应用Illumina MiSeq高通量测序技术解析梅干菜中细菌多样性
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摘要
采用Illumina Mi Seq高通量测序技术对恩施地区梅干菜中细菌多样性进行解析,同时使用传统纯培养技术对乳酸菌进行分离鉴定。结果表明,恩施地区梅干菜中含量最高的优势细菌门为硬壁菌门(Firmicutes),其平均相对含量高达60.70%。优势细菌属为乳酸杆菌属(Lactobacillus)、假单胞菌属(Pseudomonas)、葡萄球菌属(Staphylococcus)、魏斯氏菌属(Weissella)、鞘脂单胞菌属(Sphingomonas)、嗜冷杆菌属(Psychrobacter)、四联球菌属(Tetragenococcus)和棒状杆菌属(Corynebacterium),其平均相对含量分别为60.50%、8.69%、2.86%、1.73%、1.28%、1.07%、1.03%、1.00%。采用传统培养方法从梅干菜分离出5种共12株乳酸菌,经分子生物学鉴定分别为植物乳杆菌(Lactobacillus plantarum)、短乳杆菌(Lactobacillus brevis),副干酪乳杆菌副干酪亚种(Lactobacillus paracasei subsp. paracasei)、食品乳杆菌(Lactobacillus alimentarius)和融合魏斯氏菌(Weissella confusa)。恩施地区梅干菜中的细菌主要以乳酸菌为主,且乳酸杆菌多样性较高。
The bacterial diversity in preserved vegetable samples collected from Enshi was analyzed by Illumina Mi Seq high throughput sequencing technology, and the lactic acid bacteria were isolated and identified by traditional pure culture techniques. The results showed that the relative content of Firmicutes was the highest in the dominant bacteria phylum, and the average relative content of Firmicutes was 60.70%. The dominant bacterial genus were Lactobacillus, Pseudomonas, Staphylococcus, Weissella, Sphingomonas, Psychrobacter, Tetragenococcus and Corynebacterium, and the average relative contents were 60.50%, 8.69%, 2.86%, 1.73%, 1.28%, 1.07%, 1.03%, and 1.00%, respectively. 12 lactic acid bacteria belonged to 5species were isolated by traditional culture method, which were identified as Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus paracasei subsp. paracasei, Lactobacillus alimentarius, and Weissella confuse, respectively. The main bacteria in preserved vegetable collected from Enshi were lactic acid bacteria, and the diversity of Lactobacillus was higher.
The bacterial diversity in preserved vegetable samples collected from Enshi was analyzed by Illumina Mi Seq high throughput sequencing technology, and the lactic acid bacteria were isolated and identified by traditional pure culture techniques. The results showed that the relative content of Firmicutes was the highest in the dominant bacteria phylum, and the average relative content of Firmicutes was 60.70%. The dominant bacterial genus were Lactobacillus, Pseudomonas, Staphylococcus, Weissella, Sphingomonas, Psychrobacter, Tetragenococcus and Corynebacterium, and the average relative contents were 60.50%, 8.69%, 2.86%, 1.73%, 1.28%, 1.07%, 1.03%, and 1.00%, respectively. 12 lactic acid bacteria belonged to 5species were isolated by traditional culture method, which were identified as Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus paracasei subsp. paracasei, Lactobacillus alimentarius, and Weissella confuse, respectively. The main bacteria in preserved vegetable collected from Enshi were lactic acid bacteria, and the diversity of Lactobacillus was higher.
引文
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[2]程玥,徐晓兰,张宁,等.同时蒸馏萃取-气质联用分析三全梅菜扣肉的挥发性风味成分[J].食品科学,2013,34(12):147-150.
[3]卓莉,刘良凤,阮尚全.原子吸收光谱法测定梅干菜中微量元素的含量[J].安徽农业科学,2009,37,30(21):9836.
[4]黄师荣,李豪杰,戴杰辉,等.九头芥梅干菜提取物抗菌活性及其在猪肉保鲜中的应用[J].现代食品科技,2014,30(10):58-62.
[5]沈清,楼乐燕,尹培,等.五种梅干菜的酚类化合物及抗氧化能力比较分析[J].食品科学,2017,38(18):1-11.
[6]黄苇,赵玲华,李远志,等.梅菜漂洗及脱盐工艺参数优选[J].中国调味品,2009,34(6):80-82.
[7]周美琪,周其德,田赛莺,等.低盐腌制对缙云梅干菜加工品质的影响[J].核农学报,2018,32(8):1562-1571.
[8]杨翠,李祖明.分子生物学技术在肠道微生物研究中的应用进展[J].中国微生态学杂志,2017,29(2):229-233.
[9]曹荣,张井,孟辉辉,等.高通量测序与传统纯培养方法在牡蛎微生物群落分析中的应用对比[J].食品科学,2016,37(24):137-141.
[10]沈馨,王艳,代凯文,等.基于Miseq高通量测序技术的辣椒酱核心细菌类群研究[J].食品研究与开发,2018,39(10):151-167.
[11]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.
[12]张会敏,李天婵,孙美青,等.利用非培养技术初步研究古井贡酒窖泥细菌群落结构[J].食品工业科技,2014,35(13):200-206.
[13] OKA K, ASARI M, OMURA T, et al. Genotyping of 38 insertion/deletion polymorphisms for human identification using universal fluorescent PCR[J]. Mol Cell Probe, 2014, 28(1):13-18.
[14]蔡丽云,黄泽彬,须子唯,等.处理垃圾渗滤液的SBR中微生物种群与污泥比阻[J].环境科学,2018,39(2):880-888.
[15]周书楠,席修璞,董蕴,等.琚湾酸浆面浆水细菌多样性评价[J].中国酿造,2018,37(1):49-53.
[16] CAPORASO J G, KUCZYNSKI J, STOMBAUGH J, et al. QIIME allows analysis of high-throughput community sequencing data[J]. Nat Methods, 2010, 7(4):335-336.
[17] HAAS B J, GEVERS D, EARL A M, et al. Chimeric 16S r RNA sequence formation and detection in Sanger and 454-pyrosequenced PCR amplicons[J]. Genome Res, 2011, 21(3):494-504.
[18] DESANTIS T Z, HUGENHOLTZ P, LARSEN N, et al. Greengenes, a chimera-checked 16S r RNA gene database and workbench compatible with ARB[J]. Appl Environ Microbiol, 2006, 72(7):5069-5072.
[19] 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 Res, 2007, 35(1):169-172.
[20] SUN X M, CHEN X, DENG Z X, et al. A CTAB-assisted hydrothermal orientation growth of Zn O nanorods[J]. Mater Chem Phys, 2003, 78(1):99-104.