基于Illumina MiSeq高通量测序技术分析广西无盐发酵酸笋中细菌多样性
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摘要
以10年老酸水泡制酸笋样品(BSFAF)、发酵6个月块状酸笋(MBSF)和丝状酸笋样品(FBSF)为研究对象,应用Illumina Mi Seq高通量测序技术并通过生物信息统计与分析,探索广西无盐发酵酸笋样品中细菌菌群结构及多样性。结果表明:3组酸笋样品共得到148 656个有效的序列数(Tags),聚类后总共得到1 116个分类操作单元(OTUs)。根据物种注释结果,在3组酸笋样品中共注释到12个门、23个纲、37个目、98个科和190个属。在门分类水平上,3组样品中厚壁菌门(Firmicutes)占绝对优势,其次是变形杆菌门(Proteobacteria)和放线菌门(Actinobacteria);在属分类水平上,乳杆菌属(Lactobacillus)、乳球菌属(Lactococcus)和魏斯氏菌属(Weissella)是3组样品中的优势菌属。其中,丝状酸笋(FBSF)组的香农(Shannon)、辛普森(Simpson)及ACE指数均显著高于其他两组,且乳杆菌属(Lactobacillus)、乳球菌属(Lactococcus)和魏斯氏菌属(Weissella)的平均相对丰度总和最高,可达99.8%。因此,以泉水泡制发酵丝状竹笋生产酸笋可能更佳。
Using the 10-year-old mixture sample(BSFAF), 6-month-old massive(MBSF) and filamentous(FBSF) fermented sour bamboo shoot as research objects, the structure and diversity of bacterial flora in the sample of non-salt fermentation bamboo shoot in Guangxi were explored using Illumina Mi Seq high-throughput sequencing technology and biometric information. A total of 148 656 valid Tags were obtained from 3 groups of samples, and a total of 1 116 operational taxonomic units(OTUs) were obtained after clustering. According to the results of species annotation, 12 phyla,23 classes, 37 orders, 98 families and 190 genera were annotated in 3 groups of samples. At phylum level, Firmicutes was the dominant phylum, followed by Proteobacteria and Actinobacteria. At the genus classification level, the dominant bacteria in the three groups were Lactobacillus, Lactococcus and Weissella. Among them, the Shannon, Simpson and ACE indices of FBSF group were significantly higher than those of the other two groups,and the sum of average relative abundance of Lactobacillus, Lactococcus and Weissella was the highest, up to 99.8%. Therefore, the sour bamboo shoot obtained by fermenting filamentous bamboo shoots with spring water may be better.
Using the 10-year-old mixture sample(BSFAF), 6-month-old massive(MBSF) and filamentous(FBSF) fermented sour bamboo shoot as research objects, the structure and diversity of bacterial flora in the sample of non-salt fermentation bamboo shoot in Guangxi were explored using Illumina Mi Seq high-throughput sequencing technology and biometric information. A total of 148 656 valid Tags were obtained from 3 groups of samples, and a total of 1 116 operational taxonomic units(OTUs) were obtained after clustering. According to the results of species annotation, 12 phyla,23 classes, 37 orders, 98 families and 190 genera were annotated in 3 groups of samples. At phylum level, Firmicutes was the dominant phylum, followed by Proteobacteria and Actinobacteria. At the genus classification level, the dominant bacteria in the three groups were Lactobacillus, Lactococcus and Weissella. Among them, the Shannon, Simpson and ACE indices of FBSF group were significantly higher than those of the other two groups,and the sum of average relative abundance of Lactobacillus, Lactococcus and Weissella was the highest, up to 99.8%. Therefore, the sour bamboo shoot obtained by fermenting filamentous bamboo shoots with spring water may be better.
引文
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[2]杨乐,王洪新.不同可食性涂膜对方竹笋保鲜效果的影响[J].食品工业科技,2011,32(2):305-308.
[3]姚荷,谭兴和.竹笋加工方法研究进展[J].中国酿造,2017,36(11):24-27.
[4]李慧勤,彭见林,赵国华.竹笋加工中的营养成分变化及安全性研究进展[J].食品工业,2012(1):130-134.
[5]朱照华.酸笋的营养成分检测及其主要风味物质的研究[D].南宁:广西大学,2014.
[6]邱坚.傣味酸笋的加工工艺[J].林业科技,2003,28(3):56-57.
[7]解万翠,尹超,宋琳,等.中国传统发酵食品微生物多样性及其代谢研究进展[J].食品与发酵工业,2018,44(10):257-263.
[8]王越男,孙天松.不同发酵方式的益生菌酸奶代谢组学的应用及探究[J].现代食品科技,2017(4):88-94.
[9]SINGHAL P,SHUKLA L,SATYA S,et al.Scientific validation and process mechanism of traditional bamboo shoot fermentation by isolation and characterization of lactic acid[J].Curr Nutr Food Sci,2017,13(3):1-6.
[10]TAMANG B,TAMANG J P,SCHILLINGER U,et al.Phenotypic and genotypic identification of lactic acid bacteria isolated from ethnic fermented bamboo tender shoots of North East India[J].Intl J Food Microbiol,2008,121(1):35-40.
[11]THAKUR K,RAJANI C S,TOMAR S K,et al.Fermented bamboo shoots:a rich niche for bioprospecting lactic acid bacteria[J].J Bacteriol Mycol,2016,3(4):1034-1041.
[12]JEYARAM K,ROMI W,SINGH T A,et al.Bacterial species associated with traditional starter cultures used for fermented bamboo shoot production in Manipur state of India[J].Int J Food Microbiol,2010,143(1):1-8.
[13]洪秀荣.竹笋糟制工艺及其不同发酵时期微生物菌群研究[D].福州:福建农林科技大学,2014.
[14]夏雪娟,郑炯,叶秀娟,等.实时荧光定量PCR技术监测腌制麻竹笋中乳酸乳球菌动态变化[J].食品科学,2016,37(4):88-92.
[15]夏雪娟,李冠楠,叶秀娟,等.麻竹笋腌制过程中细菌群落动态变化分析[J].中国食品学报,2015,15(11):11-16.
[16]郑炯,夏雪娟,叶秀娟,等.PCR-DGGE技术分析腌制麻竹笋中微生物多样性[J].食品科学,2014,35(21):170-174.
[17]BLAXTER M,MANN J,CHAPMAN T,et al.Defining operational taxonomic units using DNA barcode data[J].Philosoph Transact Biol Sci,2005,360(1462):1935-1943.
[18]ROGNES T,FLOURI T,NICHOLS B,et al.VSEARCH:a versatile open source tool for metagenomics[J].Peer J,2016,4(10):1-28.
[19]CAPORASO JG,KUCZYNSKI J,STOMBAUGH J,et al.QIIME allows analysis of high-throughput community sequencing data[J].Nature,2010,7(5):335-336.
[20]COLE J R,CHAI B,FARRIS R J,et al.The Ribosomal Database Project(RDP-II):sequences and tools for high-throughput r RNA analysis[J].Nucl Acid Res,2005,33(Database issue):294-296.
[21]佟婷婷,田丰伟,王刚,等.基于宏基因组分析四川泡菜母水作引子的泡菜发酵过程中细菌多样性变化[J].食品工业科技,2015,36(21):173-177.
[22]LOZUPONE C A,HAMADY M,KELLEY S T,et al.Quantitative and qualitativeβdiversity measures lead to different insights into factors that structure microbial communities[J].Appl Environ Microbiol,2007,73(5):1576-1585.
[23]K簟LJALG U,LARSSON K H,ABARENKOV K,et al.UNITE:a database providing web-based methods for the molecular identification of ectomycorrhizal fungi[J].New Phytologist,2005,166(3):1063-1068.