微囊藻毒素生物合成基因及其功能研究进展

水体富营养化加剧,导致了蓝藻水华在世界范围内频发。蓝藻产生的微囊藻毒素是最常见的一种藻毒素,对人类和动物造成了很大的危害甚至导致死亡。微囊藻毒素经非核糖体合成途径由多肽合成酶合成。对微囊藻毒素的结构与性质、微囊藻毒素合成基因的功能及其生物合成、微囊藻毒素的分子生物学检测技术进行了评述,对未来的研究方向进行了展望。     

官厅水库的微囊藻毒素及其与水环境的相关性

2007年4-11月的调研结果显示,官厅水库为富营养型湖泊,TP、TN、CODMn的含量分别为0.12、1.63、6.01mg·mL-1。浮游藻类密度为1794.08×104cells.L-1,其中蓝藻占51.75%,绿藻占25.12%,硅藻占12.56%,夏秋季出现了以微囊藻(Microcystis)占优势的蓝藻水华。库区水体中微囊藻毒素(MC-LR)的平均含量为0.794μg.L-1,但8-9月份为1.231μg.L-1,超出地表饮用水安全限值23.1%。统计分析显示,除受季节制约外,总磷和硝态氮可能是影响官厅水库MC-LR生物合成的主导营养因子;可溶性铁可能是MC-LR生物合成中限制性微量元素。治理水体富营养化是降低官厅水库微囊藻毒素含量,恢复其饮用水源地功能的关键。     

蓝藻毒素的类型及其产毒基因

水体富营养化及蓝藻水华的频繁发生,严重破坏水域生态系统,有毒水华蓝藻产生的藻毒素,污染水体且危害水生动物和人类健康。本文综述了水华蓝藻的次级代谢产物藻毒素及藻毒素合成基因的国内外研究进展,介绍了蓝藻毒素的类型、不同毒素毒性机理、产毒基因以及利用产毒基因进行产毒蓝藻的分子生物学检测等,并对研究中存在的问题和未来的研究方向进行了展望,目的是为建立有毒水华蓝藻的预警机制、科学评价产毒蓝藻及其藻毒素的生态、环境危害及生态环境治理提供理论和技术支撑。     

铁胁迫对微囊藻毒素合成酶McyC和McyI表达的影响

为研究微囊藻毒素合成酶基因的蛋白表达水平与环境因子间的关系,文章以位于微囊藻毒素合成基因簇两个操纵子中的mcyC和mcyI基因为代表,利用制备的高效McyC和McyI多克隆抗体,采用Western Blot技术检测了铁胁迫对微囊藻毒素合成酶McyC和McyI蛋白表达水平的影响。研究结果表明,在铁胁迫下,铜绿微囊藻PCC 7806藻细胞内McyC和McyI的蛋白水平变化趋势一致,且与相同条件下藻细胞内毒素的合成产量变化一致,暗示铁胁迫直接通过影响微囊藻毒素合成酶的表达水平调控毒素的合成。研究为进一步了解微囊藻毒素的合成机制提供了基础材料。     

微囊藻毒素LR对大鼠肝细胞Caspase-3酶活性的影响

水体富营养化的加剧,导致藻类水华频频发生。在全球由于藻类水华所产生的毒素,对人和动物都产生了极为严重的影响,使之成为日益突出的环境问题。其中危害最严重的 是微囊藻毒素(Micnocystin,MCYST)。而微囊藻毒素LR(LR)是微囊藻毒素中存在最为普遍且毒性作用最明显的一种,是MCYST的主要代表物。       

饮用水中藻类污染及其控制技术

近年来,我国地表水体富营养化愈加严重,这导致藻类污染成为饮用水安全的重要影响因素,其中藻类及其副产物如藻毒素等的污染,是近几年造成重大供水灾害的首要原因。此文就饮用水中的藻类污染问题,研究了藻类及藻毒素的检测方式以及给水处理时藻类的控制技术等,并针对未来的发展趋势给出了建议。     

北京四校园人工湖浮游藻类与富营养化特征分析

浮游藻类作为重要的生物指标已被广泛运用于各种水体的富营养化监测与评价中.论文对北京市4个校园人工湖的浮游藻类群落组成及季节变化展开调查,探讨了浮游藻类生物指标和富营养化的关系及其季节变化规律.调查期间,镜鉴得到39个属,隶属于蓝藻,绿藻和硅藻门.4个水体生物密度的季节平均值均高于500 cell·mL^-1,多样性指数普遍介于2～3之间.TN,TP和COD显示各水体都已处于富营养化状态.综合生物指标和水质指标进行聚类分析,结果显示:4个人工湖不同季节的水体可分为绿藻型,蓝,绿藻型和硅藻型三大类富营养化水体:荷花池,荷塘浮游藻类群落相似性最高,而荷塘、未名湖夏季富营养化程度最接近.     

微囊藻毒素LR诱导大鼠肾NRK细胞Bax蛋白表达

近年来水体富营养化已成为一个全球性关注的问题,过量的氮、磷使藻类过度生长并产生水华。微囊藻毒素是蓝藻的一些属产生的次生代谢产物,在发生水华的水体中这类毒素普遍存在。微囊藻毒素产生的原因和作用目前还不是很清楚,然而它们对动物和人类都能产生极为严重的影响。       

摇蚊幼虫对底泥中氮、磷释放作用的研究

本文研究了摇蚊幼虫对底泥中氮、磷释放的影响,初步探讨了摇蚊在湖泊富营养化过程中的生态作用。结果表明,摇蚊幼虫能明显促进底泥中氮、磷的释放,而释放到水层中的氮、磷又容易被藻类吸收利用,从而促进藻类生长。这种生态效应与水体营养循环和富营养化的发生及发展过程间存在着重要的关系。     

摇蚊幼虫对底泥中氮、磷释放作用的研究

本文研究了摇蚊幼虫对底泥中氮、磷释放的影响,初步探讨了摇蚊在湖泊富营养化过程中的生态作用。结果表明,摇蚊幼虫能明显促进底泥中氮、磷的释放,而释放到水层中的氮、磷又容易被藻类吸收利用,从而促进藻类生长。这利,生态效应与水体营养循环和富营养化的发生及发展过程间存在着重要的关系。     

Fur from Microcystis aeruginosa binds in vitro promoter regions of the microcystin biosynthesis gene cluster

Promoter regions of the mcy operon from Microcystis aeruginosa PCC7806, which is responsible for microcystin synthesis in this organism, exhibit sequences that are similar to the sequences recognized by Fur (ferric uptake regulator). This DNA-binding protein is a sensor of iron availability and oxidative stress. In the presence of Fe(2+), a dimer of Fur binds the iron-boxes in their target genes, repressing their expression. When iron is absent the expression of those gene products is allowed. Here, we show that Fur from M. aeruginosa binds in vitro promoter regions of several mcy genes, which suggests that Fur might regulate, among other factors, microcystin synthesis. The binding affinity is increased by the presence of metal and DTT, suggesting a response to iron availability and redox status of the cell.     

Abundance of active and inactive microcystin genotypes in populations of the toxic cyanobacterium Planktothrix spp

To investigate the abundance of active and inactive microcystin genotypes in populations of the filamentous cyanobacterium Planktothrix spp., individual filaments were grown as clonal strains in the laboratory and analysed for microcystin synthetase (mcy) genes and microcystin. Twenty-three green-pigmented strains of P. agardhii originating mostly from shallow water bodies fell into two groups, those possessing mcyA and those lacking mcyA. In contrast, all of the 49 strains that were assigned to the red-pigmented P. rubescens contained mcyA. One strain of P. agardhii and eight strains of P. rubescens contained the total microcystin synthetase gene cluster but were found inactive in microcystin synthesis. To investigate the natural abundance of inactive mcy genotypes in P. rubescens individual filaments sampled from Lake Irrsee and Lake Mondsee (Austria) were analysed directly for the presence of mcyA and microcystin by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. All filaments assigned to P. rubescens contained mcyA. The proportion of inactive microcystin genotypes in populations with a low (Irrsee) or high density (Mondsee) of P. rubescens was 5% and 21%, each. The results of this study demonstrate that P. rubescens typically contain mcy genes whereas P. agardhii have a patchy distribution of mcy genes. In both species microcystin producers co-occur with non-microcystin producers due to the absence/inactivation of mcy genes.     

Variation between strains of the cyanobacterium Microcystis aeruginosa isolated from a Portuguese river

AIMS: The aim of this study was to investigate toxicological differences between strains of the cyanobacterium Microcystis aeruginosa isolated from a potable water supply in the north of Portugal over a 2-month period. METHODS AND RESULTS: Twenty-six strains of M. aeruginosa were isolated, grown in pure culture, and tested using a range of techniques including matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), ELISA and a PCR procedure targeting the genes implicated in the production of toxic microcystins. There was considerable variation with respect to the amounts of microcystin produced by each of the strains as measured by ELISA, with values ranging from 0.02 to 0.53% dry weight. The results of the MALDI-TOF MS analysis demonstrated the presence of several chemically distinct forms of microcystin as well as aeruginosins, anabaenopeptins and several other unidentified peptide-like compounds. CONCLUSIONS: The growth of individual strains that comprise bloom populations, with unique 'chemotypes' can potentially be an important factor affecting the toxicity of bloom populations. Molecular probes, targeting the genes responsible for microcystin production were shown to be useful for distinguishing between toxic and nontoxic strains and showed good agreement with the results obtained from the other analyses. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study show that the analysis of cyanobacterial bloom populations at the subspecies (strain) level can potentially provide important information regarding the toxin-producing potential of a cyanobacterial bloom and could be used as an 'early warning' for toxic bloom development.     

EVALUATIONS OF THE AVAILABILITY OF SOURCES OF NITROGEN AND PHOSPHORUS FOR ALGAE1

Techniques are compared for the evaluation of nutrient sources in which the potential nutrients are in contact with algae over their entire culture period versus relatively short-term exposure tests. Different nitrogen and phosphorus compounds were shown to be utilized in darkness as well as in light and were equally available at pH 7 and 9. The availability of relatively insoluble nutrients–iron-phosphorus compounds and teeth for phosphorus, hair for nitrogen iron pyrites for iron, and marble for carbon–indicates that in many cases the equilibrium between soluble and insoluble forms allows algae to compete successfully with insoluble forms for nutrients. Apparently, factors other than mere insolubility prevent algae from obtaining available nitrogen or phosphorus from phosphate rock or lake muds. Live algae or aquatic weeds with adequate or surplus nitrogen or phosphorus were shown to not share their nutrients with nitrogen- or phosphorus-limited algae, whereas the nutrients of killed algae or aquatic weeds were readily used by nutrient-limited algae. The facts–(1) that live algae and aquatic weeds do not share their adequate or surplus nutrients with nutrient-limited algae and (2) that lake muds do not provide readily available nitrogen or phosphorus–indicate that once lake waters are stripped of available nutrients by plant production, further plant production will depend upon nutrients from continuous sources of nutrients, such as wastewater effluents.     

Effects of gibberellin A(3) on growth and microcystin production in Microcystis aeruginosa (cyanophyta)

Environmental factors that affect the growth and microcystin production of microcystis have received worldwide attention because of the hazards microcystin poses to environmental safety and public health. Nevertheless, the effects of organic anthropogenic pollution on microcystis are rarely discussed. Gibberellin A(3) (GA(3)) is a vegetable hormone widely used in agriculture and horticulture that can contaminate water as an anthropogenic pollutant. Because of its common occurrence, we studied the effects of GA(3) on growth and microcystin production of Microcystis aeruginosa (M. aeruginosa) PCC7806 with different concentrations (0.001-25mg/L) in batch culture. The control was obtained without gibberellin under the same culture conditions. Growth, estimated by dry weight and cell number, increased after the GA(3) treatment. GA(3) increased the amounts of chlorophyll a, phycocyanin and cellular-soluble protein in the cells of M. aeruginosa PCC7806, but decreased the accumulation of water-soluble carbohydrates. In addition, GA(3) was observed to affect nitrogen absorption of the test algae, but to have no effect on the absorption of phosphorus. The amount of microcystin measured by enzyme-linked immunosorbent assay (ELISA) increased in GA(3) treatment groups, but the stimulatory effects were different in different culture phases. It is suggested that GA(3) increases M. aeruginosa growth by stimulating its absorbance of nitrogen and increasing its ability to use carbohydrates, accordingly increasing cellular pigments and thus finally inducing accumulation of protein and microcystin.     

微囊藻毒素对水环境的影响研究进展

微囊藻毒素是富营养化淡水水体中最常见的藻类毒素,是湖泊蓝藻产生的一类肽类毒素,它的产生受到藻类的遗传和环境因素的共同影响。由于其毒性大,分布广,结构稳定,从而成为水环境中的潜在危害物质。有关微囊藻毒素性质、毒理毒性、在环境中的迁移、转化以及控制预防已成为关注热点。在总结国内外研究的基础上,综述了微囊藻毒素的性质、产生机理以及其与水环境、水生生物(水生植物、鱼类、无脊椎动物)间的相互作用,讨论了微囊藻毒素对水生生物的影响以及水生生物对微囊藻毒素的降解作用,为水体中微囊藻毒素的防治提供科学的依据。