太湖蓝藻水华期间水体和底泥中产毒微囊藻与非产毒微囊藻种群丰度研究

运用实时荧光定量PCR(Quantitative real-time PCR,QPCR)技术研究了太湖蓝藻水华期间不同湖区水体和底泥中产毒微囊藻与总微囊藻种群丰度.结果表明,湖区间产毒微囊藻种群和总微囊藻种群丰度明显不同:在水体中,竺山湾(N5)和梅梁湾(N2)产毒微囊藻和总微囊藻种群丰度高于贡湖湾(N4)和湖心(S4...     

Effects of elevated CO2 on dynamics of microcystin-producing and non-microcystin-producing strains during Microcystis blooms

In an attempt to elucidate the effects of different CO₂ concentrations (270, 380, and 750 μL/L) on the competition of microcystin-producing (MC-producing) and non-MC-producing Microcystis strains during dense cyanobacteria blooms, an in situ simulation experimentwas conducted in the Meiliang Bay of Lake Taihu in the summer of 2012. The abundance of total Microcystis and MC-producing Microcystis genotypes was quantified based on the 16S rDNA and mcyD gene using real-time PCR. The results showed that atmospheric CO₂ elevation would significantly decrease the pH value and increase the dissolved inorganic carbon (DIC) concentration. Changes in CO₂ concentration did not show significant influence on the abundance of total Microcystis population. However, CO₂ concentrations may be an important factor in determining the subpopulation structure of Microcystis. The enhancement of CO₂ concentrations could largely increase the competitive ability of non-MC-producing over MC-producing Microcystis, resulting in a higher proportion of non-MC-producing subpopulation in treatments using high CO₂ concentrations. Concurrently, MC concentration in water declined when CO₂ concentrations were elevated. Therefore, we concluded that the increase of CO₂ concentrations might decrease potential health risks of MC for human and animals in the future.     

洪泽湖有毒和无毒微囊藻丰度及其与环境因子之间的相关分析

为了解洪泽湖有毒和无毒微囊藻丰度及其空间分布,于2014年8月对洪泽湖30个采样点水体的营养盐浓度和Calson富营养化指数(trophic state index,TSI)进行研究,同时采用实时荧光定量PCR技术测定了有毒和无毒微囊藻丰度.结果表明,洪泽湖水体的总氮和总磷浓度平均值分别为1.63 mg·L-1和0.11 mg·L-1,富营养化指数在58.1~73.6之间,洪泽湖水质呈富营养化状态;有毒微囊藻在洪泽湖广泛分布,其丰度在1.13×104~3.51×106copies·m L-1之间,总微囊藻丰度在1.06×105~1.10×107copies·m L-1之间,有毒微囊藻占总微囊藻的比例在8.5%~38.5%之间,平均值为23.6%,有毒微囊藻丰度及其比例均呈现出明显的空间差异性;相关分析结果显示,总微囊藻、有毒微囊藻和有毒微囊藻所占比例三者之间呈极显著正相关性(P0.01),总微囊藻和有毒微囊藻丰度与叶绿素a浓度和TSI有极显著正相关性(P0.01),与透明度有极显著负相关性(P0.01),有毒微囊藻所占比例与叶绿素a、总氮、总磷和TSI有极显著正相关性(P0.01),与TN/TP和透明度有极显著负相关(P0.01).因此,削减洪泽湖总氮、总磷浓度一方面可以降低水体富营养化水平,另一方面有利于抑制有毒微囊藻对无毒微囊藻的竞争优势.     

Spatial heterogeneity of cyanobacterial communities and genetic variation of Microcystis populations within large, shallow eutrophic lakes (Lake Taihu and Lake Chaohu, China)

Cyanobacteria, specifically Microcystis, usually form massive blooms in eutrophic freshwater lakes. Cyanobacterial samples were collected from eight sites of both Lake Taihu and Lake Chaohu in late summer to determine the diversity and distribution pattern of cyanobacteria and Microcystis in large, shallow, entropic lakes with significant spatial heterogeneity and long-term Microcystis bloom. Molecular methods based on denaturing gradient gel electrophoresis and clone library analysis were used. A similar heterogeneous distribution pattern of cyanobacteria in both lakes was observed. Most parts of these two lakes with high trophic level were dominated by Microcystis. However, in the regions with low trophic levels as well as low concentrations of chlorophyll a, Synechococcus occupied a considerable percentage. Different morphospecies and genotypes dominated the bloom-forming Microcystis populations in these two lakes. Microcystis viridis and Microcystis novacekii were dominant in Lake Chaohu, whereas Microcystis flos-aquae was dominant in Lake Taihu. Only 2 of the13 Microcystis operational taxonomic units were shared between these two lakes. Analysis of molecular variance based on 16S to 23S internal transcribed spacer sequences indicated the significant genetic differentiation of Microcystis between these two lakes (F_st = 0.19, p < 0.001). However, only 19.46% of the genetic variability was explained by the population variation between lakes, whereas most (80.54%) of the genetic variability occurred within the lakes. Phylogenetic analysis revealed no phylogeographic structure of Microcystis population in these two lakes, as illustrated by their cosmopolitan nature. Our results revealed that spatial heterogeneity within lakes has more impact on the cyanobacterial diversity than geographical isolation in a local scale.     

海河天津段微囊藻及其毒素的空间分布及与水环境因子的关系

采取荧光定量PCR技术及ELISA酶联免疫吸附法,以mcyD基因和PC-IGS基因为靶基因对夏季蓝藻暴发期间海河天津市区段的产毒微囊藻种群丰度和毒素含量进行了研究.结果表明：夏季海河天津市区段微囊藻种群丰度具有明显差异性：产毒微囊藻种群丰度为1.16×10⁴~2.48×10⁷copies/mL占总微囊藻种群的4.25%~28.59%.藻毒素含量最高点为8号采样点天津站,每升水体中藻细胞共含毒素195.51μg,除去藻细胞水中毒素浓度为0.97μg/L.总的来说,海河天津市区段微囊藻总基因拷贝数较高,产毒微囊藻丰度在不同采样点间差异较大,影响水体中微囊藻丰度的最主要环境因素是pH值.海河水体中毒素含量没有超过安全阈值,但是单位体积水体中藻细胞内毒素含量很高,具有较高的毒素释放潜力,可能对下游河口生态系统造成潜在威胁.     

Seasonal dynamics of water bloom-forming Microcystis morphospecies and the associated extracellular microcystin concentrations in large, shallow, eutrophic Dianchi Lake

The increasing occurrence of Microcystis blooms is of great concern to public health and ecosystem due to the potential hepatotoxic microcystins （MCs） produced by these colonial cyanobacteria. In order to interpret the relationships between variations of Microcyst/s morphospedes and extracellular MC concentrations, the seasonal dynamics of phytoplankton community composition, MC concentrations, and environmental parameters were monitored monthly from August, 2009 to July, 2010. The results indicated that Microcystis dominated total phytoplankton abundance from May to December （96%--99% of total biovolume）, with toxic Microcystis viridis and non-toxic Microcystis wesenbergii dominating after July （constituting 65%- 95% of the Microcystis population）, followed by M. viridis as the sole dominant species from November to January （49%--93%）. Correlation analysis revealed that water temperature and nutrient were the most important variables accounting for the occurrence ofM. wesenbergii, while the dominance ofM. viridis was related with nitrite and nitrate. The relatively low content of MCs was explained by the association with a large proportion of M. viriclis and M. wesenbergii, small colony size of Microcystis populations, and low water temperature, pH and dissolved oxygen. The extracellular MC （mean of 0.5 ± 0.2 μg/L） of water samples analyzed by enzyme-linked immunosorbent assay （ELISA） demonstrated the low concentrations of MC in Dianchi Lake which implied the low potential risk for human health in the basin. The survey provides the first whole lake study of the occurrence and seasonal variability of Microcyst/s population and     

A flow cytometer based protocol for quantitative analysis of bloom-forming cyanobacteria (Microcystis) in lake sediments

A quantitative protocol for the rapid analysis of Microcystis cells and colonies in lake sediment was developed using a modified flow cytometer, the CytoSense. For cell enumeration, diluted sediment samples containing Microcystis were processed with sonication to disintegrate colonies into single cells. An optimized procedure suggested that 5 mg dw (dry weight)/mL dilution combined with 200 W × 2 min sonication yielded the highest counting efficiency. Under the optimized determination conditions, the quantification limit of this protocol was 3.3×10⁴ cells/g dw. For colony analysis, Microcystis were isolated from the sediment by filtration. Colony lengths measured by flow cytometry were similar to those measured by microscopy for the size range of one single cell to almost 400 μupm in length. Moreover, the relationship between colony size and cell number was determined for three Microcystis species, including Microcystis flos-aquae, M. aeruginosa and M. wessenbergii. Regression formulas were used to calculate the cell numbers in different-sized colonies. The developed protocol was applied to field sediment samples from Lake Taihu. The results indicated the potential and applicability of flow cytometry as a tool for the rapid analysis of benthic Microcystis. This study provided a new capability for the high frequency monitoring of benthic overwintering and population dynamics of this bloom-forming cyanobacterium.     

太湖水体及表层沉积物磷空间分布特征及差异性分析

通过对水体不同程度富营养化湖泊——太湖全湖40个位点的高密度采样分析,得到太湖水体及表层沉积物各污染因子在太湖的空间分布特征图,结果表明,太湖水体中SRP、TP、TN及沉积物中TOC、TN、TP及P的各形态等在空间上表现出明显的分异性,水体中污染物主要分布于竺山湾、五里湖、梅梁湾及太湖西部等湖区,TN、TP最低值为0.05、0.88mg·L-1.沉积物中Fe-P的分布与水体中TP类似,含量在29.13～258.31mg·kg-1之间变化.Ca-P除主要分布于南部太湖及东太湖外,西北部湖区也见大量蓄积,最高值达357.68mg·kg-1.OP的高值分布于西北部湖区,最高值达371.91mg·kg-1.沉积物中IP占TP的含量高于OP,最高值高出OP含量约50%.IP中Fe-P的比例虽然低于Ca-P,但与水体中SRP、TP之间的高度相关性(R为0.49、0.64),指示Fe-P的内源释放为太湖水体中磷的重要来源之一.而沉积物中TOC与C/N、TN、TP及P的各形态之间的显著相关性,表明了高有机质含量更利于对营养盐的蓄积埋藏.太湖水体及表层沉积物各指标空间上表现出如此明显的区域性差异,除受不同湖区入湖污染源直接作用外,还和各参数不同的生物地球化学行为有关.     

Decomposition and phosphorus release from four different size fractions of Microcystis spp. taken from Lake Taihu, China

Decomposition of Microcystis is accompanied by the release of phosphorus,during bacteria play an important role.A series of experiments were undertaken to evaluate the effect of bacteria on the decomposition of Microcystis taken from Lake Taihu,China,a lake that is suffering from dense Microcystis blooms.The 16 experiments involved four size fractions of colonial Microcystis with or without the addition of lake sediment and Gram-negative bacterial inhibitor NAN_3.The highest decomposition rates were reco...     

太湖化学耗氧量和生化需氧量的时空分布特征

基于2009年2月(冬季)、5月(春季)、8月(夏季)、11月(秋季)全太湖32个站点的化学耗氧量(COD)、生化需氧量(BOD)、溶解性总氮(DTN)、溶解性总磷(DTP)、浮游植物色素数据,分析了太湖COD、BOD的时空分布特征,并探讨了影响COD、BOD时空分布的因素.结果表明,2009年太湖COD值为3.40～6.16mg·L-1,平均值为(4.38±0.72)mg·L-1;BOD为0.64～5.93mg·L-1,平均值为(1.91±1.63)mg·L-1.COD值在秋冬季节较高,其值显著大于春夏季(p0.001);而BOD值冬季较高,其它季节都较低.COD与BOD的空间分布格局相似,呈现出自竺山湾、太湖西北沿岸区向梅梁湾区、湖心区、东南湖湾区依次递减的趋势.分类统计结果显示,河口沿岸区COD、BOD值显著高于开敞水域区(p0.001).相关性分析结果表明,COD、BOD与DTN、DTP、浮游植物色素的相关性存在季节性差异.夏季浮游植物色素浓度与COD、BOD相关系数最高,说明夏季浮游植物降解对水体内COD和BOD的贡献要高于其它季节.COD与BOD的时空分布特征主要受降雨量、河流输入、"引江济太"及太湖浮游植物活动等的影响.     

Variation of cyanobacteria with different environmental conditions in Nansi Lake, China

Nansi Lake is located on the east line of the South-to-North Water Diversion Project in China. A comprehensive study was carried out to investigate the spatial and temporal distribution of cyanobacteria in the lake from June 2008 to May 2011 based on monthly sample monitoring from five stations. The effect of environmental factors on cyanobacterial abundance was also evaluated. The cyanobacterial community contained 15 genera and 23 species. The cyanobacterial abundance of each monitoring station ranged from 0 to 1.53× 10⁷ cells/L with an average of 1.45×10⁶ cells/L, which accounted for 11.66% of the total phytoplankton abundance. The dominant species of cyanobacteria were Pseudanabaena (32.94%) and Merismopedia (19.85%), not the bloom-forming algae such as Microcystis and Anabaena. In addition, the cyanobacterial community structure and water quality variables changed substantially over the survey period. Redundancy analysis (RDA) suggested that temperature and phosphorus were important environmental factors that affected cyanobacteria. Temperature was the most important factor affecting cyanobacterial abundance. The effect of phosphorus on cyanobacterial abundance was more notable in warm periods than in periods with low temperature.     

野外模拟扰动方式对太湖浮游植物群落结构的影响

浮游植物是水生生态系统的重要生产者,扰动是水体的固有特征.为探究扰动方式对浮游植物的影响,通过野外模拟试验探讨不同扰动方式（持续扰动和间歇扰动）对太湖浮游植物的影响,并在试验期间测定各扰动条件下浮游植物生长和群落结构变化情况.结果表明:①对照组、间歇扰动组和持续扰动组中浮游植物平均生物量分别为5.92、19.42、58.27 mg/L,平均细胞密度为1.06×108、1.23×108、3.48×108 L-1,持续扰动组浮游植物生物量和细胞密度显著高于对照组和间歇扰动组.②在间歇扰动组中,浮游植物优势种始终是微囊藻,其占比平均值为84.61%;对照组和持续扰动组中,浮游植物主要优势种由微囊藻逐渐转变为湖生伪鱼腥藻,其中微囊藻占比从试验开始时的77.84%到试验结束时分别降至4.72%和5.62%,而湖生伪鱼腥藻从试验开始时的0.55%到试验结束时分别升至94.40%和90.52%.③对照组、间歇扰动组和持续扰动组中Shannon-Wiener指数分别为1.02、1.03和1.63,持续扰动组显著高于对照组和间歇扰动组.研究显示,在该试验条件下,间歇扰动有利于维持微囊藻在太湖浮游植物中的优势地位,同时表明扰动方式显著影响太湖浮游植物生长和群落结构.      

不同磷质量浓度体系中pH变化对铜绿微囊藻和四尾栅藻竞争的影响

铜绿微囊藻和四尾栅藻分别是太湖中最为常见的蓝藻和绿藻.微囊藻在湖泊中形成优势是微囊藻水华发生的重要条件.在模拟太湖水的竞争实验(ρ(磷)分别为0.012,0.112,0.412和1.612 mg/L)中,四尾栅藻均形成优势,而且在稳定期时pH达到10以上.但用0.04 mol/L HCl将pH都降到5.8后,2种藻的增殖都出现明显的变化,在ρ(磷)低于0.412 mg/L时,四尾栅藻优势变得更加明显;而在ρ(磷)为1.612 mg/L时,铜绿微囊藻则形成优势.说明pH对蓝藻和绿藻竞争的影响在不同ρ(磷)的水体中是不同的.      

巢湖夏季和冬季有毒微囊藻和无毒微囊藻种群丰度研究

应用荧光定量PCR技术研究了巢湖夏季和冬季有毒微囊藻和无毒微囊藻种群丰度的空间分布,分析了有毒微囊藻和无毒微囊藻种群丰度与环境因子之间的关系,同时应用HPLC方法分析了水体中溶解性微囊藻毒素浓度.结果表明：巢湖有毒微囊藻和无毒微囊藻种群丰度存在季节和空间差异.总的来说,夏季有毒微囊藻和无毒微囊藻种群丰度高于冬季,西湖区...     

Decomposition and phosphorus release from four di erent size fractions of Microcystis spp. taken from Lake Taihu, China

Decomposition of Microcystis is accompanied by the release of phosphorus, during bacteria play an important role. A series of experiments were undertaken to evaluate the e ect of bacteria on the decomposition of Microcystis taken from Lake Taihu, China, a lake that is su ering from dense Microcystis blooms. The 16 experiments involved four size fractions of colonial Microcystis with or without the addition of lake sediment and Gram-negative bacterial inhibitor NaN3. The highest decomposition rates were recorded for the smallest size Microcystis fraction (< 25 m) with the addition of the sediment. The lowest decomposition rates were recorded for the smallest Microcystis fraction without the sediment, but with the addition of Gram-negative bacterial inhibitor NaN3. The higher decomposition rates in the treatments with NaN3 and sediment suggest that Gram-positive bacteria in the sediment are important for the decomposition process. Additionally, higher concentrations of total dissolved phosphorus (TDP) in the treatments with NaN3 suggest that more phosphorus accumulates in the Gram-negative bacterial cells around the colony, which may be an important source of phosphorus for Microcystis cells. In the no-sediment treatments, the ratios of TDP concentration to initial TP concentration were 64%–82%. The results of this experiment suggest that both Gram-negative and Gram-positive bacteria play an important role in the decomposition of Microcystis cells and the release of phosphorus from Microcystis colonies.     

太湖水华期营养盐空间分异特征与赋存量估算

基于2013年7月的空间高密度采样数据,对太湖水华期水体营养盐进行了空间分异特征分析及赋存量估算,探讨了大型浅水湖泊不同生态类型湖区水华与营养盐的相关关系及样点设置的代表性.结果发现,水华期太湖水体营养盐及叶绿素a浓度(CHL)总体上均呈现由西北向东南降低的趋势;氮主要以溶解态存在,占总氮(TN)的76.28%,磷主要以颗粒态赋存,占总磷(TP)的66.38%.采用主成分分析和聚类分析,可以将采样点分为相互之间具有显著性差异的4个区域:第一区位于西北湖区,代表水华严重的重富营养湖区;第二区主要包括梅梁湾及南太湖的入湖河口一带湖区,代表水华和富营养化程度都相对中等的湖区;第三区包括湖心区和西南湖区,代表中等污染但水华频现湖区;第四区包括贡湖湾、胥口湾和东太湖等其他区域,代表水华影响较弱、水质较好湖区.分区统计分析表明,不同湖区影响浮游藻类生长的因子也不同:从全湖来看,与CHL显著相关的营养盐指标为TP、TN、溶解性总氮(TDN)和硝态氮(NO-3-N),而在第一区则为TP和TDN,第二区为TN和TDN,第三区为TP、磷酸盐(PO3-4-P)和TDN,第四区为PO3-4-P、溶解性总磷(TDP)和亚硝酸盐(NO-2-N).基于空间插值获得调查期间太湖水体TN、TDN、TP和TDP的赋存量分别为12 800、9 800、445和150 t.研究表明,作为一个大型浅水湖泊,因蓝藻水华空间迁移积聚特征和生态类型异化等特征,太湖水华期的营养盐具有高度空间异质性,对于此类大型浅水湖泊的监测与评价,应当考虑点位的合理布设及结果的恰当解读,避免因监测布点和统计方法不当而以偏概全.     

太湖不同湖区轮虫群落结构季节变化的比较研究

2006年7月～2007年6月对太湖不同湖区(河口区、梅梁湾、太湖湖心区和贡湖湾)轮虫的季节变化进行了比较研究.整个研究期间,河口区、梅梁湾、太湖湖心区和贡湖湾轮虫种类数分别为23、15、14和21;河口区轮虫的年平均密度最高,为475个·L-1,梅梁湾最低,为164个·L-1,太湖湖心区为189个·L1-,贡湖湾为338个·L-1.4个湖区优势种不同,河口区轮虫优势种为萼花臂尾轮虫(B.cdyciflorus),梅梁湾为角突臂尾轮虫(B.angularis),太湖湖心区和贡湖湾优势种都是针簇多肢轮虫(P.trigla).食物的不同以及大型浮游甲壳动物的抑制作用,可能是太湖4个湖区轮虫群落结构不同的重要原因.相关分析表明,轮虫数量与枝角类数量、枝角类生物量和桡足类生物量极显著负相关;轮虫数量与透明度显著正相关.结果表明,太湖4个不同湖区轮虫群落结构不同.     

太湖表层底泥中产毒蓝藻群落结构和种群丰度的时空变化

为了研究太湖底泥中产毒蓝藻群落结构和种群丰度的时空变化,分别于2010年冬季(2月)、春季(4月)和夏季(6月)采集太湖不同湖区底泥样品,应用基于mcyA基因的聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)和实时定量PCR(QPCR)技术分析底泥样品.结果表明,产毒蓝藻在太湖底泥中分布广泛,其群落结构和种群丰度存在空间和季节差异性:不同湖区间群落组成相似性系数变化范围为67%～97%;冬季、春季和夏季底泥中产毒蓝藻群落结构的香农多样性指数变化范围分别为2.774～3.392、1.649～2.619和0.681～1.996,呈逐渐下降趋势;且香农多样性指数与样品中优势基因型所占比例呈极显著负相关(p<0.01,r=-0.95);从冬季到夏季,底泥中蓝藻复苏进入水体中,产毒蓝藻种群丰度逐渐降低.底泥中产毒蓝藻群落结构和种群丰度的动态变化与温度、光照、溶解氧、电导率和pH值密切相关.     

Responses of alkaline phosphatase activity to wind-driven waves in a large, shallow lake: Implications for phosphorus availability and algal blooms

Phosphorus is a vital nutrient for algal growth, thus, a better understanding of phosphorus availability is essential to mitigate harmful algal blooms in lakes. Wind waves are a ubiquitous characteristic of lake ecosystems. However, its effects on the cycling of organic phosphorus and its usage by phytoplankton remain poorly elucidated in shallow eutrophic lakes. A mesocosm experiment was carried out to investigate the responses of alkaline phosphatase activity fractions to wind waves in large, shallow, eutrophic Lake Taihu. Results showed that wind-driven waves induced the release of alkaline phosphatase and phosphorus from the sediment, and dramatically enhanced phytoplanktonic alkaline phosphatase activity. However, compared to the calm conditions, bacterial and dissolved alkaline phosphatase activity decreased in wind-wave conditions. Consistently, the gene copies of Microcystis phoX increased but bacterial phoX decreased under wind-wave conditions. The ecological effects of these waves on phosphorus and phytoplankton likely accelerated the biogeochemical cycling of phosphorus and promoted phytoplankton production in Lake Taihu. This study provides an improved current understanding of phosphorus availability and the phosphorus strategies of plankton in shallow, eutrophic lakes.