基于高通量测序的东湖蓝藻动态变化研究
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摘要
采用高通量测序(High-throughput sequencing technology)和实时荧光定量PCR技术,结合水质参数的检测,研究了东湖两个不同富营养化程度湖区蓝藻整年丰度和群落结构的变化,并统计分析蓝藻与环境参数之间的关系。结果表明:(1)两区域微囊藻在6~7月达到全年最高值,但拷贝数相差近20倍。(2)两区域蓝藻门在属水平的变化也有显著差异,H区蓝藻在4~11月主要由浮丝藻属(Planktothrix)(4~6月)、微囊藻属(Microcystis)(6~9月)、聚球藻属(Synechococcus sp. PS717)(7~11月)交替演变,L区蓝藻主要是由束丝藻属(Aphanizomenon)(4~6月)和聚球藻属(Synechococcus sp. PS717)(4~10月)组成,并且与H区在相对丰度变化上比较,L区较平稳。(3) RDA分析表明,Tw、p H、NH4-N是影响H区蓝藻属水平分类差异的主要因素,影响L区蓝藻属水平差异的主要因素是Tw、TP、p H。(4)与以往传统藻类计数相比,还发现了以往显微观察难以发现的聚球藻属和拟柱孢藻属等,且聚球藻属在东湖蓝藻门中占优势地位,应在以后的研究中得到更多的关注。
High-throughput sequencing,real-time quantitative PCR( Q-PCR) combined with the detection of water quality parameters were used to study the annual abundance and community structure of cyanobacteria in two different eutrophication regions in East Lake. The relationship between cyanobacteria and environmental parameters was analyzed statistically. The results showed that the maximum values of Microcystis in the two regions were from June to July,but their copies difference was nearly20 times. There was also a significant difference between the two regions Cyanophyta at the genus level. In the H region,the cyanobacteria were mainly Planktothrix( April to June),Microcystis( June to September),Synechococcus sp. PS717( July to November). In the L region,the cyanobacteria were mainly composed of Aphanizomenon( April to June) and Synechococcus sp.PS717( April to October),and the relative abundance in the L region was more stable than that in the H region. RDA analysis showed that Tw,p H and NH4-N were the main factors affecting the classification of cyanobacteria in the H region,and Tw,TP,p H were the main factors in the L region. In addition, compared with the traditional algae count,Synechococcus and Pseudomonas were found,which are difficult to be found by the previous microscopic observation. Synechococcus dominated in the cyanobacteria in East Lake,which should be paid more attention to in the future research.
High-throughput sequencing,real-time quantitative PCR( Q-PCR) combined with the detection of water quality parameters were used to study the annual abundance and community structure of cyanobacteria in two different eutrophication regions in East Lake. The relationship between cyanobacteria and environmental parameters was analyzed statistically. The results showed that the maximum values of Microcystis in the two regions were from June to July,but their copies difference was nearly20 times. There was also a significant difference between the two regions Cyanophyta at the genus level. In the H region,the cyanobacteria were mainly Planktothrix( April to June),Microcystis( June to September),Synechococcus sp. PS717( July to November). In the L region,the cyanobacteria were mainly composed of Aphanizomenon( April to June) and Synechococcus sp.PS717( April to October),and the relative abundance in the L region was more stable than that in the H region. RDA analysis showed that Tw,p H and NH4-N were the main factors affecting the classification of cyanobacteria in the H region,and Tw,TP,p H were the main factors in the L region. In addition, compared with the traditional algae count,Synechococcus and Pseudomonas were found,which are difficult to be found by the previous microscopic observation. Synechococcus dominated in the cyanobacteria in East Lake,which should be paid more attention to in the future research.
引文
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[2] O’Neil J M,Davis T W,Burford M A,et al. The rise of harmful cyanobacteria blooms:The potential roles of eutrophication and climate change[J]. Harmful Algae,2012,14(1):313-334.
[3]刘建康,谢平.揭开武汉东湖蓝藻水华消失之谜[J].长江流域资源与环境,1999(3):85-92.
[4] LEI A P,HU Z L,WANG J,et al. Structure of the phytoplankton community and its relationship to water quality in Donghu Lake,Wuhan,China[J]. Journal of Integrative Plant Biology,2005,47(1):27-37.
[5]余博识,吴忠兴,朱梦灵,等.水果湖湾蓝藻水华的形成及其对东湖影响的评价[J].水生生物学报,2008(2):286-289.
[6]叶艳婷,胡胜华,王燕燕,等.东湖主要湖区浮游植物群落结构特征及其与环境因子的关系[J].安徽农业科学,2011,39(23):14213-14216.
[7]裴国凤,胡江,王青,等.东湖围隔与主体湖区浮游植物群落结构的比较研究[J].中南民族大学学报:自然科学版,2016,35(3):46-51.
[8]李大命,孔繁翔,张民,等.太湖和巢湖夏季蓝藻水华期间产毒微囊藻和非产毒微囊藻种群丰度的空间分布[J].应用与环境生物学报,2011(4):480-485.
[9] Sthl-Delbanco A,Hansson L A,Gyllstr9m M. Recruitment of resting stages may induce blooms of Microcystis at low N:P ratios[J].Journal of Plankton Research,2003,25(9):1099-1106.
[10]刘建康,黄祥飞,林婉莲,等.东湖生态学研究[M].北京:科学出版社,1995.
[11]侯惠静,王素英,郭斌辉.基于DNA特征的蓝藻分类方法研究进展[J].食品研究与开发,2015(5):146-152.
[12] Zhang T,Fang H H P. Applications of real-time polymerase chain reaction for quantification of microorganisms in environmental samples[J]. Applied Microbiology and Biotechnology,2006,70(3):281-289.
[13] Gobet A,Quince C,Ramette A. Multivariate cutoff level analysis(Multi CoLA)of large community data sets[J]. Nucleic Acids Res,2010,38(15):65-73.
[14]王绍祥,杨洲祥,孙真,等.高通量测序技术在水环境微生物群落多样性中的应用[J].化学通报,2014(3):196-203.
[15] Xiao X,Sogge H,Lagesen K et al. Use of high throughput sequencing and light microscopy show contrasting results in a study of phytoplankton occurrence in a freshwater environment[J]. Plos One,2014,9(8):e106510.
[16] Pessi I S,Maalouf Pde C,Laughinghouse HDt,et al. On the use of high-throughput sequencing for the study of cyanobacterial diversity in Antarctic aquatic mats[J]. J Phycol,2016,52(3):356-368.
[17]成敏玲,刘陈飞,程丹逸,等.春夏季节东湖不同富营养化区域水体细菌群落的动态变化[J].华中农业大学学报,2017(5):52-59.
[18] Rinta-Kanto J M,Ouellette A J A,Boyer G L,et al. Quantification of toxic Microcystis spp. during the 2003 and 2004 blooms in western Lake Erie using quantitative real-time PCR[J]. Environmental Science&Technology,2005,39(11):4198-4205.
[19] Ma J,Qin B,Paerl H W,et al. The persistence of cyanobacterial(Microcystisspp.)blooms throughout winter in Lake Taihu,China[J].Limnology and Oceanography,2016,61:711-722.
[20] Liu X,Lu X H,Chen Y W. The effects of temperature and nutrient ratios on Microcystis blooms in Lake Taihu,China:An 11-year investigation[J]. Harmful Algae,2011(10):337-343.
[21]张亚,黄津辉,戚蓝,等.浅水富营养水库中藻类生物量与营养盐的关系[J].天津大学学报:自然科学与工程技术版,2014,36-41.
[22]潘畅,陈建湘,黄长红,等.洞庭湖区水环境现状调查与分析[J].人民长江,2018,49(8):20-24,48.
[23]冯丽君.太湖蓝藻多样性及群落结构时空动态研究[D].上海:华东理工大学,2016.
[24]田昌.洪泽湖浮游植物种群结构变化的水环境驱动因子分析[D].济南:山东大学,2015.
[25]秦伯强.长江中下游浅水湖泊富营养化发生机制与控制途径初探[J].湖泊科学,2002,14(3)193-202.
[26] Sundareshwar P V,Morris J T,Koepfler E K,et al. Phosphorus limitation of coastal ecosystem processes[J]. Science,2003(299):563-565.
[27] Xu D,Chen Y,Ding S,et al. Diffusive gradients in thin films technique equipped with a mixed binding gel for simultaneous measurements of dissolved reactive phosphorus and dissolved iron[J]. Environ Sci Technol,2013(47):10477-10484.
[28]龚梦丹,金增锋,王燕,等.长江中下游典型浅水湖泊沉积物水界面磷与铁的耦合关系[J].湖泊科学,2017,29(5),1103-1111.