恩施地区酸萝卜中细菌多样性研究
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摘要
采用Miseq高通量测序技术、变性梯度凝胶电泳技术以及传统微生物学手段对恩施地区酸萝卜中细菌和乳酸菌多样性进行了解析。结果发现酸萝卜中的优势菌主要为隶属于Firmicutes的Lactobacillus和Pediococcus构成,其平均相对含量分别为55.76%和34.50%。经DGGE分析发现,酸萝卜中乳酸菌主要包括L. kefiri、P. tuberculata、L. senmaizukei、L. acetotolerans、L. acetotolerans和L. kefiri。从3个样品中共分离出14株乳酸菌,其中L.plantarum subsp. plantarum 5株,E.faecium、L.paracasei subsp. paracasei、L.fermentum和L.alimentarius各2株以及L.brevis 1株。由此可见,Lactobacillus是恩施酸萝卜中的优势细菌且多样性较高。
The diversity of bacteria and lactic acid bacteria in sour radish fermentation broth collected from Enshi were analyzed by high throughput sequencing technology,denaturing gradient gel electrophoresis and traditional microbiological method. The results shown that the main dominant bacteria in samples were Lactobacillus and Pediococcus which belongs to Firmicutes,and their average relative content was 55.76% and 34.50% respectively. By DGGE analysis,it was found that lactic acid bacteria in sour radish fermentation broth mainly included L. kefiri,P. tuberculata,L. senmaizukei,L. acetotolerans,L. acetotolerans and L. kefiri. A total of 14 strains were isolated from three samples,including 2 strains of E. faecium,L. paracasei subsp. paracasei,L. fermentum and L. alimentarius,5 strains of L.plantarum subsp. plantarum and 1 strain of L.brevis. The conclusion is that Lactobacillus was the dominant bacteria in Enshi sour radish fermentation broth with high diversity.
The diversity of bacteria and lactic acid bacteria in sour radish fermentation broth collected from Enshi were analyzed by high throughput sequencing technology,denaturing gradient gel electrophoresis and traditional microbiological method. The results shown that the main dominant bacteria in samples were Lactobacillus and Pediococcus which belongs to Firmicutes,and their average relative content was 55.76% and 34.50% respectively. By DGGE analysis,it was found that lactic acid bacteria in sour radish fermentation broth mainly included L. kefiri,P. tuberculata,L. senmaizukei,L. acetotolerans,L. acetotolerans and L. kefiri. A total of 14 strains were isolated from three samples,including 2 strains of E. faecium,L. paracasei subsp. paracasei,L. fermentum and L. alimentarius,5 strains of L.plantarum subsp. plantarum and 1 strain of L.brevis. The conclusion is that Lactobacillus was the dominant bacteria in Enshi sour radish fermentation broth with high diversity.
引文
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[2]Zhao N,Cai J,Zhang C,et al.Suitability of various DNAextraction methods for a traditional Chinese paocai system[J].Bioengineered,2017,8(5):642-650..
[3]罗菊英,闫永才,李灿,等.恩施自治州气候资源分析及旅游适宜性区划[J].长江流域资源与环境,2013,22(1):39-45.
[4]裴乐乐,罗青春,孟霞,等.不同原料四川发酵泡菜的细菌多样性分析[J].中国调味品,2016,41(2):39-43.
[6]朱琳,高凤,曾椿淋,等.萝卜泡菜细菌多样性的高通量测序分析[J].现代食品科技,2018,34(2):225-231.
[6]Xiong T,Guan Q,Song S,et al.Dynamic changes of lactic acid bacteria flora during Chinese sauerkraut fermentation[J].Food control,2012,26(1):178-181.
[7]Xiong T,Song S,Huang X,et al.Screening and identification of functional Lactobacillus specific for vegetable fermentation[J].Journal of food science,2013,78(1):84-89.
[8]沈馨,尚雪娇,董蕴,等.基于MiSeq高通量测序技术对3个孝感凤窝酒曲细菌多样性的评价[J].中国微生态学杂志,2018,30(5):525-544.
[9]Tom V,Vicky D P,Evie D B,et al.Molecular monitoring of the fecal microbical of healthy human subjects during administration of lactulose and saccharomyces boulardii[J].Environ microbiol,2006,72(9):5990-5997.
[10]Andreoni V,Benedetti A,Canzi E,et al.Seleniumenriched biomass,method for preparing thereof and probiotic and nutraceutical products including said biomass:U.S.Patent7,666,638[P].2010-2-23.
[11]Oka K,Asari M,Omura T,et al.Genotyping of 38insertion/deletion polymorphisms for human identification using universal fluorescent PCR[J].Molecular and cellular probes,2014,28(1):13-18.
[12]蔡宏宇,王艳,沈馨,等.常规食用调味面制品对青年志愿者肠道菌群多样性影响[J].食品工业科技,2017,38(21):289-294.
[13]葛英亮,时文歆.应用Illumina Miseq测序分析饮用水源水中病毒多样性[J].食品科学,2018,39(2):287-292.
[14]Caporaso J G,Kuczynski J,Stombaugh J,et al.QIIMEallows analysis of high-throughput community sequencing data[J].Nature methods,2010,7(4):335-336.
[15]Edgar R C.Uparse:Highly accurate OTU sequences from microbial amplicon reads[J].Nature methods,2013,10(10):996-998.
[16]Desantis T Z,Hugenholtz P,Larsen N,et al.Greengenes,a chimera-checked 16S rRNA gene database and workbench compatible with ARB[J].Applied and environmental microbiology,2006,72(7):5069-5072.
[17]Cole J R,Chai B,Farris R J,et al.The ribosomal database project(RDP-II):introducing myRDP space and quality controlled public data[J].Nucleic acids research,2007,35(1):169-172.
[18]夏围围,贾仲君.高通量测序和DGGE分析土壤微生物群落的技术评价[J].微生物学报,2014,54(12):1489-1499.
[19]刘石泉,胡治远,赵运林.变性梯度凝胶电泳法初步解析茯砖茶渥堆发酵过程中细菌群落结构[J].食品科学,2014,35(15):172-177.
[20]Burgmann H,Pesaro M,Widmer F,et al.A strategy for optimizing quality and quantity of DNA extracted from soil[J].Journal of microbiological methods,2001,45(1):7-20.
[21]曹佳璐.传统四川泡菜盐水乳酸菌多样性的研究[D].北京:中国农业大学,2017:39-41.
[22]刘鹏飞,赵丹,宋刚,等.变性梯度凝胶电泳技术在微生物多样性研究中的应用[J].微生物学杂志,2013,33(6):88-92.