微囊藻群体形成影响因子及机理

富营养化导致的湖泊、水库蓝藻水华带来了一系列环境和生态问题。微囊藻群体形成,在水体表面的聚集是形成水华的重要策略之一。微囊藻群体形成有效抵御了草食性动物摄食、病毒、细菌的侵害,耐受不良环境因子(紫外辐射、高光强、重金属等)能力显著增强。文章探讨了野外微囊藻群体占优势的机制并综述了影响微囊藻群体形成的外源因子及作用机理,包括非生物因子(营养、温度、光照、重金属、微囊藻毒素、乙醛酸)、生物因子(草食性动物、细菌、鱼类、藻类)。基于现有研究,本综述还对未来研究进行展望:(1)非生物因子与生物因子协同效应对藻类形态影响研究; (2)人类活动对浮游藻类形态及动态的影响研究;(3)藻类形态对沉水植物的响应在未来淡水生态系统修复中的应用; (4)促进藻类群体形成的胞外多糖分泌在未来工业生产中的作用。     

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.     

一株枯草芽孢杆菌的生长特性及抑藻效果研究

溶藻微生物能引起水华蓝藻的快速消亡,从而达到控制水华之目的。为了探讨枯草芽孢杆菌(Bacillus subtilis)对铜绿微囊藻(Microcystis aeruginosa)及小球藻(Chlorell)的抑制效果,在实验室条件下研究了枯草芽孢杆菌对两种藻类生长的影响、抑制藻类生长的作用方式以及发酵液处理下铜绿微囊藻丙二醛(MDA)及谷胱甘肽(GSH)含量、超氧化物歧化酶(SOD)及过氧化氢酶(CAT)活性。试验结果表明,筛选出的菌株具有较好的环境适应性和抑制藻类生长的能力,其抑制效果是通过分泌胞外物质实现的。经过枯草芽孢杆菌处理后,铜绿微囊藻的叶绿素a及藻体蛋白含量均显著低于对照组,而胞内MDA及GSH含量显著升高,SOD及CAT活性也有增高的趋势。     

萼花臂尾轮虫培养滤液对铜绿微囊藻、斜生栅藻和小球藻群体形成及生长的影响

为探索浮游动物和藻类之间可能存在的信息传递,研究了萼花臂尾轮虫培养滤液对铜绿微囊藻、斜生栅藻和小球藻的生长及群体形成的影响.把萼花臂尾轮虫按1000个·L^-1的初始密度置于小球藻中培养24h后,用孔径0.15μm的微孔滤膜抽滤,得到轮虫培养滤液,此滤液中含有轮虫在生活过程中释放的一些信息化学物质.将轮虫培养滤液以20%的比例分别加入纯培养的铜绿微囊藻、斜生栅藻和小球藻中,进行为期7d的试验.结果表明,萼花臂尾轮虫培养滤液能显著地促进斜生栅藻的群体形成,而对铜绿微囊藻和小球藻在群体形成方面没有显著作用.另外,该滤液能显著提高小球藻种群的增长,对铜绿微囊藻和斜生栅藻的生长无明显影响.3种藻类对萼花臂尾轮虫的潜在牧食采取了不同的生态策略:斜生栅藻形成群体,增大摄食阻力,从而降低被摄食的风险;小球藻通过提高增长率来抵消被取食的损失;铜绿微囊藻是通过其它方式来降低被牧食(例如毒素).这些方式分别是这些藻类维持种群规模的反牧食防御策略之一.     

光强对微囊藻群体形态的影响及其生理机制研究

为了探讨光照对微囊藻形态的影响,研究了6株不同种的群体微囊藻在不同光强下群体形态的变化及其响应机制。研究发现,随着光强的增加,6株群体微囊藻的群体尺寸变大。当光强为80200 mol/（m2s）时,群体微囊藻DH-M1和DC-M2的比生长速率显著增大,而另4株在高光强下比生长速率无显著性差异;对多糖含量分析发现,高光强对群体微囊藻TH-M2、DC-M1、FACHB1174和FACHB1027胞外及胶被多糖的分泌与释放有显著的促进效果,而DH-M1和DC-M2多糖含量增加不明显。对于不同的微囊藻株,高光强促进群体形态变化的作用机理不同:光饱和点低的微囊藻是通过分泌大量的胞外及胶被多糖使群体尺寸变大,而光饱和点高的微囊藻是通过生长来促进群体尺寸的增大。此外,对产毒藻株在不同光强下的毒素基因表达及胞内毒素测定发现,高光强组的群体微囊藻mcyB和mcyD表达量升高,且胞内微囊藻毒素含量增加显著,推测微囊藻毒素也可能是影响微囊藻群体形态及大小的作用因子之一。       

光强对微囊藻群体形态的影响及其生理机制研究简

为了探讨光照对微囊藻形态的影响,研究了6株不同种的群体微囊藻在不同光强下群体形态的变化及其响应机制。研究发现,随着光强的增加,6株群体微囊藻的群体尺寸变大。当光强为80—200μmol/(m2·s)时,群体微囊藻DH-M1和DC-M2的比生长速率显著增大,而另4株在高光强下比生长速率无显著性差异;对多糖含量分析发现,高光强对群体微囊藻TH-M2、DC-M1、FACHB1174和FACHB1027胞外及胶被多糖的分泌与释放有显著的促进效果,而DH-M1和DC-M2多糖含量增加不明显。对于不同的微囊藻株,高光强促进群体形态变化的作用机理不同:光饱和点低的微囊藻是通过分泌大量的胞外及胶被多糖使群体尺寸变大,而光饱和点高的微囊藻是通过生长来促进群体尺寸的增大。此外,对产毒藻株在不同光强下的毒素基因表达及胞内毒素测定发现,高光强组的群体微囊藻mcyB和mcyD表达量升高,且胞内微囊藻毒素含量增加显著,推测微囊藻毒素也可能是影响微囊藻群体形态及大小的作用因子之一。     

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。     

棕鞭藻及其培养滤液对铜绿微囊藻生长及生理特性的影响

为探究藻类之间的可能存在的信息传递, 研究了棕鞭藻(Ochromonas sp.)及其培养滤液对铜绿微囊藻的生长及生理特性的影响。结果发现, 3种不同接种比例(1﹕4、1﹕1和4﹕1)的棕鞭藻与微囊藻共培养下, 微囊藻细胞密度到第4天均下降到最低值, 而棕囊藻细胞密度则显著增加。同时, 棕鞭藻培养滤液能够抑制微囊藻的生长、导致丙二醛(MDA)含量和过氧化氢酶(CAT)活性。此外, 棕鞭藻培养滤液也能促进微囊藻胞外多糖(EPS)含量显著增加。这表明棕鞭藻不仅能吞噬微囊藻, 而且可能释放某些化感物质抑制微囊藻生长及生理参数。这暗示了棕鞭藻可作为潜在的藻类水华控制生物, 抑制早期藻类大量增殖。     

三株铜绿微囊藻对外源无机碳利用的研究

本文研究了三株铜绿微囊藻在不同条件下光合放氧对外源溶解无机碳的响应。当温度从20℃上升到30℃时,三株铜绿微囊藻的光合放氧速率都显著增加。随着反应介质pH值从7．0上升到9．0,铜绿微囊藻的光合放氧逐渐增强,其最大光合放氧速率显著增大。胞外碳酸酐酶抑制剂乙酰唑胺（acetazolamide,AZ）对铜绿微囊藻的光合放氧无显著影响,而碳酸酐酶抑制剂乙氧苯丙噻唑磺胺（ethoxyzolamide,EZ）则抑制其光合放氧。通过酶动力学方程拟合,得出这三株铜绿微囊藻的表观无机碳亲和力常数K0.5（DIC）均低于50Lmol／L,显示出铜绿微囊藻对外源无机碳的亲和力较高。在20-30℃范围内,温度的变化对微囊藻无机碳亲和力无显著影响,表明环境温度不影响其对外源无机碳利用的能力。当反应介质pH值升高,铜绿微囊藻FACHB905和PCC7806的K0.5（DIC）增大,而铜绿微囊藻FACHB469的K0.5（DIC）值却减小。在加入AZ和EZ后铜绿微囊藻的K0.5（DIC）值均无显著变化。本文还比较了铜绿微囊藻与莱氏衣藻在介质pH值变化和加入碳酸酐酶抑制剂条件下光合放氧的不同响应。结果表明,莱氏衣藻的光合放氧随反应介质pH值的升高而减弱,而AZ和EZ均抑制莱氏衣藻的光合放氧。当反应介质pH值升高以及加入AZ或EZ后,莱氏衣藻的K0.5（DIC）值均减小。     

铜绿微囊藻的某些生长特性及毒性的研究

藻类毒素的研究随着有毒藻类对人类的危害加剧,越来越受到各国的重视和深入研究。铜绿微囊藻(Microcystis aeruginosaMicrocystis aeruginosa)是世界许多地方报到最多的有毒藻类,对导致其毒素产生的因子及产毒机理的研究,几十年来在温度、pH值、生长年龄、光照强度、质粒、伪空胞与铜绿微囊藻毒性关系做了大量工作,但是对营养元素       

Variation of microcystin content of microcystis aeruginosa relative to medium N:P ratio and growth stage

Changes in the microcystin content of Microcystis aeruginosa UTEX 2388 were investigated at several N:P ratios of the medium and various growth stages. Under the P-fixed condition, the microcystin content of the cells changed with different medium N:P ratios, with the highest at 2748 microg g-1 at a N:P ratio of 16 after incubation for 7 d. The microcystin content of M. aeruginosa exhibited a high correlation with the total N content regardless of an N-fixed or P-fixed culture. When the N:P ratio of the medium was fixed to 16 : 1, the microcystin content of M. aeruginosa at various growth stages was highest at 2191 microg g-1 after an incubation of 4 d and the chlorophyll-a content showed a similar tendency. There was a highly significant relationship between the microcystin content of M. aeruginosa and the chlorophyll-a concentration in the culture during the incubation. Accordingly, the microcystin content of M. aeruginosa during incubation can be easily estimated and monitored by measuring the in vivo fluorescence changes in the culture.     

Nitrogen Effect on Water-Soluble Polysaccharide Accumulation in <Emphasis Type="Italic">Streblonema</Emphasis> sp. (Ectocarpales,Phaeophyceae)

The water-soluble polysaccharides of brown algae attract the increasing attention of researchers as an important class of polymeric materials of biotechnological interest. The sole source for production of these polysaccharides has been large brown seaweeds such as members of Laminariales and Fucales. A new source of water-soluble polysaccharides is suggested here: it is a filamentous brown alga Streblonema sp., which can be cultivated under controlled conditions in photobioreactors that allow obtaining algal biomass with reproducible content and quality of polysaccharides. The accumulation of water-soluble polysaccharides can be stimulated by macronutrient limitation. In response to nitrogen deficiency, Streblonema sp. accumulated water-soluble polysaccharides (WSPs) rich in laminaran. WSP accumulation started after 3–4 days following nitrate depletion and reached a plateau at around day 7. Polysaccharide accumulation was related to cellular nitrogen content. The critical internal N level that triggered the onset of polysaccharide accumulation was 2.3% dry weight (DW); at a cellular N concentration less than 1.4% DW, the polysaccharide synthesis stopped. Upon nitrate re-supply, mobilization of WSP occurred after 3 days. These results suggest that a two-stage cultivation process could be used to obtain large algal biomass with high water-soluble polysaccharide production: a first cultivation stage using nitrate-supplemented medium to accumulate algal biomass followed by a second cultivation stage in a nitrate-free medium for 3 to 7 days to enhance polysaccharide content in the alga.     

光强和氮源对念珠藻胞外多糖分泌的影响

胞外多糖（EPS）是结皮蓝藻形成生物结皮的胶结剂,为了理解常球状存在的丝状蓝藻Nostoc胶结沙粒的机理,探讨了光强40、80 E/（m2s）和氮源（气态氮,硝态氮）对结皮优势种Nostoc sp.分泌EPS（包括荚膜多糖CPS和释放多糖RPS）的影响规律及其内在机理。结果发现:Nostoc sp.在气态氮和硝态氮下都有相似的快速生长,但其分泌的RPS、CPS及EPS量,在硝态氮下均随光强的增加而增加,在气态氮下却与光强没有关系。相关代谢研究发现,在硝态氮下细胞内有更高含量的可溶性糖和蔗糖。进一步的相关分析发现,在两种氮源下,蔗糖量与RPS量或CPS量间的显著正相关都只发生在80 E/（m2s）下,在气态氮中,两光强下的胞内总糖量都与CPS量显著负相关。以上结果说明,Nostoc sp.在氮源利用和光强适应方面都有明显优势,它即使在快速生长的对数期,也可同时分泌相当量的EPS,这使其在球状藻殖段形成之前胶结沙粒成为可能。由此可推知,Nostoc sp.在贫瘠沙土表面的最初生长过程中,其胞外的EPS均来自胞内的固碳产物,在高光强下,蔗糖很可能是其EPS合成的原料。       

黑暗条件下不同氮源对铜绿微囊藻(Microcystis aeruginosa)生长和pH的影响

在黑暗条件下,利用不同形态的氮源(硝酸盐氮,氨氮,有机氮和硝酸盐氮,有机氮)培养蓝藻水华优势种铜绿微囊藻(Microcystis aeruginosa),分析其氮代谢和对水体pH的影响.研究结果表明,在不同氮源的培养液中铜绿微囊藻密度在最初的24 h内出现波动,之后下降.培养液中pH值在试验最初的24 h显著下降,之后趋于稳定,在硝态氮培养液中pH值下降最大,从8.18下降到7.19,其反硝化作用产生的NO-2浓度也最大.不同氮源培养液中总氮含量都有所下降,以混合氮源培养液中总氮减少量最大,说明化合态氮经过反硝化作用生成了氮气并溢出培养液,因此,在黑夜条件下藻华水体中存在反硝化作用.     

Effects of macrograzers and light on periphyton stoichiometry

Ecological stoichiometry describes the biochemical constraints of trophic interactions emerging from the different nutrient content and nutrient demand of producers and consumers, respectively. Most research on this topic originates from well-mixed pelagic food webs, whereas the idea has received far less attention in spatially structured habitats. Here, we test how light as well as grazing and nutrient regeneration by consumers affects growth and biomass of benthic primary producers. In the first laboratory experiment, we manipulated grazer presence (two different snail species plus ungrazed control), in the second experiment we factorially combined manipulation of grazer presence and light intensity. We monitored snail and periphyton biomass as well as dissolved and particulate nutrients (nitrogen and phosphorus) over time. Grazers significantly reduced algal biomass in both experiments. Grazers affected periphyton nutrient content depending on the prevailing nutrient limitation and their own body stoichiometry. In the nitrogen (N-) limited first experiment, grazers increased N both in the periphyton and in the water column. The effect was stronger for grazers with lower N-content. In the phosphorus (P-) limited second experiment, grazers increased the P-content of the periphyton, but the grazer with lower N-content had additionally positive effects on algal N. Light reduction did not affect periphyton biomass, but increased chlorophyll-, N- and P-content of the periphyton. These experiments revealed that the indirect effects of grazers on periphyton were bound by stoichiometric constraints of nutrient incorporation and excretion.     

氮浓度对铜绿微囊藻、大型溞和金鱼藻三者相互作用的影响

为了解氮浓度对生物操纵和草-藻竞争的影响, 选取铜绿微囊藻、大型溞和金鱼藻分别作为浮游植物、浮游动物和沉水植物的代表, 在温度25℃, 光强2600 lx, 光暗比14h﹕10h, 磷浓度1.5 mg/L时, 研究5种氮浓度(0.5、2、4、8和16 mg/L, 用KNO₃溶液配制)下, 溞-藻, 草-藻和溞-草-藻共培养时各自的增长率和培养液中氮磷削减率的变化。结果表明: 在单独培养铜绿微囊藻时, 氮浓度控制在1.97 mg/L以下, 可有效降低培养液中藻的增长率。在溞-藻共培养时, 大型溞有效控藻的氮浓度范围为0.5—4 mg/L; 在草-藻共培养时, 有效控藻的氮浓度范围为0.5—2 mg/L, 对应氮浓度下(0.5和2 mg/L), 实验末期铜绿微囊藻细胞密度分别是溞-藻共培养的23.89%和21.51%, 控藻效果更好; 在溞-草-藻三者共培养时, 有效控藻的氮浓度范围为0.5—16 mg/L, 且氮浓度为0.5—4 mg/L时, 大型溞和金鱼藻的增长率均显著大于铜绿微囊藻, 铜绿微囊藻的增长率均为负值, 控藻效果最好。大型沉水植物的加入, 可以有效提高生物操纵的控藻效果, 减少水中氮磷含量, 长期有效地改善水质。     

Microalgae-mediated chemicals production and wastes removal

Biotechnology of microalgae has gained importance in recent years due to the development of new production and environmental technologies. Because their growth requires unexpensive substrates such as solar light and CO₂, microalgae can be used as cheap and effective biocatalysts to obtain high added-value compounds, from simple metabolites to complex molecules, i.e., chemicals, vitamins, carotenoids, pigments, or polysaccharides. During productive processes, the algal biomass formed may be used as a food source like proteins. On the other hand, microalgae can also be employed in contaminant bioelimination processes especially for nitrogen, phosphorus, or sulfur compounds. Particularly relevant is their use for heavy metal removal from wastewaters; upon enriching the biomass in the metal, they can be recovered, thereby providing economic advantages.

The use of immobilized microalgae in these processes is very adequate and offers significant advantages in bioreactors.     

施肥对短枝木麻黄幼苗总酚和可溶性缩合单宁含量的影响

研究了施用氮肥和磷肥对短枝木麻黄幼苗总酚(total phenolics,TP)和可溶性缩合单宁(extractable condensed tannin,ECT)含量的影响,探讨短枝木麻黄单宁形成的养分效应.结果表明:施加氮肥使短枝木麻黄幼苗小枝的TP和ECT含量显著降低,支持碳氮平衡假说和生长分化平衡假说,但对氮含量没有显著影响,从而导致TP/N和ECT/N降低;施加磷肥对TP和ECT含量没有显著影响;随着处理时间的延长,短枝木麻黄幼苗小枝TP含量升高了9.91%～14.32%,而ECT含量降低了14.32%～298.88%;TP或ECT与有机物质含量的关系则相反,表明不同类型单宁的合成途径不同,但由于TP和ECT均与氮含量无显著相关性,故不支持蛋白质竞争模型;在贫瘠土壤条件下,TP/N和ECT/N的水平较高,有利于提高短枝木麻黄的防御水平,降低凋落物的分解率,减少养分损失,从而保持较高的生产力.     

The ecological stoichiometry of toxins produced by harmful cyanobacteria: an experimental test of the carbon-nutrient balance hypothesis

The elemental composition of primary producers reflects the availability of light, carbon and nutrients in their environment. According to the carbon-nutrient balance hypothesis, this has implications for the production of secondary metabolites. To test this hypothesis, we investigated a family of toxins, known as microcystins, produced by harmful cyanobacteria. The strain Microcystis aeruginosa HUB 5-2-4, which produces several microcystin variants of different N:C stoichiometry, was cultured in chemostats supplied with various combinations of nitrate and CO₂. Excess supply of both nitrogen and carbon yielded high cellular N:C ratios accompanied by high cellular contents of total microcystin and the nitrogen-rich variant microcystin-RR. Comparable patterns were found in Microcystis -dominated lakes, where the relative microcystin-RR content increased with the seston N:C ratio. In total, our results are largely consistent with the carbon-nutrient balance hypothesis, and warn that a combination of rising CO₂ and nitrogen enrichment will affect the microcystin composition of harmful cyanobacteria.     

Differences in growth,pigment composition and photosynthetic rates of two phenotypes Microcystis aeruginosa strains under high and low iron conditions

This paper provided insight into the influence of iron on the growth of Microcystis aeruginosa strains related to different phenotypes of this species. In this research it was intended to compare the growth, pigment composition, photosynthetic efficiency and extracellular polysaccharides production of unicellular and colonial strains of M. aeruginosa. A significantly growth inhibition under iron-limited condition on unicellular M. aeruginosa was noted, whereas the colonial strain could maintain a steady growth along with the culture time. This observation was reconfirmed by the content of chlorophyll a. Compared with unicellular strain; the colonial strain exhibited a higher PSII maximum light energy transformation, photosynthetic oxygen evolution and extracellular polysaccharides (EPS) production in iron-limited condition. Further, in order to gain more information about the accessibility of iron in the two phenotypic Microcystis, we found the two strains could produce hydroxamate-type siderophores, the content of siderophores produced by the colonial strain was more than those in unicellular strain under the iron-limited condition. It was interpreted as an adaptation to the dilute environment. Our results demonstrated that the colonial phenotypes possessed stronger ability to endure iron-limited condition than unicellular strain by higher pigment contents, higher photosynthetic activities, higher EPS production and higher siderophores secretion. It might elucidate that the colonial M. aeruginosa bloom can sustain in eutrophic reservoirs and lakes.