文献类型：期刊文献

英文题名：Specific and effective detection of anammox bacteria using PCR primers targeting the 16S rRNA gene and functional genes

作者：Yang, Yuchun[1,8];Li, Meng[2];Li, Hui[3];Li, Xiao-Yan[4];Lin, Jih-Gaw[5];Denecke, Martin[6];Gu, Ji-Dong[7,8]

机构：[1]Univ Hong Kong, Sch Biol Sci, Lab Environm Microbiol & Toxicol, Pokfulam Rd, Hong Kong, Peoples R China;[2]Shenzhen Univ, Shenzhen Key Lab Marine Microbiome Engn, Inst Adv Study, Shenzhen 518060, Peoples R China;[3]East China Univ Sci & Technol, Sch Resource & Environm Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[4]Univ Hong Kong, Dept Civil & Environm Engn, Pokfulam Rd, Hong Kong, Peoples R China;[5]Natl Chiao Tung Univ, Inst Environm Engn, 1001 Univ Rd, Hsinchu 30010, Taiwan;[6]Univ Duisburg Essen, Dept Urban Water & Waste Management, Univ Str 15, D-45141 Essen, Germany;[7]Guangdong Ind Polytech, Sch Food & Biotechnol, Guangzhou 510300, Guangdong, Peoples R China;[8]Guangdong Technion Israel Inst Technol, Environm Engn, 241 Daxue Rd, Shantou 515063, Guangdong, Peoples R China

年份：2020

卷号：734

外文期刊名：SCIENCE OF THE TOTAL ENVIRONMENT

收录：;EI(收录号：20202108701579);WOS:【SCI-EXPANDED(收录号:WOS:000540272000001)】；

基金：Wewould like to thank the anonymous reviewers for their constructive comments to improve the earlier version of this manuscript; and the language editing by Professor Leslie H.G. Morton to earlier versions of this manuscript. This study was supported by the Shenzhen Shenzhen Science and Technology Program (No. JCYJ20170818091727570), the CAS Interdisciplinary Innovation Team (No. JCTD-2018-16), and the Key Project of Department of Education of Guangdong Province (No. 2017KZDXM071) (ML); a Post-graduate Ph.D. Studentship (YY); and RGC GRF grant No. 701913 (J-DG), and TRS Project T21-711/16-R (X-YL).

语种：英文

外文关键词：Anammox bacteria; PCR primer; 16S rRNA gene; Functional gene; Efficiency; Specificity

摘要：Anaerobic ammonium-oxidizing (anammox) bacteria play an important role in the nitrogen cycle by coupling ammonium and nitrite to produce dinitrogen gas (N-2). Polymerase chain reaction (PCR) is a fast, simple, and sensitive method that is widely used to assess the diversity, abundance, and activity of the slow-growing bacteria. In this review, we summarize and evaluate the wide variety of PCR primers targeting the 16S rRNA gene and functional genes (hzo, nir, and hzs) of anammox bacteria for their effectiveness and efficiencies in detecting this group of bacteria in different sample types. Furthermore, the efficiencies of different universal high-throughput sequencing 16S rRNA gene primers in anammox bacteria investigations were also evaluated to provide a reference for primer selection. Based on our in silico evaluation results, none of the 16S rRNA gene primers could recover all of the known anammox bacteria, but multiple hzo and hzs gene primers could accomplish this task. However, uncertain copies (13 copies) of hzo genes were identified in the genomes, and the hydrazine oxidation reaction catalyzed by hydrazine oxidoreductases (HZOs) can also be catalyzed by other hydroxylamine oxidoreductases (HAOs) in anammox bacteria, which can potentially result in large deviations in hzo-based qPCR and RT-qPCR analyses and results. Therefore, the use of optimal primers targeting unique hzs genes are recommended, although the efficiencies of these newly designed primers need further verification in practical applications. This article provides comprehensive information for the effective and specific detection of anammox bacteria using specific primers targeting the 16S rRNA gene and functional genes and serves as a basis for future high-quality primer design.