四川不同地区浓香型大曲微生物群落结构比较
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摘要
利用高通量测序技术分析四川不同地区浓香型大曲微生物群落结构差异。结果表明:大曲中优势细菌菌群为葡萄球菌属(Staphylococcus)、魏斯氏菌属(Weissella)、芽孢杆菌属(Bacillus)、乳杆菌属(Lactobacillus)、Kroppenstedtia、高温放线菌属(Thermoactinomyces)、糖多孢菌属(Saccharopolyspora)、短杆菌属(Brevibacterium),其中在品温较高的宜宾、泸州曲样中,魏斯氏菌属、高温放线菌属比例均高于遂宁曲样,而遂宁两个曲样的优势菌属均为葡萄球菌属和芽孢杆菌属,其中葡萄球菌属远高于宜宾和泸州曲样。另一方面,4种曲样的真菌主要以嗜热真菌属(Thermomyces)、曲霉属(Aspergillus)、嗜热子囊菌属(Thermoascus)、丝衣霉属(Byssochlamys)、节担菌属(Wallemia)为主,在品温较高的宜宾、泸州曲样中,嗜热真菌属和嗜热子囊菌属为优势菌属,其比例远高于遂宁曲样,而遂宁两个曲样的优势菌属均为曲霉属,其比例远高于宜宾和泸州的曲样。遂宁1、遂宁2曲样的细菌和真菌多样性均高于宜宾和泸州曲样。通过主成分分析发现,遂宁1和遂宁2曲样微生物群落结构较相似,而宜宾和泸州曲样的微生物群落结构则更接近。
In this study, we analyzed the microbial community structure of Luzhou-flavor liquor Daqu samples collected from different regions of Sichuan province by high throughput sequencing technology. The results showed that the dominant bacteria in the Daqu were Staphylococcus, Weissella, Bacillus, Lactobacillus, Kroppenstedtia,Thermoactinomyces, Saccharopolyspora, and Brevibacterium. The proportions of Thermoactinomyces and Weissella in Yibin Daqu and Luzhou Daqu, with higher temperature, were higher than those in two samples of Suining Daqu.The dominant strains in Suining Daqu were Staphylococcus and Bacillus, and Staphylococcus was much higher than in Yibin Daqu and Luzhou Daqu. On the other hand, the fungal communities in all four Daqu samples were predominated mainly by Thermomyces, Aspergillus, Thermoascus, Byssochlamys, and Wallemia. In Yibin and Luzhou samples, Thermomyces and Thermoascus were the dominant species, and their proportions were much higher than those in Suining Daqu. Aspergillus was the dominant species in Suining Daqu, and its proportion was much higher than those in Yibin Daqu and Luzhou Daqu. The diversities of bacteria and fungi in Suining Daqu were higher than in Yibin Daqu and Luzhou Daqu. Principal component analysis revealed that the microbial community structures of the Suining samples were more similar to each other while the microbial community structure of Yibin Daqu was more similar to Luzhou Daqu.
In this study, we analyzed the microbial community structure of Luzhou-flavor liquor Daqu samples collected from different regions of Sichuan province by high throughput sequencing technology. The results showed that the dominant bacteria in the Daqu were Staphylococcus, Weissella, Bacillus, Lactobacillus, Kroppenstedtia,Thermoactinomyces, Saccharopolyspora, and Brevibacterium. The proportions of Thermoactinomyces and Weissella in Yibin Daqu and Luzhou Daqu, with higher temperature, were higher than those in two samples of Suining Daqu.The dominant strains in Suining Daqu were Staphylococcus and Bacillus, and Staphylococcus was much higher than in Yibin Daqu and Luzhou Daqu. On the other hand, the fungal communities in all four Daqu samples were predominated mainly by Thermomyces, Aspergillus, Thermoascus, Byssochlamys, and Wallemia. In Yibin and Luzhou samples, Thermomyces and Thermoascus were the dominant species, and their proportions were much higher than those in Suining Daqu. Aspergillus was the dominant species in Suining Daqu, and its proportion was much higher than those in Yibin Daqu and Luzhou Daqu. The diversities of bacteria and fungi in Suining Daqu were higher than in Yibin Daqu and Luzhou Daqu. Principal component analysis revealed that the microbial community structures of the Suining samples were more similar to each other while the microbial community structure of Yibin Daqu was more similar to Luzhou Daqu.
引文
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[17]WOLF M,MULLER T,DANDEKAR T,et al.Phylogeny of Firmicutes with special reference to mycoplasma(Mollicutes)as inferred from phosphoglycerate kinase amino acid sequence data[J].International Journal of Systematic&Evolutionary Microbiology,2004,54(3):871-875.DOI:10.1099/ijs.0.02868-0.
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[19]王彩虹.基于克隆文库法研究不同香型大曲微生物群落结构[D].自贡:四川理工学院,2014:27-28.
[20]郭新梅.葡萄球菌属的鉴定及分类方法研究[D].贵阳:贵州大学,2006:6-8.
[21]李中华,段雄波,李金钟.魏斯菌属细菌的分类与鉴定新进展[J].国际检验医学杂志,2006,27(8):721-723.DOI:10.3969/j.issn.1673-4130.2006.08.019.
[22]王海燕,张晓君,徐岩,等.浓香型和芝麻香型白酒酒醅中微生物菌群的研究[J].酿酒科技,2008(2):86-89.DOI:10.13746/j.njkj.2008.02.020.
[23]李德林.浓香型白酒酒醅微生物分子生物学研究[D].自贡:四川理工学院,2013:43-44.
[24]罗俊成.浓香型白酒糟醅中芽孢杆菌属细菌的分类鉴定和16SrDNA序列系统发育分析[D].成都:四川大学,2007:21-24.
[25]职晓阳,蔡曼,杨玲玲,等.建立放线菌门的证据[J].微生物学通报,2006,33(3):181-183.DOI:10.13344/j.microbiol.china.2006.03.039.
[26]程古月.高温放线菌CDF芽胞表面蛋白酶及胞外丝氨酸蛋白酶的性质和功能研究[D].武汉:武汉大学,2010:4-7.
[27]叶光斌,董瑞丽,王彩虹,等.基于免培养法的中高温大曲细菌群落结构研究[J].江苏农业科学,2013,41(8):333-336.DOI:10.15889/j.issn.1002-1302.2013.08.005.
[28]孟镇,熊正河,钟其顶,等.应用P C R-D G G E技术解析白酒大曲细菌群落结构[J].食品与发酵工业,2010,36(10):159-162.DOI:10.13995/j.cnki.11-1802/ts.2010.10.031.
[29]李大和,李国红.浓香型曲酒酿造生产工艺要素:二[J].食品与发酵科技,2010,37(4):102-105.
[30]刑来君,李明春.普通真菌学[M].北京:高等教育出版社,1999:83-101.
[31]罗惠波,杨晓东,杨跃寰,等.浓香型大曲中可培养真菌的分离鉴定与系统发育学分析[J].现代食品科技,2013,29(9):2047-2052.DOI:10.13982/j.mfst.1673-9078.2013.09.028.
[32]赵春青,李多川.嗜热子囊菌两个中国新记录种[J].菌物研究,2005,3(1):27-29.DOI:10.13341/j.jfr.2005.01.006.
[33]王彩虹,罗惠波,卓毓崇,等.基于ITS基因文库法研究中高温大曲和超高温大曲真菌群落结构[J].食品科技,2014,39(7):41-46.DOI:10.13684/j.cnki.spkj.2014.07.009.
[34]董瑞丽,罗惠波,叶光斌.浓香型大曲中的嗜热真菌[J].中国酿造,2011,30(4):75-77.DOI:10.3969/j.issn.0254-5071.2011.04.022.
[2]沈怡方.白酒生产技术全书[M].北京:中国轻工业出版社,1998:54-75.
[3]张会敏,王杰,何宏魁,等.利用非培养技术初步解析古井贡酒大曲真核微生物群落结构[J].食品工业,2014(6):158-162.
[4]夏玙,罗惠波,周平,等.不同处理方式的大曲真菌群落差异分析[J].食品科学,2018,39(22):166-172.DOI:10.7506/spkx1002-6630-201822026.
[5]施思,彭智辅,乔宗伟,等.浓香型大曲贮藏过程中糖化力发酵力变化及真菌多样性分析[J].食品与发酵工业,2017,43(5):76-79.DOI:10.13995/j.cnki.11-1802/ts.201705013.
[6]LIPTHAY J R D,ENZINGER C,JOHNSEN K,et al.Impact of DNA extraction method on bacterial community composition measured by denaturing gradient gel electrophoresis[J].Soil Biology&Biochemistry,2004,36(10):1607-1614.DOI:10.1016/j.soilbio.2004.03.011.
[7]SHOKRALLA S,PORTER T M,GIBSON J F,et al.Massively parallel multiplex DNA sequencing for specimen identification using an Illumina MiSeq platform[J].Scientific Reports,2015,5:9687.DOI:10.1038/srep09687.
[8]LUO C,TSEMENTZI D,KYRPIDES N,et al.Direct comparisons of Illumina vs.Roche 454 sequencing technologies on the same microbial community DNA sample[J].PLoS ONE,2012,7(2):e30087.DOI:10.1371/journal.pone.0030087.
[9]QUAIL M A,MIRIAM S,PAUL C,et al.A tale of three next generation sequencing platforms:comparison of ion torrent,pacific biosciences and Illumina MiSeq sequencers[J].BMC Genomics,2012,13(1):341.DOI:10.1186/1471-2164-13-341.
[10]李晓娇.蜡蚧轮枝菌(Verticillum lecanii)对柑橘粉虱(Diaelorude citri)基因表达的影响[D].重庆:西南大学,2014:22-23.
[11]SCHMIDT T S,MATIAS RODRIGUES J F,VON M C.Limits to robustness and reproducibility in the demarcation of operational taxonomic units[J].Environmental Microbiology,2015,17(5):1689-1706.DOI:10.1111/1462-2920.12610.
[12]AMATO K R,YEOMAN C J,KENT A,et al.Habitat degradation impacts black howler monkey(Alouatta pigra)gastrointestinal microbiomes[J].ISME Journal,2013,7(7):1344-1353.DOI:10.1038/ismej.2013.16.
[13]WANG Y,SHENG H F,HE Y,et al.Comparison of the levels of bacterial diversity in freshwater,intertidal wetland,and marine sediments by using millions of Illumina tags[J].Applied&Environmental Microbiology,2012,78(23):8264-8271.DOI:10.1128/aem.01821-12.
[14]SCHLOSS P D,GEVERS D,WESTCOTT S L.Reducing the effects of PCR amplification and sequencing artifacts on 16S rRNA-based studies[J].PLoS ONE,2011,6(12):e27310.DOI:10.1371/journal.pone.0027310.
[15]MCCALL M N,KENT O A,YU J,et al.MicroRNA profiling of diverse endothelial cell types[J].BMC Medical Genomics,2011,4(1):78-78.DOI:10.1186/1755-8794-4-78.
[16]陈玲,袁玉菊,曾丽云,等.16S rDNA克隆文库法与高通量测序法在浓香型大曲微生物群落结构分析中的对比研究[J].酿酒科技,2015(12):33-36.DOI:10.13746/j.njkj.2015145.
[17]WOLF M,MULLER T,DANDEKAR T,et al.Phylogeny of Firmicutes with special reference to mycoplasma(Mollicutes)as inferred from phosphoglycerate kinase amino acid sequence data[J].International Journal of Systematic&Evolutionary Microbiology,2004,54(3):871-875.DOI:10.1099/ijs.0.02868-0.
[18]关统伟.塔里木盆地土壤放线菌区系及抗植物病原真菌的研究[D].乌鲁木齐:新疆农业大学,2007:2-3.
[19]王彩虹.基于克隆文库法研究不同香型大曲微生物群落结构[D].自贡:四川理工学院,2014:27-28.
[20]郭新梅.葡萄球菌属的鉴定及分类方法研究[D].贵阳:贵州大学,2006:6-8.
[21]李中华,段雄波,李金钟.魏斯菌属细菌的分类与鉴定新进展[J].国际检验医学杂志,2006,27(8):721-723.DOI:10.3969/j.issn.1673-4130.2006.08.019.
[22]王海燕,张晓君,徐岩,等.浓香型和芝麻香型白酒酒醅中微生物菌群的研究[J].酿酒科技,2008(2):86-89.DOI:10.13746/j.njkj.2008.02.020.
[23]李德林.浓香型白酒酒醅微生物分子生物学研究[D].自贡:四川理工学院,2013:43-44.
[24]罗俊成.浓香型白酒糟醅中芽孢杆菌属细菌的分类鉴定和16SrDNA序列系统发育分析[D].成都:四川大学,2007:21-24.
[25]职晓阳,蔡曼,杨玲玲,等.建立放线菌门的证据[J].微生物学通报,2006,33(3):181-183.DOI:10.13344/j.microbiol.china.2006.03.039.
[26]程古月.高温放线菌CDF芽胞表面蛋白酶及胞外丝氨酸蛋白酶的性质和功能研究[D].武汉:武汉大学,2010:4-7.
[27]叶光斌,董瑞丽,王彩虹,等.基于免培养法的中高温大曲细菌群落结构研究[J].江苏农业科学,2013,41(8):333-336.DOI:10.15889/j.issn.1002-1302.2013.08.005.
[28]孟镇,熊正河,钟其顶,等.应用P C R-D G G E技术解析白酒大曲细菌群落结构[J].食品与发酵工业,2010,36(10):159-162.DOI:10.13995/j.cnki.11-1802/ts.2010.10.031.
[29]李大和,李国红.浓香型曲酒酿造生产工艺要素:二[J].食品与发酵科技,2010,37(4):102-105.
[30]刑来君,李明春.普通真菌学[M].北京:高等教育出版社,1999:83-101.
[31]罗惠波,杨晓东,杨跃寰,等.浓香型大曲中可培养真菌的分离鉴定与系统发育学分析[J].现代食品科技,2013,29(9):2047-2052.DOI:10.13982/j.mfst.1673-9078.2013.09.028.
[32]赵春青,李多川.嗜热子囊菌两个中国新记录种[J].菌物研究,2005,3(1):27-29.DOI:10.13341/j.jfr.2005.01.006.
[33]王彩虹,罗惠波,卓毓崇,等.基于ITS基因文库法研究中高温大曲和超高温大曲真菌群落结构[J].食品科技,2014,39(7):41-46.DOI:10.13684/j.cnki.spkj.2014.07.009.
[34]董瑞丽,罗惠波,叶光斌.浓香型大曲中的嗜热真菌[J].中国酿造,2011,30(4):75-77.DOI:10.3969/j.issn.0254-5071.2011.04.022.