高寒草甸魏斯氏乳酸菌的分离鉴定及理化特性研究

doi:10.11686/cyxb20140132

摘要/Abstract

摘要： 本试验对青藏高原藏北嵩草附着乳酸菌进行了分离培养,并利用传统培养法和16S rRNA序列分析对分离乳酸菌进行了鉴定,以期探讨高寒地区乳酸菌的发酵生物学特性并分离筛选优良发酵乳酸菌。试验共分离得到17株乳酸菌,其中有6株菌为食窦魏斯氏乳酸菌(Weissella confusa),其余均为融合魏斯氏乳酸菌(Weissella cibaria)。分离出的融合魏斯氏乳酸菌菌株均能在4℃条件下生长,其中有5个菌株能在pH值为3的培养基中生长,且分离的食窦魏斯氏乳酸菌均能利用半乳糖,10株融合魏斯氏菌均能够发酵利用阿拉伯糖。体现出高寒地区分离出的融合魏斯氏乳酸菌菌株具有耐低温,耐酸性强和发酵利用碳源广泛的特征。

Abstract: To isolate and screen optimal lactic acid bacteria (LAB) in alpine zones,conventional identification methods and 16S rRNA sequence analysis were used to identify isolated lactic acid bacteria attached to Kobresia littledalei in the Qinghai-Tibetan Plateau. A total number of 17 lactic acid bacterial strains were isolated in this study,of which 6 strains were identified as Weissella confusa and 11 strains as Weissella cibaria. The isolated strains grew well at 4℃. From 17 strains,five were able to grow at pH 3. With the exception of one strain,the other W. confusa strains could utilize arabinose for fermentation,and all W. cibaria could ferment galactose. The characteristics of Weissella in alpine zones included low temperature resistance,high acid resistance and utilization of a wide variety of carbon sources during fermentation.

中图分类号:

S812.3

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.

YANG Yang,SHI Chao,GUO Xu-sheng. Characterization and identification of Weissella species isolated from Kobresia littledalei growing in alpine meadows[J]. Acta Prataculturae Sinica, 2014, 23(1): 266-275.

参考文献

Reference:
[1]Shang Z H, Ji Q M, Duo J D Z, et al. Discussion on the development of crop-grass system in the “Three Rivers” region of Tibet[J]. Pratacultural Science, 2009, 26(8): 141-146.
[2]Zhao J X, Qi B, Duo J D Z, Effects of short-term enclose on the community characteristics of three types of degraded alpine grasslands in the north Tibet[J]. Pratacultural Science, 2011, 28(1): 59-62. 
[3]Li Y K, Ran F, Bao S K, et al. Physiological response of Kobresia humilis to nitrogenous fertilizer in an alpine meadow[J]. Acta Prataculturae Sinica, 2010, 19(3): 240-244. 
[4]Liu F, Yang L J, Hou J C, et al. Isolation and Identification of Lactic Acid Bacteria from Silage[J]. Jiangxi Feed, 2007, 1: 22-25.
[5]Chen M X, Liu Q H, Zhang J G. Characteristics of new forage rice and effects of additives on its silage quality[J]. Acta Prataculturae Sinica, 2011, 20(5): 201-206.
[6]Hong M, Diao Q Y, Jiang C G, et al. Review for effect of Lactobacillus buchneri on the silage[J]. Acta Prataculturae Sinica, 2011, 20(5): 266-271. 
[7]Chen Z L, Cheng C, Ma K, et al. Isolation and Identification of Lactic Acid Bacteria from Fermented Yak Milk Products in Tibet Area[J]. Food Science, 2008, 29(12): 408-412.
[8]Duan Y H, Tan Z F, Wang Y P, et al. Identification and characterization of lactic acid bacteria isolated from Tibetan Qula cheese[J]. Journal of General and Applied Microbiology, 2008, 54(1): 51-60. 
[9]Yu J, Sun Z H, Liu W J. Rapid identification of lactic acid bacteria isolated from home-made fermented milk in Tibet[J]. Journal of General and Applied Microbiology, 2009, 55(3): 181-190.
[10]Wu X H, Luo Z, Yu L, et al. A survey on composition and microbiota of fresh and fermented yak milk at different Tibetan altitudes[J]. Dairy Science & Technology, 2009, 89(2): 201-209. 
[11]Chen Y F, Sun T S, Wang J C, et al. Comparison of nutrition and microbiological compositions between two types of fermented milk from Tibet in China[J]. International Journal of Food Sciences and Nutrition, 2009, 60: 243-250. 
[12]Airidengcaicike, Chen X, Du X H. Isolation and identification of cultivable lactic acid bacteria in traditional fermented milk of Tibet in China[J]. International Journal of Dairy Technology, 2010, 63(3): 437-444. 
[13]Tan Z F, Pang H L, Duan Y H, et al. 16S ribosomal DNA analysis and characterization of lactic acid bacteria associated with traditional Tibetan Qula cheese made from yak milk[J]. Animal Science Journal, 2010, 81(6): 706-713. 
[14]Zhang D W. Isolation, identification of lactic acid bacteria and its application on silage for corn stalk[D]. Zhengzhou: Zhengzhou University, 2007.
[15]Liu H L. The effect of epiphytic lactic acid bacteria on gramineous grass silage fermentation characteristics and dairy cows performance[D]. Hohhot: Inner Mongolia Agricultural University, 2008.
[16]Zhao Z F. Isolation and seening of lctic aid bcteria fom crn slage and the efect of ading it on crn slage qality and arobic sability[D]. Hohhot: Inner Mongolia Agricultural University, 2009.
[17]Zhang L, De K J. Experiment of Quality Evaluation of Special Silage in Qinghai-Xizang Plateau[J]. Chinese Journal of Animal and Veterinary Sciences, 2007, 1: 15-16.
[18]Li M H. Research and application of lactic acid bacteria silage additive in southeastern Tibet[D]. Lhasa: Tibet University, 2009. 
[19]Chen Q G, Zhang P J, Wang C X. Tibet's Nagqu Grassland Animal Husbandry Resources[M]. Lanzhou: Gansu Science and Technology Press, 1991: 26-38.
[20]Wu H Y, Ma Y S, Dong Q M. Seasonal dynamics of aboveground biomass and nutrients of Kobresia tibetica meadow in Yellow River headwater area[J]. Pratacultural Science, 2009, 26(1): 8-12. 
[21]Ling D W. Identification of Lactic Acid Bacteria and Experimental Methods[M]. Beijing: China Light Industry Press, 1999: 1-25. 
[22]Yang J B, Guo X H, Ling D W. Lctic Acid Bacteria - Basic and Applied Biology[M]. Beijing: China Light Industry Press, 1991: 1-3.
[23]Dong X Z, Cai M Y. Common Bacterial Identification System Manual[M]. Beijing: China Light Industry Press, 2001.
[24]Li Z H, Duan X B. Weiss genus classification and identification of bacteria progress[J]. International Journal of Laboratory Medicine, 2006, 27(8): 721-723.
[25]Cai Y M, Yoshimi B, Masuhiro O, 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: 2982-2987. 
[26]Olano A, Chua J, Schroeder S, et al. Weissella confusa(basonym: Lactobacillus confusus) bacteremia: a case report[J]. Journal of Clinical Microbiol, 2001, 39(4): 1604-1607. 
[27]Flaherty J D, Levett P N, Dewhirst F E, et al. Fatal case of endocarditis due to Weissella confusa[J]. Journal of Clinical Microbiology, 2003, 41(5): 2237-2239.

参考文献:
[1]尚占环, 姬秋梅, 多吉顿珠, 等. 西藏“一江两河地”农区草业发展探讨[J]. 草业科学, 2009, 26(8): 141-146. 
[2]赵景学, 祁彪, 多吉顿珠，等. 短期围栏封育对藏北3类退化高寒草地群落特征的影响[J]. 草业科学, 2011, 28(1): 59-62. 
[3]李以康, 冉飞, 包苏科，等. 高寒草甸矮嵩草对氮肥的生理响应[J]. 草业学报, 2010, 19(3): 240-244. 
[4]刘飞, 杨丽杰, 侯俊财，等. 青贮饲料中的分离鉴定优良乳酸菌[J]. 江西饲料, 2007, 1: 22-25. 
[5]陈明霞, 刘秦华, 张建国. 饲料稻新材料的特性及添加物对其青贮品质的影响[J]. 草业学报, 2011, 20(5): 201-206. 
[6]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. 
[7]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. 
[8]Duan Y H, Tan Z F, Wang Y P,et al. Identification and characterization of lactic acid bacteria isolated from Tibetan Qula cheese[J]. Journal of General and Applied Microbiology, 2008, 54(1): 51-60. 
[9]Yu J, Sun Z H, Liu W J. Rapid identification of lactic acid bacteria isolated from home-made fermented milk in Tibet[J]. Journal of General and Applied Microbiology, 2009, 55(3): 181-190.
[10]Wu X H, Luo Z, Yu L,et al. A survey on composition and microbiota of fresh and fermented yak milk at different Tibetan altitudes[J]. Dairy Science & Technology, 2009, 89(2): 201-209. 
[11]Chen Y F, Sun T S, Wang J C,et al. Comparison of nutrition and microbiological compositions between two types of fermented milk from Tibet in China[J]. International Journal of Food Sciences and Nutrition, 2009, 60: 243-250. 
[12]Airidengcaicike, Chen X, Du X H. Isolation and identification of cultivable lactic acid bacteria in traditional fermented milk of Tibet in China[J]. International Journal of Dairy Technology, 2010, 63(3): 437-444. 
[13]Tan Z F, Pang H L, Duan Y H,et al. 16S ribosomal DNA analysis and characterization of lactic acid bacteria associated with traditional Tibetan Qula cheese made from yak milk[J]. Animal Science Journal, 2010, 81(6): 706-713. 
[14]张大伟. 乳酸菌的分离鉴定及其在玉米秸秆青贮中的应用[D]. 郑州: 郑州大学, 2007. 
[15]刘晗璐. 禾本科牧草乳酸菌发现及发酵制品检测与动物生产性能影响研究[D]. 呼和浩特: 内蒙古农业大学, 2008. 
[16]赵子夫. 乳酸菌的分离筛选及其对玉米青贮品质和有氧稳定性的影响[D]. 呼和浩特: 内蒙古农业大学, 2009. 
[17]张琳, 德科加. 青藏高原特种青贮牧草的品质评定试验[J]. 青海畜牧兽医, 2007, 1: 15-16. 
[18]李梦寒. 藏东南乳酸菌青贮添加剂的研究与应用[D]. 拉萨: 西藏大学, 2009. 
[19]陈全功, 张普金, 王春喜. 西藏那曲地区草地畜牧业资源[M]. 兰州: 甘肃科学技术出版社, 1991: 26-38. 
[20]吴海艳, 马玉寿, 董全民. 黄河源区藏嵩草沼泽化草甸地上生物量及营养季节动态研究[J]. 草业科学, 2009, 26(1): 8-12. 
[21]凌代文. 乳酸菌分类鉴定及实验方法[M]. 北京: 中国轻工业出版社, 1999: 1-25. 
[22]杨洁彬, 郭兴华, 凌代文. 乳酸菌—生物学基础及应用[M]. 北京: 中国轻工业出版社, 1991: 1-3. 
[23]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. 
[24]李中华, 段雄波. 魏斯菌属细菌的分类与鉴定新进展[J]. 国际检验医学杂志, 2006, 27(8): 721-723. 
[25]Cai Y M, Yoshimi B, Masuhiro O,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: 2982-2987. 
[26]Olano A, Chua J, Schroeder S,et al. Weissella confusa (basonym: Lactobacillus confusus) bacteremia: a case report[J]. Journal of Clinical Microbiol, 2001, 39(4): 1604-1607. 
[27]Flaherty J D, Levett P N, Dewhirst F E,et al. Fatal case of endocarditis due to Weissella confusa[J]. Journal of Clinical Microbiology, 2003, 41(5): 2237-2239.