复合垂直流人工湿地基质理化性质对nirS和nirK型反硝化菌多样性的影响
Effects of Substrate Physical and Chemical Properties on the Diversity of nirS and nirK Denitrifying Bacteria in an Integrated Vertical-Flow Constructed Wetland
投稿时间:2022-02-01  修订日期:2022-03-29
DOI:10.15928/j.1674-3075.202202010030
中文关键词:nirS和nirK型  反硝化菌  Illumina高通量测序  冗余分析  复合垂直流人工湿地
英文关键词:nirS and nirK types  denitrifying bacteria  Illumina high-throughput sequencing  redundancy analysis  the integrated vertical-flow constructed wetland
基金项目:2018年贵州省科技支撑计划项目(黔科合支撑[2018]2807);2015年国家自然科学基金(31560107)
作者单位
赵 伟 贵州民族大学生态环境工程学院贵阳 550025国家民委喀斯特环境地质灾害防治重点实验室贵阳 550025 
朱四喜 贵州民族大学生态环境工程学院贵阳 550025国家民委喀斯特环境地质灾害防治重点实验室贵阳 550025 
夏国栋 贵州民族大学生态环境工程学院贵阳 550025国家民委喀斯特环境地质灾害防治重点实验室贵阳 550025 
高 宁 贵州民族大学生态环境工程学院贵阳 550025国家民委喀斯特环境地质灾害防治重点实验室贵阳 550025 
赵 斌 贵阳人文科技学院, 贵阳 550025 
王 众 国家海洋局秦皇岛海洋环境监测中心站河北 秦皇岛066000 
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中文摘要:
      深入研究复合垂直流人工湿地基质理化性质及其对相关脱氮反硝化微生物的影响,可为解析人工湿地污水处理及提高脱氮效率提供理论依据。于2020年12月采集贵州特地区复合垂直流人工湿地4个池子(一级垂直流下行池、一级垂直流上行池、二级垂直流下行池、二级垂直流上行池)的基质样品,用Illumina高通量测序技术获得反硝化功能基因序列,分析其物种组成和种群多样性,并采用冗余分析揭示反硝化菌群落与基质理化性质之间的相关性。结果表明,复合垂直流人工湿地中4个池子的基质理化性质存在显著差异。其中,pH在4个池子中均呈中性,溶解氧(DO)呈先下降后升高趋势,下行池的基质电导率(EC)均比上行池要高;总氮(TN)、总磷(TP)含量随着人工湿地梯级呈显著下降趋势,未出现厌氧现象;NO3--N和NH4+-N含量在二级垂直流下行池中大量积累并达到峰值。在多样性分析中,nirS、nirK型反硝化菌丰度随着人工湿地梯级呈先降低后升高的趋势,多样性随人工湿地级数逐渐降低,且下行池丰度多样性显著高于上行池。基质中nirS型反硝化菌共5门9属,nirK型为6门14属,变形菌门是nirS、nirK型反硝化菌群的共同优势菌门,相对丰度为55.36%~65.72%、30.06%~45.66%。由韦恩图可知其nirS、nirK型反硝化菌共有物种数分别为392、245,并与反硝化菌群OTUs数目趋势一致,从多角度表明nirS型反硝化菌群丰度明显高于nirK型,且共有物种占有率高达41.6%和41.7%,表明各级池中反硝化菌群相似度较高。冗余分析表明基质中nirS和nirK反硝化菌群的构建主要受pH、NH4+-N、DO、TP、TN影响,且nirS型对pH,以及nirK型对NH4+-N的响应更为强烈。
英文摘要:
      In this study, we explored the effects of substrate physical and chemical properties on the community diversity of nirS and nirK denitrifying bacteria in an integrated vertical-flow constructed wetland in the karst area of south China. In December 2020, substrate samples were collected in four pools (the first-level vertical flow descending pool, the first-level vertical flow ascending pool, the second-level vertical flow descending pool, and the second-level vertical flow ascending pool) of the integrated vertical-flow constructed wetland, and the?sequences?of?the nirS and nirK denitrifying functional?bacteria?were obtained by Illumina Qualcomm sequencing technique for species composition and community diversity analysis. The relationship between the denitrifying bacteria and the substrate physical and chemical properties was analyzed by redundancy analysis. Results show that there were significant differences in substrate physical and chemical properties of the four pools. Among them, pH was neutral in all four ponds, DO decreased initially and then increased, and substrate conductivity (EC) of the descending pool was higher than that of the ascending pool. The contents of TN and TP decreased significantly while cascading through the constructed wetlands, and there was no anaerobic activity. The contents of NO3--N and NH4+-N accumulated and peaked in the secondary vertical flow descending pool. The abundance of nirS and nirK denitrifying bacteria decreased initially and then increased in the four cascaded pools, while community diversity of the denitrifying bacteria gradually decreased. Diversity in the descending pools was significantly higher than in the ascending pools. The nirS and nirK denitrifying bacteria identified in the substrate included, respectively, 9 genera and 5 phyla, and 14 genera and 6 phyla. Proteus was the common dominant phylum of nirS and nirK denitrifying bacteria, with a relative abundance of 55.36%-65.72%, 30.06%-45.66%, respectively. The dominant bacteria genera were Rhodobacter, Dechloromonas, Rhodopseudomonas, Bradyrhizobium and Bosea. The Wayne diagram shows that the common OTUs of nirS and nirK denitrifying bacteria were 392 (41.6%) and 245 (41.7%), respectively, consistent with the total OTU composition of the denitrifying?bacteria, and the abundance of denitrifying bacteria of nirS was significantly higher than nirK, indicating that the similarity of denitrifying bacteria in all levels of pools was high. Redundancy analysis shows that the construction of nirS and nirK denitrifying bacteria in the substrate was primarily affected by pH, DO, TP, TN, NH4+-N, and the responses of nirS to pH and nirK to NH4+-N were stronger. Our study provides a theoretical basis for efficient nitrogen removal in integrated vertical flow constructed wetlands.
赵 伟,朱四喜,夏国栋,高 宁,赵 斌,王 众.2024.复合垂直流人工湿地基质理化性质对nirS和nirK型反硝化菌多样性的影响[J].水生态学杂志,45(2):132-140.
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