基于PCR-DGGE技术辅助筛选氨氮降解菌株

doi:10.3969/j.issn.1004-1524.2017.02.15

浙江农业学报 ›› 2017, Vol. 29 ›› Issue (2): 286-291.DOI: 10.3969/j.issn.1004-1524.2017.02.15

基于PCR-DGGE技术辅助筛选氨氮降解菌株

张智超¹, 孙宏², 吴逸飞², 王新², 姚晓红², 柳永², 汤江武^{2, *}, 葛向阳^{1, *}

1.华中农业大学微生物学国家重点实验室,湖北武汉 430030;
2.浙江省农业科学院植物保护与微生物研究所,浙江杭州 310021

收稿日期:2016-05-30 出版日期:2017-02-15 发布日期:2017-03-06
通讯作者: 葛向阳,E-mail:gxy@mail.hzau.edu.cn;汤江武,E-mail:tangjiangwu@gmail.com
作者简介:张智超(1989—),男,河南驻马店人,硕士研究生,从事环境污染处理方面的研究。E-mail:116113258@qq.com
基金资助:
浙江省重点研发计划项目(2015C03013); 浙江省农科院青年人才培养项目

PCR-DGGE assisted selection of ammonia degrading bacteria

ZHANG Zhichao¹, SUN Hong², WU Yifei², WANG Xin², YAO Xiaohong², LIU Yong², TANG Jiangwu^{2, *}, GE Xiangyang^{1, *}

1. State Key Laboratory of Microbiology, Huazhong Agricultural University, Wuhan 430030, China;
2. Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2016-05-30 Online:2017-02-15 Published:2017-03-06

摘要/Abstract

摘要： 将PCR-变性梯度凝胶电泳(DGGE)与传统梯度浓度富集手段相结合,从不同氨氮浓度养殖污水中筛选高活性氨氮降解菌株。结果表明,经过不同氨氮浓度的驯化,初筛得到22株菌株,分别测定其氨氮降解能力,最终得到3株氨氮降解能力较强的菌株,分别命名为ZZC-3、ZZC-4和ZZC-14。经16S rDNA鉴定后,分别为戈登氏菌属、反硝化菌属和红球菌属细菌。PCR-DGGE结果表明,反硝化杆菌属的微生物在富集液中的含量最高,因此选择ZZC-4为目标菌株。经过污水降解效果验证,添加ZZC-4菌株的处理在144 h后水中氨氮和化学需氧量降解率分别达到90.8%和94.7%,显示该菌株具有良好的应用开发前景。

关键词: PCR-DGGE, 氨氮降解, 反硝化菌属

Abstract: In the present study, strains able to efficiently degrade ammonia in livestock waste water were screened with traditional enrichment selection method combined with PCR-denaturing gradient gel electrophoresis (DGGE). Twenty-two strains were screened out after domestication under different ammonia concentrations. Among them, 3 strains exhibited relatively higher ammonia degradation abilities, and were named as ZZC-3, ZZC-4 and ZZC-14, respectively, which were identified as Gordonia sp., Denitrobacter sp. and Rhodococcus sp. by 16S rDNA sequence analysis. The PCR-DGGE results confirmed that the Denitrobacter sp. had the highest concentrations in the enriched solution of waste water. Therefore, strain ZZC-4 was selected as the target strain. After incubation for 144 h, the treatment with strain ZZC-4 reached the highest degradation efficiency, which was 90.8% for ammonia nitrogen and 94.7% for COD, respectively.

Key words: PCR-DGGE, ammonia degradation, Denitrobacter sp.

中图分类号:

X703

张智超, 孙宏, 吴逸飞, 王新, 姚晓红, 柳永, 汤江武, 葛向阳. 基于PCR-DGGE技术辅助筛选氨氮降解菌株[J]. 浙江农业学报, 2017, 29(2): 286-291.

ZHANG Zhichao, SUN Hong, WU Yifei, WANG Xin, YAO Xiaohong, LIU Yong, TANG Jiangwu, GE Xiangyang. PCR-DGGE assisted selection of ammonia degrading bacteria[J]. , 2017, 29(2): 286-291.

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基于PCR-DGGE技术辅助筛选氨氮降解菌株

PCR-DGGE assisted selection of ammonia degrading bacteria

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