兼性固CO_2、N_2菌BHJ的分子生物学鉴定及生长条件的确定  

Molecular Biological Identification and Determination of Growth Conditions of Facultative fixed Carbon and Nitrogen bacteria BHJ

作　　者：周盛[1,2] 王小敏[1] ZHOU Sheng;WANG Xiao-min(College of Biology ＆ Pharmacy, Yulin Normal University, Yulin, Guangxi 537000;Guangxi University of Science ＆ Technology, Eiuzhou, Guangxi 545000)

机构地区：[1]玉林师范学院,广西玉林537000 [2]广西科技大学,广西柳州546200

出　　处：《玉林师范学院学报》2017年第5期83-89,104,共8页Journal of Yulin Normal University

基　　金：广西自然科学基金资助项目(2014GXNSFAA118092);广西高校科研项目资助(KY2015YB172);广西科技大学博士科研启动经费资助项目(15Z02)

摘　　要："温室效应"日趋严重,生物固碳特别是微生物固碳将发挥独特的作用.CO_2、N_2是温室气体主要贡献者.目的:研究对一种命名为BHJ的兼性固CO_2、N_2菌(能同时固定CO_2和N_2)进行分子生物学鉴定及生长条件确定.方法:用细菌基因组DNA提取试剂盒提取兼性固CO_2、N_2菌BHJ总DNA,再用PCR试剂盒对其16Sr DNA进行扩增,最后把16SrDNA测序后序列在Geneank比对进行分子生物学鉴定;用无碳、氮培养基(配方:KH_2PO_40.2g,CaCl_20.2g,NaCl0.2g,MgSO_40.3g,VB0.0625g,微量元素溶液0.5ml,蒸馏水:1000ml)进行单一变量培养,通过分光光度计测其OD值变化确定兼性固CO_2、N_2菌BHJ的最佳温度、CO_2和N_2浓度比、pH.兼性固CO_2、N_2菌BHJ的16Sr DNA序列与在Genebank登记的Burkholderiacepacia的16SrDNA序列的同源性达到99%以上,结果:最佳生长温度为28℃,最佳生长CO_2和N_2浓度比为1:99,最佳生长pH7~9.Currently,greenhouse effect becomes more and more serious,and biological carbon-fixing,especially microbial biological carbon-fixing,can play a unique role in curbing the greenhouse effect.CO_2 and N_2 are major contributors to greenhouse gases. In this study, a facultative fixed Carbon and Nitrogen bacteria named BHJ（which can fix CO_2 and N_2 simultaneously） was identified by molecular biology and the growth conditions were determined. the facultative fixed Carbon and Nitrogen bacteria BHJ DNA total was extracted by DNA bacterial genomic DNA Extraction Kit, then PCR kit of 16S rDNA were amplified, Finally, the 16S rDNA sequence was identified by molecular biology at the Geneank alignment; Medium without carbon and nitrogen（KH2 PO4 0.2 g, CaCl_2 formula： 0.2 g, Na Cl 0.2 g, MgSO_4 0.3 g, VB 0.0625 g, trace element solution 0.5 ml, distilled water： 1000 ml） were cultured by a single variable. Spectrophotometer to measure the OD changes determine the optimum temperature, the concentration of CO_2 and N_2 ratio, p H of the facultative fixed Carbon and Nitrogen bacteria BHJ. The 16S rDNA sequence of the facultative fixed Carbon and Nitrogen bacteria BHJ was above 99% Homology with Burkholderia cepacia in Genebank; the optimum growth temperature was 28 ℃; the best growth of concentration ratio of CO_2 and N_2 was 1：99; the optimum growth was p H7-9.

关 键 词：兼性固CO2、N2菌 无碳氮培养基 分子生物学鉴定 最佳生长条件 

分 类 号：Q936[生物学—微生物学]