葡萄藤叶青贮中乳酸菌的分离与鉴定
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摘要
为了筛选出性状优良的乳酸菌菌株,本试验以实验室纯培养方式从葡萄藤叶自然发酵的青贮中分离鉴定乳酸菌。经过菌落形态、细胞形态、生理生化鉴定和16S r DNA基因序列以及系统发育树的分析,从葡萄藤叶青贮中分离出5株乳酸菌,其中菌株P-1. 2和P-2. 2为戊糖乳杆菌(Lactobacillus pentosus),菌株P-1. 7为戊糖片球菌(Pediococcus pentosaceus),菌株P-2. 1为粪肠球菌(Enterococcus faecium),菌株P-2. 3为短乳杆菌(Lactobacillus brevis)。测定产酸速率和生长速率,发现菌株P-2. 1的产酸能力和生长性能最好。
In order to screen out lactic acid bacteria with excellent characters,the laboratory pure culture method was used to isolate and identify the bacteria from the silage of grape vine leaves. Through the colony morphology,cell morphology,physiological and biochemical identification,16 S r DNA gene sequence analysis and phylogenetic tree analysis,five strains were isolated from the silage of grape vine leaves. Strain P-1. 2 and strain P-2. 2 were Lactobacillus pentose,strain P-1. 7 was Pediococcus pentosaceus,strain P-2. 1 was Enterococcus faecium and strain P-2. 3 was Lactobacillus brevis. By measuring the rate of acid production and growth rate,it was found that strain P-2. 1 had the best acid producing ability and growth performance.
In order to screen out lactic acid bacteria with excellent characters,the laboratory pure culture method was used to isolate and identify the bacteria from the silage of grape vine leaves. Through the colony morphology,cell morphology,physiological and biochemical identification,16 S r DNA gene sequence analysis and phylogenetic tree analysis,five strains were isolated from the silage of grape vine leaves. Strain P-1. 2 and strain P-2. 2 were Lactobacillus pentose,strain P-1. 7 was Pediococcus pentosaceus,strain P-2. 1 was Enterococcus faecium and strain P-2. 3 was Lactobacillus brevis. By measuring the rate of acid production and growth rate,it was found that strain P-2. 1 had the best acid producing ability and growth performance.
引文
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[2]Yang Z Z,Aikeber,Ding M,et al.The advantages of silage and silage making[J].Grass-feeding Livestock,2005(1):57-58.杨志忠,艾克拜尔,丁敏,等.青贮饲料的优点及制作技术[J].草食家畜,2005(1):57-58.
[3]Zhou D B.Research,development and actuality for corn stover silage[J].Amino Acids&Biotic resources,2004,26(2):32-34.周德宝.青贮饲料的研究、发展及现状[J].氨基酸和生物资源,2004,26(2):32-34.
[4]Pholsen S,Khota W,Pang H,et al.Characterization and application of lactic acid bacteria for tropical silage preparation[J].Anim Sci J,2016,87(10):1202-1211.
[5]Liu H L,Ta N,Gui R.Silage characteristics[J].Hubei Agricultural Sciences,2009,48(10):2584-2586.刘晗璐,塔娜,桂荣.青贮饲料的特性[J].湖北农业科学,2009,48(10):2584-2586.
[6]Lin Z.Technology and research situation of silage[J].Fujian Journal of Animal Husbandry and Veterinary Medicine,2016,38(2):17-21.林芝.青贮技术及其研究状况[J].福建畜牧兽医,2016,38(2):17-21.
[7]Lin D W,Dong X Z.Classification and identification of lactic acid bacteria and method[M].Beijing:China Light Industry Press,1999:117-127.凌代文,东秀珠.乳酸细菌分类鉴定及实验方法[M].北京:中国轻工业出版社,1999:117-127.
[8]Yin X,Guo X F,Patiguli M,et al.Isolation and identification of Aspergillus flavus inhibiting lactic acid bacteria[J],Journal of Northwest A&F University(Nat.Sci.Ed.),2018,46(5):1-6.尹雪,郭雪峰,帕提古丽·毛拉红,等.抑制黄曲霉菌乳酸菌的分离及鉴定[J].西北农林科技大学学报(自然科学版),2018,46(5):1-6.
[9]Liu H.Modern food microbiology experiment technology[M].Beijing:China Light Industry Press,2011:260-284.刘慧.现代食品微生物学实验技术[M].北京:中国轻工业出版社,2011:260-284.
[10]Dong X Z,Cai M Y.Common bacteria manual system identification[M].Beijing:Science Press,2001:364-370.东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001:364-370.
[11]Zhang G.Lactic acid bacteria-based technology and application[M].Beijing:Chemical Industry Press,2007:40-43.张刚.乳酸细菌--基础、技术和应用[M].北京:化学工业出版社,2007:40-43.
[12]Li D X,Ni K K,Pang H L,et al.Identification and antimicrobial activity detection of lactic acid bacteria isolated from corn stover silage[J].Asian-Australas J Anim Sci,2015,28(5):620-631.
[13]Yin X,Guo X F,Patiguli M,et al.Isolation and identification of lactic acid bacteria from Chinese traditional sourdough[J].Science and Tenchnology of Food Industry,2017,38(14):141-145.尹雪,郭雪峰,帕提古丽·毛拉红,等.传统发面面肥中乳酸菌的分离与鉴定[J].食品工业科技,2017,38(14):141-145.
[14]Liu F.Characterization and identification of lactic acid bacteria species isolated from silage and their application for silage preparation[D].Harbin:Northeast Agricultural University,2005.刘飞.青贮饲料中优良乳酸菌的分离鉴定及其应用[D].哈尔滨:东北农业大学,2005.
[15]Yang J S,Tan H S,Cai Y M.Characteristics of lactic acid bacteria isolates and their effect on silage fermentation of fruit residues[J].J Dairy Sci,2016,99(7):5325-5334.
[16]Yin X,Guo X F,Liu J F,et al.Isolation and identification of lactic acid bacteria from Halostachys caspica silage[J].Scientia Agricultura Sinica,2018,51(14):2825-2834.尹雪,郭雪峰,刘俊峰,等.盐穗木青贮中乳酸菌的分离及筛选[J].中国农业科学,2018,51(14):2825-2834.