冬季太湖表层底泥产毒蓝藻群落结构和种群丰度

应用荧光定量PCR对冬季太湖不同湖区底泥表面有毒微囊藻和总微囊藻种群丰度进行调查,同时基于PCR-DGGE技术对底泥中有毒微囊藻群落结构进行分析。结果表明:微囊藻在太湖底泥表面分布广泛,所有采样点都检测到有毒微囊藻存在,且不同湖区有毒微囊藻和总微囊藻种群丰度存在显著差异,有毒微囊藻和微囊藻基因型丰度范围分别为1.23×10⁴-3.75×10⁶拷贝数/g干重和2.56×10⁴-1.07×10⁷ 拷贝数/g干重,有毒微囊藻与微囊种群丰度的比例为4.8％-35.2％;DGGE指纹图谱显示,冬季太湖不同湖区表层底泥中有毒微囊藻群落结构相似性较高,相似性系数为70.2％-96.0％。虽然不同湖区基因型组成存在差异,但所有样品中占优势的基因型是一致的。同时发现,优势基因型所占的比例与样品的香农多样性指数呈负相关。序列分析表明,mcyA序列长度为291bp,序列相似性超过97％。综合定量PCR结果和底泥中叶绿素a和藻蓝素浓度的测定结果,可以得出2010年冬季太湖蓝藻越冬主要集中在梅梁湾、竺山湾、贡湖湾和湖心。通过建立荧光定量PCR分析方法,为研究湖泊底泥中蓝藻种群丰度动态变化奠定了基础。     

越冬和复苏时期太湖水体蓝藻群落结构的时空变化

为研究太湖蓝藻在越冬与复苏时期群落结构的时空变化规律,于2008年11月,2009年2月,2009年4月,在太湖富营养化较严重的湖区选取8个采样点 (梅梁湾、竺山湾、贡湖湾、大浦、西太湖、南太湖、湖心和湖湾交汇处),分3层采水样,过滤并提取样品DNA经PCR扩增蓝藻16S rDNA序列,采用T-RFLP(末端标记的限制性酶切片段长度多样性)技术分析蓝藻群落结构和多样性变化。共得到87个不同的T-RFs(末端限制性酶切片段),表明太湖蓝藻具有丰富的基因多样性。T-RF相对丰度和聚类分析结果表明,太湖蓝藻群落结构在垂直空间上相似性较高,相似度 > 50%;在水平空间,与Microcystis spp.对应的信号峰在8个采样点均为最强峰(相对丰度为17.7% 47.5%)。竺山湾蓝藻多样性最低,西太湖最高,但其余采样点间蓝藻群落和Shannon多样性指数没有显著差异(P>0.05)。Microcystis相对丰度与Shannon多样性指数呈显著负相关(皮尔逊相关系数为-0.958)。在时间尺度上,相似性分析(Analysis of similarity,ANOSIM)结果显示太湖蓝藻群落结构存在极显著差异( P<0.01) 。春季复苏时蓝藻多样性最高,秋季衰亡时最低。聚类分析表明样品聚成两大特征类群,秋季衰亡时样品独自聚为1支,而春季复苏期和冬季越冬期样品彼此混杂。     

太湖沉积物中氮循环菌的微生态

采用荧光原位杂交(FISH)技术,对梅梁湾和贡湖湾湖区沉积物中微生物的主要种群,及氮循环功能微生物的数量和分布进行了研究,发现随着深度增加,细菌和古菌数量均逐渐减少,但古菌在总菌数中所占比例有所增加;梅梁湾沉积物中氨氧化细菌及亚硝酸氧化细菌的数量均高于贡湖湾,随深度增加,两者数量均逐渐减少,但在贡湖湾其占总菌数的比例高于梅梁湾,表明水生植物可能对沉积物中微生物组成及氮循环有重要影响。泉古菌在太湖沉积物中普遍存在且数量高于氨氧化细菌,说明其在淡水湖泊中对氮循环可能有重要作用。     

基于盐分梯度的荒漠植物多样性与群落、种间联接响应

土壤盐分是影响干旱区荒漠植物群落动态的决定因素之一。基于样方调查和不同土壤盐分梯度下植物多样性指数及群落与种间关联的计算结果,分析干旱区荒漠群落植物多样性、群落联接性和种间关联对土壤盐分梯度的响应动态。结果表明,在土壤盐含量为0.03%-0.55% (S1)、0.61%-1.24% (S2)和1.41%-1.79% (S3)的盐分梯度上,(1)随土壤盐含量升高,群落生活型结构改变,草本比例减少,乔木比例增加;(2)植物多样性指数随土壤盐分增加而下降,低盐分梯度下,二者极显著正相关(P<0.01),中盐梯度下二者间呈部分显著负相关(P<0.05),高盐梯度下则转为以正相关为主(P>0.05);(3)群落联接性随土壤盐分梯度转变,在0.61%-1.24%的中度盐含量时正联接性最强(VR=1.89),高盐度含量下群落转为不显著的负联接,稳定性降低;(4)沿盐分升高梯度,种间的负相关种对数增加,正相关种对数减少,种间关联(相关)强度提高,正负相关种对比(PNR)与多样性指数呈显著正相关(P<0.05)。综上可知,干旱区荒漠植物多样性在土壤盐含量达到1.41%-1.79%水平时总体显著降低;土壤盐分水平显著影响植物群落和种间联接性,种间互利性随盐分梯度增加下降,物种趋于独立分布,并最终导致荒漠植物多样性降低。     

秦岭天然次生油松林冠层降雨再分配特征及延滞效应

为了研究秦岭典型地带性植物油松林冠层降雨再分配特征及延滞效应,选择陕西宁陕县秦岭森林生态系统国家野外科学观测研究站55龄天然次生油松林,从2006-2008年(5-10月份)对林外降水、穿透降雨和树干茎流进行定位观测。利用其中100次实测数据进行分析研究,结果表明:总降雨量为1576.4 mm,穿透降雨量为982.9 mm,树干茎流量为69.5 mm,冠层截留量为524. 0mm,分别占总降雨量的62.4%、4.4%和33.2%。降雨分配与降雨量级密切相关,降雨量级增大,穿透降雨率和茎流率呈增大趋势,截留率呈降低趋势,变化幅度分别为46.6%-68.9%、0.8%-9.2%、53.4%-22.0%。穿透降雨量、树干茎流量和林冠截留量与林外降雨量之间的关系分别为:TF=0.6548P-0.4937,R²＝0.9596;SF=-0.2796+0.0452P+0.0005P²,R²＝0.8179;I=0.5958P^0.8175,R²＝0.8064。降雨事件发生后,穿透降雨和树干茎流出现的时间与降雨发生的时间并不同步,均表现出一定的延滞性,随着降雨量级增大,滞后时间表现出逐渐缩短的趋势((78.5±8.8)-(16.0±0.0) min,(111.0±33.0)-(41.2±0.0) min)。降雨终止时,特别是当降雨量>10.0 mm,穿透降雨终止时间也存在一定的延滞性((3.2±2.6)-(12.0±0.0) min)。但树干茎流终止时间先于降雨终止时间,降雨量级越小,树干茎流终止时间愈早((-58.3±21.5)-(-9.8±0.0) min)。     

太湖大型底栖动物群落结构及多样性

为揭示现阶段太湖大型底栖动物群落现状及其对生态环境变化的响应, 于2007年2月至2008年11月对太湖大型底栖动物进行为期两周年的季度调查。30个采样点共记录底栖动物3门7纲19科40种, 底栖动物平均密度和生物量空间差异较大, 平均密度高值出现在梅梁湾、竺山湾及河口, 主要为寡毛纲颤蚓类; 平均生物量高值出现在贡湖湾、西湖区、东太湖和东部湖湾, 主要为软体动物。霍甫水丝蚓(Limnodrilus hoffmeisteri)、中华河蚓(Rhyacodrilus sinicus)、河蚬(Corbicula fluminea)、铜锈环棱螺(Bellamya aeruginosa)、中国长足摇蚊(Tanypus chinensis)和钩虾属一种(Gammarus sp.)是现阶段太湖大型底栖动物的优势种。聚类分析将30个采样点分成3组, 相似性分析检验表明各聚类组大型底栖动物群落具有显著差异(P<0.05)。多样性分析结果表明, 东太湖和东部湖湾物种多样性、丰富度和均匀度最高, 优势种为腹足纲螺类; 梅梁湾、竺山湾及河口多样性最低、密度最高, 霍甫水丝蚓和中华河蚓在该区占据绝对优势; 贡湖湾、湖心和西湖区多样性处于中等水平, 其优势种为河蚬。研究结果表明营养水平、底质类型以及水生植被的分布是决定太湖大型底栖动物群落结构及多样性的关键因子。     

太湖沉积物微生物生物量及其与碳、氮、磷的相关性

对太湖沉积物中微生物生物量碳(MBc)、氮(MBN)、磷(MBp),以及沉积物总有机碳(TOC)、总氮(TN)、总磷(TP)进行测定并进行相关性分析,揭示太湖沉积物微生物对太湖沉积物营养盐的响应及反馈特征.结果表明:沉积物微生物生物量(MB)在湖体沿岸地区大于湖心区,平均值为184.66 mg·kg-1,MBc在西部沿岸区以及竺山湾和梅梁湾区域较高,平均值为127.57 mg·kg-1;MBN在梅梁湾、贡湖部分区域以及靠近梅梁湾和贡湖的湖心区域和东部沿岸区较高,平均值为19.25 mg·kg-1;MBp在东部沿岸区及其附近的湖心区最高,平均值为19.09 mg·kg-1;沉积物TOC高值区(≥2.30 g· kg-1)主要集中在竺山湾、西部沿岸区、梅梁湾、贡湖地区,平均值为1.59 g·kg-1;沉积物TN高值区(≥0.30g· kg-1)主要集中在贡湖、梅梁湾、竺山湾部分地区以及西部沿岸区,平均值为0.21 g·kg-1;沉积物TP高值区(≥1.20g·kg-1)主要集中在东部沿岸区以及湖心部分区域,平均值为0.55 g·kg-1;太湖沉积物TOC/TN在7～19,平均值为8.97,表明太湖沉积物中的有机质具有明显的双重来源,其中陆源有机质主要集中在西部沿岸区;太湖沉积物MB与沉积物TOC和TN呈显著正相关,与沉积物TP相关性不显著;沉积物MBc/MBN与沉积物TOC/TN显著相关.太湖沉积物微生物主要受沉积物TOC、TN影响,且沉积物TOC/TN的变化显著影响微生物群落结构.     

洋山港潮间带大型底栖动物群落结构及多样性

2009-2010年在洋山港海域大洋山岛和圣姑礁进行四个季度潮间带大型底栖动物生态学研究。共采集到大型底栖动物61种,以广布种为主,部分为河口低盐种,其中软体动物22种,环节动物16种,节肢动物12种,苔藓动物5种,腔肠动物4种,星虫动物和棘皮动物各1种。优势种为短滨螺(Littorina brevicula)、多齿围沙蚕(Perinereis nuntia)、日本笠藤壶(Tetraclita japonica)、齿纹蜒螺(Nerita yoldii)和特异大权蟹(Macromedaeus distinguendus)。丰度和生物量在不同季节明显不同(P<0.05):丰度的最高值出现在春季,为(3204.9±837.84)个/m²,最低值出现在秋季,仅为(2213.2±731.27)个/m²;生物量的最高值则出现在夏季(2233.2±1493.42)g/m²,冬季最少,仅为(819.95±484.80) g/m²。大洋山岛和圣姑礁的丰度和生物量具有较大差异(P<0.05):年均丰度以大洋山断面较高,为(3090±742.74)个/m²,圣姑礁断面较低,为(2133±372.51)个/m²;而年均生物量则以圣姑礁断面较高,为(1711.1±1180.76) g/m²,大洋山断面较低,仅为(1028.5±627.61) g/m²。运用ABC曲线、等级聚类和MDS对大型底栖动物群落结构分析发现:大洋山潮带间大型底栖动物群落尚未受到干扰或干扰较轻,群落结构相对稳定;而圣姑礁断面的潮间带大型底栖动物群落受到了中度的干扰,群落结构稳定性下降。与15年前的研究相比,圣姑礁大型底栖动物的密度和生物量均有大幅度的降低,这与洋山港海域盐度的升高、生态环境变化和人为干扰强度增加等有关。     

太湖水体叶绿素a含量与氮磷浓度的关系

基于太湖水体1993—2002年5—9月监测资料,进行了叶绿素a含量与总氮(TN)、总磷(TP)浓度关系的分区统计分析,探讨了太湖藻类生长的TN、TP适宜浓度。结果表明:太湖叶绿素a含量与TN、TP浓度的关系存在显著的空间差异;在梅梁湾和西北区,当TN、TP浓度较低时,叶绿素a含量与TN、TP浓度呈正相关;当TN、TP浓度较高时(梅梁湾TN、TP浓度分别超过5.4和0.31mg.L-1;西北区分别超过4.5和0.27mg.L-1),叶绿素a含量与TN、TP浓度呈负相关;在湖心区和贡湖区,叶绿素a含量与TN、TP浓度呈正相关,尤其当TP浓度超过0.1mg.L-1时,叶绿素a含量随TP浓度增加而上升;在东太湖和湖东滨岸区,随TN、TP浓度的升高,叶绿素a含量变化较小;在西南区,叶绿素a含量与TN浓度无显著相关关系,与TP浓度呈正相关;太湖藻类生长的适宜浓度是TN<5.4mg.L-1,TP为0.1～0.31mg.L-1。     

丰水期长江感潮河口段网采浮游植物的分布与长期变化

于2009年6、8月对长江口门至江阴的河口段浮游植物进行了拖网采集,共检出浮游植物6门99属239种。其中:硅藻123种,甲藻19种,绿藻和蓝藻各42种,裸藻9种,黄藻4种。河口段网采浮游植物丰度以蓝藻占绝对优势,硅藻次之,两者合计在群落中的比例超过了95%。优势种也主要以蓝藻(水华鱼腥藻Anabaena flos-aquae、柔软腔球藻Coelosphaerium kuetzingiarum、微囊藻Microcystis spp.、颤藻Oscillatoria spp.和席藻Phorimidium spp.)构成,硅藻仅有2种(骨条藻Skeletonema spp.和颗粒直链藻Aulacoseira granulata)。口门内盐度均<0.5,群落基本以淡水类群为主,口门附近则以半咸水类群为主,海水类群主要位于口门外(盐度>13)。随着水温和营养盐水平的升高,8月浮游植物平均丰度(347.75×10⁴ 个/m³)明显高于6月(204.19×10⁴ 个/m³)。根据多维尺度和相似性分析,丰水期长江河口段浮游植物群落组成与分布存在显著(P<0.01)的时空差异。对比20世纪80年代以来的历史资料发现,长江口门内网采浮游植物丰度显著升高,且优势种也从硅藻(骨条藻、直链藻和圆筛藻)转变为蓝藻(颤藻、鱼腥藻和微囊藻)。     

Lake Erie Microcystis: Relationship between microcystin production, dynamics of genotypes and environmental parameters in a large lake

Cyanobacteria of genus Microcystis sp. have been commonly found in Lake Erie waters during recent summer seasons. In an effort to elucidate relationships between microcystin production, genotypic composition of Microcystis community and environmental parameters in a large lake ecosystem, we collected DNA samples and environmental data during a three-year (2003–2005) survey within Lake Erie and used the data to perform a series of correlation analyses. Cyanobacteria and Microcystis genotypes were quantified using quantitative real-time PCR (qPCR). Our data show that Microcystis in Lake Erie forms up to 42% of all cyanobacteria, and that Microcystis exists as a mixed population of potentially toxic and (primarily) non-toxic genotypes. In the entire lake, the total abundance of Microcystis as well as the abundance of microcystin-producing Microcystis is strongly correlated with the abundance of cyanobacteria suggesting that Microcystis is a significant component of the cyanobacterial community in Lake Erie during summer seasons. The proportion of total Microcystis of all cyanobacteria was strongly linked to the microcystin concentrations, while the percentage of microcystin-producing genotypes within Microcystis population showed no correlation with microcystin concentrations. Correlation analysis indicated that increasing total phosphorus concentrations correlate strongly with increasing microcystin concentrations as well as with the total abundance of Microcystis and microcystin-producing Microcystis.     

Estimating spatial variation in the abundance of potential microcystin-producing Microcystis spp. using real-time PCR during summer bloom in Lake Taihu

Lake Taihu, which is the third largest freshwater lakes in China, is a hypertrophic shallow lake in eastern China that has experienced lake-wide cyanobacterial blooms annually during the last few decades. In this study, quantitative real-time PCR assays targeting on phycocyanin intergenic spacer (PC-IGS) and a microcystin synthetase gene mcyD were established, respectively. Water samples collected from eight sampling sites (including Zhushan Bay (N5), Meiliang Bay (N2), Gonghu Bay (N4), West lake areas (W2 and W4), south-middle lake areas (S2, S4 and S5)) in August of 2009 and 2010 were analyzed using real time PCR for the distribution and abundance of toxic and total Microcystis populations. The results showed that Microcystis exists as a mixed population of potential toxic and non-toxic genotypes, and there was significant spatial changes in the abundance of potential toxic Microcystis on the basis of quantification by quantitative real-time PCR analysis: the abundance of toxic Microcystis population in 2009 and 2010 varied from 4.08 × 10⁴ to 8.28 × 10⁶ copies mL^?1, from 4.45 × 10⁵ to 5.22 × 10⁷ copies mL^?1, respectively. Meanwhile the ratio of the mcyD subpopulation to the total Microcystis varied considerably, from 5.7% to 41.1% in 2009 and from 10.3% to 65.8% in 2010 in all sampling sites, and the value is high in Zhushan Bay and Meiliang Bay with the high level of eutrophication. Correlation analysis showed the abundance of toxic and total Microcystis being strongly related (P < 0.01). However, there is different effects of environmental factors on the abundance of toxic and non-toxic Microcystis populations. The abundance of toxic and total Microcystis populations were positively correlated with chlorophyll-a (Chl-a) concentration (P < 0.01) suggesting that Microcystis is dominated genera of cyanobacterial bloom in Lake Taihu. It was also found that the abundance of toxic Microcystis and the proportion of toxic subpopulation to the total Microcystis were positively correlated with total phosphorus and orthophosphate concentrations (P < 0.01), whereas there was no significant correlation with total nitrogen and nitrate concentration (P > 0.05). All data suggest that phosphorus concentration is a critical factor for determining the abundance of toxic Microcystis population.     

Microcystis Genotype Succession and Related Environmental Factors in Lake Taihu during Cyanobacterial Blooms

From spring to autumn, heavy Microcystis blooms always occur in Lake Taihu, although environmental conditions vary markedly. We speculated that Microcystis genotype succession could play an important role in adaptation to environmental changes and long-term maintenance of the high Microcystis biomass. In this study, we investigated Microcystis genotype succession pattern and the related environmental variables in Lake Taihu during cyanobacterial blooms. Denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction -amplified the genus-specific cpcBA and mcyJ gene fragments was used to monitor the variations of Microcystis genotype and potential microcystin (MC)-producing Microcystis genotype compositions during blooms biweekly in three sites (Meiliang Bay, lake center, and Gonghu Bay) and CANOCO 4.5 for Windows were used for the multivariate statistical analysis of their relationships to environmental variables. DGGE patterns indicated that the number of dominant cpcBA genotype per sample increased from spring to autumn. Principal component analysis ordination plots of DGGE profiles showed clear temporal distribution pattern, but not spatial distribution pattern based on both cpcBA and mcyJ genotype compositions. These results indicated there were relatively gradual successions of Microcystis cpcBA and mcyJ genotype compositions in each site, and no distinct spatial difference among the three sites. Redundancy analyses of the gel patterns showed that, in all the three sites, three environmental factors (nitrate, pH, and chemical oxygen demand) were correlated significantly to successions of both cpcBA and mcyJ genotypes except for mcyJ genotype in the lake center. Spearman’s correlations indicated that the three environmental variables were also strongly correlated with chl a and MC concentrations. These results suggested that the environmental factors affecting succession of Microcystis community composition might also influence the growth of Microcystis and MC production.     

Temporal evolution and vertical stratification of Microcystis toxic potential during a first bloom event

In order to study the setup of a Microcystis bloom and the evolution of its toxic potential, we studied the temporal and vertical variations in Microcystis aeruginosa abundance, microcystins (MC) concentrations (intracellular and extracellular), and the relative proportion of potentially microcystin-producing cells (MC-producing cells) in relation to physicochemical parameters in the recently setup Moroccan reservoir “Yaacoub Al Mansour.” The Microcystis bloom appeared relatively late in the season and was associated with a low proportion of MC-producing cells in the water surface layer, probably related to non-limiting nutrient concentrations. Interestingly, the setup of the bloom leads to a vertical gradient, showing a decrease in Microcystis cell abundance inversely coupled with an increase in the proportion of MC-producing cells. Thus, this can be the result of the growth where non-MC-producing cells remain in the lighted water layer easier than MC-producing ones. Nevertheless, parameters other than light intensity may influence the toxic potential of bloom as no vertical pattern was observed concerning microcystins cellular quotas. The high microcystins concentrations measured in the deep water layer have also proved the importance of considering the deep part of aquatic ecosystem in the management of health risks associated with cyanobacterial proliferations.     

The abundance of microcystin-producing genotypes correlates positively with colony size in Microcystis sp. and determines its microcystin net production in Lake Wannsee

The working hypotheses tested on a natural population of Microcystis sp. in Lake Wannsee (Berlin, Germany) were that (i) the varying abundance of microcystin-producing genotypes versus non-microcystin-producing genotypes is a key factor for microcystin net production and (ii) the occurrence of a gene for microcystin net production is related to colony morphology, particularly colony size. To test these hypotheses, samples were fractionated by colony size with a sieving procedure during the summer of 2000. Each colony size class was analyzed for cell numbers, the proportion of microcystin-producing genotypes, and microcystin concentrations. The smallest size class of Microcystis colonies (<50 microm) showed the lowest proportion of microcystin-producing genotypes, the highest proportion of non-microcystin-producing cells, and the lowest microcystin cell quotas (sum of microcystins RR, YR, LR, and WR). In contrast, the larger size classes of Microcystis colonies (>100 microm) showed the highest proportion of microcystin-producing genotypes, the lowest proportion of non-microcystin-producing cells, and the highest microcystin cell quotas. The microcystin net production rate was nearly one to one positively related to the population growth rate for the larger colony size classes (>100 microm); however, no relationship could be found for the smaller size classes. It was concluded that the variations found in microcystin net production between colony size classes are chiefly due to differences in genotype composition and that the microcystin net production in the lake is mainly influenced by the abundance of the larger (>100- microm) microcystin-producing colonies.     

Dominant genera of cyanobacteria in Lake Taihu and their relationships with environmental factors

Cyanobacterial blooms in freshwaters have become one of the most widespread of environmental problems and threaten water resources worldwide. Previous studies on cyanobacteria in Lake Taihu often collected samples from one site (like Meiliang Bay or Zhushan Bay) and focused on the variation in patterns or abundance of Microcystis during the blooming season. However, the distribution of cyanobacteria in Lake Taihu shows differing pattern in various seasons. In this study, water samples were collected monthly for one year at five sites in Lake Taihu with different trophic status and a physicochemical analysis and denaturing gradient gel electrophoresis (DGGE) were conducted. DGGE fingerprint analysis showed that Microcystis (7/35 bands) and Synechococcus (12/35 bands) were the two most dominant genera present during the study period at all five sites. Cyanobium (3/35 bands) was the third most common genus which has seldom been previously reported in Lake Taihu. Redundancy analysis (RDA) indicated that the cyanobacterial community structure was significantly correlated with NO₃^--N, COD_Mn, and NH₄⁺-N in the winter and spring, whereas it was correlated with water temperature in the summer and autumn. Limiting the nutrient input (especially of N and C loading) in Lake Taihu would be a key factor in controlling the growth of different genera of cyanobacteria.     

Growth response of Microcystis spp. to iron enrichment in different regions of Lake Taihu, China

Iron (Fe) is an essential micronutrient for algal growth and can be a potential limiting nutrient in aquatic system, especially regions that exhibits nitrogen (N) limitation. Using short-term nutrient addition bioassays, we evaluated the potential role that iron might play in modifying the response of Microcystis spp. to the anthropogenic phosphorus (P) and N enrichment in hypereutrophic Lake Taihu, the third largest freshwater lake in China. Three nutrient enrichment experiments involving additions of N (as NO₃ ^?) and P (as PO₄ ^3?) with and without Fe were conducted during 2009?C2010 in Meiliang Bay, a region characterized by summer cyanobacterial (Microcystis spp.) blooms, and East Taihu, a region largely free of cyanobacterial blooms and dominated by macrophytes. In Meiliang Bay, Fe addition alone did not significantly increase Microcystis spp. biomass. However, Fe addition occasionally increased the stimulatory effect of N and P additions on Microcystis spp., indicating that Fe was not a primary limiting nutrient for Microcystis spp. growth. Occasionally Fe was co-limiting with N and P in this region. In East Taihu, the addition of Fe alone significantly stimulated Microcystis spp. growth, while addition of N and/or P had no effects on growth, indicating that Fe was a primary limiting nutrient in East Taihu. The combined addition of Fe and N resulted in a growth response similar to Fe alone, while combined addition of Fe and P yielded greater biomass increases than the addition of Fe alone. This indicated that in East Taihu, N was not limiting and Fe and P supplies facilitated Microcystis spp. growth. These results reflect differential availabilities and limitations of N, P, and Fe in distinct regions of Taihu. The potential role of Fe in eutrophication dynamics of large, regionally complex lakes like Taihu requires further attention.     

Quantitative Real-Time PCR for Determination of Microcystin Synthetase E Copy Numbers for Microcystis and Anabaena in Lakes

Cyanobacterial mass occurrences in freshwater lakes are generally formed by Anabaena, Microcystis, and Planktothrix, which may produce cyclic heptapeptide hepatotoxins, microcystins. Thus far, identification of the most potent microcystin producer in a lake has not been possible due to a lack of quantitative methods. The aim of this study was to identify the microcystin-producing genera and to determine the copy numbers of microcystin synthetase gene E (mcyE) in Lake Tuusulanjärvi and Lake Hiidenvesi in Finland by quantitative real-time PCR. The microcystin concentrations and cyanobacterial cell densities of these lakes were also determined. The microcystin concentrations correlated positively with the sum of Microcystis and Anabaena mcyE copy numbers from both Lake Tuusulanjärvi and Lake Hiidenvesi, indicating that mcyE gene copy numbers can be used as surrogates for hepatotoxic Microcystis and Anabaena. The main microcystin producer in Lake Tuusulanjärvi was Microcystis spp., since average Microcystis mcyE copy numbers were >30 times more abundant than those of Anabaena. Lake Hiidenvesi seemed to contain both nontoxic and toxic Anabaena as well as toxic Microcystis strains. Identifying the most potent microcystin producer in a lake could be valuable for designing lake restoration strategies, among other uses.     

Development and application of a multiplex qPCR technique to detect multiple microcystin-producing cyanobacterial genera in a Canadian freshwater lake

The emergence and persistence of complex blooms comprising multiple toxigenic cyanobacteria genera pose significant challenges for water quality management worldwide. The co-occurrence of morphologically indistinguishable toxic and non-toxic strains makes monitoring and control of these noxious organisms particularly challenging. Conventional monitoring approaches are not only incapable of discriminating toxic from non-toxic strains but also have proven to be less sensitive and specific. In this study, a multiplex quantitative real-time polymerase chain reaction (qPCR) approach was developed and tested for its sensitivity and specificity at detecting, differentiating and estimating potentially toxic Anabaena, Microcystis and Planktothrix genotype compositions in environmental samples. The oligonucleotide primers and probes utilized were designed to target portions of the microcystin synthetase (mcy) E gene that encode synthesis of the unique 3-amino-9-methoxy-2,6,8-trimethyl-10-phenyldeca-4,6-dienoic acid (ADDA) moiety of microcystins in the three target genera. Laboratory evaluation showed the developed assay to be highly sensitive and specific at detecting and quantifying targeted genera. Indeed, the assay standards for the Anabaena, Microcystis and Planktothrix reactions attained efficiencies above 90 %, with coefficients of determination consistently above 0.99. Analysis of water samples from Missisquoi Bay, Quebec, Canada, resulted in successful detection and quantification of target toxigenic cyanobacteria even when cell numbers were below the detection limit for the conventional microscopy methods. Furthermore, toxigenic Microcystis spp. were found to be the main putative microcystin-producing cyanobacteria in the study lake. The qPCR technique developed in this study therefore offers simultaneous detection, differentiation and quantification of multiple toxigenic cyanobacteria that otherwise cannot be accomplished by current monitoring approaches.