无磷条件诱导铜绿微囊藻(Microcystis aeruginosa)释放挥发性有机化合物对莱茵衣藻(Chlamydomonas reinhardtii)的影响

为了揭示磷(P)营养缺乏对蓝藻释放挥发性有机化合物(VOCs)的影响及其对其他藻类的化感作用,以形成蓝藻水华的主要种类铜绿微囊藻(Microcystis aeruginosa)为材料,在无P培养条件下对其释放的VOCs进行分析,同时测定VOCs对莱茵衣藻(Chlamydomonas reinhardtii)生长、光合色素含量和光合性能的影响.结果表明,采用无P培养基培养铜绿微囊藻24 h后,其释放的VOCs种类和含量均明显增加,与标准培养基培养相比,VOCs总释放量增加了73.4%,并出现7种新化合物.将铜绿微囊藻释放的VOCs通入莱茵衣藻溶液中,在标准培养基中铜绿微囊藻释放的VOCs对莱茵衣藻生长无显著影响,而无P条件下释放的VOCs则明显抑制莱茵衣藻生长,其响应指数(RI)为-0.25.此外,莱茵衣藻光合色素含量、光系统II(PSII)最大光化学量子产量(Fv/Fm)、有效光化学量子产量[Y(II)]、光化学淬灭系数(q P)和光合电子传递速率(ETR)也明显降低,而非光化学淬灭系数(NPQ)则明显升高,其RI为0.26.由此可见,蓝藻在富营养化水体中大量繁殖以及P自身沉降特性导致的P缺乏会促进蓝藻释放VOCs,同时这些VOCs在保持蓝藻营养竞争优势和水体藻类多样性减少中具有化感抑制作用.     

有毒铜绿微囊藻对鱼和溞的毒性

用有毒铜绿微囊藻细胞及其从毒细胞中提取的毒素喂养和腹腔注射鱼类，结果表明：（1）微囊藻毒素MCYST－LR对所有实验过的鱼有毒害作用（LD_(50)为110－500μg／kg）．注射毒素24h后，鱼肝电镜超微结构观察均表现出肝损伤．（2）在受控条件下，当用有毒铜绿微囊藻作为食物喂养日鲢和罗非鱼时，鱼类能在以毒藻为唯一食物的环境中生活20d以上．白鲢和罗非鱼粪便的毒性比喂食藻类的毒性分别降低约2—4．5倍，其肝的超微结构现察仍有轻度损伤．（3）在培养液中加入15μg／mLMCYST－LR和用实验室纯培养的铜绿微囊藻作为食物培养枝角类时，均可使它们在48h死亡．以无毒微囊藻培养时则生长不良亦不怀卵，表明它不是枝角类合适的食物．     

不同温度下水流对铜绿微囊藻生长的影响模拟

在蓝藻暴发机理研究中,水动力是一个非常重要的参数,目前多数研究在模拟各种流速环境的同时,未能实现实验过程中恒定的光、温条件控制.本研究采用小型环形槽模拟不同水体流速,在恒定的光、温条件下研究铜绿微囊藻(Mi-crocystis aeruginosa905)在流动水体中的生长情况,并对比了两种典型温度(15℃和25℃)条件下的差异.结果表明,水流使微囊藻生长滞后,改变其对数生长期持续时间和比增长率,直接影响微囊藻的最大生物量;不同流速的水流对微囊藻生长的影响与一定温度范围内的温度条件有关,15℃时,水体流动不利于微囊藻的生长繁殖;而在25℃时,微囊藻细胞生理活性增加,微囊藻对水体流动环境适应能力增强,水体流动更有利于微囊藻的生长繁殖,且流速在15cm/s时微囊藻获得最大生物量.     

有毒铜绿微囊藻对鱼和溞的毒性

用有毒铜绿微囊藻细胞及其从毒细胞中提取的毒素喂养和腹腔注射鱼类,结果表明:(1)微囊藻毒素MCYST-LR对所有实验过的鱼有毒害作用(LD₅₀为110-500μg/kg).注射毒素24h后,鱼肝电镜超微结构观察均表现出肝损伤.(2)在受控条件下,当用有毒铜绿微囊藻作为食物喂养日鲢和罗非鱼时,鱼类能在以毒藻为唯一食物的环境中生活20d以上.白鲢和罗非鱼粪便的毒性比喂食藻类的毒性分别降低约2-4.5倍,其肝的超微结构现察仍有轻度损伤.(3)在培养液中加入15μg/mLMCYST-LR和用实验室纯培养的铜绿微囊藻作为食物培养枝角类时,均可使它们在48h死亡.以无毒微囊藻培养时则生长不良亦不怀卵,表明它不是枝角类合适的食物.     

不同藻类对温度与磷叠加作用的响应模式

气候变暖对富营养化引起的蓝藻水华扩张具有叠加作用,为探索不同藻类对气候变暖与富营养化叠加作用的响应模式,选用富营养化水体常见藻类铜绿微囊藻(Microcystis aeruginosa)、水华鱼腥藻(Anabaena flos-aquae)和斜生栅藻(Scenedesmus obliquus)作为材料,采用室内培养方法,模拟春末不同升温幅度(20+3、23+3和20+6℃),5个磷浓度水平(0、0.05、0.15、0.30和0.50 mg/L),通过生长速率、光化学参数等的测定,分析不同藻类对温度升高和营养盐添加的生长与光化学响应.结果表明:3种藻的生长速率对温度和磷的叠加作用呈现不同的响应模式:随着磷浓度的升高,铜绿微囊藻的生长速率受温度的叠加作用不断增加,26℃时差异最大;水华鱼腥藻生长速率除在0.30 mg/L磷浓度时受温度叠加作用比较明显外,其他磷浓度水平均没有显著的叠加效应;斜生栅藻在不同磷浓度水平下受温度叠加作用影响有较大波动,但无明显差异.铜绿微囊藻对温度的叠加作用随着磷浓度的增加呈现逻辑斯蒂模式增长,而水华鱼腥藻和斜生栅藻均是在特定温度或特定营养盐浓度时呈现最大的叠加作用,叠加作用的趋势不明显.从Fv/Fm结果可知,铜绿微囊藻下降幅度明显高于水华鱼腥藻和斜生栅藻,随温度升高,磷浓度越高,铜绿微囊藻的Fv/Fm下降幅度越大,这可能是铜绿微囊藻在高生长速率下色素无法积累所致;水华鱼腥藻的Fv/Fm基本保持稳定,各组间差异较小;斜生栅藻的Fv/Fm在低磷浓度组明显低于高磷浓度组,并且随温度升高下降趋势增加,3种藻的光学特性响应反映了藻类在叠加作用下生长的变化.综上所述,铜绿微囊藻对温度上升与磷浓度的升高有更强的响应,导致其产生更高的生长速率,从而促进了蓝藻水华的扩张.     

温度对铜绿微囊藻(Microcystis aeruginosa)和鱼腥藻(Anabaena sp.)生长及胞外有机物产生的影响

以巢湖优势种铜绿微囊藻(Microcystis aeruginosa)和鱼腥藻(Anabaena sp.)为研究对象,研究不同温度(35、25和10℃)对这两种藻生长特性和胞外有机物产生的影响.结果表明,温度对铜绿微囊藻和鱼腥藻的藻细胞密度、碱性磷酸酶活性和胞外有机物浓度影响显著.25℃是铜绿微囊藻和鱼腥藻最适宜的生长温度,最高细胞密度分别达到3.12×107cells/ml和2.03×107 cells/ml.不同温度下两种藻的碱性磷酸酶活性特征,证实了高温对鱼腥藻生长的抑制和低温对铜绿微囊藻生长的抑制.胞外有机物释放总量受蓝藻生物量和单位细胞有机物释放速率的影响.铜绿微囊藻的溶解性有机碳和胞外总多糖释放量在25℃最高,最大值分别为49.28和38.46 mg/L;而鱼腥藻在35℃时释放量最高,最大值分别为45.82和40.60 mg/L;10℃条件抑制了两种藻的生长及胞外有机物的释放.鱼腥藻胞外多糖含量在35℃培养条件下最高,而铜绿微囊藻在10℃条件下最高,说明不利的生长条件会促进蓝藻胞外多糖的分泌.三维荧光图谱分析结果表明,铜绿微囊藻和鱼腥藻胞外有机物以类蛋白质和类腐殖酸为主,温度主要影响藻细胞胞外有机物浓度,而对有机物种类组成没有影响.     

微囊藻和栅列藻吸收与散射特性的实验研究

利用分光光度计,分别检测不同浓度梯度的微囊藻和栅列藻的光束衰减系数、吸收系数等。根据线性加和关系计算出2种藻的散射系数。结果表明,400～650 nm光束衰减系数大致呈下降趋势,之后又逐渐上升.在675 nm附近出现由于浮游植物吸收而形成的峰值,微囊藻、栅列藻的c*(675)分别为(0.075±0.007)m~(-1)、(0.079±0.007)m~(-1)。微囊藻、栅列藻吸收系数、比吸收系数在440、675 nm存在明显的吸收峰值,PAR波段积分平均比吸收系数分别为0.0172、0.0178 m~2/(mgCh1a)。散射系数在440、675 nm一般出现谷值,而在550、700 nm一般为峰值。微囊藻、栅列藻PAR波段积分平均比散射系数分别为0.0686、0.0737 m~2/ (mgCh1a),比散射系数明显大于比吸收系数。另外,栅列藻比散射系数均要大于微囊藻.而其比吸收系数则差异不明显。微囊藻、栅列藻吸收系数与叶绿素a浓度存在非常好的线性关系,显示其不存在色素包裹效应。微囊藻、栅列藻平均Q*.值分别为1.13、1.19.均大于1.包襄因子的分析也证明不存在色素包裹效应。实验条件下得到比吸收系数、比散射系数可以用于计算浮游植物的吸收和散射系数。     

Effects of rice straw on the cell viability,photosynthesis, and growth of Microcystis aeruginosa

Rice straw is supposed to be an environment-friendly biomaterial for inhibiting the growth of harmful blooms of the cyanobacterium Microcystis aeruginosa. However, its potential mechanism is not well known. To explore this mechanism, the growth, cell viability （esterase activity, membrane potential, and membrane integrity）, photosynthesis, and cell size ofM. aeruginosa were determined using flow cytometry and Phyto-PAM after exposure to rice straw extracts （RSE）. The results show that doses from 2.0 to 10.0 g/L of RSE efficiently inhibited the alga for 15 days, while the physiologic and morphologic responses of the cyanobacteria were time-dependent. RSE interfered with the cell membrane potential, cell size, and in vivo chlorophyll-a fluorescence on the first day. After 7 days of exposure, RSE was transported into the cytosol, which disrupted enzyme activity and photosynthesis. The cyanobacteria then started to repair its physiology （enzyme activity, photosynthesis） and remained viable, suggesting that rice straw act as an algistatic agent.     

Abiotic factors in colony formation: effects of nutrition and light on extracellular polysaccharide production and cell aggregates of Microcystis aeruginosa

Colony morphology is important for Microcystis to sustain a competitive advantage in eutrophic lakes. The mechanism of colony formation in Microcystis is currently unclear. Extracellular polysaccharide (EPS) has been reported to play an important role in cell aggregate formation of some phytoplankton. Microcystis aeruginosa was cultivated under varied abiotic conditions, including different nutrient, light, and temperature conditions, to investigate their effects on EPS production and morphological change. The results show that nutrient concentration and light intensity have great effects on EPS production in M. aeruginosa. There was a considerable increase in EPS production after M. aeruginosa was cultivated in adjusted culture conditions similar to those present in the field (28.9mg C/L, 1.98mg N/L, 0.65mg P/L, light intensity:100 μmol/(m2s)). These results indicate that abiotic factors might be one of the triggers for colony formation in Microcystis.     

Laboratory studies on zooplankton‐cyanobacteria interactions

Laboratory studies on cyanobacteria‐zooplankton interactions have largely focused on the inadequacy of cyanobacteria as a food source. Some features of cyanobacteria can be regarded as anti‐herbivore defences. Large colonies of Aphanizomenon, Anabaena and Microcystis cannot be handled by zooplankton, but do not interfere seriously with the filtering process. Small colonies and filaments, however, may cause severe inhibition of the feeding process by mechanical interference. This reduces zooplankton growth, reproduction, and survival. Copepods, rotifers, and Bosmina are less affected by mechanical disturbance than cladocerans. If ingested, some cyanobacteria may be poorly digested or may not provide essential nutrients. Some cyanobacteria are reported to be toxic to zooplankton. Several strains of Microcystis produce an endotoxin, but the amount of toxin produced differs among strains and with the condition of Microcystis. Zooplankton encountering toxic cells cease feeding. Some evidence of an extra‐cellular herbivore deterrent was found for Anabaena. Thus, cyanobacteria may either be individually protected or may create an environment that is unfavourable for efficient grazers (e.g., Daphnia).