绿汁发酵液中的微生物多样性及其对菌糠发酵品质的影响
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摘要
本试验旨在通过高通量测序技术,测定分析5种不同原料制备的绿汁发酵液的微生物分布情况,通过Alpha多样性、主成分分析等解明其微生物群落结构。同时,为探究其微生物与发酵品质的关联性,本试验调制了对照组(无添加)和添加2%绿汁发酵液的5个菌糠青贮处理组,每个处理5个重复。结果表明:5种材料制作的绿汁发酵液样本通过Illumina Miseq测序平台共获得548 266条高质量有效序列,经分类学鉴定分属23门,129属,其中丰度大于1%的有9属。聚类分析结果表明甘蔗梢、玉米秸秆和葛藤绿汁发酵液归为一类,红象草和菌糠绿汁发酵液各自单独一类。5种绿汁发酵液处理组的菌糠青贮料乳酸含量显著(P<0.05)升高,丁酸含量显著(P<0.05)降低,能够提高青贮料的发酵品质。其中,红象草绿汁发酵液处理菌糠的发酵效果最佳,原因是象草绿汁发酵液中与青贮发酵密切相关菌属的分布比例最高。
The objective of this experiment was to analyze microbial diversity of fermented juice using epiphytic lactic acid bacteria(FJLB).There were 5 kinds of FJLB in this experiment including corn stalk FJLB,red elephant grass FJLB,fungus chaff FJLB,kudzu FJLB and sugarcane top FJLB.Alpha diversity and principal component analysis were carried out to investigate the microbial diversity of 5 kinds of FJLB.In order to investigate the fermentation effect of 5 kinds of FJLB,the control group(without adding)and adding 5 kinds of 2% FJLB to fungus chaff silage treatments were made.Each treatment had 5 repeats.The results showed that a total of 548 266 high-quality effective sequences were obtained by Illumina Miseq sequencing platform,and totally 23 phyla,129 genera of bacteria were identified in samples of 5 kinds of FJLB.And there were 9 genera with abundance greater than 1%.Cluster analysis results showed that sugarcane FJLB,corn stalk FJLB and kudzu FJLB were classified into one group,red elephant grass FJLB and fungus chaff FJLB fell into one category separately.The lactic acid was significantly increased(P<0.05),the butyric was significantly decreased(P<0.05)when treated with 5 kinds of FJLB.What's more,the lowest pH value and the highest lactic acid content were achieved by treated with red elephant grass FJLB.By Miseq high-throughput sequencing technology,we found out that the highest proportion of genera of bacteria which were closely related to silage fermentation quality were red elephant grass FJLB.
The objective of this experiment was to analyze microbial diversity of fermented juice using epiphytic lactic acid bacteria(FJLB).There were 5 kinds of FJLB in this experiment including corn stalk FJLB,red elephant grass FJLB,fungus chaff FJLB,kudzu FJLB and sugarcane top FJLB.Alpha diversity and principal component analysis were carried out to investigate the microbial diversity of 5 kinds of FJLB.In order to investigate the fermentation effect of 5 kinds of FJLB,the control group(without adding)and adding 5 kinds of 2% FJLB to fungus chaff silage treatments were made.Each treatment had 5 repeats.The results showed that a total of 548 266 high-quality effective sequences were obtained by Illumina Miseq sequencing platform,and totally 23 phyla,129 genera of bacteria were identified in samples of 5 kinds of FJLB.And there were 9 genera with abundance greater than 1%.Cluster analysis results showed that sugarcane FJLB,corn stalk FJLB and kudzu FJLB were classified into one group,red elephant grass FJLB and fungus chaff FJLB fell into one category separately.The lactic acid was significantly increased(P<0.05),the butyric was significantly decreased(P<0.05)when treated with 5 kinds of FJLB.What's more,the lowest pH value and the highest lactic acid content were achieved by treated with red elephant grass FJLB.By Miseq high-throughput sequencing technology,we found out that the highest proportion of genera of bacteria which were closely related to silage fermentation quality were red elephant grass FJLB.
引文
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[15]张洁,原现军,郭刚,等.添加剂对西藏燕麦和箭筈豌豆混合青贮发酵品质的影响[J].草业学报,2014,23(5):359-364
[16]闫艳红,李君临,郭旭生,等.多花黑麦草与大豆秸秆混合青贮发酵品质的研究[J].草业学报,2014,23(4):94-99
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[20]杜淑清,李志.青贮饲料添加剂的种类和使用[J].兽药与饲料添加剂,2002,7(3):26-27
[21]陈鹏飞,白史且,杨富裕,等.添加剂和水分对光叶紫花苕青贮品质的影响[J].草业学报,2013,22(2):80-86
[22]郑丹,下条雅敬,邵涛.凋萎和添加绿汁发酵液对杂交狼尾草青贮发酵品质的影响[J].草地学报,2011,19(2):273-276
[23]Shao Tao,Zhang L,Shimojo M,et al.Fermentation quality of Italian Ryegrass(Lolium multiflorum Lam.)silages treated with encapsulated-glucose,glucose,sorbic acid and pre-fermented juices[J].Asian-Australasian Journal of Animal Sciences,2007,20(11):1699-1704
[24]许庆方,周禾,玉柱,等.贮藏期和添加绿汁发酵液对袋装苜蓿青贮的影响[J].草地学报,2006,14(2):129-146
[25]叶杭,贺宇,张国轶,等.绿汁发酵液和纤维素酶对香蕉叶青贮品质的影响[J].福建农业学报,2014,29(7):628-632
[26]Cai Y M,Benno Y,Ogawa M,et al.Influence of Lactobacillus spp.from an inoculant and of Weissella and Leuconostoc spp.from forage crops on silage fermentation[J].Applied and environmental microbiology,1998,64(8):2982-2987
[27]陈东科,孙长贵.实用临床微生物学检验与图谱[M].北京:人民卫生出版社,2011
[28]第二届微生物学名词审定委员会.微生物学名词[M].北京:科学出版社,2012
[2]朱玉环,廉美娜,郭旭生.藏嵩草绿汁发酵液提高苜蓿青贮发酵品质[J].农业工程学报,2013,29(5):199-206
[3]孙璀,王育群,许留兴,等.饲草青贮添加剂研究进展[J].贵州畜牧兽医,2014,38(6):15-19
[4]Naoki N,Senji U.Labotatory evaluation of previously fermented juice as a fermentation stimulant for Lucerne silage[J].Science of Food and Agriculture,1999,79(10):1285-1288
[5]Ohshima M,Ohshima E,Kimura H.Fermentation quality of alfalfa and Italian ryegrass silages treated with previously fermented juices prepared from both the herbages[J].Journal of Animal Feed Science and Technology,1997,66:41-44
[6]Lin C J,Bolsen K K,Brent B E,et al.Epiphytic lactic acid bacteria succession during the pre-ensiling and ensiling periods of alfalfa and maize[J].Journal of Applied Microbiology,2010,73(5):375-387
[7]Jiang Y,Ogunade I M,Qi S,et al.Effects of the dose and viability of Saccharomyces cerevisiae.Diversity of ruminal microbes as analyzed by Illumina MiSeq sequencing and quantitative PCR[J].Journal of Dairy Science,2017,100(1):325-342
[8]Bureenok S,Namihira T,Tamaki M,et al.Fermentative quality of gineagrass silage by using fermented juice of the epiphytic lactic acid bacteria(FJLB)as a silage additive[J].AsianAust.Journal of animal science,2005,18(6):807-811
[9]张丽英.饲料分析及饲料质量检测技术[M].北京:中国农业科技出版社,2003:46-75
[10]Playne M J,McDonald P.The buffering constituents of herbage and of silage[J].Journal of the Science of Food and Agriculture,1966,17(6):264-268
[11]Van Soest P J,Robertsom J B,Lewis B A.Methods for dietary fiber,neutral detergent fiber and non-starch polysaccharides in relation to animal nutrition[J].Journal of Dairy Science,1991,74(10):3583-3597
[12]Murphy R P.A method for the extraction of plant samples and the determination of total soluble carbohydrates[J].Journal of the Science of Food and Agriculture,1958,9(11):714-717
[13]Dennis K L,Wang Y W,Blatner N R,et al.Adenomatous polyps are driven by microbe-instigated focal inflammation and are controlled by IL-10-producing T cells[J].Cancer Research,2013,73(19):5905-5913
[14]李君风,孙肖慧,原现军,等.添加乙酸对西藏燕麦和紫花苜蓿混合青贮发酵品质和有氧稳定性的影响[J].草业学报,2014,23(5):271-278
[15]张洁,原现军,郭刚,等.添加剂对西藏燕麦和箭筈豌豆混合青贮发酵品质的影响[J].草业学报,2014,23(5):359-364
[16]闫艳红,李君临,郭旭生,等.多花黑麦草与大豆秸秆混合青贮发酵品质的研究[J].草业学报,2014,23(4):94-99
[17]PM.The biochemistry of silage[Z].Chalcombe publication:Cambrian Printer,199181-136
[18]Bureenok S,Namihira T,Kawamoto Y,et al.Additives effects of fermented juice of epiphytic lactic acid bacteria on the fermentative quality of guineagrass(Panium maximumJacq.)silage[J].Grassland Science,2005,51(3):243-248
[19]王建福,雷赵民,成述儒,等.添加乳酸菌制剂和麸皮对去穗玉米秸秆青贮质量的影响[J].草业学报,2018,27(4):162-169
[20]杜淑清,李志.青贮饲料添加剂的种类和使用[J].兽药与饲料添加剂,2002,7(3):26-27
[21]陈鹏飞,白史且,杨富裕,等.添加剂和水分对光叶紫花苕青贮品质的影响[J].草业学报,2013,22(2):80-86
[22]郑丹,下条雅敬,邵涛.凋萎和添加绿汁发酵液对杂交狼尾草青贮发酵品质的影响[J].草地学报,2011,19(2):273-276
[23]Shao Tao,Zhang L,Shimojo M,et al.Fermentation quality of Italian Ryegrass(Lolium multiflorum Lam.)silages treated with encapsulated-glucose,glucose,sorbic acid and pre-fermented juices[J].Asian-Australasian Journal of Animal Sciences,2007,20(11):1699-1704
[24]许庆方,周禾,玉柱,等.贮藏期和添加绿汁发酵液对袋装苜蓿青贮的影响[J].草地学报,2006,14(2):129-146
[25]叶杭,贺宇,张国轶,等.绿汁发酵液和纤维素酶对香蕉叶青贮品质的影响[J].福建农业学报,2014,29(7):628-632
[26]Cai Y M,Benno Y,Ogawa M,et al.Influence of Lactobacillus spp.from an inoculant and of Weissella and Leuconostoc spp.from forage crops on silage fermentation[J].Applied and environmental microbiology,1998,64(8):2982-2987
[27]陈东科,孙长贵.实用临床微生物学检验与图谱[M].北京:人民卫生出版社,2011
[28]第二届微生物学名词审定委员会.微生物学名词[M].北京:科学出版社,2012