Two virus-like particles that cause lytic infections in freshwater cyanobacteria

Dear Editor, We report the results of the preliminary isolation of two virus-like particles (VLPs) that are infectious to freshwater cyanobacteria from Lake Donghu,the largest urban lake in China,located in Wuhan City,Hubei Province.By light and transmission electron microscopy,we observed VLPs causing lytic infections in freshwater bloom-forming cyanobacteria,and we detected their infections by exposing the VLPs to 12 cyanobacterial strains,including Microcystis aeruginosa HAB 1801 and Anabaena spiroides HAB 1211.They were termed ‘Anabaena spiroides geminivirus-like (AsGV-L)' particles and ‘Microcystis aeruginosa corticoviruslike (MaCV-L)' particles,based on their ultrastructural morphological characteristics and host specificities.     

西江颗粒直链藻种群生态特征

西江肇庆段是珠江干流汇入珠三角河网水域的咽喉通道,结合长期定点连续采样和空间季节性调查,重点对2009年西江干流肇庆段的颗粒直链藻(Aulacoseira granulata)的种群生态学特征进行系统分析。结果显示:全年中,颗粒直链藻在总种群中所占百分比均值为25.37%,与总种群丰度存在极显著正相关关系,这表明颗粒直链藻是西江浮游植物群落的重要优势种。调查期间发现了颗粒直链藻的4种形态,包括原变种(var.granulata)、极狭变种(var.angustissima)、弯曲变种(var.curvata)和极狭变种螺旋变型(var.angustissima f.spiralis)。其中,原变种占有绝对优势地位,其相对百分组成的年均值为91.47%,推测为原变种的形态结构比其它3种形态更容易适应外界环境变化,进而形成优势群体。颗粒直链藻总密度的周年变化呈现明显的双峰型,两个峰值分别出现在8月和11月,主要得益于适宜的水温、相对稳定的水体和较高的营养盐水平,这些均有助于颗粒直链藻的生长繁殖。第一个高峰之前,颗粒直链藻总密度未出现明显的波动,这很可能是由于水体径流量增大导致的稀释作用和频繁降雨导致的水体剧烈波动对颗粒直链藻增长的负作用掩盖了水温上升所带来的益处。从空间分布来看,颗粒直链藻总丰度的最大值一般出现在S1、S5和S6站点,这与营养盐最大值的空间分布格局是一致的。     

Phytoplankton assemblages in a Brazilian subtropical cascading reservoir system

Variations in the structure and dynamics of a phytoplankton community were determined during two periods in 2001, along five cascading reservoirs of the Iguaçu River, Paraná State (Brazil). The vertical dynamics of the phytoplankton was related to underwater light availability and the water column-mixing regime. Correlations between physical and chemical variables and phytoplankton abundance were evaluated by Canonical Correspondence Analysis (CCA). No significant cascading effect throughout the serial reservoirs was obtained. Thus, the main factor affecting the structure of the phytoplankton assemblages of each reservoir was the hydrodynamic pattern. The instability of the water column during the winter favored the development of R-strategist species such as Aulacoseira granulata, Urosolenia longiseta and U. eriensis in most reservoirs. In the period of the highest hydraulic stability and temperature, the greatest contribution to biomass came from Cyanophyceae S-strategists such as Aphanocapsa sp.1, Anabaena planctonica and A. crassa. The phytoplanktonic associations recorded were important indicators of the environmental conditions of these reservoirs.     

Morphological variability in selected heterocystous cyanobacterial strains as a response to varied temperature, light intensity and medium composition

The effect of temperature, light and nutrient composition on morphological traits was determined in seven nostocacean cyanobacteria (Anabaena planctonica, A. sphaerica var. conoidea, A. spiroides, Aphanizomenon gracile, Nostoc sp., Scytonema sp., and Tolypothrix sp.). Their morphological variability was high but only some of the features showed changes reflecting varied growth conditions. The frequency of heterocyst occurrence decreased with increasing nitrogen concentration. Within the range studied, the effect of temperature on heterocyst frequency of Tolypothrix sp. and planktonic Anabaena strains could be fitted by a normal curve with a clear optimum while linear correlation was found in Aphanizomenon gracile. T-and S-type branching was observed in both Scytonema sp. and Tolypothrix sp. strains. T-type branching was found to be markedly dependent on nitrogen concentration. The abundance of necridic cells of Tolypothrix sp. increased linearly with temperature and light intensity. Regularity of trichome coiling of A. spiroides depended on culture medium, suggesting that nutrient composition may be the main controlling factor. In contrast, the effect of the experimental conditions on the dimensions of vegetative cells and heterocysts was weak. Their variability was markedly higher within each experimental treatment than between treatments.     

洱海螺旋鱼腥藻生长生理特性的初步研究

近年来,洱海正处于中营养水平向富营养湖泊的过渡阶段,蓝藻水华也频繁发生。本文作者在洱海大规模水华暴发期间,分离、纯化了水华优势种螺旋鱼腥藻,并对其生长生理特性进行了初步研究,以期为探讨洱海鱼腥藻水华发生的环境影响因素提供基础的参考资料。实验结果表明,氮含量1.5mmol/L、磷含量12µmol/L、光照强度30E/ m·s、pH值8－10及温度25℃时,螺旋鱼腥藻生长状况最好,生物量及相对生长速率较高。不同氮、磷浓度下的氮磷代谢活性表明：氮浓度在0－0.36mmol/L时,硝酸还原酶活性随着氮浓度的增加而增强;氮浓度在0.36－6 mmol/L时,酶活性由其生长状况决定,生长越好,酶活性越高。碱性磷酸酶受磷影响较大,随着磷浓度的增加其活性逐渐减弱,磷充足时,氮对其活性并无显著影响。此外,洱海螺旋鱼腥藻可在低的氮、磷浓度下生长,这与其氮磷代谢活性的调节作用有关。     

Abundance and diversity of phytoplankton in the Paraná River (Argentina) 220 km downstream of the Yacyretá reservoir.

Patterns in the temporal composition, abundance and diversity of the phytoplankton community of the Paraná river prior to and after the initial filling phase of the Yacyretá reservoir are analyzed. The study site is located 220 km downstream from the Yacyretá reservoir and 30 km downstream from the confluence with the Paraguay river. Because both rivers remain separate and unmixed at the study site, we compared the possible effects of the impoundment on both river banks (left and right banks) in hydrological periods with similar duration and magnitude of the low and high water phases. Physical and chemical conditions measured on the right bank (water from the Paraguay river) were similar at both periods (pre and post-impoundment) whereas conductivity, pH and orthophosphate concentration increased on the left bank (water from the High Paraná river and Yacyretá reservoir) after the impoundment. Changes in phytoplankton density and diversity were observed only in samples collected from water flowing from the reservoir (left bank). The density of Chlorophyceae (Chloromonas acidophila, Chlamydomonas leptobasis, Choricystis minor, Chlorella vulgaris, Scenedesmus ecornis, Monoraphidium minutum, M. contortum and M. pusillum) and Cryptophyceae (Rhodomonas minuta, Cryptomonas marssonii and C. ovata) increased while Cyanophyceae (Cylindrospermopsis raciborskii, Raphidiopsis mediterranea and Planktolyngbya subtilis) and Bacillariophyceae (Aulacoseira granulata and its bioforms) decreased compared to previous studies conducted on the left bank of the Paraná river. Phytoplankton collected from the right bank of the river did not differ in pre and post- impoundment samples because they originate from the Paraguay river, which remains relatively unaffected by human activities.     

珠江下游颗粒沟链藻Aulacoseira granulata (Ehrenberg) Simonsen及形态相近种赫组基沟链藻Aulacoseira herzogii (Lemmerman) Simonsen的形态学研究

采用扫描电镜(SEM)技术,对采自珠江下游的沟链藻属2个物种颗粒沟链藻Aulacoseira granulata(Ehrenberg)Simonsen和赫组基沟链藻A.herzogii(Lemmerman)Simonsen的形态特征进行了观察和描述,并与其他相近种的形态异同点进行了比较分析。     

Methane oxidation by methanotrophs and its effects on the phosphate flux over the sediment-water interface in a eutrophic lake

The effect of methane oxidation in aerobic sediment on oxygen consumption and phosphate flux was investigated in diffusion chambers. The diffusion chambers consisted of two compartments separated by a Teflon membrane. In the upper chamber a thin sediment layer was present and the lower chamber was continuously flushed with gas. The hydrophobic membrane allowed for diffusion of gases from the lower chamber through the sediment layer toward the headspace of the upper chamber. In experiments with a methane oxidation rate of 9.8 mmol m^–2 day^–1, the oxygen consumption rate increased by a factor of two compared with controls without methane oxidation (8.6 vs 17.7 mmol m^–2 day^–1). Methane oxidation significantly decreased oxygen penetration depth (2.5–4.0 vs 1.0–2.0 mm). However, despite the shrinkage of the oxidized microlayer, no differences were found in phosphate flux across the sediment water interface. Batch experiments with standard additions of methane revealed that the growth of methanotrophic bacteria contributes to the phosphate uptake of aerobic sediment. From the batch experiments a molar ratio of carbon to phosphate of 45 mol:mol was calculated for the growth of methanotrophs. Results suggest that a decrease in chemical phosphate adsorption caused by a decrease in the oxygen penetration depth could be compensated for entirely by the growth of methanotrophic bacteria. Send offprint requests to: A.J.C. Sinke     

洋河水库浮游植物组成及优势种演替规律研究

在洋河水库设置6个采样点, 对浮游植物进行了周年研究, 并在夏季进行了每周一次的加密采样, 以揭示水华期间藻类优势种演替规律。结果表明洋河水库全年共检测到浮游植物8门41属49种, 群落季节变化与温度密切相关。春季隐藻门的啮齿隐藻(Cryptomonas erosa)为优势, 夏季初期表层水温在25℃以下时, 绿藻门的波吉卵囊藻(Oocystis borgei)占主要优势; 当表层水温升至25℃以上, 微囊藻(Microcystis spp.)迅速取代其成为绝对优势。秋季硅藻门的克洛脆杆藻(Fragiaria crotomensis)和隐藻门的啮齿隐藻(C. erosa)为优势。空间分布上水库北部浅水区域隐藻和硅藻生物量普遍高于南部; 受东南风影响, 蓝藻生物量在西洋河口S2点位最大。CCA分析表明夏季水华主体微囊藻的生物量与氮浓度正相关, 螺旋鱼腥藻在夏季仅作为第二优势种短暂出现于西洋河口处, 其出现与否受到磷营养盐的限制。       

The structural nif genes of the cyanobacteria Gloeothece sp. and Calothrix sp. share homology with those of Anabaena sp., but the Gloeothece genes have a different arrangement.

Probes carrying the Anabaena sp. strain PCC 7120 nitrogenase reductase (nifH) and nitrogenase (nifK and nifD) genes were hybridized to Southern blots of DNA from the unicellular, aerobic nitrogen-fixing cyanobacterium Gloeothece sp. strain PCC 6909 and from the filamentous cyanobacterium Calothrix sp. strain PCC 7601. These data suggest that the Gloeothece sp. nif structural proteins must be similar to those of other diazotrophs and that the ability for aerobic nitrogen fixation does not reside in the nif protein complex. We also found that the nif structural genes of Gloeothece sp. are clustered, whereas those of Calothrix sp. are arranged more like those of Anabaena sp.     

19F-nuclear magnetic resonance: measurements of [O2] and pH in biological systems.

19F-nuclear magnetic resonance (NMR) has been used to determine both intracellular pH and oxygen concentrations in cell suspensions. Oxygen concentrations in Paracoccus denitrificans and insulinoma cells, RINm5F, in the NMR probe can be monitored directly by 1/T1 measurements of perfluorotripropylamine (FTPA)/lecithin emulsion added to the suspensions. With FTPA oxygen monitoring, we investigated the relative aeration capabilities of two types of NMR chambers. Both normal and transformed eucaryotic cells can be maintained in either chamber for at least 1-2 h at cytocrits of up to 20-25%, with 30% oxygen saturation and cell viabilities of 90-95%. Similar concentrations of procaryotes were maintained aerobic with high FTPA concentrations in the more efficient of the two NMR chambers. A new precursor molecule for the 19F-NMR pH indicator difluoromethylalanine, the para-chlorophenyl ester, has been tested and used in RINm5F cells and P. denitrificans, neither of which hydrolyzes methyl esters.     

Activity and thermostability of the small self-splicing group I intron in the pre-tRNA(lle) of the purple bacterium Azoarcus.

The 205-nt group I intron located in the pre-tRNA(lle) from the bacterium Azoarcus sp.BH72 is the smallest self-splicing group I intron identified to date. Comparative sequence analysis has placed this intron and the Anabaena pre-tRNA(Leu) intron into the same subgroup, IC3; we now compare their activity and stability. Unlike the Anabaena intron, the Azoarcus intron has two transitions in the kinetics of the first step of splicing. The faster transition occurs with a larger k(cat)/K(m) than that of the Anabaena or other group I introns, due to a rapid K(cat) (5 min(-1) at 32 degrees C) and a low K(m) for guanosine (17 microM). The excised intron circularizes by releasing a trinucleotide from the 5' end of the intron, another property unlike the Anabaena intron. Although it is smaller in size, the Azoarcus intron retains activity at higher temperatures, higher concentrations of urea, and higher pH than the Anabaena intron. Melting curves show that tertiary structure is disrupted at a lower temperature in the Anabaena intron. Some structural features that may explain the unusual stability of the Azoarcus intron include a G-C rich secondary structure and the presence of two 11-nt motifs, which are known to interact strongly with GAAA loops in group I and group II introns. The disruption of one of these interactions by substituting the Anabaena structural element in fact lowered the thermal stability of the Azoarcus intron. Thus, even superficially similar group I introns from the same structural subgroup can differ significantly in activity and stability.     

Stoichiometry of aerobic mineralization (O/C) of aquatic macrophytes leachate from a tropical lagoon (São Paulo –Brazil)

The temporal variation of stoichiometry between consumed oxygen and oxidized carbon was investigated for the aerobic mineralization of leachates from aquatic macrophytes. Seven species of aquatic plants, viz. Cabomba piauhyensis, Cyperus giganteus, Egeria najas, Eichhornia azurea, Salvinia auriculata, Scirpus cubensisand Utricularia breviscapa, were collected from Òleo lagoon located in the floodplain of Mogi-Guacu river (São Paulo State, Brazil). After being collected, the plants were washed, oven-dried and triturated. In order to obtain the leachate, the fragments were submitted to an aqueous extraction (cold). Mineralization chambers were incubated at 20 °C containing leachates dissolved in water samples from Òleo lagoon to a final concentration of ca. 200 mg l^–1on carbon basis. The chambers were maintained under aerobic conditions; the concentrations of the organic carbon (particulate and dissolved) and the dissolved oxygen were measured during approximately 80 days. Elemental analysis of the detritus and the concentrations of the remaining material (DOC and POC) were used to determine the amounts of mineralized organic carbon. The data were analyzed with first-order kinetics models, from which the daily rates of consumption (carbon and oxygen) and the stoichiometry (O/C) were determined. In the early phase of mineralization the O/C rates increased before reaching a maximum, after which they tended to decrease. For the mineralization of leachates from C. giganteus, S. auriculata and U. breviscapa, the decrease was relatively slow. For all substrata the initial values were smaller than 1, and ranged from 0.42 (S. cubensis) to 0.81 (C. piauhyensis). The maximum values were within the range from 0.58 (U. breviscapa) to 23.1 (E. najas) and at their highest 26th (C. piauhyensis) and 106th (C. giganteus) days. These variations are believed to be associated with the chemical composition of the leachates, with their transformations and alterations of metabolic pathways involved in the mineralization.     

Physiological adaptations in response to environmental stress during an n(2)-fixing anabaena bloom

Anabaena spiroides has the ability to maintain intense biomass production for extensive periods in the epilimnion of a small eutrophic lake characterized by conditions shown to cause photooxidative death in a number of other phytoplankton. By the enhancement of carotenoid synthesis chlorophyll a was protected from photooxidation and prevented from catalyzing other photooxidative reactions within the cells. By temporally separating CO(2) and N(2) fixation, maximum utilization of photosynthetically active radiation was achieved. Because CO(2) fixation was more sensitive than N(2) fixation to a high oxygen concentration, the former was maximized during morning hours, before the afternoon buildup of dissolved oxygen. The diurnal partitioning of carbon and N(2) fixation has two additional advantages; possible competition for reductant-generating compounds is minimized, and adequate endogenous pools of carbon skeletons are assured to accept newly fixed ammonia. Hence, Anabaena, far from undergoing photooxidative death, appears to utilize a physiological strategy which allows optimization of radiant energy use for reductive processes and dominance of surface waters and shading of deeper phytoplankton during summer blooms.     

The oxygen uptake of Sarotherodon niloticus L. and the oxygen binding properties of its blood and hemolysate (Pisces: Cichlidae)

The oxygen consumption of Sarotherodon niloticus L. was found to decline below a critical oxygen concentration of about 2 mg O2/l. An important influence of CO2 on the oxygen affinity of whole blood was observed at all temperatures between 20 and 35 degrees C for gas mixtures containing 5.6% CO2. Purified hemolysate showed extremely high oxygen affinities (p50 = 1.08 mmHg at pH 8.2 and 20 degrees C). Low cooperativity was observed at all temperatures from 20 to 35 degrees C, and pH values between 6.5 and 8.2. The Bohr effect proved to be important at pH values lower than pH 7.5 (phi = delta log P50/delta pH = -0.58 between pH 6.5 and 7.0 at 35 degrees C). The oxygen affinities show high thermal sensitivity without a marked pH influence (delta H value for overall oxygenation at pH was -71.7 kJ/mol). The obtained results are interpreted as adaptations to diurnal variations in ambient temperature and oxygen availability.     

Prolonged diving and recovery in the freshwater turtle, Pseudemys scripta--IV. Effects of profound acidosis on O2 consumption in turtle vs rat (mammalian) brain and heart slices

The oxygen consumption of rat versus turtle brain and heart slices was compared as a function of extracellular pH and temperature. At pH = 6.20 rat (mammalian) brain and heart slices show a significant depression of oxygen consumption as compared to pH = 7.50 at temperatures of both 24 degrees and 37 degrees C. In the turtle oxygen consumption in brain and heart slices was not depressed at pH = 6.20 compared to pH = 7.50 at 24 degrees C and brain oxygen consumption was not significantly different at the two pH values at 37 degrees C. Turtle heart QO2 was depressed at 37 degrees C. The results suggest that extracellular acidosis depresses mitochondrial O2 uptake in mammalian brain and heart, playing a role in the bioenergetic manifestations of O2 depletion. Turtle brain mitochondria do not show a depression of QO2 at the acidotic pH. The resistance to acidosis of turtle brain mitochondria presumably enhances the possibility of survival following prolonged diving by maintaining ATP generation during the early diving period and during recovery.     

Purification and characterization of glutamine synthetase from the archaebacterium Methanobacterium ivanovi.

Glutamine synthetase (GS) was purified to electrophoretic homogeneity from the obligate anaerobic archaebacterium Methanobacterium ivanovi. The 130-fold-purified enzyme was obtained by heat treatment, ion-exchange chromatography, and gel filtration. Like all other eubacterial GSs known so far, the GS of M. ivanovi was found to be a dodecamer of about 600,000 daltons composed of a single type of subunit. The enzyme was stable at 63 degrees C for 10 min and was not sensitive to oxygen. The isoelectric point was 4.6, and the optimum pH of gamma-glutamyltransferase activity was 8.0. The Km values for hydroxylamine, glutamine, and ADP in the transferase reaction were 6.8, 22.7, and 0.35 mM, respectively. L-Methionine-DL-sulfoximine strongly inhibited the activity. Like the GS from gram-positive bacteria, Anabaena sp., several yeasts, and mammals, the enzyme from M. ivanovi was not regulated by adenylylation as demonstrated by snake venom phosphodiesterase treatment. Inhibition of the transferase activity by L-alanine, glycine, L-histidine, and L-tryptophan was observed. L-Glutamine alone or in the presence of AMP did not inhibit the GS synthetic activity. The GS of Methanobacterium ivanovi did not cross-react with a variety of antisera against GS from Escherichia coli, Anabaena strain 7120, or Bacillus megaterium. Archaebacterial GS appears to be structurally and functionally similar to eubacterial GS in gram-positive bacteria.     

Novel DNA-binding proteins in the cyanobacterium Anabaena sp. strain PCC 7120

As an approach towards elucidation of the biochemical regulation of the progression of heterocyst differentiation in Anabaena sp. strain PCC 7120, we have identified proteins that bind to a 150-bp sequence upstream from hepC, a gene that plays a role in the synthesis of heterocyst envelope polysaccharide. Such proteins were purified in four steps from extracts of vegetative cells of Anabaena sp. Two of these proteins (Abp1 and Abp2) are encoded by neighboring genes in the Anabaena sp. chromosome. The genes that encode the third (Abp3) and fourth (Abp4) proteins are situated at two other loci in that chromosome. Insertional mutagenesis of abp2 and abp3 blocked expression of hepC and hepA and prevented heterocyst maturation and aerobic fixation of N(2).     

四种浮游生物的碎屑形成过程

本文就不同学者的观点,讨论了有机碎屑的具体定义问题。从有机碎屑产生的具体途径来看,细菌侵入已死亡的有机体这一时刻,就是碎屑形成的开始。根据Golterman的藻类矿化概念,以细菌侵入藻细胞这一时相作为有机碎屑的开始期;作者将隆线溞,水华束丝藻、螺旋鱼腥藻和铜绿微囊藻的碎屑开始期分别定为:心跳停止和细菌入侵(枝角类)、藻丝体断裂期(水华束丝藻及螺旋鱼腥藻)、灰蓝色细胞质显现期(铜绿微囊藻)。碎屑开始期及其后的各种形态均属碎屑范畴。       

Identification of ten Anabaena sp. genes that under aerobic conditions are required for growth on dinitrogen but not for growth on fixed nitrogen

Heterocysts are specialized cells required for aerobic fixation of dinitrogen by certain filamentous cyanobacteria. Numerous genes involved in the differentiation and function of heterocysts in Anabaena sp. strain PCC 7120 have been identified by mutagenizing and screening for mutants that require fixed nitrogen for growth in the presence of oxygen. We have verified that 10 Anabaena sp. genes, all1338, all1591, alr1728, all3278, all3520, all3582, all3850, all4019, alr4311, and all4388, identified initially by transposon mutagenesis, are such genes by complementing or reconstructing the original mutation and by determining whether the mutant phenotype might be due to a polar effect of the transposon. Elucidation of the roles of these genes should enhance understanding of heterocyst biology.