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Chinese Agricultural Science Bulletin ›› 2015, Vol. 31 ›› Issue (26): 60-65.doi: 10.11924/j.issn.1000-6850.casb15030114

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Isolation and Identification of a Nitrogen-fixing Bacillus Strain

Liu Yingying1,2, Wang Jiwen1,2, Yue Dandan1,2, Gong Tao1,2, Liu Li1,2, Li Guanjie1,2, Zhen Jing1,2, Zhao Junjie1,2, Yang Wenling1,2, Mu Qi2, Chen Guocan1,2   

  1. (1Key Laboratory of Microbial Engineering of Henan Province, Zhengzhou 450008; 2Limited Liability Company of Institute of Biology, Henan Academy of Science, Zhengzhou 450008)
  • Received:2015-03-13 Revised:2015-08-05 Accepted:2015-07-06 Online:2015-09-23 Published:2015-09-23

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Abstract: The purpose of this study was to obtain a strain of Bacillus with high nitrogen fixing activity isolated from soil and provide excellent strain for the development of nitrogen fixing microorganism fertilizer. Strains of Bacillus sp. which could grow on culture medium without nitrogen were isolated from wheat rhizosphere soil, and the Touch-Up PCR (increased PCR) method was used to amplify nifH structure gene. Strains with nifH structural gene and high nitrogen fixing efficiency were rescreened by total nitrogen colorimetry. Then morphological analysis, physiological and biochemical analysis, and 16S rDNA sequence analysis were conducted to identify the strains. The results showed that 5 strains of Bacillus sp. were screened on free nitrogen medium from wheat rhizosphere soil, from which a strain C2 with strong resistance, nifH structural gene and high nitrogen fixing efficiency was rescreened and identified as Bacillus megaterium.