基于Illumina HiSeq技术分析浆水中细菌多样性及理化性质
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摘要
基于Illumina HiSeq测序平台,对陕西、甘肃浆水中细菌16S rRNA基因V3～V4区进行测序,解析其细菌群落结构多样性。结果表明,各地浆水细菌菌群结构相似,多样性较高,其中,乳酸杆菌属丰度较高,占主导地位,发酵乳杆菌、乙醇片球菌和Lactobacillus parafarraginis为优势菌,相对丰度占比分别为33.39%、21.13%、3.28%。理化性质显示,浆水液盐度范围为0.09%～0.34%,pH值为3.32～4.00,酸度为0.194～0.872 g/L,亚硝酸盐含量为5.123～10.246 mg/kg,亚硝酸盐含量均低于我国酱腌菜卫生标准。浆水中乳酸含量最高,也含有甲酸、乙酸、丙酸、丁酸等多种短链脂肪酸,蔗糖、葡萄糖、果糖含量较少。本研究揭示陕西、甘肃不同地区浆水的细菌多样性及其理化性质,为传统发酵食品研究提供一定理论基础。
In this study, the bacterial community structure and diversity in Jiangshui samples from Shaanxi and Gansu provinces was analyzed through Illumina HiSeq sequencing of the V3-V4 region of the 16 S rRNA gene. The results showed that the bacterial community structure in all samples was similar with high diversity. The relative abundance of Lactobacillus was high and Lactobacillus, L. fermentum, Pediococcus ethanolidurans and L. parafarraginis were the dominant species,whose relative abundance was 33.39%, 21.13% and 3.28%, respectively.The physicochemical properties showed that salinity ranged from 0.09% to 0.34%, pH values from 3.32 to 4.00,acidity from 0.194 to 0.872 g/L and nitrite content from 5.123 to 10.246 mg/kg(below the maximum limit of the national standard for preserved vegetables). In all Jiangshui samples, the most abundant organic acid was lactic acid followed by short-chain fatty acids such as formic acid, acetic acid,propionic acid and butyric acid. Lesser amounts of sucrose, glucose and fructose were found in the samples. This study can provide a theoretical basis for research on traditional fermented foods.
In this study, the bacterial community structure and diversity in Jiangshui samples from Shaanxi and Gansu provinces was analyzed through Illumina HiSeq sequencing of the V3-V4 region of the 16 S rRNA gene. The results showed that the bacterial community structure in all samples was similar with high diversity. The relative abundance of Lactobacillus was high and Lactobacillus, L. fermentum, Pediococcus ethanolidurans and L. parafarraginis were the dominant species,whose relative abundance was 33.39%, 21.13% and 3.28%, respectively.The physicochemical properties showed that salinity ranged from 0.09% to 0.34%, pH values from 3.32 to 4.00,acidity from 0.194 to 0.872 g/L and nitrite content from 5.123 to 10.246 mg/kg(below the maximum limit of the national standard for preserved vegetables). In all Jiangshui samples, the most abundant organic acid was lactic acid followed by short-chain fatty acids such as formic acid, acetic acid,propionic acid and butyric acid. Lesser amounts of sucrose, glucose and fructose were found in the samples. This study can provide a theoretical basis for research on traditional fermented foods.
引文
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[19]张晓辉,杨靖鹏,王少军,等.浆水中细菌多样性分析及乳酸菌的分离鉴定[J].食品科学,2017,38(4):70-76.DOI:10.7506/spkx1002-6630-201704012.
[20]卫生和计划生育委员会,国家食品药品监督管理总局.食品中亚硝酸盐与硝酸盐的测定:GB/T 5009.33-2016[S].北京:中国标准出版社,2016.
[21]卫生和计划生育委员会.酱腌菜:GB 2714-2015[S].北京:中国标准出版社,2015.
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[23]CHAO A,SHEN T J.Nonparametric estimation of Shannon’s index of diversity when there are unseen species in sample[J].Environmental and Ecological Statistics,2003,10(4):429-443.DOI:10.1023/A:1026096204727.
[24]SIMPSON E H.Measure of diversity[J].Nature,1949(163):688.
[25]CHAO A.Non-parametric estimation of the classes in a population[J].Scandinavian Journal of Statistics,1984,11(4):265-270.
[26]LUNDBERG D S,YOURSTONE S,MIECZKOWSKI P,et al.Practical innovations for high-throughput amplicon sequencing[J].Nature Methods,2013,10(10):999-1002.DOI:10.1038/nmeth.2634.
[27]EDGAR R C.UPARSE:highly accurate OTU sequences from microbial amplicon reads[J].Nature Methods,2013,10(10):996-998.DOI:10.1038/nmeth.2604.
[28]盛海圆.传统泡菜中乳酸菌的分离鉴定及其多样性分析[D].合肥:安徽农业大学,2010.
[29]赵楠.四川泡菜的主要特性及其成因分析[D].无锡:江南大学,2017.
[30]YU J,GAO W,QING M,et al.Identification and characterization of lactic acid bacteria isolated from traditional pickles in Sichuan,China[J].Journal of General&Applied Microbiology,2012,58(3):163.DOI:10.2323/jgam.58.163.
[2]杜鹏,霍贵成.传统发酵食品及其营养保健功能[J].中国酿造,2004,132(6):6-8.DOI:10.13982/j.mfst.1673-9078.2004.01.039.
[3]张轶,王玉丽,陈晓前,等.传统发酵食品:桨水中微生物的分离与初步鉴定[J].食品科学,2007,28(1):219-222.
[4]侯智勇,黄文刚,孙晋康,等.川北地区浆水菜传统发酵工艺的初步研究[J].中国调味品,2015,40(7):100-103.DOI:10.3969/j.issn.1000-9973.2015.07.22.
[5]赵树欣.应重视对我国传统发酵食品的研究:兼论发酵食品中的功能成分[J].中国食物与营养,2004(1):28-30.
[6]李良凤.浆水中益生菌群的研究及浆水发酵工艺的研究[D].西安:陕西科技大学,2013.
[7]张丽珂,周鸽鸽,孟宪刚.传统发酵食品浆水中厌氧微生物分离鉴定初探[J].食品科技,2010,35(4):39-41.DOI:10.13684/j.cnki.spkj.2010.04.049.
[8]辛博.浆水接种发酵中亚硝酸盐的控制研究[D].西安:陕西科技大学,2014.
[9]何玲.浆水芹菜营养功能成分与亚硝酸盐变化规律及加工机理研究[D].杨凌:西北农林科技大学,2007.
[10]侯智勇,黄文刚,孙晋康,等.川北地区浆水菜传统发酵工艺的初步研究[J].中国调味品,2015,40(7):100-103.DOI:10.13684/j.cnki.spkj.2010.04.049.
[11]吕嘉枥,李良凤.传统浆水菜发酵工艺条件的研究[J].陕西科技大学学报(自然科学版),2013,31(3):89-92.
[12]李宏珍,贠建民,贾亚莉,等.浆水传统酿制过程中挥发性物质的动态变化[J].食品与机械,2016,32(6):29-33.DOI:10.13652/j.issn.1003-5788.2016.06.007.
[13]何玲,杨公明.浆水芹菜自然和接种发酵过程中有机酸的变化[J].北方园艺,2011(4):179-181.
[14]李雪萍.浆水中可培养微生物多样性及特性的研究[D].兰州:兰州交通大学,2014.
[15]LI R,ZHU H,RUAN J,et al.De novo assembly of human genomes with massively parallel short read sequencing[J].Genome Research,2010,20(2):265-272.DOI:10.1101/gr.097261.109.
[16]YANG F,ZENG X,NING K,et al.Saliva microbiomes distinguish caries:active from health human populations[J].The ISME Journal,2012,6(1):1-10.DOI:10.1038/ismej.2011.71.
[17]雷振河.采用高通量测序技术分析清香型白酒酿造微生物[J].食品与发酵工业,2015,41(9):164-167.DOI:10.13995/j.cnki.11-1802/ts.201509032.
[18]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(1):49-53.DOI:10.11882/j.issn.0254-5071.2018.01.011.
[19]张晓辉,杨靖鹏,王少军,等.浆水中细菌多样性分析及乳酸菌的分离鉴定[J].食品科学,2017,38(4):70-76.DOI:10.7506/spkx1002-6630-201704012.
[20]卫生和计划生育委员会,国家食品药品监督管理总局.食品中亚硝酸盐与硝酸盐的测定:GB/T 5009.33-2016[S].北京:中国标准出版社,2016.
[21]卫生和计划生育委员会.酱腌菜:GB 2714-2015[S].北京:中国标准出版社,2015.
[22]LI B,ZHANG X,GUO F,et al.Characterization of tetracycline resistant bacterial community in saline activated sludge using batch stress incubation with high-throughput sequencing analysis[J].Water Research,2013,47(13):4207-4216.DOI:10.1016/j.watres.2013.04.021.
[23]CHAO A,SHEN T J.Nonparametric estimation of Shannon’s index of diversity when there are unseen species in sample[J].Environmental and Ecological Statistics,2003,10(4):429-443.DOI:10.1023/A:1026096204727.
[24]SIMPSON E H.Measure of diversity[J].Nature,1949(163):688.
[25]CHAO A.Non-parametric estimation of the classes in a population[J].Scandinavian Journal of Statistics,1984,11(4):265-270.
[26]LUNDBERG D S,YOURSTONE S,MIECZKOWSKI P,et al.Practical innovations for high-throughput amplicon sequencing[J].Nature Methods,2013,10(10):999-1002.DOI:10.1038/nmeth.2634.
[27]EDGAR R C.UPARSE:highly accurate OTU sequences from microbial amplicon reads[J].Nature Methods,2013,10(10):996-998.DOI:10.1038/nmeth.2604.
[28]盛海圆.传统泡菜中乳酸菌的分离鉴定及其多样性分析[D].合肥:安徽农业大学,2010.
[29]赵楠.四川泡菜的主要特性及其成因分析[D].无锡:江南大学,2017.
[30]YU J,GAO W,QING M,et al.Identification and characterization of lactic acid bacteria isolated from traditional pickles in Sichuan,China[J].Journal of General&Applied Microbiology,2012,58(3):163.DOI:10.2323/jgam.58.163.
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