青藏高原川西北高寒牧区黑麦青贮饲料中耐低温乳酸菌的分离与鉴定
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摘要
在低温条件下对青藏高原川西北高寒牧区黑麦青贮饲料中的乳酸菌进行了培养分离,用16S rDNA序列分析法对乳酸菌进行鉴定,得到植物乳杆菌、清酒乳杆菌、弯曲乳杆菌和屎肠球菌4个耐低温菌株;设置9个温度梯度培养4个菌株,研究其从低温到高温的生长活性。结果表明:4个乳酸菌株最适生长温度为20～37℃,在20℃以下低温范围培养也有较好的生长表现,均为耐低温菌株。
The objectives of this study were to identify lactic acid bacteria(LAB)from rye forage silages on the alpine pastoral area of northwestern Sichuan and screen the low temperature resistant LAB strains suitable for the preparation of silage in this area.LAB strains were cultured and isolated by low temperature conditions and identified with 16 SrDNA sequence analysis to obtain L.plantarum,L.sakei,L.curvatus and E.faecium.The four strains were cultured with nine temperature gradients to investigate their growth activity from low to high temperatures.The results showed that the optimal growth temperature of the four lactic acid strains ranged from 20℃ to 37℃.Below 20℃ low temperature range,the four strains had the better growth performance,which also showed that they were the low temperature resistant strains.
The objectives of this study were to identify lactic acid bacteria(LAB)from rye forage silages on the alpine pastoral area of northwestern Sichuan and screen the low temperature resistant LAB strains suitable for the preparation of silage in this area.LAB strains were cultured and isolated by low temperature conditions and identified with 16 SrDNA sequence analysis to obtain L.plantarum,L.sakei,L.curvatus and E.faecium.The four strains were cultured with nine temperature gradients to investigate their growth activity from low to high temperatures.The results showed that the optimal growth temperature of the four lactic acid strains ranged from 20℃ to 37℃.Below 20℃ low temperature range,the four strains had the better growth performance,which also showed that they were the low temperature resistant strains.
引文
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[18]尚天翠.温度及pH条件对乳酸菌生长影响的研究[J].伊犁师范学院学报(自然科学版),2011(3):32-36.
[19]王夫杰,鲁绯,渠岩,等.3株风味乳酸菌生长及产酸特性的研究[J].中国酿造,2010(9):58-61.
[2]李平,李世洪,沈益新,等.川西北高寒牧区青贮调制技术研究现状[J].草学,2017(6):4-11.
[3]洪梅,刁其玉,姜成钢,等.布氏乳杆菌对青贮发酵及其效果的研究进展[J].草业学报,2011,20(5):266-271.
[4]张丽霞,李英,毛中丽.川西北牧区应积极推广牧草捆裹青贮[J].草业与畜牧,2012(12):16.
[5]储徐建,李长慧,刘明灿,等.耐低温乳酸菌的分离与优化培养[J].草业科学,2014,31(7):1380-1388.
[6]陈芝兰,程池,马凯,等.西藏地区牦牛发酵乳制品中乳酸菌的分离与鉴定[J].食品科学,2008,29(12):408-412.
[7]杨杨,石超,郭旭生.高寒草甸魏斯氏乳酸菌的分离鉴定及理化特性研究[J].草业学报,2014,23(1):266-275.
[8]许冬梅,曹蕾,荆佩欣,等.青藏高原几种牧草附着耐低温乳酸菌的筛选[J].微生物学杂志,2017,37(3):87-94.
[9]崔棹茗,郭刚,原现军,等.青稞秸秆青贮饲料中优良乳酸菌的筛选及鉴定[J].草地学报,2015,23(3):608-615.
[10]李剑楠.青贮技术及其研究的进展[J].饲料广角,2014(24):30-34.
[11]赵士萍,周敏,蒋林树.青贮饲料添加剂的研究进展[J].中国农学通报,2016,32(20):6-10.
[12] Escalante A,Wacher C,Farres A.Lactic acid bacterial diversity in the traditional Mexican fermented dough pozol as determined by 16SrDNA sequence analysis[J].International Journal of Food Microbiology,2001,(64):21-31.
[13]王红梅,孙启忠,屠焰,等.呼伦贝尔草原野生牧草青贮中优良乳酸菌的分离及鉴定[J].草业学报,2016,25(8):189-196.
[14] Cai Y M.Identification and characterization of enterococcus species isolated from forage crops and their influence on silage fermentation[J].Journal of Dairy Science,1999,82:2466-2471.
[15] Pahlow G,Muck R E,Driehuis F,et al.Microbiology of ensiling[C]∥Buxton D R,Muck R E,Harrison J H.Silage Science and Technology.Madison:American Society of Agronomy,Crop Science Society of America,Soil Science Society of America,2003:38-41.
[16]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001:364-417.
[17] Woo P C,Lau S K,Teng J L,et al.Then and now:use of16SrDNA gene sequencing for bacterial identification and discovery of novel bacteria in clinical microbiology laboratories[J].Clin Microbiol Infect,2008,14(10):908934.
[18]尚天翠.温度及pH条件对乳酸菌生长影响的研究[J].伊犁师范学院学报(自然科学版),2011(3):32-36.
[19]王夫杰,鲁绯,渠岩,等.3株风味乳酸菌生长及产酸特性的研究[J].中国酿造,2010(9):58-61.