植物化感抑藻的作用机理

对植物化感物质抑藻机理,即化感物质影响藻细胞的光合作用、破坏细胞膜、改变细胞酶活性、影响细胞超微结构及基因表达等几个方面进行了综合评述。研究表明,化感物质主要通过降低叶绿素a含量,破坏光合系统Ⅱ,使细胞膜结构裂解,酶活性改变,抗氧化系统破坏,细胞膜内超微结构受损,基因表达异常等几个方面来抑制藻细胞的分裂或直接杀死藻细胞,从而减少藻细胞数量,抑制藻类的生长。未来应该从更微观角度,即基因表达、分子和遗传机制等方面对化感抑藻机理进行深入研究,以期为开发安全、高效的抑藻化感物质提供理论基础。     

红树植物木榄有机溶剂提取物对球形棕囊藻的抑制效应

基于化感作用原理,利用有机溶剂对红树植物木榄(Bruguiera gymnorrhiza)叶片中的活性物质进行连续提取分离,分别得到正己烷相、乙酸乙酯相、正丁醇相和水相提取物。利用这些提取物进行抑藻实验,通过测定球形棕囊藻(Phaeocystis globosa)的细胞密度发现:四种组分提取物对球形棕囊藻均具有显著的化感抑制作用;正己烷相和乙酸乙酯相提取物对球形棕囊藻的抑制效果明显优于正丁醇相和水相提取物的抑制效果。正己烷相提取物和乙酸乙酯相提取物对球形棕囊藻的48hEC₅₀分别为14.90mg/L和12.18mg/L。研究表明,藻细胞初始接种密度影响提取物的抑藻效应,低接种密度时抑制率极高,而随着接种密度的升高抑制率下降;接种密度极高时,提取物不但不会抑制甚至还会促进藻细胞的生长。     

抑藻菌株Bacillus sp. hsn03分离鉴定及其对铜绿微囊藻的抑制效果与特征

【背景】蓝藻常形成水华,严重破坏水生环境及威胁水生生物。微生物能够专一性地抑制蓝藻的生长,从而可以被用于蓝藻水华的控制。【目的】分离筛选一株对有害水华藻——铜绿微囊藻(Microcystisaeruginosa)7806有高效抑藻能力的菌株,对其进行生理生化和分子鉴定,并研究其抑藻效果与特征。【方法】通过API试剂条确定其生理生化特征,对细菌的16SrRNA基因进行测序并构建系统进化树,通过探究细菌菌体、上清、发酵液的抑藻效果确定该菌的抑藻方式,通过测定不同温度、pH、蛋白酶K、有机溶剂萃取和透析条件下的抑藻效果及傅里叶红外光谱来确定胞外抑藻物质的性质。【结果】菌株hsn03与芽孢杆菌属(Bacillus sp.)的Bacillus sonorensis NBRC101234有最高相似性,达99.59%;通过分泌胞外抑藻物质对铜绿微囊藻生长表现出抑制效果,且最佳抑藻添加量为7.0%(体积比);胞外抑藻物质为一类分子量大于500 Da且小于1 kD,耐高温,耐酸碱,含有叁键和累积双键,非蛋白、多糖类的小分子物质。【结论】抑藻菌株的挖掘与鉴定对于丰富有害藻华藻的抑藻菌质资源有非常大的促进作用,通过对抑藻方式、效果和特征的研究为进一步将抑藻菌应用于铜绿微囊藻的治理奠定基础。     

大型海藻内共生真菌次生代谢产物中抑制水华蓝藻物质的分离与活性

为获得能明显抑制水华蓝藻的化感物质,本研究从28株大型海藻内共生真菌中筛选出了4株菌株,其粗提物能显著抑制水华鱼腥藻（Anabaenaflos-aquae）和铜绿微囊藻（Microcystis aeruginosa）的生长。对这4株真菌扩大培养后,检测了从其次生代谢产物中分离出的32种单体化合物的抑藻活性。 其中, 化合物sterigmatocystin、（2’E,4’E,6’E）-6-（1’-carboxyocta-2’,4’,6’-triene）-11,12-epoxy-9,11-dihydroxydrim-7-ene、 6,8-di-O-methylnidurufin和aversin对水华鱼腥藻的抑制率显著大于50%（P 〈 0.05）, 24 h半数有效浓度（EC50,24h）分别为6.865、4.302、1.824、2.319 μg/mL;sterigmatocystin、indol-3-carboxaldehyde和（2’E,4’E,6’E）-6-（1’-carboxyocta-2’,4’,6’-triene）-11,12-epoxy-9,11-dihydroxydrim-7-ene对铜绿微囊藻的抑制率显著大于50%（P 〈 0.05）, EC50,24h 分别为4.167、5.374、8.386 μg/mL。     

基于化感物质释放特性的沉水植物抑藻作用模式研究进展

沉水植物对藻类的化感抑制作用,是沉水植物获取竞争优势和维持清水稳态的重要机制之一。化感物质是由植物产生并释放到水环境中的次生代谢产物,化感物质的有效释放和作用是实现沉水植物化感抑藻作用的关键环节。因此,在化感物质释放水平阐明沉水植物化感抑藻的作用模式、过程和机制具有重要意义。通过比较沉水植物化感物质释放到水环境中的种类、含量和常规急性毒性测试中化感物质的抑藻效果,发现沉水植物化感物质在释放水平上的作用模式不同于常规急性毒性试验中的单次作用。为了回答沉水植物化感物质在释放水平如何高效抑藻的问题,结合化感物质的释放特性,重点从化感物质的联合作用和持续作用等角度探讨沉水植物化感抑藻的作用模式,提出沉水植物可能通过多种化感物质低剂量持续释放的方式,实现对目标藻类的持续协同控制。今后有必要进一步结合沉水植物与目标藻类的共存系统与原位实验,借助分析化学、植物化学、细胞和分子生物学的技术手段,从生态学水平加强沉水植物化感抑藻作用机制研究。     

化感作用抑制有害藻类生长的研究进展

利用植物化感作用抑制有害藻类生长的方法具有生态安全性好、快速高效等优点, 为有效控制有害藻类的爆发提供了新思路。论文从化感作用的概念、化感抑藻机理、陆生植物对藻类的化感抑制作用、水生植物对藻类的化感抑制作用等方面进行了论述, 并对植物化感作用抑藻技术的研究方向进行了展望, 提出对化感物质在环境中的降解特性、化感物质选择性抑藻机理及具有活性的化感物质的人工合成等方面需要做更深入的研究。     

大漂提取物抑制铜绿微囊藻的试验研究

为了确定高效脱氮除磷的水生漂浮植物大漂的主要化感活性物质, 本文通过乙醇超声波提取获得大漂的化感粗提物, 利用溶剂萃取法从粗提物中分离得到4种能抑制铜绿微囊藻(FACHB-911)生长的化感萃取物,当其浓度均为0.1 g/L时共培养5d后抑藻率分别为40%、69%、40%、35%。将抑藻率最高的化感萃取物通过酸性氧化铝柱层析法进一步分离纯化, 分离得到4组抑藻作用较弱的(5d后抑藻率分别为12%、11%、33%、41%)和1组抑藻作用较强的化感层析流分(5d后抑藻率高达80%)。将抑藻作用最强的化感层析流分通过气相色谱-质谱联用(GC-MS)方法进行成分检测, 发现大漂提取物中含有对甲苯磺酸甲酯、硬脂酸酰胺两种具有化感潜力的化合物。     

岩兰草油对淡水藻类的抑制作用

利用植物分泌的化感物质进行藻类控制已成为国内外生态环境领域研究的热点之一。首次探讨了岩兰草油的抑藻效应,建立了岩兰草油对铜绿微囊藻和斜生栅藻两种藻共培养时抑制作用的数学模型,采用GC-MS分析方法对岩兰草油中主要成分进行了鉴定。结果表明:岩兰草油对铜绿微囊藻和斜生栅藻在单独和共培养状态下均有良好的抑制作用,对单独培养的铜绿微囊藻和斜生栅藻抑制作用的EC50分别为0.20和0.30 m L/L,说明岩兰草油对蓝藻(铜绿微囊藻)的抑制效果明显好于对绿藻(斜生栅藻)。岩兰草油的主要成分为具有良好抑藻作用的倍半萜类物质,而所建立的岩兰草油对两藻共培养状态下抑制作用的数学模型可以求出在任何时间节点上铜绿微囊藻和斜生栅藻的种群密度以及要达到抑藻最佳状态的岩兰草油浓度。该研究对水华藻类的控制有较重要的理论与应用价值。     

海菖蒲浸提液对锥状斯氏藻的抑制效应研究

海菖蒲(Enhalus acodoides)是典型热带亚热带沿海常见大型海草, 但关于海菖蒲对赤潮藻的抑制作用研究少见报道。研究选取海菖蒲为实验材料, 利用人工海水和两种有机溶剂提取海菖蒲中抑藻活性物质并对锥状斯氏藻进行抑藻试验研究, 结果表明, 三种提取物对锥状斯氏藻均有显著抑制作用, 在试验设定浓度下, 水浸物在 2.5 g·L^–1 时具有最高抑藻率 95.34%, 乙醇提取物在浓度为 12.5 mg·L^–1 时即达到 78.55%, 正己烷提取物在浓度为 50 mg·L^–1 时最高抑制率达到58.28%。研究表明, 海菖蒲水提液抑制效果最好, 具有应用于近海水体富营养化的治理和开发新型抑藻剂的潜力。     

N-苯基-2-萘胺对拟柱胞藻生长、抗氧化酶及光合系统Ⅱ的影响

为探究化感物质对入侵水华蓝藻-拟柱胞藻(Cylindrospermopsis raciborskii)的影响, 研究对不同浓度(0、0.05、0.10、0.50、1.00和2.00 mg/L)N-苯基-2-萘胺处理下拟柱胞藻的生长、抗氧化酶活性及叶绿素荧光诱导动力学进行了测定。结果表明, 藻叶绿素a含量随N-苯基-2-萘胺浓度升高而显著降低, 浓度大于0.50 mg/L时, 拟柱胞藻叶绿素a含量显著低于对照。72h, N-苯基-2-萘胺对拟柱胞藻的EC50为1.02 mg/L。叶绿素荧光诱导动力学参数表明, 浓度小于0.50 mg/L时, 拟柱胞藻的最大光化学效率(P0)、单位面积上有活性的反应中心(RC/CS0)、以吸收光能为基础的性能指数(PIABS)及以单位面积为基础的性能指数(PICS)均显著高于对照, 而反映2ms时有活性的反应中心的开放程度的0则显著低于对照组。当N-苯基-2-萘胺浓度大于0.50 mg/L时, 以上参数均呈现相反趋势。这表明低浓度N-苯基-2-萘胺能提高拟柱胞藻的光合效率, 而在高浓度时, 拟柱胞藻PSⅡ有活性的反应中心减少, 电子传递受到阻碍, 光合效率下降。与对照相比, 在浓度高于0.50 mg/L条件下, 拟柱胞藻的过氧化氢酶(CAT)、过氧化物酶(SOD)及谷胱甘肽过氧化物酶(GPX)的活性显著增加, 表明拟柱胞藻能产生大量的抗氧化酶来减少N-苯基-2-萘胺的损伤。这些结果表明N-苯基-2-萘胺可能通过抑制PSⅡ的电子传递和减少其有活性的反应中心来影响拟柱胞藻的生长, 也暗示了N-苯基-2-萘胺能作为潜在的抑藻物质来控制拟柱胞藻水华。       

球等鞭金藻对氟苯尼考胁迫的响应研究

研究以球等鞭金藻(Isochrysis galbana)为研究对象, 分析在不同浓度氟苯尼考暴露下, 球等鞭金藻的生长、叶绿素、类胡萝卜素和脂肪酸含量的变化。结果表明, 氟苯尼考对球等鞭金藻的生长呈“低促高抑”作用, 其72h-EC₅₀为17.12 mg/L; 随着暴露浓度(≥1 mg/L)增加, 细胞内叶绿素含量显著下降, 光合作用受到抑制, 而类胡萝卜素含量显著上升。脂肪酸和类胡萝卜素共同参与藻细胞的防御机制, 氟苯尼考(< 1 mg/L)暴露引起细胞内总脂肪酸含量显著降低。研究探究了水产养殖环境中抗生素残留的生态风险, 为水产养殖过程中抗生素的科学合理使用提供了理论依据。     

Joint action of phenolic acid mixtures and its significance in allelopathy research

Although the ecological significance of mixtures of phytotoxins is recognized in research on chemical plant interference (allelopathy), few studies convincingly demonstrate the joint action of phytotoxin mixtures, key to understanding the ecological impact of these materials, using established models from other biological disciplines, e.g. toxicology and pharmacology. Addressing this need, the present study investigates the joint action of the phenolic acids, p -hydroxybenzoic, p -coumaric and ferulic acids on root growth inhibition of perennial ryegrass ( Lolium perenne L). The Additive Dose Model (ADM) isobole and estimated concentration of phenolic acid mixture were calculated on ED₂₀, ED₅₀ and ED₈₀ from the dose–response curves for the phenolic acids applied alone or in mixtures of fixed ratios. The binary combination of three selected phenolic acids is generally antagonistic relative to the ADM. No evidence for synergistic activities of phenolic acids in the mixture was noted. Since allelopathic activities in nature are largely due to the presence of several compounds in a mixture, the present study advances understanding of the joint action of binary combination of allelochemicals in a mixture.     

敌百虫对中肋骨条藻生长的影响

研究了有机磷农药敌百虫对中肋骨条藻Skeletonema costatum生长的影响。相对于水生甲壳类和鱼类,敌百虫对中肋骨条藻毒性较低,72 h半抑制浓度(EC₅₀)为84.8 mg/L。在31 d的亚慢性暴露下,低浓度组(≤10 mg/L)藻细胞生长趋势与对照组相似;50 mg/L的敌百虫在暴露初期对骨条藻的生长具有明显抑制作用,但在暴露15 d后,该浓度组藻细胞密度仍保持增长,最大细胞密度为2.77×10⁷ cells/mL,是对照组的137.7%。100 mg/L则能完全抑制藻细胞的生长。本实验结果说明敌百虫等有机磷农药的广泛使用可能会刺激浮游植物的生长,从而增加引发赤潮的风险。     

A freshwater bacterial strain,Shewanella sp. Lzh-2, isolated from Lake Taihu and its two algicidal active substances,hexahydropyrrolo[1,2-a]pyrazine-1,4-dione and 2, 3-indolinedione

Cyanobacterial blooms have become a serious problem in Lake Taihu during the last 20 years, and Microcystis aeruginosa and Synechococcus sp. are the two dominant species in cyanobacterial blooms of Lake Taihu. A freshwater bacterial strain, Shewanella sp. Lzh-2, with strong algicidal properties against harmful cyanobacteria was isolated from Lake Taihu. Two substances with algicidal activity secreted extracellularly by Shewanella sp. Lzh-2, S-2A and S-2B, were purified from the bacterial culture of strain Lzh-2 using ethyl acetate extraction, column chromatography, and high performance liquid chromatography (HPLC) in turn. The substances S-2A and S-2B were identified as hexahydropyrrolo[1,2-a]pyrazine-1,4-dione and 2, 3-indolinedione (isatin), respectively, based on liquid chromatography-mass spectrometry (LC-MS), gas chromatography-mass spectrometry (GC-MS), and hydrogen-nuclear magnetic resonance (H-NMR) analyses, making this the first report of their algicidal activity toward cyanobacteria. S-2A (hexahydropyrrolo[1,2-a]pyrazine-1,4-dione) had no algicidal effects against Synechococcus sp. BN60, but had a high level of algicidal activity against M. aeruginosa 9110. The LD50 value of S-2A against M. aeruginosa 9110 was 5.7 μg/ml. S-2B (2, 3-indolinedione) showed a potent algicidal effect against both M. aeruginosa 9110 and Synechococcus sp. BN60, and the LD50 value of S-2B against M. aeruginosa 9110 and Synechococcus sp. BN60 was 12.5 and 34.2 μg/ml, respectively. Obvious morphological changes in M. aeruginosa 9110 and Synechococcus sp. BN60 were observed after they were exposed to S-2A (or S-2B) for 24 h. Approximately, the algicidal activity, the concentration of S-2A and S-2B, and the cell density of Lzh-2 were positively related to each other during the cocultivation process. Overall, these findings increase our knowledge about algicidal substances secreted by algicidal bacteria and indicate that strain Lzh-2 and its two algicidal substances have the potential for use as a bio-agent in controlling cyanobacterial blooms in Lake Taihu.     

Use of a rapid bioluminescence assay for detecting cyanobacterial microcystin toxicity

The recent rise in the awareness of the occurrence of toxic cyanobacterial blooms in aquatic environments, with associated human health problems and animal deaths, has increased the need for rapid, reliable and sensitive methods of determining cyanobacterial toxicity. A luminescent bacterial toxicity test was assessed as a complement to the established mouse bioassay. Seventeen samples including pure cyanobacterial microcystin-LR hepatotoxin, laboratory isolates and natural blooms of cyanobacteria were tested and toxicity data compared with mouse LD50 values. Microcystin-LR and all five microcystin-containing cyanobacterial samples, hepatotoxic by mouse test gave EC50 values of less than 0.46 mg/ml in bioluminescence-based Microtox assays. Of 11 samples non-toxic by mouse bioassay, only two gave an EC50 of less than 0.98 mg/ml by bioluminescence assay. It is suggested that the Microtox bioluminescence assay may prove useful in the preliminary screening of cyanobacterial blooms for microcystin-based toxicity.     

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。     

Allelopathic control of cyanobacterial blooms by periphyton biofilms

Periphyton biofilms are natural mixtures comprised of photoautotrophic and heterotrophic complex microorganisms. In this work, the inhibition effects of periphyton biofilms on cyanobacterial blooms were studied in pilot and field trials. Results show that the cyanobacterial species responsible for the blooms had an upper nutrient concentration threshold, below which it could not effectively compete with other organisms in the periphyton. The disappearance of the cyanobacterial blooms was due to the allelopathy between the cyanobacteria and periphyton biofilm. In particular, it was found that the periphyton biofilm could produce water-soluble allelochemicals such as indole and 3-oxo-α-ionone to significantly inhibit the growth of the cyanobacteria. These allelochemicals are able to damage the thylakoid membranes of the cyanobacteria, interrupt the electron transport in photosystem II, decrease effective quantum yields, and eventually lead to the failure of photosynthesis. A comprehensive discussion on the ecological consequences of these findings is also presented. This work demonstrates the potential of periphyton biofilm to be used as an environmentally friendly ecological engineering solution for (i) the control of cyanobacterial blooms and (ii) a transitional means for the construction of beneficial conditions for ecosystem restoration. In addition, this work provides significant insights into the competitive relationships between algae and biofilms.     

Isolation and Identification of Algicidal Compound from Streptomyces and Algicidal Mechanism to Microcystis aeruginosa

The biological control of cyanobacterial harmful algal blooms (cyanoHABs) is important to promote human health, environmental protection, and economic growth. Active algicidal compounds and algicidal mechanisms should be identified and investigated to control cyanoHABs. In this study, the algicidal actinobacterium Streptomyces sp. L74 was isolated from the soil of a nearby pond which located in the center lake of Guanghzou Higher Education Mega Center. Results showed that the algicidal activities of cyanoHABs are mainly achieved via an indirect attack by producing algicidal compounds. All active algicidal compounds are hydrophilic substances that are heat and pH stable. In the present study, an active compound (B3) was isolated and purified by high-performance liquid chromatography and identified as a type of triterpenoid saponin (2-hydroxy-12-oleanene-3, 28-O-D-glucopyranosyl) with a molecular formula of C₄₂H₇₀O₁₃ as determined by infrared spectrometry, electrospray ionization mass spectrometry, and nuclear magnetic resonance. Active algicidal compounds from Streptomyces sp. L74 were shown to disrupt the antioxidant systems of Microcystis aeruginosa cells.     

基于叶绿素荧光探讨链霉素对念珠藻生长及光合毒性效应

为了探究不同浓度链霉素对念珠藻生长和光合的毒性效应, 研究对不同浓度(0、0.05、0.1、0.2、0.5和1.0 mg/L)链霉素处理下念珠藻的生长、叶绿素荧光进行了测定。结果发现, 培养至96h, 链霉素对念珠藻的EC₅₀(96h-EC₅₀)为(0.13±0.037) mg/L。与对照组相比, 各链霉素浓度处理下叶绿素a的含量显著降低。最大光化学效率(φP₀)、可变荧光(F_v)、PSⅡ的潜在活性(F_v/F₀)、单位面积上有活性的反应中心(RC/CS₀)、以吸收光能为基础的性能指数(PIabs)及反应中心吸收的光能用于电子传递的量子产额(φE₀)均随着链霉素浓度的升高而降低。然而, 单位叶绿素最大荧光(F_m/Chl. a)、每个有活性的反应中心吸收的光能(ABS/RC)和热耗散(DI₀/RC)则随着链霉素浓度的升高而增加。当链霉素浓度大于0.1 mg/L时, 叶绿素荧光动力学曲线出现K点的出现。结果表明链霉素可能通过抑制光合系统Ⅱ(PSⅡ)的电子传递和减少有活性的反应中心来影响念珠藻的生长, 也暗示了高浓度链霉素对藻类的生长具有抑制作用。     

邻苯二甲酸二乙酯和壬基酚联合暴露对杜氏盐藻生长的影响

为了探讨环境激素类物质邻苯二甲酸二乙酯(DEP)和壬基酚(NP)对海洋微藻的联合毒性效应,选取杜氏盐藻(Dunaliella salina)为受试生物,以环境激素对杜氏盐藻单一暴露的96h EC₅₀的毒性效应作为一个毒性单位(IU),采用毒性单位法比较研究了DEP和NP单一暴露以及两者以三种不同混合比例(毒性单位比:1:1、1:4和4:1)暴露对杜氏盐藻的细胞生长、叶绿体色素含量、可溶性蛋白含量、SOD活性以及最大光能转化效率(Fv/Fm)的影响.实验结果表明:DEP和NP单一暴露对杜氏盐藻的96h EC₅₀分别为69.54 mg/L和1.47 mg/L,两种环境激素对杜氏盐藻均有抑制作用,且NP较DEP对杜氏盐藻的毒性更强.DEP和NP联合暴露较单一暴露对杜氏盐藻的细胞生长、叶绿体色素和可溶性蛋白的合成有较强的抑制作用,两种环境激素在毒性单位比为1:1、1:4、4:1三个比例水平上的联合毒性效应均表现为协同效应,其中比例为1:1的协同效应最强.