生物群落的系统发育树并确定属种，进而分析和掌握不同地区和不同发酵阶段样品中微生物群落的组成和分布情况，试图全面、准确地揭示东北地区自然发酵酸菜中微生物的多样性及其变化规律。项目采用的技术方法快速、精确，所得数据较为客观全面，对于进一步深入研究、挖掘和保护我国传统发酵酸菜中宝贵的微生物资源，具有重要的意义。

采用454 FLX+平台进行测序，以期揭示东北地区传统自然发酵酸菜中优势菌群的组成，以及不同纬度省份间酸菜中微生物群落变化情况，为探明菌群多样性的进化规律奠定基础。同时，采用PCR-DGGE 分子生物学技术,结合16S rDNA 同源性分析及生物信息学手段,对传统自然发酵酸菜中乳酸菌进行全面系统研究，揭示不同时期的乳酸菌菌落结构的真实状态。