白屈菜红碱对铜绿微囊藻生长和光合系统的影响

为了阐明白屈菜红碱(Chelerythrine)对铜绿微囊藻(Microcystis aeruginosa)生长及光合系统的影响, 研究了白屈菜红碱胁迫下铜绿微囊藻M. aeruginosa NIES-843生长、细胞色素含量及叶绿素光诱导荧光动力学变化. 结果显示, 当白屈菜红碱浓度10 g/L时, M. aeruginosa NIES-843的生长受到显著抑制. 通过线性回归分析, 白屈菜红碱对M. aeruginosa NIES-843生长50%抑制浓度(EC50)为(30.621.32) g/L. 当白屈菜红碱浓度为160 g/L时, M. aeruginosa NIES-843单位细胞内叶绿素a (Chl. a)和类胡萝卜素含量均显著低于对照. Chl. a光诱导荧光动力学分析结果显示, 白屈菜红碱胁迫下单位反应中心吸收的光能(ABS/RC)、单位反应中心捕获的用于还原QA的能量(TR0/RC)及单位反应中心捕获的用于电子传递的能量(ET0/RC)均受到显著抑制. 光合系统Ⅱ(PSⅡ)能量分配比率参数分析结果显示, 白屈菜红碱能显著抑制光合系统反应中心电子供体侧电子传递.       

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。     

铜绿微囊藻与小球藻对低温和黑暗的响应与恢复

研究以水华蓝藻铜绿微囊藻(Microcystis aeruginosa PCC 7806)与绿藻小球藻(Chlorella sp. FACHB-31)为研究对象, 探讨低温和黑暗对其生长、色素含量、最大光化学效率(F_v/F_m)、丙二醛(MDA)含量及过氧化氢酶活性的变化。结果表明, 30d的低温和黑暗处理, 显著降低了铜绿微囊藻和小球藻的叶绿素a浓度, 增加了单位细胞类胡萝卜素含量。在低温黑暗条件下, 铜绿微囊藻的MDA含量及CAT活性均显著增加, 而小球藻变化不明显。30d低温黑暗处理, 铜绿微囊藻的存活率为54.6%, 显著高于小球藻的31.3%。当恢复正常温度与光照, 2种藻均迅速生长。这些结果表明低温黑暗影响了微囊藻和小球藻的生理特性。在低温黑暗处理下, 微囊藻的F_v/F_m显著降低, 而小球藻则保持较为恒定的F_v/F_m, 表明微囊藻通过降低自身光合活性来渡过冬季低温黑暗的条件, 而小球藻在低温黑暗条件下仍保持较高的光合活性。     

生长素吲哚乙酸对铜绿微囊藻生理生化及产毒特性的影响

为了探究生长素吲哚乙酸(IAA)对产毒铜绿微囊藻(Microcystis aeruginosa)的影响, 从生长、光合色素含量、叶绿素光诱导荧光特征、脂质氧化和微囊藻毒素合成特性等方面, 研究了IAA对M. aeruginosa CHAB6301生理生化及产毒的影响。结果表明, 在低浓度IAA(0.04和0.2 mg/L)条件下, 铜绿微囊藻生长、叶绿素含量、光合系统(PSⅡ)电子传递效率及藻毒素含量均无明显变化, 藻蓝蛋白、别藻蓝蛋白和丙二醛(MDA)含量均低于对照。高浓度IAA(1和5 mg/L)能够促进细胞生长, 提高叶绿素含量, 但是抑制藻蓝蛋白和别藻蓝蛋白含量, 降低膜脂过氧化程度和细胞内藻毒素合成。综合各指标测定结果, 低浓度IAA对M. aeruginosa CHAB6301生长和光合作用影响不明显, 而高浓度IAA可促进藻细胞生长和光合作用, 增加微囊藻水华形成几率。     

高大气CO2浓度下氮素对小麦叶片光能利用的影响

关于氮素对高大气CO₂浓度下C₃植物光合作用适应现象的调节机理已有较为深入的研究, 但对其光合作用适应现象的光合能量转化和分配机制缺乏系统分析。该文以大气CO₂浓度和施氮量为处理手段, 通过测定小麦(Triticum aestivum)抽穗期叶片的光合作用-胞间CO₂浓度响应曲线以及荧光动力学参数来测算光合电子传递速率和分配去向, 研究了长期高大气CO₂浓度下小麦叶片光合电子传递和分配对施氮量的响应。结果表明, 与正常大气CO₂浓度处理相比, 高大气CO₂浓度下小麦叶片较多的激发能以热量的形式耗散, 增施氮素可使更多的激发能向光化学反应方向的分配, 降低光合能量的热耗散速率; 大气CO₂浓度升高后小麦叶片光化学淬灭系数无明显变化, 高氮叶片的非光化学猝灭降低而低氮叶片明显升高, 施氮促进PSII反应中心的开放比例, 降低光能的热耗散; 高大气CO₂浓度下高氮叶片通过PSII反应中心的光合电子传递速率(J_F)较高, 而且参与光呼吸的非环式电子流速率(J₀)显著降低, 较正常大气CO₂浓度处理的高氮叶片下降了88.40%, 光合速率增加46.47%; 高大气CO₂浓度下小麦叶片J_F-J₀升高而J₀/J_F显著下降, 光呼吸耗能被抑制, 更多的光合电子分配至光合还原过程。因此, 大气CO₂浓度增高条件下, 小麦叶片激发能的热耗散速率增加, 但增施氮素后小麦叶片PSII反应中心开放比例提高, 光化学速率增加, 进入PSII反应中心的电子流速率明显升高, 光呼吸作用被抑制, 光合电子较多地进入光化学过程, 这可能是高氮条件下光合作用适应性下调被缓解的一个原因。     

水深对苦草生长及叶片PSⅡ光化学特性的影响

利用植物效率仪(Handy-PEA)测定水深0.6、1.3、2.0 m下苦草叶片的快速荧光诱导动力学曲线(OJIP曲线),并采用JIP test方法分析和处理数据,研究水深对苦草生长和叶片光合机理的影响.结果表明: 随着水深增加,水下光强显著衰减,苦草生物量、无性系分株数、叶片数、根系总长度、根系表面积等形态指标显著降低,而最大叶长、平均叶长、最大叶宽无显著变化,2.0 m水深对苦草的生长产生了负面影响;单位反应中心吸收、捕获、电子传递、传递到电子传递末端的量子效率(ABS/RC、TR₀/RC、ET₀/RC、RE₀/RC)显著降低,单位反应中心的耗散量子效率(DI₀/RC)也显著下降,导致3种水深处理下单位反应中心吸收的能传递电子链末端的效率(φ_R0)以及用于电子传递的能量成功传递到电子链末端的效率(δ_R0)差异不显著,表明水深梯度对单位反应中心光合效率影响不显著;单位面积反应中心数(RC/CS₀)显著增加,相同受光面积时水深2.0 m处叶片光合作用显著强于水深0.6 m处;性能参数PI_abs、PI_cs和PI_abs,total显著提高,表明低光胁迫有利于光能向活跃化学能转变.苦草叶片通过激活未激活的反应中心,而不是提高单位反应中心光能利用效率来适应弱光强,且水深1.3 m较适合苦草生长.
     

水深对苦草生长及叶片PSⅡ光化学特性的影响

利用植物效率仪(Handy-PEA)测定水深0.6、1.3、2.0 m下苦草叶片的快速荧光诱导动力学曲线(OJIP曲线),并采用JIP test方法分析和处理数据,研究水深对苦草生长和叶片光合机理的影响.结果表明: 随着水深增加,水下光强显著衰减,苦草生物量、无性系分株数、叶片数、根系总长度、根系表面积等形态指标显著降低,而最大叶长、平均叶长、最大叶宽无显著变化,2.0 m水深对苦草的生长产生了负面影响;单位反应中心吸收、捕获、电子传递、传递到电子传递末端的量子效率(ABS/RC、TR₀/RC、ET₀/RC、RE₀/RC)显著降低,单位反应中心的耗散量子效率(DI₀/RC)也显著下降,导致3种水深处理下单位反应中心吸收的能传递电子链末端的效率(φ_R0)以及用于电子传递的能量成功传递到电子链末端的效率(δ_R0)差异不显著,表明水深梯度对单位反应中心光合效率影响不显著;单位面积反应中心数(RC/CS₀)显著增加,相同受光面积时水深2.0 m处叶片光合作用显著强于水深0.6 m处;性能参数PI_abs、PI_cs和PI_abs,total显著提高,表明低光胁迫有利于光能向活跃化学能转变.苦草叶片通过激活未激活的反应中心,而不是提高单位反应中心光能利用效率来适应弱光强,且水深1.3 m较适合苦草生长.
     

CO2浓度升高对苦草(Vallisneria natans)叶绿素荧光特性的影响

为了探究大气CO₂升高对沉水植物光合生理的影响,利用便携式植物效率分析仪（Handy PEA）,在无损的情况下测定不同CO₂浓度处理下的苦草（Vallisneria natans）叶绿素荧光诱导曲线,并采用JIP-test分析方法分析数据,研究CO₂浓度对苦草叶片叶绿素荧光特性的影响。结果表明在实验进行60 d后,与对照相比,高CO₂浓度处理下的苦草叶片PSⅡ反应中心受体侧荧光参数V_j、M_o显著升高,S_m、ψ_o、φ_Eo显著降低,叶片电子传递能力减弱;K相相对可变荧光W_k显著提高,PSⅡ反应中心供体侧放氧复合体OEC受到伤害;ABS/RC、DI_o/RC、TR_o/RC、DI_o/CS_o显著升高,ET_o/RC、RE_o/RC、ET_o/CS_o、RE_o/CS_o显著降低,苦草叶片用于热耗散的能量显著增加,导致用于电子传递及传递到电子链末端的能量显著减少;性能参数F_v/F_m、PI_abs显著降低,苦草叶片PSⅡ潜在活性和光合作用原初反应过程受到抑制。以上结果表明,在长期高CO₂浓度处理下,苦草叶片光合机构功能受到抑制,PSⅡ反应中心活性降低,光合功能下调,发生光适应现象。     

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。     

雪衣藻叶绿素荧光特性及抗氧化系统对汞胁迫的响应

文章选用对汞(Hg)具有较强耐受性的雪衣藻(Chlamydomonas nivalis)为对象,探究藻细胞对汞胁迫的响应及其耐受机制。结果表明, Hg²⁺对藻细胞的胁迫具有明显的时间依赖的量效关系, 48h半数效应浓度EC₅₀为1.44 mg/L。与对照组相比, 1.0和1.5 mg/L Hg²⁺处理组能维持生长和光合活性; 2.0 mg/L Hg²⁺处理组生物量和色素含量在胁迫6h时无显著变化(P>0.05), 48h时均显著降低(P<0.05)。随着Hg²⁺浓度和暴露时间的增加,藻细胞最大光化学量子产率(F_v/F_m)、最大电子传递速率(ETR_max)、半饱和光强(I_k)及光能利用效率(α)均降低,OJIP快速荧光诱导动力学曲线和Q_A^-再氧化变化明显,光合电子传递受阻。Hg²⁺胁迫导致能量流的变化,藻细胞部分反应...     

黑暗限气条件下铜绿微囊藻细胞死亡的形态结构和生理生化变化

【目的】研究铜绿微囊藻细胞死亡过程中形态和生理生化变化,探讨蓝藻细胞死亡机制。【方法】通过黑暗限气处理模拟水华爆发后期水体环境,在处理后不同时间取样,对藻液的OD值,溶氧含量和pH值进行监测,使用透射电镜对细胞形态结构变化进行观察,通过胱天蛋白酶(Cysteine-dependent aspartate specificprotease,Caspase)活性检测、活性氧含量测定、末端脱氧核糖核酸转移酶介导的dUTP缺口末端标记(Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling,TUNEL)染色和琼脂糖凝胶电泳对处理后藻细胞的死亡生理进行研究。【结果】黑暗限气处理后,藻培养液pH值和溶解氧含量下降,处理12 h后藻液开始变黄,48 h后藻细胞全部死亡。电镜观察结果表明,藻细胞在黑暗限气处理所导致的死亡过程中出现空泡和类囊体、核糖体等内部结构解体但细胞壁仍保持完整等现象。活性氧含量和caspase活性检测表明,在藻细胞死亡过程中活性氧含量和caspase活性上升。TUNEL染色和琼脂糖凝胶电泳分析发现,藻细胞在死亡过程中DNA发生断裂和降解。【结论】铜绿微囊藻细胞在黑暗和限气处理中表现出和真核生物细胞程序性死亡相类似的死亡特征,这说明细胞死亡机制是保守的,原核细胞和真核细胞一样具有程序性死亡机制。     

Functional analysis of PilT from the toxic cyanobacterium Microcystis aeruginosa PCC 7806

The evolution of the microcystin toxin gene cluster in phylogenetically distant cyanobacteria has been attributed to recombination, inactivation, and deletion events, although gene transfer may also be involved. Since the microcystin-producing Microcystis aeruginosa PCC 7806 is naturally transformable, we have initiated the characterization of its type IV pilus system, involved in DNA uptake in many bacteria, to provide a physiological focus for the influence of gene transfer in microcystin evolution. The type IV pilus genes pilA, pilB, pilC, and pilT were shown to be expressed in M. aeruginosa PCC 7806. The purified PilT protein yielded a maximal ATPase activity of 37.5 +/- 1.8 nmol P(i) min(-1) mg protein(-1), with a requirement for Mg(2+). Heterologous expression indicated that it could complement the pilT mutant of Pseudomonas aeruginosa, but not that of the cyanobacterium Synechocystis sp. strain PCC 6803, which was unexpected. Differences in two critical residues between the M. aeruginosa PCC 7806 PilT (7806 PilT) and the Synechocystis sp. strain PCC 6803 PilT proteins affected their theoretical structural models, which may explain the nonfunctionality of 7806 PilT in its cyanobacterial counterpart. Screening of the pilT gene in toxic and nontoxic strains of Microcystis was also performed.     

Immunogold localisation of microcystins in cryosectioned cells of Microcystis

Insights into the origins, function(s), and fates of cyanobacterial toxins may be obtained by an understanding of their location within cyanobacterial cells. Here, we have localised microcystins in laboratory cultures of Microcystis PCC 7806 and PCC 7820 by immunogold labelling. Cryosectioning was used for immunoelectron microscopy since microcystins were extracted during the ethanol-based dehydration steps routinely used for sample preparation. Microcystins were specifically localised in the nucleoplasm and were associated with all major inclusions of the microcystin-producing strains Microcystis PCC 7806 (MC(+)) and Microcystis PCC 7820, and labelling was preferentially associated with the thylakoids and around polyphosphate bodies. A mutant strain of Microcystis PCC 7806 (MC(-)) which does not produce microcystins was used as a control. Distribution of total gold label within each cell region or associated with inclusions indicated that most of the cells' microcystin pool was associated with the thylakoids (69%, PCC 7806 (MC(+)); 78%, PCC 7820), followed by the nucleoplasmic region (19%, PCC 7806 (MC(+)); 12%, PCC 7820). Cryosectioning is a useful technique since it reduces the extraction of microcystins during sample preparation for electron microscopy.     

Identification of pilus-like structures and genes in Microcystis aeruginosa PCC7806

Four putative type IV pilus genes from the toxic, naturally transformable Microcystis aeruginosa PCC7806 were identified. Three of these genes were clustered in an arrangement which is identical to that from other cyanobacterial genomes. Type IV pilus-like appendages were also observed by electron microscopy.     

A kinetic completion of the cyclic photosynthetic electron pathway of Rhodopseudomonas sphaeroides: Cytochrome b-cytochrome c2 oxidation-reduction

In Rhodopseudomonas sphaeroides, following a single-turnover flash of light, cytochrome c₂ is oxidized by reaction center bacteriochlorophyll, and a cytochrome b is reduced by the primary electron acceptor, probably via ubiquinone. In this report we show that, in the uncoupled state, the rate of re-oxidation of the cytochrome b is identical to the rate of reduction of the cytochrome c₂, a kinetic completion of the cyclic photosynthetic electron transport system.     

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.     

阴离子表面活性剂LAS对产毒和无毒微囊藻生长及产毒特性的影响

文章研究了低浓度范围内不同浓度梯度的阴离子表面活性剂直链烷基苯磺酸盐(LAS)对产毒微囊藻(Microcystis aeruginosa, FACHB905)和无毒微囊藻(Microcystis wesenbergii, FACHB908)生长、光合特性、种间竞争及毒素合成的影响。结果表明,在0.05—5.0 mg/L LAS浓度梯度处理下,产毒微囊藻的生物量、产毒基因mcyD表达量和每细胞MCs含量均在培养12d后显著增加。产毒微囊藻胞内和胞外MCs含量在各LAS浓度处理后分别为0.069、0.052、0.061、0.038和0.037 fg/fg Chl.a及107.1、103.7、127.1、99.6和113.7 ng/L。即使在0.5 mg/L低浓度LAS处理条件下,上述3个参数也分别比对照组显著增加了24.2%、12.4倍和10.4%。浓度为0—0.2 mg/L LAS对无毒微囊藻的生物量无明显影响,而较高浓度的LAS(0.5—5.0 mg/L)明显抑制了无毒微囊藻的生长。在两株微囊藻混合培养时, 0.2—1.0 mg/L LAS处理组的产毒铜绿微囊藻mcy D的表达对LAS...