俄罗斯卡尔梅克共和国发酵蔬菜中细菌多样性研究

Study on bacteria diversity in fermented vegetable of Russian Republic of Kalmykia Elista

通过纯培养的方法结合16S rRNA测序技术对采集自俄罗斯卡尔梅克共和国埃利斯塔市郊的2份发酵蔬菜样品中的乳酸菌进行了分离、鉴定,并构建了系统发育树。经过鉴定,共从2份样品中获得了19株乳酸菌,分属于Lactobacillus(14株)、Pediococcus(3株)和Leuconostoc(2株),其中包括7株Lactobacillus plantarum;1株Lactobacillus brevis;2株Lactobacillus delbrueckii;1株Lactobacillus helveticus;1株Lactobacillus kefiranofaciens;2株Lactobacillus kefiri;1株Leuconostoc lactis;1株Leuconostoc mesenteroides;2株Pediococcus ethanolidurans;1株Pediococcus pentosaceus。此外,该研究以细菌16S rRNA的V1-V3区为靶序列,采用454焦磷酸测序技术从宏基因组角度对2份样品中细菌的丰度和多样性进行了分析,分别从2份样品中获得了10869和12204条高质量序列。经过与RDP(Ribosomal Database)数据库比对,发现隶属于硬壁菌门(Firmicutes)的乳杆菌属(Lactobacillus)和乳球菌属(Lactococcus)分别占到样品中细菌总数的91.66%和3.81%,成为绝对优势菌属。此外,还鉴定到了弧菌属(Vibrio)、片球菌属(Pediococcus)、肠球菌属(Enterococcus)、克鲁维菌属(Kluyvera)等。

In this study, the lactic acid bacteria of two fermented vegetable samples collected from the Russian Republic of Kalmykia Elista were isolated and identified by a combinatory approach of laboratory cultivation and 16S rRNA sequence analysis. Besides, phylogenetic dendrograms were constructed based on these isolates. A total of 19 strains of lactic acid bacteria were identified from these two samples. These bacteria belonged to the genera Lactobacil- lus （ 14 strains）, Pediococcus （ 3 strains） and Leuconostoc （2 strains）, including Lactobacillus plantarum （ 7 strains）, Lactobacillus brevis （1 strain）, Lactobacillus delbrueckii （2 strains）, Lactobacillus helveticus （1 strain）, Lactobacillus kefiranofaciens （1 strain）, Lactobacillus kefiri （2 strains）, Leuconostoc lactis （1 strain ）, Leuconostoc mesenteroides （ 1 strain） , Pediococcus ethanolidurans （2 strains） , and Pediococcus pentosaceus （ 1 strain）. In addi- tion, the bacterial abundance and diversity at the metagenomics level were analyzed using barcoded pyrosequencing targeting at the V1-V3 variable regions of 16S rRNA. A total of 10869 and 12204 unique sequences were analyzed from these two samples. After comparing with the Ribosomal Database, Lactobacillus and Lactococcus, which belonged to the phylum Firmicutes, were the dominant genera in the samples and accounted for 91.66% and 3.81% of the to- tal number of bacteria, respectively. In addition, other genera such as Vibrio, Pediococcus, Enterococcus and Kluyveromyces in the samples were also identified herein.