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中国农学通报 ›› 2015, Vol. 31 ›› Issue (26): 60-65.doi: 10.11924/j.issn.1000-6850.casb15030114

• 生物技术科学 • 上一篇    下一篇

一株固氮芽孢杆菌的分离与鉴定

刘莹莹1,2,王继雯1,2,岳丹丹1,2,巩 涛1,2,刘 莉1,2,李冠杰1,2,甄 静1,2,赵俊杰,杨文玲1,2,慕 琦2,陈国参1,2   

  1. (1河南省微生物工程重点实验室,郑州 450008;2河南科学院生物研究所有限责任公司,郑州 450008)
  • 收稿日期:2015-03-13 修回日期:2015-08-05 接受日期:2015-07-06 出版日期:2015-09-23 发布日期:2015-09-23
  • 通讯作者: 王继雯
  • 基金资助:
    2014年河南省转制机构研究发展专项“新型多功能微生物肥料的研制及应用”[豫财政(2014)383号];2012年河南省重大科技专项“新型复合微生物肥料关键技术研究及产业化”(121100110100);2013年转制专项基金项目“高效降解有机磷农药复合菌剂的技术研究”(132104810018);2013年省级科技基础平台建设专项经费项目“新型农业微生物菌剂的研制及应用”[豫财政(2013)435号]。

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

摘要: 为从土壤中分离得到高固氮效能的芽孢杆菌,为固氮微生物肥料的开发提供优良的菌株,从小麦的根际土壤中分离得到能在无氮培养基上生长的芽孢杆菌株,然后采用Touch-Up PCR(上升PCR)的方法从菌株中扩增固氮酶的nifH结构基因和用全氮比色法测定固氮效能的方法,复筛出具有nifH结构基因同时具有高固氮酶活的菌株,进一步从形态、生理生化、16S rDNA序列分析等方面对其进行鉴定。结果表明从小麦的根际土壤中初筛得到能在无氮培养基上生长的5株芽孢杆菌,然后复筛得到1株抗逆性强、具有nifH结构基因同时具有高固氮效能的菌株C2,经鉴定为巨大芽孢杆菌(Bacillus megaterium)。

关键词: 循环水养殖, 循环水养殖, 水质, 主成分分析

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.