广西师范大学学报（自然科学版） ›› 2015, Vol. 33 ›› Issue (4): 137-143.doi: 10.16088/j.issn.1001-6600.2015.04.022

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一株产酸克雷伯氏菌的分离鉴定及其药敏检测

江绍锋1,2, 李云飞1,2, 蓝运华1,2, 刘祎1,2, 陆祖军1,2,3   

  1. 1.广西师范大学生命科学学院,广西桂林541006;
    2.珍稀濒危动植物生态与环境保护省部共建教育部重点实验室,广西桂林541006;
    3.广西高校野生动植物生态学重点实验室,广西桂林541006
  • 收稿日期:2015-05-20 出版日期:2015-12-25 发布日期:2018-09-21
  • 通讯作者: 陆祖军(1963—),男,广西百色人,广西师范大学教授,博士。E-mail: luzujun2002@aliyun.com.cn
  • 基金资助:
    国家自然科学基金资助项目(31060120);广西壮族自治区自然科学基金资助项目(2013GXNSFAA019059);广西研究生教育创新计划项目基金资助项目(YCSZ2014090)

Isolation and Identification of a Nitrogen-fixing Bacteria Klebsiella oxytoca and Screening of Sensitive Antimicrobials

JIANG Shao-feng1,2, LI Yun-fei1,2, LAN Yun-hua1,2, LIU Yi1,2, LU Zu-jun1,2,3   

  1. 1.College of Life Science, Guangxi Normal University, Guilin Guangxi 541006, China;
    2. Key Laboratory of Ecologyof Rare and Endangered Species and Environmental Protection(Ministry of Education), Guangxi Normal University,Guilin Guangxi 541006, China;
    3. Key Ecological Laboratory of Wild Animal and Plant Species of Guangxi Collegesand Universities, Guilin Guangxi 541006, China
  • Received:2015-05-20 Online:2015-12-25 Published:2018-09-21

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摘要： 采集崇左市江门林场10年桉树林地土壤并从中分离自生固氮菌,通过常规细菌鉴定、16S rRNA基因可变区(V1~V3区)序列测序及Biolog自动微生物鉴定系统确定其种属,并利用药敏纸片琼脂扩散法(K-B法)测定其耐药性,结果表明:从所采土壤中分离得到1株自生固氮菌KO108,提交到Genbank获得登陆号为KM460926.1,与GenBank中产酸克雷伯氏菌基因序列相似性达到99%以上;Biolog自动微生物鉴定系统相似值分别为0.509和0.610,大于0.5,因此鉴定该菌株为革兰氏阴性产酸克雷伯氏菌Klebsiella oxytoca,属于肠杆菌科Enterbateriaceae克雷伯氏菌属Klebsiella;药敏实验结果显示该菌对呋喃妥因、链霉素、头孢曲松、卡那霉素、氧氟沙星、强力霉素、环丙沙星、氯霉素敏感;对复方新诺明中度敏感;对多粘菌素、青霉素G、四环素、红霉素耐药。本实验所获得菌株可作为基因改造的工程菌,为林业生产中的固氮微生物的改造和利用提供了实验依据。

关键词: 产酸克雷伯氏菌, 药敏检测, 分子鉴定, Biolog微生物鉴定

Abstract: The aim of this study was to isolate bacterial strains of azotobacter from the soil of Eucalyptus plantations with 10-year-old stands in Dongmen town, Fusui county, Chongzuo city, Guangxi Zhuang Autonomous Region, China, and to identify its genus by conventional methods, sequencing the Variable region(V1~V3) of gene of 16S rRNA and Biolog Automatic Microbial Identification System. The Variable region sequences of 16S rRNA gene was cloned by PCR method, KO108 is submitted to NCBI database with the accession number of KM460926.1,and the similarity values of 16S rRNA sequences between KO108 and Klebsiella oxytoca was above 99%. The similarity is greater than 0.5,which were 0.509 and 0.610 by using Biolog automatic microbial identification system respectively. The strain KO108 was indentified as Gram-negative acid-producing Klebsiella oxytoca belonging to the family of Enterbateriaceae genus Klebsiella. In addition, drug resistance of them was also detected by the disc agar diffusion test. The results showed that it was sensitive to nitrofurantoin, streptomycin, ceftriaxone, kanamycin, ofloxacin, doxycycline, ciprofloxacin, chloromycetin; and moderately sensitive to cotrimoxazole but resistant to polymyxin, penicillin G, tetracycline and erythromyc. The strain could be used as an object for genetic engineering, which laid the foundation for further study and transformation of nitrogen fixation microorganisms in forestry.

Key words: Klebsiella oxytoca, sensitive antimicrobials, molecular identification, Biolog system identi-fication

中图分类号: 

  • X53
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[1] 江绍锋, 黄金清, 于晓宇, 李云飞, 陆祖军. 产酸克雷伯氏菌基因组文库的构建及鉴定[J]. 广西师范大学学报（自然科学版）, 2016, 34(2): 151-157.
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