铜绿微囊藻和斜生栅藻非稳态营养盐限制条件下的生长竞争特性

采用“脉冲”添加方法进行了非稳态条件下铜绿微囊藻（M.）和斜生栅藻（S.）分别在氮磷单营养盐和双营养盐限制时的共培养试验。试验结果显示：当添加频率为1d时,无论何种营养盐限制,M.均成为优势藻种。氮限制条件下,氮时均浓度范围在0.3—2.4 mg/L时,M.始终具有竞争优势。磷限制条件下,磷浓度范围在0.018—0.035 mg/L时,S.只在生长初期阶段占优。氮磷双营养盐限制条件下,添加液的氮磷质量比为35:1（设定为最优比）,添加频率为8 d时,两种藻表现出共生特征;而偏离最优比时（N:P=70:1,17:1）,在不同的添加频率下均未出现共生现象,且氮的时均浓度为0.6—4.8 mg/L时（70:1）,M.具有竞争优势,而降低为0.15—0.3 mg/L时（17:1）,S.占优。随着添加频率的变化,两种藻的细胞大小也会随之改变,S.随着营养盐浓度的降低而增大,且在双营养盐限制条件下变化更显著。上述试验结果分析表明：两种藻竞争能力与添加频率相关,在藻种浓度的变化上,按照‘拾遗－机会’交替竞争理论,M.表现出机会主义者特征,而S.则表现出拾遗者的特征,两者的共生特征也符合‘中度干扰’假说。藻细胞大小变化表明,两种藻均可以改变大小实现最大限度争夺受限营养盐。在低浓度时,S.细胞大小的变化同样也变现出了“拾遗者”的特征。     

铜绿微囊藻和四尾栅藻在不同温度下的生长特性及竞争参数计算

在实验室条件下,对铜绿微囊（Microcystis aeruginosa）和四尾栅藻（Scendesmus quadricauda）分别在温度22℃、26℃和30℃下进行了纯培养和混合培养,实验结果显示温度对两种藻类的生长和竞争都有显著影响。微囊藻在22℃、26℃和30℃纯培养下的最大生物量及平均特定增长率（μ）分别为444、1180和998（×104cells/mL）及0.33、0.38和0.37/d,说明微囊藻在26℃和30℃下生长较好;混合培养下的最大生物量及平均特定增长率分别为270、778和647（×104cells/mL）及0.34、0.43和0.46/d,显示微囊藻在混合培养下受到了栅藻一定程度的竞争抑制。栅藻在22℃、26℃和30℃纯培养下的最大生物量及平均特定增长率分别为830、984和464（×104cells/mL）及0.36、0.34和0.32/d;混合培养下的最大生物及平均特定增长率分别为538、554和387（×104cells/mL）及0.43、0.40和0.39/d,说明栅藻在温度22℃、26℃下生长较好,混合培养下栅藻的生长受微囊藻影响较大。各温度下两种藻类的生长都可以用Logistic方程拟合。微囊藻对栅藻的抑制参数α分别为1.68（22℃）、0.65（26℃）和0.76（30℃）,而栅藻对微囊藻的抑制参数β依次为0.43（22℃）、0.51（26℃）和0.25（30℃）,三个温度下α均大于β,产生这一结果的原因可能是由于微囊藻与栅藻在竞争过程中不仅表现为资源竞争,同时还存在着明显的他感作用,且可推测微囊藻对栅藻的他感作用大于栅藻对微囊藻的,且在22℃时为最大。     

水生花卉对铜绿微囊藻、斜生栅藻和小球藻生长的影响

选择黄菖蒲(Iris pseudacorus)、溪荪(I.sanguinea)、梭鱼草(Pontederia cordata)、白花水龙(Jussiaea repens)、水罂粟(Hydrocleys nymphoides)和大藻(Pistia stratiotes)6种具有较高观赏价值的水生花卉,通过将植物种植水与藻类共同培养的方式研究了不同种植时间的种植水对铜绿微囊藻(Microcystis aeruginosa)、斜生栅藻(Scenedesmus obliqnus)和小球藻(Chlorella vulgaris)生长的影响.结果表明:6种水生花卉种植水对3种藻类的化感作用具有选择性.通过6d的处理,种植水对铜绿微囊藻生长的抑制率为31.22％ ～ 96.53％,除白花水龙外,其余5种花卉的种植水对铜绿微囊藻生长的抑制率均超过70％,表现出很好的抑藻效果;种植水对斜生栅藻生长的抑制率为23.15％～77.25％;而种植水对小球藻有抑制也有促进,抑制率为-26.07％ ～75.70％,大藻、梭鱼草和溪荪抑制小球藻的生长,黄菖蒲、白花水龙表现为低促高抑,水罂粟表现为促进作用.随着种植时间的延长,种植水对3种藻类的抑制作用增强.6种水生花卉种植水对铜绿微囊藻生长的抑制作用由大到小依次为水罂粟＞黄菖蒲＞梭鱼草＞大藻＞溪荪＞白花水龙;对斜生栅藻生长的抑制作用由大到小依次为梭鱼草＞溪荪＞黄菖蒲＞水罂粟＞白花水龙＞大藻;对小球藻生长的抑制作用由大到小依次为大藻＞梭鱼草＞溪荪＞黄菖蒲、白花水龙＞水罂粟.试验表明,6种水生花卉在控制城市景观水体中的藻类水华有一定的推广价值.     

微囊藻和栅列藻光吸收特性的实验研究

在室内模拟槽中,以人工灯为光源,分别检测不同密度的微囊藻和栅列藻细胞悬液的光谱衰减,检测各样品的叶绿素含量和过滤后清液的溶解性有机碳(DOC)含量,并在紫外分光光度计上检测DOC光谱吸收,然后换算成吸收系数。再根据近似的线性加和关系计算出细胞悬液中各组分(藻细胞、水、DOC)的吸收系数。结果表明,微囊藻和栅列藻光衰减的波谱特征相似,在3个较高密度下的衰减系数的变化趋势相近,其衰减系数贡献率的平均值分别为84.95%和88.15%。将衰减系数除以各自的叶绿素浓度即可得到叶绿素的比吸收系数,两种藻3个较高密度的比吸收系数的平均值具有相似的波谱特征,在380～500、660～690nm两个波段有较明显的峰值,栅列藻和微囊藻在435nm的比吸收系数峰值分别为0.1118、0.0413(m2·mg-1chla),在670nm处的峰值分别为0.075、0.032(m2·mg-1chla)。但两者的比吸收系数差异较大,三个密度栅列藻在340～800nm的平均比吸收系数是0.0573(m2·mg-1chla),而微囊藻仅为0.0234(m2·mg-1chla),且微囊藻的所有波长的比吸收系数均小于栅列藻。根据这些参数来讨论两种藻类在分光衰减中的贡献率和光谱特征方面的差异及其生态学意义,以及在浮游植物的遥感监测方面的参考价值。     

角突网纹溞培养滤液对铜绿微囊藻和栅藻形态及生长率的影响

在藻类中,随着环境条件的变化而产生相应的单细胞和群体之间的形态转化是一种广泛存在的现象,称为表型可塑性。例如,栅藻属成员就是这些藻类的代表,在野外具有典型的群体特征,但在室内纯培养条件下栅藻常较少形成群体而是保持以单细胞形态为主。     

绿藻栅藻对微囊藻的抑制效应及评价

为筛选具有控制微囊藻水华潜力的绿藻, 研究基于藻类之间的化感作用, 对34株绿藻开展筛选及评价工作。结果发现, 栅藻FACHB-1229的抑制率最高, 其滤液对微囊藻FACHB-3550和FACHB-905的抑制率分别为53.95%和48.39%; 通过对滤液中的物质进行GC-MS检测, 推测该藻株的效应物质可能为邻苯二甲酸二(2-甲氧基乙基)酯。进一步对FACHB-1229生长和光合放氧、氨氮耐受等指标进行评价, 结果发现: 该藻株在孔板中的生长速率(0.38±0.06)/d高于微囊藻生长速率(0.13±0.03)/d; 栅藻FACHB-1229的光合放氧速率高于同期测定的其他绿藻光合放氧速率, 可达(229.91±10.49) μmol O₂/(mg Chl.a·h); 同时该藻株对氨氮的耐受能力最强, 在氨氮浓度为1888.60 mg/L时其生长速率可达(0.30±0.08)/d。此外, 在与微囊藻FACHB-3550进行共培养时, 栅藻FACHB-1229所占比例持续升高。综上, 栅藻FACHB-1229具备与微囊藻竞争的优势, 有望成为微囊藻水华生物控制的优选材料。     

微囊藻和栅列藻的垂直迁移及生态学意义

本研究在自制沉降柱中,检测铜绿微囊藻(Microcystis aeroginosa)和斜生栅列藻(Scenedesmus obliquus)不同生长时期的细胞群体在不同光照条件下的垂直迁移。本文首次提出藻密度衰减系数(K)的概念、拟合方法和生态学意义,并根据等细胞密度沉降面的沉降速率来计算细胞沉降速率(Vc),以这两个参数来衡量细胞的沉降特性,对细胞垂直迁移研究的方法进行了的探索。结果表明,两种藻细胞群体在三个生长时期和四种光照条件的沉降特性表现已定的差异。栅列藻细胞密度的衰减系数(K)最小值为-0.0021cm^-1(上浮),最大值为0.0065cm^-1(下沉);微囊藻的K最小值为-0.0029cm^-1(上浮),最大值为0.0036cm^-1(下沉)。两种藻沉降曲线的指数拟合的R²大多在0.6以上,说明K值是一个较好的衡量细胞群体沉降特性的参数,细胞密度与深度的关系较好地服从指数规律。细胞沉降速度V的最大值(4.681cm·h^-1,下沉)出现在栅列藻的指数期,最大上浮速率-1.790cm·h^-1,出现在微囊藻的稳定期。本文得到的活体藻细胞群体的垂直迁移特性差异的实验数据,说明该实验方法的适用性和两种藻沉降特性差异的可检测性。本文讨论了水华优势种形成与藻细胞群体迁移特性之间的关系。     

从种群竞争的角度初步研究微囊藻的产毒机理

采用将微囊藻和栅藻混合培养的方法,从种群竞争的角度初步探讨了微囊藻毒素的产毒机理,结果显示：当起始接种浓度相同时,混合培养组比纯微囊藻培养组产生更多的毒素,由于其它培养条件完全一致,所以推论是由于栅藻的存在,增加了微囊藻的生存压力。当起始接种浓度微囊藻：栅藻为10：1时,此混合培养组比纯微囊藻产生的毒素少,并且毒素降解也更快,推论原因是微囊藻在种群数量上远远超过栅藻,竞争压力较小,同时由于栅藻的存在,增加了培养液中色素的含量,加快了光降解的速度。     

普生轮藻浸提液对两种淡水藻类的化感抑制作用及其数学模型

利用大型水生植物的化感作用抑制水华藻类是水域生态学研究的热点课题之一。探讨了不同浓度普生轮藻浸提液对产毒铜绿微囊藻和斜生栅藻(单纯以及混合藻类)的抑制作用,并根据实验过程中得到的数据和数据特征,在传统的Logistic模型和Lotka-Volterra模型基础上,通过微元法建立了普生轮藻浸提液对单纯产毒铜绿微囊藻、单纯斜生栅藻抑制的数学模型以及两藻混合时抑制的数学模型。结果表明,(1)普生轮藻浸提液无论对单独的毒性铜绿微囊藻或斜生栅藻还是共生状态的毒性铜绿微囊藻和斜生栅藻均有很强抑制作用,且对毒性铜绿微囊藻的抑制作用要显著高于对斜生栅藻;(2)所建立的抑藻模型可有效表征和预测在一定范围内的产毒铜绿微囊藻、斜生栅藻及其混合藻在普生轮藻浸提液胁迫下藻密度随时间变化的规律;通过这些模型可方便地计算出实验期间任何时间节点上普生轮藻浸提液的半抑制浓度(EC50)、最小有效浓度(MIC)等指标的预测值、混合藻在小生境中相对稳定时的预测值等等。该研究可为实际抑藻的方案制定和实施提供有价值的数据支撑和参考,具有一定的理论与应用意义。     

蓖齿眼子菜对栅藻和微囊藻的他感作用及其参数

主要报道篦齿眼子菜 (俗称红线草 )的种植水抽滤液对栅藻、微囊藻生长的他感作用试验。应用修正的逻辑斯谛 (Logistic)方程和Lotka Volterra种群竞争模型来计算篦齿眼子菜对藻类的他感作用参数。纯培养试验结果表明 :种植水抽滤液对栅藻生物量和生长率都有一定的促进作用。当浓度大于等于 5 0 %时 ,对栅藻生长的影响则是有抑制效应 ,他感作用效应与种植水的浓度没有明显的相关性 ,他感作用参数平均为 - 0 0 2 6。所有浓度设对照。微囊藻的生长都有明显的抑制作用 ,他感作用参数平均为 - 0 2 37。共培养试验结果表明 :篦齿眼子菜种植水抽滤液对纯培养试验结果有放大效应。所有的种植水抽滤液浓度对栅藻生长都起促进作用 ,对栅藻的生物量效应、生长率存在明显的对数正相关 ,但与他感作用参数的相关不明显 ,共培养的他感作用参数平均为 0 0 2 0 ;对微囊藻生长抑制作用更加明显 ,共培养的他感作用参数平均为 - 0 4 2 0。不管是纯培养还是共培养中 ,种植水抽滤液对栅藻的他感作用系数值都很低 ;而种植水抽滤液浓度对微囊藻的生物量效应、生长率或他感作用的参数都存在明显的线性负相关 ,并且他感作用系数值约高一个数量级     

Heptapeptide toxin production during the batch culture of two Microcystis species (Cyanobacteria)

Changes in the content of cyclic heptapeptide hepatotoxins called microcystins were investigated during batch culture of two Microcystis species using high performance liquid chromatography. After adsorption to ODS-silica gel cartridges and elution with methanol, the toxins were analyzed and quantified by HPLC. 35 μg per 100 mg dry cells of microcystin-RR, 34 μg of -YR and 43 μg of -LR were present at the beginning of the exponential growth phase of M. viridis. Microcystin-RR increased markedly towards the end of the exponential phase with the maximum content of 112 μg per 100 mg cells was measured at the late stage of the exponential phase. A remarkable increase of microcystin-YR from 130 μg per 100 mg cells to 1020 μg was observed during the exponential phase of a highly toxic strain of M. aeruginosa. However no clear differences were found in the pattern of change among the three toxins during the growth course.     

Effect of dilution rate on competitive interactions between the cyanobacterium Microcystis novacekii and the green alga Scenedesmus quadricauda in mixed chemostat cultures

We examined the competition between the cyanobacterium Microcystisnovacekii (Kom.) Comp. and the green alga Scenedesmus quadricauda(Turpin) Brébisson using unialgal and mixed chemostatcultures with various supply rates of culture medium where limited algal growth. In unialgal cultures, bothspecies grew at all of the dilution rates examined (0.1, 0.3and 0.8 day^-1): steady-state cell densities were 1 x 10⁴ to8 x 10⁴ cells mL^-1 for M. novacekii and 0.5 x 10⁵ to 2.1 x 10⁵cells mL^-1 for S. quadricauda. Microcystis novacekii was dominantin mixed cultures at a dilution rate of 0.1 day^-1, where thesteady-state cell density was 1 x 10⁴ to 7 x 10⁴ cells mL^-1for M. novacekii and 1 x 10² to 5 x 10² cells mL^-1 for S. quadricauda.Scenedesmus quadricauda was dominant in mixed cultures at thehigher dilution rates (0.3 and 0.8 day^-1), where the final celldensity was 0.5 x 10² to 6.4 x 10² cells mL^-1 for M. novacekiiand 0.2 x 10⁵ to 7 x 10⁵ cells mL^-1 for S. quadricauda. Thisresult indicates that the dilution rate affects the competitiveinteraction. We conclude that it is necessary to consider waterexchange in the study of mechanisms of cyanobacterial blooms.     

First report on the allelopathic effect of Tychonema bourrellyi (Cyanobacteria) against Microcystis aeruginosa (Cyanobacteria)

The allelopathic effect of the cyanobacterium Tychonema bourrellyi against the cyanobacterium Microcystis aeruginosa is reported for the first time in this paper. The filtrate of T. bourrellyi CHAB663 culture showed strong inhibitory effect on M. aeruginosa NIES-843, but the inhibitory effect was weakened by shaking culture, and such results implied that the allelopathic effect was probably mediated by the volatile substances secreted by T. bourrellyi. β-Ionone was identified as a major ingredient in the volatile substances in the cultures of T. bourrellyi, and it may act as an important allelochemical responsible for this allelopathic activity. The filtrates of T. bourrellyi culture were shown to decrease the maximum electron transport rate (ETR_max) and elevate the reactive oxygen species (ROS) levels in the cells of M. aeruginosa NIES-843.     

Role of Cell Hydrophobicity on Colony Formation in Microcystis (Cyanobacteria)

Colony formation is highly import ant for the competitive advantage of the cyanobacterium Microcystis over other phytoplankton species. The laboratory‐grown colonial Microcystis strains isolated from Lake Taihu (China) maintained colonial forms under the low light condition (10 μE m^–2 s^–1). The cell surface hydrophobicities of the Microcystis colonies were measured by cyanobacterial adherence to xylene in comparison with unicellular Microcystis strains. The cells of the tested colonial strains were all hydrophobic, while the cells of the tested unicellular strains were all hydrophilic. Incubation under the higher light condition (75 μE m^–2 s^–1) leaded to the significant decrease in the cell hydrophobicities of the colonial Microcystis and the transition from colonial forms to unicellular forms. These findings indicated that the cell hydrophobicity of Microcystis may play a role in cell‐cell adherence and colony formation. Phosphate‐limitation, nitrate‐limitation and pH did not affect cell hydrophobicities of colonial Microcystis. Treatment with proteolytic enzymes had no effect on the cell hydrophobicity, indicating that cell surface proteins did not contribute to high cell hydrophobicity. (© 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Urease of blue-green algae (Cyanobacteria)Anabaena doliolum andAnacystis nidulans

Urease fromAnabaena doliolum andAnacystic nidulans showed maximum activity at pH 7.0–7.4 at 40°C when measured in cell-free, phosphate-buffered extracts. It is a soluble enzyme located in cytoplasm. The apparent K_m forA. doliolum urease was 120 M. Anacystis nidulans urease exhibited biphasic kinetics (K_m=250 M and 1.66 mM). Enzyme, fully expressed in cells grown with urea, nitrate, or N₂, was repressed in ammonia-grown cells, but ammonia did not inhibit the activity in vitro. Incubation of algal cells in N₂ medium with chloramphenicol for 12 h caused degradation of urease. Cu²⁺ at 1 M inhibited the enzyme activity by 50%, whereas Co²⁺ and Ni²⁺ up to 20 M had no effect.p-Hydroxymercuribenzoate appeared to be a more powerful inhibitor of urease than acetohydroxamic acid.Address reprint requests to: c/o Prof. Robert Tabita, Department of Microbiology, Experimental Science Building #319, The University of Texas at Austin, Austin, TX 78712, USA.     

Development of a Solid Medium for Growth and Isolation of Axenic Microcystis Strains (Cyanobacteria)

Solid media on a base of B-12 or CB medium with agarose or agarose of low melting temperature were developed for the cultivation of Microcystis species. The media with 0.4% gel showed the highest number of CFU, and increasing the gel concentration resulted in a reduction of the number of CFU. There was no difference in the numbers of CFU between pour and spread plates made of the solid media. By using the solid media, 31 clones of Microcystis species were isolated from natural blooms in Lake Kasumigaura, and 5 axenic strains (1 of M. wesenbergii and 4 of M. aeruginosa) were established from the clones.     

Genetic Diversity： Geographical Distribution and Toxin Profiles of Microcystis Strains （Cyanobacteria） in China

Twenty strains of Microcystis Kütz were isolated from different freshwater bodies in China to analyze the diversity, geographical distribution and toxin profiles. Based on whole-cell polymerase chain reaction of cpcBA-IGS nucleotlde sequence, the derived neighbor-joining （NJ） and maximum parsimony （MP） trees Indicate that these strains of Microcystis can be divided into four clusters. The strains from south, middle and north region of China formed distinct lineages, suggesting high diversity and a geographical distribution from south to north locations. Moreover, the results being indicating high variable genotypes of the strains of the Microcystis strains from the same lake show that there Is high diversity of Microcystis within a water bloom population. Comparing the results of the present study with those reported for compared with 43 strains of Microcystis from other locations, also reveals Chinese strains have high similarity with those from regions in the North Hemispherical. This suggests that the Microcystis strains In the world might have a geographical distribution. Analysis of 30 strains using the primers MCF/TER and TOX2P/TOX2M showed that there was no correlation between the gene of cpcBA-IGS and the presence of racy. Toxic strains were founded to be predominant in different water bodies throughout China.