盐穗木根际耐盐碱细菌的分离筛选及鉴定
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摘要
将采集于新疆和田地区于田县盐碱地的2种盐穗木根际土壤样品制备成菌悬液,经适当稀释后,涂布于牛肉膏蛋白胨培养基中,经分离纯化获得20株菌株。经过分别或同时提高培养基的盐、碱浓度进行菌株耐盐碱性能的筛选,获得1株耐盐碱菌株D8,其在pH值12且盐(NaCl)浓度达到120 g/L的培养基中仍可生长,该菌株既属于极端嗜盐微生物又属于极端嗜碱微生物。通过对筛选出来的菌株进行革兰氏染色制片、生理生化检测、电镜观察、Biolog鉴定、16S rRNA基因测序及系统发育树的建立来鉴定种属关系。鉴定结果是该菌为太平洋盐单胞菌(Halomonas pacifica)。
Two kinds of rhizosphere soil were collected from Halostachys caspica in Yutian county of Hetian region in Xinjiang.The rhizosphere soil samples were prepared by appropriate dilution of the bacterial suspension and coating in beef extract peptone medium,20 strains were isolated.After respectively or simultaneously improving the salt and alkali concentration of the medium for screening the performance of salt tolerant strains,resistant strain D8 was obtained,which could survive in pH12 and salt concentration(NaCl) 120 g/L.The strain belongs to extreme halophilic and extreme alkalophiles microorganisms.Based on the selected strain's leather blue staining,detection,physiological and biochemical electron microscopy,Biolog identification,16 S rRNA gene sequencing and phylogenetic tree system to identify species,the identification result was the pathogen for the Halomonas pacifica.
Two kinds of rhizosphere soil were collected from Halostachys caspica in Yutian county of Hetian region in Xinjiang.The rhizosphere soil samples were prepared by appropriate dilution of the bacterial suspension and coating in beef extract peptone medium,20 strains were isolated.After respectively or simultaneously improving the salt and alkali concentration of the medium for screening the performance of salt tolerant strains,resistant strain D8 was obtained,which could survive in pH12 and salt concentration(NaCl) 120 g/L.The strain belongs to extreme halophilic and extreme alkalophiles microorganisms.Based on the selected strain's leather blue staining,detection,physiological and biochemical electron microscopy,Biolog identification,16 S rRNA gene sequencing and phylogenetic tree system to identify species,the identification result was the pathogen for the Halomonas pacifica.
引文
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[3]Sneath P H A,Mair N S,Sharpe M E,et al.Bergey's manual of systematic bacteriology.Volume 2.[M].Bergey's Manual of Systematic Bacteriology,1986.
[4]刘延吉,王靖,田晓艳,等.耐盐菌株的筛选及其促生作用研究[J].沈阳农业大学学报,2009,40(3):360-362.
[5]李彬,王志春,孙志高,等.中国盐碱地资源与可持续利用研究[J].干旱地区农业研究,2005,(2):154-158.
[6]张广志,周红姿,杨合同,等.碱土壤中耐盐细菌的分离与鉴定[J].山东农业科学,2008,(9):49-50,54.
[7]赵航,贾富强,张富春,等.盐胁迫下盐穗木差异表达基因的转录组信息分析[J].生物信息学,2014,(2):90-98.
[8]陆雅海,张福锁.根际微生物研究进展[J].土壤,2006,38(2):113-121.
[9]张为艳.新疆阿克苏盐矿嗜盐微生物多相分类和基因组研究[D].杭州:浙江大学,2015.
[10]沈萍,陈向东.微生物学实验(第四版)[M].北京:高等教育出版社,2007.111-116.
[11]车振明.微生物学实验[M].北京:科学出版社,2011.
[12]蔡妙英,东秀珠.常见细菌系统鉴定手册[M].北京:科学出版社,2001.
[13]郑华,欧阳志云,方治国,等.biolog在土壤微生物群落功能多样性研究中的应用[J].土壤学报,2004,41(3):456-461.
[14]孟素香,曹健.极端微生物对极端环境的适应机理及应用研究进展[J].现代农业科技,2014,(9):249-250.
[15]龙雯,陈存社.16S r RNA测序在细菌鉴定中的应用[J].北京工商大学学报(自然科学版),2006,24(5):10-12.
[16]冯思玲.系统发育树构建方法研究[J].信息技术,2009,(6):38-40.
[17]王建明,罗晓霞,贺江舟.塔里木盆地荒漠盐碱生境嗜盐碱细菌的初步研究[J].微生物学杂志,2009,29(1):79-86.