基于微生物群落结构相关的窖泥品质理化指标的筛选
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摘要
为了解窖泥理化指标与微生物群落的相关性,筛选能反映窖泥品质的理化指标,通过高通量测序分析了不同品质窖泥微生物群落结构,并与理化指标进行了相关性分析。结果发现:水分质量分数、总氮和氨态氮含量随窖泥品质的提高而升高,pH值随窖泥品质的提高而接近于7;主成分分析表明氨态氮、总氮、有效磷含量、pH值、腐殖酸质量分数和铁元素含量对窖泥品质影响较大。总氮含量分别和厚壁菌门(Firmicutes)、互养菌门(Synergistetes)、放线菌(Actinobacteria)比例呈正相关,与绿弯菌门(Chlorflexi)、变形菌门(Proteobacteria)比例呈负相关。氨态氮含量分别和Firmicutes、Synergistetes、Actinobacteria比例呈正相关,和Proteobacteria、螺旋体门(Spirochaetes)比例呈负相关。有效磷含量分别和Synergistetes和Actinobacteria比例呈正相关。腐殖酸质量分数和Proteobacteria比例呈正相关。pH值分别和Firmicutes、Actinobacteria比例呈正相关,和Chlorflexi、Spirochaetes比例呈负相关。其相关性均达到了显著或极显著水平(P<0.05,P<0.01)。从冗余分析图中可以看出氨态氮、总氮、有效磷含量和pH值对微生物群落的影响最大,其次为水分质量分数、腐殖酸质量分数、钙和铁元素含量。综上所述,从水分质量分数、总氮、氨态氮、有效磷、腐殖酸质量分数、pH值、钙和铁元素含量中初步筛选到总氮、氨态氮含量和pH值3个指标,即在一定范围内,窖泥品质越好,总氮和氨态氮含量越高,pH值越接近于7,其可作为窖泥质量的初步评定标准。
In order to understand the correlation between physicochemical indexes and microbial communities of pit mud and further to screen out the physicochemical indexes that reflect the quality of pit mud, the microbial community structures in pit muds with different qualities were investigated by high-throughput sequencing, and correlation analysis was carried out between microbial community structure and physicochemical indexes. The results showed that the contents of water, total nitrogen and ammonium nitrogen showed an increasing trend with improved quality of pit mud, and the pH approached 7. Principal component analysis of physicochemical indicators showed that the contents of ammonium nitrogen, total nitrogen, available phosphorus, humic acid, pH and Fe had great influence on pit mud quality. Total nitrogen was positively correlated with Firmicutes, Synergistetes and Actinobacteria, but negatively correlated with Chlorflexi and Proteobacteria. Ammonium nitrogen content was positively correlated with Firmicutes, Synergistetes and Actinobacteria, but negatively correlated with Proteobacteria and Spirochaetes. There was a positive correlation between available phosphorus and Synergistetes or Actinobacteria. Humic acid and Proteobacteria were positively correlated. pH was positively correlated with Firmicutes and Proteobacteria, but negatively correlated Chlorflexi with Spirochaetes. All correlations were statistically significant(P < 0.05 or P<0.01). Redundancy analysis showed that ammonium nitrogen, total nitrogen, available phosphorus and pH had the greatest influence on microbial communities, followed by water, humic acid, calcium and iron contents. To sum up, total nitrogen, ammonium nitrogen and pH were screened out as quality indicators of pit mud. The better the quality of pit mud, the higher the contents of total nitrogen and ammonium nitrogen and the closer the pH was to 7.
In order to understand the correlation between physicochemical indexes and microbial communities of pit mud and further to screen out the physicochemical indexes that reflect the quality of pit mud, the microbial community structures in pit muds with different qualities were investigated by high-throughput sequencing, and correlation analysis was carried out between microbial community structure and physicochemical indexes. The results showed that the contents of water, total nitrogen and ammonium nitrogen showed an increasing trend with improved quality of pit mud, and the pH approached 7. Principal component analysis of physicochemical indicators showed that the contents of ammonium nitrogen, total nitrogen, available phosphorus, humic acid, pH and Fe had great influence on pit mud quality. Total nitrogen was positively correlated with Firmicutes, Synergistetes and Actinobacteria, but negatively correlated with Chlorflexi and Proteobacteria. Ammonium nitrogen content was positively correlated with Firmicutes, Synergistetes and Actinobacteria, but negatively correlated with Proteobacteria and Spirochaetes. There was a positive correlation between available phosphorus and Synergistetes or Actinobacteria. Humic acid and Proteobacteria were positively correlated. pH was positively correlated with Firmicutes and Proteobacteria, but negatively correlated Chlorflexi with Spirochaetes. All correlations were statistically significant(P < 0.05 or P<0.01). Redundancy analysis showed that ammonium nitrogen, total nitrogen, available phosphorus and pH had the greatest influence on microbial communities, followed by water, humic acid, calcium and iron contents. To sum up, total nitrogen, ammonium nitrogen and pH were screened out as quality indicators of pit mud. The better the quality of pit mud, the higher the contents of total nitrogen and ammonium nitrogen and the closer the pH was to 7.
引文
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[30]李大和,刘念,李国红.浓香型大曲酒酿造中酯化菌研究的现状与展望[J].酿酒科技,2008(2):92-98.DOI:10.13746/j.njkj.2008.02.022.
[31]WEIMER P J,STEVENSON D M.Isolation,characterization,and quantification of Clostridium kluyveri from the bovine rumen[J].Applied Microbiology&Biotechnology,2012,94(2):461-466.DOI:10.1007/s00253-011-3751-z.
[2]吴树坤,刘梅,邓杰,等.不同品质浓香型窖泥的酶活与微生物群落的相关性[J].食品与发酵工业,2018,44(1):19-24.DOI:10.13995/j.cnki.11-1802/ts.015418.
[3]钟姝霞,邓杰,卫春会,等.不同窖龄窖泥微生物群落结构与理化指标的相关分析[J].生物技术通报,2016,32(7):119-125.DOI:10.13560/j.cnki.biotech.bull.1985.2016.07.018.
[4]邓杰.基于高通量测序的浓香型白酒窖泥微生物群落结构研究[D].自贡:四川理工学院,2015:54-56.
[5]王福荣.酿酒分析与检测[M].北京:化学工业出版社,2012:25.
[6]刘晔,柳小明,胡兆初,等.ICP-MS测定地质样品中37个元素的准确度和长期稳定性分析[J].岩石学报,2007(5):1203-1210.DOI:10.3969/j.issn.1000-0569.2007.05.034.
[7]MARGULIES M,EGHOLM M,ALTMAN W E,et al.Genome sequencing in microfabricated high-density picolitre reactors[J].Nature,2005,437:376-380.DOI:10.1038/nature03959.
[8]XU C L,SUN R,QIAO X J,et al.Protective effect of glutamine on intestinal injury and bacterial community in rats exposed to hypobaric hypoxia environment[J].World Journal of Gastroenterology,2014,20(16):4662-4674.DOI:10.3748/wjg.v20.i16.4662.
[9]OBERAUNER L,ZACHOW C,LACKNER S,et al.The ignored diversity:complex bacterial communities in intensive care units revealed by 16S pyrosequencing[J].Scientific Reports,2013,3:1413.DOI:10.1038/srep01413.
[10]邓杰,卫春会,边名鸿,等.浓香型白酒不同窖龄窖池窖泥中古菌群落结构分析[J].食品科学,2017,38(8):37-42.DOI:10.7506/spkx 1002-6630-201708007.
[11]VENABLES W N,SMITH D M.An introduction to R:a programming environment for data analysis and graphics[J].Economic Journal,2009,401:650-652.
[12]WILLIAMS G J.Rattle:a data mining GUI for R[J].The R Journal,2009,1(2):45-55.
[13]闫朝晖.R软件在多元统计分析教学中的应用研究[J].科技创新导报,2011(1):157-158.DOI:10.16660/j.cnki.1674-098x.2011.01.122.
[14]韩光,张宿义,卢中明.窖泥微生物、窖泥酶活性与窖泥养分相关性研究[J].酿酒科技,2011(3):48-51;54.DOI:10.13746/j.njkj.2011.03.023.
[15]张强,沈才洪,刘清斌,等.窖泥质量评价研究进展[J].酿酒科技,2013(7):84-86.DOI:10.13746/j.njkj.2013.07.051.
[16]景晓卫,唐玉明,任道群,等.人工老窖发展及其研究现状[J].酿酒科技,2010(9):77-80.DOI:10.13746/j.njkj.2010.09.018.
[17]胡晓龙.浓香型.白酒窖泥中梭菌群落多样性与窖泥质量关联性研究[D].无锡:江南大学,2015:23-24.
[18]廖昶,吴生文,黄小晖,等.特香型酒功能窖泥和普通窖泥理化指标对比分析[J].酿酒科技,2010(2):86-90.DOI:10.13746/j.njkj.2010.02.021.
[19]景晓卫,唐玉明,任道群,等.人工老窖发展及其研究现状[J].酿酒科技,2010(9):77-80.DOI:10.13746/j.njkj.2010.09.018.
[20]张世伟.酿酒葡萄微生物群落多样性及其氮代谢通量研究[D].北京:中国矿业大学,2018:19.
[21]王春阳,周建斌,董燕婕,等.黄土区六种植物凋落物与不同形态氮素对土壤微生物量碳氮含量的影响[J].生态学报,2010,30(24):7092-7100.
[22]程伟.土壤有效磷含量对土壤微生物量及代谢活性的影响[D].长春:吉林农业大学,2013:22-25.
[23]宋海燕,尹友谊,宋建中.不同来源腐殖酸的化学组成与结构研究[J].华南师范大学学报(自然科学版),2009(1):61-66.
[24]陶勇,徐占成,刘孟华,等.剑南春窖泥理化特性及矿质元素变化趋势研究[J].酿酒科技,2011(7):51-54.DOI:10.13746/j.njkj.2011.07.029.
[25]张艳,刘弘,刘中利,等.诗仙太白窖泥矿质元素变化趋势研究[J].食品工业,2015,36(3):262-265.
[26]赵长青,杨秦欢,邓静,等.窖泥的评定指标检测[J].生物技术进展,2012,2(3):212-216.DOI:10.3969/j.issn.2095-2341.2012.03.10.
[27]PINDER R S,PATTERSON J A.Growth of acetogenic bacteria in response to varying pH,acetate or carbohydrate concentration.agriculture[J].Food and Analytical Bacteriology,2013(3):6-16.
[28]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.
[29]熊亚.泸州老窖不同窖龄窖泥中细菌及古菌种群多样性和系统发育研究[D].雅安:四川农业大学,2014:93-99.
[30]李大和,刘念,李国红.浓香型大曲酒酿造中酯化菌研究的现状与展望[J].酿酒科技,2008(2):92-98.DOI:10.13746/j.njkj.2008.02.022.
[31]WEIMER P J,STEVENSON D M.Isolation,characterization,and quantification of Clostridium kluyveri from the bovine rumen[J].Applied Microbiology&Biotechnology,2012,94(2):461-466.DOI:10.1007/s00253-011-3751-z.