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机构地区:[1]石河子大学生命科学学院,新疆石河子832003 [2]新疆大学生命科学与技术学院,乌鲁木齐830046 [3]新疆农业科学院微生物应用研究所,乌鲁木齐830091
出 处:《新疆农业科学》2012年第4期723-729,共7页Xinjiang Agricultural Sciences
基 金:新疆特殊环境微生物实验室开放课题(XJYS0203-2008-02)
摘 要:【目的】了解乌鲁木齐10号冷泉水中可培养细菌群落结构组成,以及嗜盐微生物的分离鉴定。【方法】梯度稀释乌鲁木齐10号冷泉水,采用TSA、R2A、0.1×LB、HM等4种培养基进行分离纯化。其中采用0.1%、0.5%、1%、2%、5%和10%NaCl的HM培养基分离嗜盐微生物。CTAB法提取所得菌株DNA,用细菌通用引物27F/1492R扩增16SrRNA基因,构建系统发育树。【结果】分离得到33株不同基因型细菌,16SrRNA测序、BLAST比对及系统发育分析将其归为假单胞菌属、金黄杆菌属、红杆菌属、芽孢杆菌属等,其中假单胞菌属占优势。R2A培养基分离效果较好,HM培养基在NaCl 5%~20%区间分离到的菌株数较多。【结论】乌鲁木齐10号冷泉水可培养细菌多样性较低,而中度嗜盐菌数量较丰富。【Objective】In order to investigate the composition of bacteria in a cold spring in Xinjiang faulting zone,separate and identify halophilic bacteria.【Method】Bacteria of the water of the No.10 cold spring were obtained by dilution plate and spread plate methods,and culture medium were TSA,R2A,0.1 ×LB,HM for separation and purification.Halophilic bacteria were isolated by HM medium containing 1%-10% sodium chloride.The DNA of different strains was extracted from the water of the No.10 cold spring by the CTAB method.The 16S rRNA genes were amplified from the total DNA by PCR with bacteria-specific primers.Phylogenetic trees based on 16S rRNA gene sequences were constructed by using the neighbour-joining.【Result】A total of 33 strains were obtained.Phylogenetic analysis of the bacterial 16S rRNA gene showed that all isolates fell into one of the following four bacterial lineages: Pseudomonas,Chryseobacterium,Erythrobacillus and Bacillus.The separation effect of R2A medium was the best,and the number of the isolated strains was abundant by HM medium of salt concentration range between 5% and 20%.【Conclusion】The bacterial population diversity was low,and the number of moderately halophilic bacteria was abundant in the Urumqi NO.10 spring water.
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