基于人工湿地微生物群落结构与功能分析的大气颗粒物生态效应研究

Recently, regional atmospheric Particulate contamination has become an important issue affecting environmental quality, people’s health and the sustainable economic and social development. As the most harmful pollutant to human health, it has attracted attentions of scholars worldwide. However, previous research mainly focuses on the health and environmental effects of atmospheric particulates, while the microbial ecological effects were largely ignored. In this study, we investigate the particulate matter effects to the microbial biomass and diversity of air, soil, and plant system in constructed wetlands using SEM, constructing 16S/18SrDNA gene library, next-generation sequencing technology and BIOLOG methods, comparing and analyzing microbial aerosol morphology, particle size distribution，biotoxicity, and structural and functional diversities of bioaerosols from aspects of morphology, gene, and overall metabolic levels, to determine the correlation between morphology, symbiotic relationship, diversity changes and atmospheric particulates. By using the RT-PCR method, the composition and biological activity of indicator microorganisms and pathogenic microorganisms and their correlation can be determined. Through laboratory simulation experiments, we study the effects of atmospheric particulates on soil, plant system microbial biomass, and microbial community and functional diversity. The microbial ecological effects of atmospheric particulates will be elucidated systematically. The study results should help to broaden and advance the research on environment safety of constructed wetlands.

大气颗粒物不仅使能见度降低，还可引发健康、环境、生态等深层次问题，但现有研究大多偏重其健康、环境效应，而忽略生态效应方面的研究。本研究从大气颗粒物对人工湿地空气、土壤、植物系统微生物群落结构与功能影响着手，采用SEM、构建16S/18S rDNA 基因文库、高通量测序和Biolog方法，从形态、基因和整体代谢水平，研究大气颗粒物对人工湿地空气微生物形貌、粒径分布、生物毒性、微生物群落结构与功能的影响，确定空气微生物在形态、共生关系和多样性上与大气颗粒物的相关性；采用RT-PCR技术，分析大气颗粒物对人工湿地空气微生物中指示与病原微生物数量、活性、分布规律的影响及相关性；研究大气颗粒物对土壤、植物系统群落结构及代谢功能多样性的影响。研究结果一方面可望解决大气颗粒物生态效应研究领域存在的诸多问题， 同时也可以进一步深化人工湿地环境卫生安全领域的研究内容。

大气颗粒物不仅使能见度降低，还可引发健康、环境、生态等深层次问题，但现有研究大多偏重其健康与环境效应，而忽略生态效应方面的研究。本项目从大气颗粒物对人工湿地空气、土壤、植物系统微生物群落结构与功能影响着手，在以下4个方面开展研究工作：（1）以自由表面流人工湿地为研究对象，采用KC-1000型大流量空气采样器采集TSP、PM10和PM2.5样品，分析TSP中细菌群落结构及多样性的月变化、PM10和PM2.5中细菌和真菌群落结构组成及多样性的季节变化、TSP、PM10和PM2.5中致病菌和条件致病菌丰度变化，确定其与环境因子的相关性。（2）采集青岛市市区街道和人工湿地大气颗粒物样品，选取细叶小羽藓（Haplocladium microphyllum）与大灰藓（Hypnum plumaeforme）作为研究对象，比较分析颗粒物（TSP、PM10和PM2.5）对苔藓生理及叶表面细菌群落结构的影响；采用气体交换和叶绿素荧光测定技术、Farquhar光合模型、叶肉细胞导度（gm）等方法，研究大气颗粒物处理下的细叶小羽藓和大灰藓光合特征、叶片内部氮分配、CO2叶肉细胞导度的适应性差异。（3）分别采用实时荧光定量PCR和限制性片段长度多态性分析（RFLP）方法，分析自由表面流人工湿地基质、根际土壤、污水、植物叶际和空气中马赛菌属（Massilia）数量和组成的月变化，确定马赛菌属的产生与传播过程。（4）采用SAS SUPER ISO 100空气浮游菌采样仪采集空气微生物样品，应用BIOLOG法对不同季节雾霾期与非雾霾期空气微生物群落多样性进行比较研究；采用安德森FA-I型六级空气微生物采样器采集微生物气溶胶样品，比较分析冬季雾霾期与非雾霾期生物气溶胶粒径变化。研究结果不仅明确了人工湿地中大气颗粒物与空气、土壤、植物系统微生物间的相互关系，同时也深化了人工湿地环境卫生安全领域的研究内容。