铜绿微囊藻在不同供磷水平下对砷胁迫的响应

采用磷饥饿培养后的微囊藻细胞进行不同供磷水平的五价砷(As(V))暴露实验,考察单一胞外磷变化的情况下As(V)对滇池分离出的铜绿微囊藻FACHB905生长及产毒的影响.结果表明胞外磷浓度变化不会影响铜绿微囊藻对As(V)的耐受阈值(-10-7 mol/L).少磷条件下的半数抑制浓度(IC50)为10-2.79 mol/L,比无磷条件下的IC50(10-5.81 mol/L)高3个数量级,并且少磷条件下As(V)与细胞活性位点的结合常数要远低于无磷条件,因此胞外磷在As(V)对微囊藻的毒性效应上具有关键的作用.As(V)对微囊藻单细胞的叶绿素含量没有显著影响,但是对毒素产量具有剂量效应.在少磷条件下,As(V)浓度大于10-7 mol/L可促进微囊藻FACHB905的胞内产毒量;而在无磷条件下,所有As(V)处理组的胞内产毒量均上升78%左右.由上可知,微囊藻在产毒方面与As(V)具有协同效应,这对于全面了解滇池水华暴发期间毒素的变化规律具有一定的参考价值.     

太湖不同解有机磷菌株胞外碱性磷酸酶活性对蓝藻碎屑的响应

浮游细菌胞外碱性磷酸酶在湖泊磷循环过程中具有关键作用, 但其与生物可利用性磷和颗粒态有机质之间的关系尚未得到充分的研究。研究以从太湖沉积物分离的3 株解有机磷细菌(OPB)和东湖水中OPB菌群为实验对象, 以蓝藻干物质为颗粒态有机质, 系统分析了模拟条件下OPB 的数量、不同大小颗粒所表现的胞外碱性磷酸酶活性(APA)与不同形态磷的浓度及其相互关系。从总体上讲, APA 与OPB 数量显著正相关, 且明显反比于溶解反应性磷(SRP)浓度, 在模拟实验过程中, 不同菌株(群)所对应的SRP 浓度均显著不同, 故其解磷能力各异。从绝对活性及其在总活性中所占的比例来看, 各处理间小颗粒(0.22—3.0 μm)APA亦明显不同, 而大颗粒(>3.0 μm)APA和溶解态(8 CFU/mL), SRP 浓度逐渐增加。实验后期(20—33 d)细菌数量大幅度减少, 且伴随SRP 和溶解态有机磷浓度的显著升高。因此, 在缺磷且富含颗粒态有机质的条件下, OPB 将产生胞外碱性磷酸酶, 分解有机磷, 进而满足其大量生长的需要, 同时有效改变溶解态磷的形态与生物可利用性。       

池塘水华与底层磷营养状态的关系

比较了武汉东郊相邻池塘间隙水、上覆水与沉积物中磷形态、水相中与不同大小颗粒相联系的碱性磷酸酶活性 (APA)、溶解态与沉积物APA动力学参数及沉积物有机质的含量。铜绿微囊藻 (Microcystisaeruginosa)出现或繁盛的池塘上覆水中正磷酸盐 (O—P)或间隙水中不同形态磷的浓度较高 ,而底层水相不同形态磷浓度较低的池塘叶绿素的浓度亦低 ,且未见铜绿微囊藻 ,故水华与底层磷营养之间联系紧密。各塘沉积物磷主要以铁结合态形式存在 ,铜绿微囊藻繁盛的池塘沉积物中铁结合态磷含量较低 ,而有机质含量较高 ,底层APA亦表现出较高的底物亲和能力 (较小的Km 值 )和反应速度 (较大的Vmax值 ) ,上述结果说明 ,有机质可能在上覆水、间隙水与沉积物诱导具不同动力学特征的高APA ,同时引发厌氧状态 ,这双重因素均有利于O—P的迅速释放 ,从而促进水华的形成     

斜生栅藻对振荡和磷胁迫的生理生化响应

为了探讨水文条件的变化对常见绿藻水华发生的潜在影响,实验以斜生栅藻为材料,研究了不同水流状态及磷浓度对其生长及磷利用策略的影响。实验分别设置静止条件(0 r/min)、低振荡条件(90 r/min)和高振荡条件(120 r/min),同时设置磷限制(0.2 mg/L)和磷充足(2 mg/L)两组磷浓度,整个实验过程为3周。实验过程测定的指标为:比生长速率、胞外碱性磷酸酶活性、叶绿素a含量以及磷吸收动力学参数。实验结果表明:(1)同静止条件相比,低振荡和高振荡条件均能显著降低斜生栅藻的比生长速率,但是两种不同磷浓度下其比生长速率却无显著性差异。(2)在实验第21天时,振荡条件培养下,斜生栅藻的胞外碱性磷酸酶活性显著高于静止条件培养,与此相反,胞内磷浓度却显著高于静止条件。(3)在磷限制条件下斜生栅藻的叶绿素a含量显著降低,同时两种振荡条件培养均使其在磷充足条件下叶绿素a含量降低。(4)通过磷吸收动力学参数的比较,静止条件培养的斜生栅藻对磷的亲和力高于其在振荡条件下。由此可见,斜生栅藻适应于静止无扰动且磷营养丰富的水体,随着水体中磷浓度进一步升高,静止和水流缓慢的水体存在导致像斜生栅藻这一类绿藻发生水华的风险。       

改性纳米TiO2 对蓝藻的生理生态影响

在自然光照条件和日光灯光照条件下, 用掺1%Ag纳米TiO2 、掺5%Fe2O3纳米TiO2、掺10%ZnO纳米TiO2和纯纳米TiO2对铜绿微囊藻(Synechocystis sp.)PCC6803和集胞藻(Microcystis aeruginosa)进行处理, 并对2种藻样的叶绿素含量、类胡萝卜素含量、体外超氧自由基含量及超氧化物歧化酶(superoxidase dismutase, SOD)活性进行测定。结果表明, 自然光照条件比日光灯光照条件更利于纳米材料光催化反应的进行; 改性后的3种纳米材料对蓝藻的抑制效果要好于纯纳米TiO2,其中效果最好的为掺1%Ag纳米TiO2; 2种藻样在适应由光催化剂引发光催化而产生的自由基胁迫时, 集胞藻的适应能力比铜 绿微囊藻更强。     

北京市各种水体蓝细菌物种多样性研究

2016至2018年的5月和9月研究了北京市五大水系53个水体蓝细菌的物种多样性。结果表明,共发现蓝细菌4目4科22属63种,优势种(IRI)为小型色球藻(Chroococcus minor(Kütz.) N?g.)、湖泊鞘丝藻(Lyngbya limentica Lemm.)、水华束丝藻(Aphanizomenon flos-aquae(L.) Rafls.)、小席藻(Phormidium tende(Menegh.) Gom.)、钝顶节旋藻(Arthrospira platensis)、小颤藻(Oscillatoria tenuis Ag.)、铜绿微囊藻(Microcystis aeruginosa)、水华微囊藻(Microcystis flos-aquae)。通过对北京各种水体的蓝藻优势种进行CCA分析,得出大部分优势种与活性磷、叶绿素a、温度、溶解氧、总碱度、总硬度、pH呈正相关,与TN、TP呈负相关,与蓝藻的特性较符合,其中活性磷是限制蓝细菌生长的重要因素。各水体蓝藻平均密度为1.32×10~6 ind/L,平均生物量为0.003 2 mg/L,多样性指数平均为0.45,2018年较2017年多数水体的密度和生物量下降,多样性指数上升,说明北京市水环境治理有所改善,但仍需继续加强治理。     

阴离子表面活性剂LAS对产毒和无毒微囊藻生长及产毒特性的影响

文章研究了低浓度范围内不同浓度梯度的阴离子表面活性剂直链烷基苯磺酸盐(LAS)对产毒微囊藻(Microcystis aeruginosa, FACHB905)和无毒微囊藻(Microcystis wesenbergii, FACHB908)生长、光合特性、种间竞争及毒素合成的影响。结果表明,在0.05—5.0 mg/L LAS浓度梯度处理下,产毒微囊藻的生物量、产毒基因mcyD表达量和每细胞MCs含量均在培养12d后显著增加。产毒微囊藻胞内和胞外MCs含量在各LAS浓度处理后分别为0.069、0.052、0.061、0.038和0.037 fg/fg Chl.a及107.1、103.7、127.1、99.6和113.7 ng/L。即使在0.5 mg/L低浓度LAS处理条件下,上述3个参数也分别比对照组显著增加了24.2%、12.4倍和10.4%。浓度为0—0.2 mg/L LAS对无毒微囊藻的生物量无明显影响,而较高浓度的LAS(0.5—5.0 mg/L)明显抑制了无毒微囊藻的生长。在两株微囊藻混合培养时, 0.2—1.0 mg/L LAS处理组的产毒铜绿微囊藻mcy D的表达对LAS...     

氮、磷对小新月菱形藻无机碳利用与碳酸酐酶活性的影响

在实验室条件下研究了氮磷浓度变化对小新月菱形藻无机碳利用与碳酸酐酶活性的影响,结果显示小新月菱形藻随培养液中氮、磷浓度的升高比生长速率明显提高。低氮浓度导致胞外碳酸酐酶活性丧失,但胞内碳酸酐酶活性依然存在。高氮浓度下胞内、外碳酸酐酶活性均明显升高。胞内碳酸酐酶活性在高磷浓度下明显升高,但胞外碳酸酐酶活性并没有受到磷浓度变化的影响。高氮、磷浓度培养下的小新月菱形藻的最大光合作用速率(Vmax)、对CO2亲和力(K0.5(CO2))和光系统II最大光化学效率(Fv/Fm)均明显提高。以上结果表明小新月菱形藻可以通过改变胞内、外碳酸酐酶活性调节无机碳利用以适应不同氮磷浓度的环境。     

水华束丝藻对磷的生理响应研究

束丝藻(Aphanizomenon Morr. ex Born. et Flah.)是我国淡水水体常见的水华蓝藻之一, 由其引发的水华已产生了严重的环境及生态安全问题。然而, 目前对束丝藻的研究仍相对较少。为了揭示环境因子对束丝藻的影响, 研究从淡水水体限制因子?磷入手, 探讨其对束丝藻的生理生态效应, 研究了不同磷浓度(0.00、0.02、0.05、0.50、1.00 mg/L)对水华束丝藻的生长、光合作用及碱性磷酸酶变化的影响。结果表明: 水华束丝藻在磷浓度低于0.50 mg/L 条件下, 其比生长速率()、最大光合反应(Pm)、饱和光强(Ik)、PSⅡ光化学效率(Fv/Fm)和最大电子传递速率(ETRmax)均下降, 而暗呼吸(Rd)显著增加, 这表明培养基磷浓度低于0.50 mg/L 时,水华束丝藻产生磷营养胁迫, 导致其光合作用受到抑制, 呼吸作用增强, 进而抑制其生长。为了应对这种胁迫, 束丝藻显著增加了其碱性磷酸酶活性(APA), APA 的增加, 使得水华束丝藻能够分解有机形态磷物质转化为其可利用无机磷来缓解磷胁迫。当磷浓度高于0.50 mg/L 时, 水华束丝藻各种参数并没显著性差异, 表明磷浓度高于0.50 mg/L 能够保证水华束丝藻的正常生理特征。这些结果揭示了在低磷条件下, 水华束丝藻能通过调节光合作用和APA 等生理响应, 使其保持生存和竞争优势。       

细菌DC10的溶藻作用及环境因子对该作用的影响

从云南滇池分离获得溶藻细菌DC₁₀, 该菌属于假单胞菌属(Pseudomonas sp.), 它能强烈地溶解绿色微囊藻(Microcystis viridis, FACHB 102)等6种蓝藻及月芽藻 (Selenastrum capricornutum, FACHB 271) 等2种绿藻. 试验发现, 该菌通过分泌某种物质来溶解藻类, 较低的温度以及黑暗条件有利于细菌溶藻, 不同浓度的氯化钙、硝酸钠对该菌的溶藻作用有一定影响; 不同pH条件下, 溶藻作用的强弱依次为: pH 4 > pH 9 > pH 7 > pH 5.5. 发展利用该菌进行水华控制的技术具有现实意义.     

Effects of experimentally induced cyanobacterial blooms on crustacean zooplankton communities

SUMMARY 1. Large in situ enclosures were used to study the effects of experimentally induced cyanobacterial blooms on zooplankton communities. A combination of N and P was added to shallow (2 m) and deep enclosures (5 m) with the goal of reducing the TN : TP ratio to a low level (∼5 : 1) to promote cyanobacterial growth. After nutrient additions, high biomass of cyanobacteria developed rapidly in shallow enclosures reaching levels only observed during bloom events in eutrophic lakes.
2. In the shallow enclosures, particulate phosphorus (PP) was on average 35% higher in comparison with deep enclosures, suggesting that depth plays a key role in P uptake by algae. Phytoplankton communities in both deep and shallow enclosures were dominated by three cyanobacteria species – Aphanizomenon flos-aquae , Anabaena flos-aquae and Microcystis aeruginosa – which accounted for up to 70% of total phytoplankton biomass. However, the absolute biomass of the three species was much higher in shallow enclosures, especially Aphanizomenon flos-aquae . The three cyanobacteria species responded in contrasting ways to nutrient manipulation because of their different physiology.
3. Standardised concentrations of the hepatotoxic microcystin-LR increased as a result of nutrient manipulations by a factor of four in the treated enclosures. Increased biomass of inedible and toxin producing cyanobacteria was associated with a decline in Daphnia pulicaria biomass caused by a reduction in the number of individuals with a body length of >1 mm. Zooplankton biomass did not decline at moderate cyanobacteria biomass, but when cyanobacteria reached high biomass large cladocerans were reduced.
4. Our results demonstrate that zooplankton communities can be negatively affected by cyanobacterial blooms and therefore the potential to use herbivory to reduce algal blooms in such eutrophic lakes appears limited.     

东湖蓝藻水华毒性的研究 Ⅱ.季节变化及微囊藻的毒性

1984年至1986年间武汉东湖(包括湖边池塘)的水华有7种,即铜绿微囊藻、大型铜绿微囊藻、边缘微囊藻、水华鱼腥藻、卷曲鱼腥藻、美丽颤藻和束丝藻。经生物测定,除束丝藻未测、卷曲鱼腥藻和美丽颤藻未测出毒性外,其余4种皆为有毒水华。东湖的水华随着季节变化而有不同类型的更迭,其出现格局为:卷曲鱼腥藻、微囊藻、颤藻、微囊藻。微囊藻水华的毒性在不同季节也有较大差异,毒性最低在8—9月份,最高在12月份(LD_(50)=24mg/kg鼠重),随着温度的降低而提高。讨论了某些环境参数与微囊藻水华形成及其毒性变化的关系。此外,还用脑内注射和腹腔注射方法,研究了微囊藻毒素的毒性表现。     

Occurrence of isopropylthio compounds in the aquatic ecosystem (Lake Neusiedl, Austria) as a chemical marker for Microcystis flos-aquae

Abstract Volatile organic sulfur compounds occuring during a bloom of different species of Microcystis in Lake Neusiedl, Austria, were analyzed by gas chromatography and mass spectrometry. In open water diisopropyl disulfide and diisopropyl tri-sulfide were the only sulphur compounds to be found. It was shown that Microcystis flos-aquae was the causative agent for the generation of these sulphur compounds, since high concentrations of these substances were found both in the floating scum of cyanobacteria taken from open lake and in axenic cultures of five isolated strains of M. flos-aquae . Strains isolated from colonies of Microcystis aeruginosa were not able to synthesize isopropylthio compounds. Alternatively, methylthio compounds were released. The rather unusual formation of the isopropylthio group can be used as a chemical marker to differentiate between M. flos-aquae and M. aeruginosa as two separate species which hitherto have been regarded as formae. In a canal passing through the reed belt of Lake Neusiedl where Microcystis was missing, these compounds were not detected. Different sulfur compounds (dimethyl disulfide, dimethyl trisulfide, dibutyl sulfide and bis(methylthio) methane) which in part have not yet been reported for freshwater ecosystems occurred at this site. Their origin, however, remains obscure.     

PROPERTIES OF MICROCYSTIS AERUGINOSA AND M. FLOS-AQUAE (CYANOPHYTA) IN CULTURE: TAXONOMIC IMPLICATIONS1

Cultures were cloned from a sample containing Microcystis aeruginosa, M. flos-aquae and a few morphological intermediates. The M. aeruginosa cultures remained distinct from the M. flos-aquae cultures in (a) cell size, (b) cell aggregation pattern, (c) width of the mucilage surrounding the multicellular colonies, (d) sharpness of the mucilage boundary, (e) efect of 0.1–1.0 μM calcium chloride on the disaggregation of multicellular colonies, (f) frequency of mucilage mutants and (g) colony morphology on agar media. No M. flos-aquae culture produced morphs resembling M. aeruginosa, inconsistent with proposals that M. flos-aquae is a developmental stage or environmentally-induced variant of M. aeruginosa. After longterm cultivation, but not soon after origanal isolation, several M. aeruginosa cultures contained mutants with diminished mucilage production and an altered colony shape.     

Different physiological responses of cyanobacteria to ultraviolet‐B radiation under iron‐replete and iron‐deficient conditions: Implications for underestimating the negative effects of UV‐B radiation

Iron deficiency has been considered one of the main limiting factors of phytoplankton productivity in some aquatic systems including oceans and lakes. Concomitantly, solar ultraviolet‐B radiation has been shown to have both deleterious and positive impacts on phytoplankton productivity. However, how iron‐deficient cyanobacteria respond to UV‐B radiation has been largely overlooked in aquatic systems. In this study, physiological responses of four cyanobacterial strains (Microcystis and Synechococcus), which are widely distributed in freshwater or marine systems, were investigated under different UV‐B irradiances and iron conditions. The growth, photosynthetic pigment composition, photosynthetic activity, and nonphotochemical quenching of the different cyanobacterial strains were drastically altered by enhanced UV‐B radiation under iron‐deficient conditions, but were less affected under iron‐replete conditions. Intracellular reactive oxygen species (ROS) and iron content increased and decreased, respectively, with increased UV‐B radiation under iron‐deficient conditions for both Microcystis aeruginosa FACHB 912 and Synechococcus sp. WH8102. On the contrary, intracellular ROS and iron content of these two strains remained constant and increased, respectively, with increased UV‐B radiation under iron‐replete conditions. These results indicate that iron‐deficient cyanobacteria are more susceptible to enhanced UV‐B radiation. Therefore, UV‐B radiation probably plays an important role in influencing primary productivity in iron‐deficient aquatic systems, suggesting that its effects on the phytoplankton productivity may be underestimated in iron‐deficient regions around the world.     

Potassium Salts Inhibit Growth of the Cyanobacteria Microcystis spp. in Pond Water and Defined Media: Implications for Control of Microcystin-Producing Aquatic Blooms

Ten metals were assayed in 21 Indian ponds which comprised three groups: (i) eutrophic alkaline ponds containing <2.5 mM potassium and thick growths of Microcystis aeruginosa or Microcystis flos-aquae during most of the year, (ii) equally eutrophic alkaline ponds containing >2.8 mM potassium and no detectable Microcystis growth, and (iii) oligo- or mesotrophic ponds with various potassium and hydrogen ion concentrations and no persistent Microcystis blooms. The effects of potassium on Microcystis growth were examined in filter-sterilized pond water and in defined culture media. A 50% reduction in the 10-day yield of cultured M. aeruginosa was observed in DP medium and pond water supplemented with 1 and 3 mM KCl, respectively. In contrast, the addition of 2 to 30 mM NaCl did not suppress the growth of M. aeruginosa in either DP medium or pond water. Both 5 mM KCl and 20 mM KHCO(inf3) in J medium strongly inhibited the growth of M. flos-aquae C3-9, whereas 5 to 30 mM NaCl had no effect and 20 mM NaHCO(inf3) was stimulatory. For pond water cultured with a mixture of M. aeruginosa and the duckweed Wolffia arrhiza, M. aeruginosa dominated in unsupplemented water and W. arrhiza dominated in water supplemented with 4.8 mM KCl. Implications for the ecology and control of Microcystis blooms are discussed.     

The amino acid sequence of low-potential cytochrome c550 from the cyanobacterium Microcystis aeruginosa

The low-potential cytochrome c550 has been purified from the cyanobacterium Microcystis aeruginosa and its amino acid sequence has been determined. The protein contains 135 amino acid residues with the Cys-X-X-Cys-His heme binding site at residues 37 to 41. The sequence from residue 28 to 45 shows similarity to cytochrome c553 residues 1 to 18 when the heme binding sites are aligned. Another region of similarity is in the carboxyl-terminal regions of these two proteins. The two aligning regions of cytochrome c553 correspond to helical segments in other related cytochromes. A partial sequence of cytochrome c550 from Aphanizomenon flos-aquae was obtained and showed a 48% identity to the sequence of the M. aeruginosa cytochrome. The single methionine residue in cytochrome c550 of M. aeruginosa occurs at position 119 but there is no methionine in this region in the A. flos-aquae cytochrome, indicating that methionine is not the sixth ligand to the heme iron atom. Histidine 92 is a possible sixth ligand in M. aeruginosa cytochrome c550. The far-uv circular dichroism spectrum indicates that this protein is approximately 17% alpha helix, 42% beta-pleated sheet, and 41% random coil.     

Growth, Nitrogen Fixation, and Nodularin Production by Two Baltic Sea Cyanobacteria

In late summer, nitrogen-fixing cyanobacteria Nodularia spumigena and Aphanizomenon flos-aquae form blooms in the open Baltic Sea. N. spumigena has caused several animal poisonings, but Baltic A. flos-aquae is not known to be toxic. In this laboratory study, performed with batch cultures, the influences of environmental conditions on the biomass and nitrogen fixation rate of N. spumigena and A. flos-aquae were compared and the toxin (nodularin) concentration produced by N. spumigena was measured. Several differences in the biomasses and nitrogen fixation rates of N. spumigena and A. flos-aquae were observed. A. flos-aquae preferred lower irradiances, salinities, and temperatures than N. spumigena. The biomass of both species increased with high phosphate concentrations and with accompanying bacteria and decreased with unnaturally high inorganic nitrogen concentrations. Nodularin concentrations in cells and growth media, as well as nitrogen fixation rates, were generally highest under the conditions that promoted growth. Intracellular nodularin concentrations increased with high temperature, high irradiance, and high phosphate concentration and decreased with low and high salinities and high inorganic nitrogen concentrations. Nodularin concentrations in growth media increased with incubation time, indicating that intracellular nodularin was released when cells lysed. The different responses of A. flos-aquae and N. spumigena to changes in salinity, irradiance, and temperature may explain the different spatial and temporal distribution of these species in the Baltic Sea. According to the results, toxic N. spumigena blooms may be expected in late summer in areas of the Baltic Sea with high phosphorus concentrations and moderate salinity.     

Effects of UVB Radiation on competition between the bloom‐forming cyanobacterium Microcystis aeruginosa and the Chlorophyceae Chlamydomonas microsphaera1

The growth, photosynthetic characteristics, and competitive ability of three algal strains were investigated under different doses of ultraviolet‐B (UVB) radiation (0, 0.285, and 0.372 W · m^?2). The organisms were the toxic bloom‐forming cyanobacterium Microcystis aeruginosa FACHB 912, nontoxic M. aeruginosa FACHB 469, and the green microalga Chlamydomonas microsphaera FACHB 52. In monocultures, the growth of all three strains was inhibited by UVB. In mixed cultures, enhanced UVB radiation resulted in decreased percentages of the two M. aeruginosa strains (19%–22% decrease on d 12 of the competition experiment). UVB radiation resulted in increased contents of chlorophyll a, b, and carotenoids (CAR) in C. microsphaera, and decreased contents of allophycocyanin (APC) or phycocyanin in the two Microcystis strains. All three strains showed increased levels of UVabsorbing compounds and intracellular reactive oxygen species under 0.372 W · m^?2 UVB radiation, and decreased light compensation points, dark respiratory rates, and maximal quantum efficiency of PSII. After a 20 h recovery, the photosynthetic oxygen evolution of C. microsphaera was restored to its maximum value, but that of Microcystis strains continued to decrease. Nonphotochemical quenching was increased by UVB radiation in C. microsphaera, but was unaffected in the two M. aeruginosa strains. Our results indicated that C. microsphaera has a competitive advantage relative to Microcystis during exposure to UVB irradiation.