高通量测序技术研究辽河真核浮游藻类的群落结构特征  

Community Structure Characteristics of Eukaryotic Planktonic Algae in Liaohe River Through High-throughput Sequencing

作　　者：王靖淇[1,2,3] 王书平[2,3] 张远[2,3] 林佳宁[2,3] 高欣[2,3] 臧小苗 赵茜[2,3] 

机构地区：[1]辽宁大学环境学院,沈阳110036 [2]中国环境科学研究院环境基准与风险评估国家重点实验室,北京100012 [3]中国环境科学研究院流域水生态保护技术研究室,北京100012

出　　处：《环境科学》2017年第4期1403-1413,共11页Environmental Science

基　　金：国家水体污染控制与治理科技重大专项(2012ZX07501-001-04)

摘　　要：真核浮游藻类在河流生态系统中发挥着重要生态功能,本研究对辽河水环境样本的18S rRNA基因的V4可变区进行454焦磷酸测序,对获得的OTU代表性序列进行物种注释.比较注释后的生物种类信息与光学显微镜观察结果的异同,并将OTUs的代表性序列与NCBI中真核浮游藻类的18S rRNA基因序列进行系统发育分析,此外研究了真核浮游藻类群落结构特征及其环境影响指标.研究共获得约167 901条18S rRNA基因V4可变区的高质量序列,注释为424个OTUs,涵盖了134种真核浮游藻类OTUs.与光学显微镜鉴别结果类似,高通量测序结果以硅藻门和绿藻门为优势类群,此外检测到轮藻门、隐藻门、定鞭藻门和金藻纲等显微镜观察结果中未有的类群.系统发育分析可在门这一分类阶元对隐藻门和甲藻门聚类,并将隐藻门在属水平进行聚类和区分,甲藻门可以在部分科或属水平聚类和区分.RDA结果显示,氨氮、活性磷和硝态氮是影响真核浮游藻类群落结构特征最重要的环境因子.本研究为认识辽河水环境中真核浮游藻类的多样性、群落结构及其环境影响因子提供了新的视角,同时也预示着高通量测序技术在辽河流域浮游植物监测和水环境质量指示中的应用潜力.Eukaryotic phytoplankton plays an important ecological function in river ecosystem.The 18 S rRNA gene V4 variable region of the environmental samples in Liaohe river was sequenced by using 454 pyrosequencing technology.The reprehensive sequences of OTUs were annotated,and then the OTUs list was compared to the species list obtained by microscope observation.The phylogenetic tree was constructed based on the reprehensive sequences of OTUs and 18 S rRNA genes of eukaryotic phytoplankton algae obtained from Gen Bank.The community structure characteristics of eukaryotic phytoplankton and its environmental impact indicators were further studied.In this study,we got 167 901 high-quality sequences of 18 S rRNA gene V4 region.These sequences were annotated to 424 OTUs,including 134 eukaryotic algae.The results of the high-throughput sequencing showed that bacillariophyta and chlorophyta were the dominant groups of the total phyla.The high-throughput sequencing also revealed the presence of Charophyta,Cryptophyta,Haptophyta and Chrysophyceae which were not observed by optical microscope.The phylogeny analysis clustered the Cryptophyta and Pyrroptata at phylum level.The different genera of Cryptophyta and part of families or genera of Pyrroptata were clustered and distinguished by phylogeny analysis.The results of RDA showed that NH＋4-N,PO34-P and NO-3were the most important environmental factors that affected the community structure.This study provided a new perspective to understand the diversity,community structure and influence factors of eukaryotic phytoplankton in Liaohe river.The study also indicated that high-throughput sequencing has great application potential in investigation of phytoplankton and the water environment quality assessment in Liaohe river.

关 键 词：18S RRNA 高通量测序 真核浮游藻类 多样性 群落结构 

分 类 号：X171[环境科学与工程—环境科学]