栅藻LX1和雨生红球藻的胞外产物产生特性及其对藻细胞生长的影响

基于微藻在资源化大规模培养过程中,其胞外产物对藻细胞生长影响的不确定性,对比研究了两株高含油脂藻种,栅藻LX1（Scenedesmus sp.LX1）和雨生红球藻（Haematococcus pluvislis）的生长特征、胞外产物的产生特性,以及胞外产物对藻细胞生长的影响。研究结果表明,在相同培养条件下,栅藻LX1与雨生红球藻的内禀生长速率基本相当,分别为0.16和0.17 d-1。两株藻在不同生长期的胞外产物（以DOC值表征）产生速率不同,其中栅藻LX1胞外产物的对数期产生速率（1.4 mg·L-1·d-1）﹥稳定期后期产生速率（0.56 mg·L-1·d-1）﹥稳定期产生速率（0.48 mg·L-1·d-1）;雨生红球藻胞外产物的稳定期后期产生速率（2.3 mg·L-1·d-1）﹥稳定期产生速率（0.88 mg·L-1·d-1）﹥对数期产生速率（0.66 mg·L-1·d-1）。栅藻LX1胞外产物对其整个生长过程都有明显抑制;雨生红球藻胞外产物则在稳定期后期才对其生长产生明显抑制作用。     

雨生红球藻3个株系的生长及对光盐反应初探

对雨生红球藻3个株系（No.3、No.712和No.797)的生长、对诱导虾青素形成的光盐因子的反应进行了比较,结果表明：No.3、No.712和No.797的生长速率分别为0．128d^-1,0.137d^-1和0.144d^-1;外加盐的量较大（4gL^-1)时导致雨生红球藻细胞数的减少,而外加盐浓度较低（2gL^-1)时,细胞数的反应因株系和光照的不同而有所差异;盐和强光照均导致静细胞比例显著增加;盐的加入仅在弱光照下诱导雨生红球藻3个株系的虾青素累积,在强光照下却导致件株系虾青素含量的下降,强光照下不外加盐时No.3、NO.712和No.797藻液的虾青素含量无显著差异,分别为2.15、2.77和2．45mg L^-1。图1表2参12     

焦酚对共培养铜绿微囊藻和雨生红球藻影响的初步研究

以单株藻为对象的纯培养抑藻测试体系被广泛用于化感抑藻活性物质筛选和作用机理研究,但自然水体中藻类常常相伴而生并相互作用,共存藻类对化感物质抑藻效果的影尚不清楚。为探讨藻类共存状态下对化感物质的响应,选择沉水植物穗花狐尾藻(Myriophyllum spicatum)的典型化感抑藻物质焦酚,以有害蓝藻铜绿微囊藻(Microcystis aeruginosa)和经济绿藻雨生红球藻(Haematococcuspluvialis)为受试藻,同时设置混合共培养体系和纯培养体系,比较焦酚对不同培养体系中两株藻的影响。结果显示,纯培养和共培养体系中,焦酚对铜绿微囊藻细胞增长的抑制率分别为96.82%和93.18%,而对雨生红球藻细胞增长的抑制率显著降低,分别为29.39%和45.40%。焦酚处理的纯培养和共培养体系中铜绿微囊藻胞外藻毒素质量浓度分别为3.23、2.00μg·L~(-1),雨生红球藻单个细胞内虾青素质量浓度分别为0.82、1.21 pg·cell-1。与纯培养相比,共培养体系中焦酚对铜绿微囊藻生长的抑制作用减弱,微囊藻毒素释放量显著降低(P0.05),而对雨生红球藻生长的抑制作用增强(P0.05),单个细胞内虾青素积累量最大(P0.05),表明两者共存减弱了焦酚对铜绿微囊藻的抑制效应,却增强了焦酚对雨生红球藻的影响。这些结果初步说明共存藻类会影响化感物质对目标藻株的抑制效应,在后续化感抑藻作用研究中,充分考虑藻类所处生物和非生物环境,将有助于深入揭示水生植物化感抑藻作用生态机制,明确化感作用和化感物质的生态学价值。     

有效微生物群(EM)中抑藻酵母及其活性物质特性

采用选择性培养基从有效微生物群(EM)中富集出酵母菌,通过向对数增长期的混合藻液中加入酵母菌、葡萄糖、酵母菌+葡萄糖、经酵母菌发酵除菌后的葡萄糖溶液分别考察了其对水华藻的抑制效应.结果表明,混合酵母菌自身对于水华藻没有抑制作用,单独添加葡萄糖的水华藻甚至出现快速增殖;而在添加葡萄糖+酵母菌或酵母菌发酵液的实验中水华藻近似被完全抑制,最大抑制率达97%.据此可知,抑制水华藻增殖的并非酵母菌自身,而是具有高热稳定性的胞外分泌物.HPLC-SEC与三维荧光光谱分析表明,酵母菌发酵液中含有明显的芳环类羧酸及蛋白质物质,但由于其在高温灭菌后仍有良好的抑藻活性,因而进一步确定其抑藻活性物质为含苯环的有机酸.此外,依据对实验中水华藻细胞形态的显微镜观察,该酸性胞外物可能是通过破坏藻细胞的叶绿体结构来达到抑藻效果的.     

混合培养对城市污水厂二级出水培养能源微藻的生长促进作用

利用城市污水培养能源微藻可以实现水质净化和生物质生产的耦合,备受关注。生物质生产效率较低是限制其大规模应用的主要因素之一,混合培养是提高微藻生物质产率的一种潜在方法。为筛选出城市二级出水条件下合适的能源微藻混合藻种,考察了二级出水条件下3株高含油脂藻种栅藻LX1（Scenedesmussp. LX1）、椭圆小球藻YJ1（Chlorella ellipsoidea YJ1）和雨生红球藻（Haematococcus pluvislis）单一藻种和两两混合培养时的生长特性,比较了各微藻单一藻种及两两混合培养时的生长特性参数及生物质产量。研究结果表明,3种微藻的两两混合组合均能在二级出水条件下正常生长,各微藻干物质量浓度在第10天左右均能达到100 mg·L^-1左右。与栅藻LX1和椭圆小球藻YJ1相比,雨生红球藻在两两混合培养条件下表现出更高的藻细胞干物质量浓度增长趋势。3种微藻的内禀生长速率均显著高于各自的单一培养,栅藻 LX1和雨生红球藻混合培养时分别达到最高内禀增长速率（分别为1.36 d^-1、0.97 d^-1）。栅藻LX1与雨生红球藻混合培养时比生长速率分别为0.59 d^-1和0.42 d^-1,分别比栅藻LX1和雨生红球藻的单一藻种培养提高了36%、9.0%。与单一藻种相比,混合培养促进了微藻的生物质产量,栅藻LX1和雨生红球藻混合藻种的生物质产量（277 mg·L^-1）分别比栅藻LX1、雨生红球藻的单一藻种培养提高了64%和42%。栅藻LX1与雨生红球藻藻种组合具备作为二级出水条件下能源微藻培养合适混合藻种的潜力。     

纳米MgO对斜生栅藻的毒性效应及致毒机理

为探究纳米氧化镁(MgO)对微藻的毒性效应及致毒机制,以斜生栅藻(Scenedesmus obliquus)为实验材料,测定了纳米MgO对斜生栅藻细胞形态、生长、叶绿素含量及SOD和POD酶活性的影响.结果表明,低浓度的纳米MgO(0.8 mg·L-1)即可对斜生栅藻的生长及叶绿素的合成有抑制作用,说明纳米MgO对斜生栅藻具有明显的毒性.100 mg·L-1纳米MgO处理时,完全抑制了栅藻的生长和叶绿素的合成,藻细胞在4 d内死亡.通过亚甲基蓝还原法测定培养体系中活性氧(ROS)含量发现,随着纳米MgO浓度的升高,ROS大量增加;且纳米MgO暴露条件下,POD酶活性相比于对照组显著增强,但不同浓度纳米MgO处理组之间POD酶活性无明显差异.纳米MgO处理96 h后,当其浓度高于0.8 mg·L-1时,SOD酶活性随浓度升高而降低,表明长时间暴露于纳米MgO后,SOD酶活性受高浓度纳米MgO抑制,而POD酶则作为主要的抗氧化酶清除产生的各种自由基.扫描电镜观察发现,纳米MgO(20 mg·L-1)处理时,斜生栅藻细胞变形,甚至裂解.纳米MgO发生团聚,附着在栅藻细胞表面;团聚的纳米MgO对藻细胞鞭毛有缠绕作用,使细胞聚集成团,限制了藻细胞的游动,并导致细胞间相互遮弊,不利于藻细胞吸收光能.透射电镜观察发现,纳米MgO没有进入藻细胞内部.通过测定纳米MgO解离出的Mg2+的含量和对藻细胞部分生理生化指标的影响发现,其对藻细胞没有毒性效应.因此,纳米MgO对斜生栅藻的致毒机理可归纳为:通过释放大量ROS对藻细胞产生氧化胁迫抑制其生长;高浓度的纳米MgO引起藻细胞的接触性物理损伤,导致藻细胞质壁分离,裂解;同时纳米MgO团聚并覆盖在藻细胞表面,通过与藻细胞鞭毛的相互作用使细胞聚集成团,影响细胞的正常游动及其对光能、营养物质的吸收利用和气体的交换.     

从绿色巴夫藻超微结构观察硒对锌毒性的保护效应

利用透射电镜研究绿色巴夫藻硒对锌毒性的保护效应 .从细胞超微结构特性比较对照和两个实验组 ,实验组 1培养基加入 15mgL-1Zn (ZnSO4·7H2 O) ,实验组 2培养基中加入 15mgL-1Se (Na2 SeO3 ·5H2 O)和 15mgL-1Zn .实验结果表明 ,在该浓度下单独加Zn ,藻细胞的叶绿体结构严重受损 ,而加入Se后可以阻止这种效应的产生 .藻细胞受Zn的影响 ,在细胞质和叶绿体基质中产生不同体积的电子致密的颗粒体 ,相反用Zn和Se处理的藻细胞则在液泡包含体中出现电子致密的颗粒体 .因此假设 :当单独加Zn时 ,藻细胞以形成电子致密的颗粒体的特殊分子形式分隔Zn ;而当形成锌硒化合物时 ,这一过程被阻止 ,此时Zn无活性 ,因而可保护细胞抵抗Zn的毒性 .图 3表 1参 2 8     

纳米银和银离子对2种微藻的急性毒性效应

为了探讨AgNPs对典型微藻的急性毒性效应及其机制,采用柠檬酸钠还原法制备AgNPs,以摇瓶实验法评估了不同浓度的AgNPs和Ag+对铜绿微囊藻和普通小球藻叶绿素a含量、形态结构和叶绿素荧光参数的影响。实验结果表明:AgNPs对普通小球藻和铜绿微囊藻的96 h-EC50分别为1.113 mg·L-1和0.697 mg·L-1,而Ag+对2种藻的96 h-EC50分别为0.106 mg·L-1和0.032 mg·L-1。扫描电镜结果表明:AgNPs处理使普通小球藻细胞表面出现褶皱,细胞变形甚至向内塌陷。对铜绿微囊藻部分细胞出现变形变得不规则,且出现某些胞外物质使细胞粘附在一起。透射电镜观察发现,高浓度Ag+处理使2种藻的细胞均发生质壁分离,部分细胞转变为孢子。而AgNPs处理使普通小球藻细胞蛋白核增大,蛋白核与类囊体区无明显连接通道。铜绿微囊藻拟核区膨大,类囊体和色素体被推向四周,部分类囊体断裂,同时,发现该藻可以分泌胞外物质在细胞周围吸附AgNPs颗粒。对于普通小球藻,0.6 mg·L-1AgNPs处理后细胞光系统Ⅱ的最大光化学量子产率ΦP0相对于CK没有显著差异,但0.09 mg·L-1Ag+处理使ΦP0显著增加。在高浓度AgNPs或Ag+处理时,ΦP0均显著降低。AgNPs未对普通小球藻光系统II性能参数PI_Abs造成影响,但不同浓度Ag+处理均使得该参数显著升高。对于铜绿微囊藻,2种毒物均使其ΦP0显著降低。而PI_Abs仅在2种毒物的最高浓度处理时显著降低。综上,AgNPs对2种藻的急性毒性远小于Ag+,而两者对铜绿微囊藻的毒性均大于普通小球藻。AgNPs胁迫使2种藻叶绿素a含量显著降低,并诱导2种藻在形态结构和光合生理方面发生了显著变化,造成不同程度的损伤,但与Ag+的毒性效应存在一定的差异。提高光吸收能通量补偿耗散能量和分泌胞外物质结合Ag+是微藻2种重要的解毒机制。     

纳米氧化锌对羊角月牙藻毒性效应及其在藻细胞内外的分布

为研究纳米氧化锌(ZnO NP)的毒性效应及其在细胞内外分布,以羊角月牙藻(Selenastrum capricornutum)为模型藻类,研究了不同浓度ZnO NP对羊角月牙藻生长、叶绿素含量、可溶性蛋白含量、超氧化物岐化酶(SOD)及过氧化物酶(POD)活性、丙二醛(MDA)含量及细胞内外ZnO NP含量变化。结果表明,ZnO NP对羊角月牙藻的生长抑制与处理浓度呈现正相关。在45 mg·L~(-1)ZnO NP暴露24 h后,其生长抑制率已达到95%。当ZnO NP处理藻细胞72 h后,羊角月牙藻细胞的叶绿素含量与处理浓度之间存在剂量-效应关系。低浓度(0.5 mg·L~(-1))ZnO NP处理后藻细胞可溶性蛋白质含量、SOD和POD活性明显下降,MDA含量升高,其产生的毒性效应高于高浓度组(5 mg·L~(-1)、45 mg·L~(-1))。细胞培养液溶出Zn2+量及藻细胞外吸附的ZnO NP量与ZnO NP处理浓度成正比,但是藻细胞内ZnO NP量与ZnO NP浓度没有相关性,胞内积累量基本维持不变。研究表明,各浓度组对藻细胞毒性的差异,不仅与细胞内Zn2+量有关,还与细胞外粘附的ZnO NP有关。     

FDA-PI双色荧光法检测蓝藻细胞活性的研究

对FDA-PI双色荧光法检测水华鱼腥藻和铜绿微囊藻的活性进行了研究,用荧光显微镜对染色结果进行测定.结果表明,在蓝色光激发下(495nm),活细胞被双醋酸荧光素FDA染成亮绿色,死亡细胞被碘化丙锭PI染成红色.染色效率与原植体类型和细胞密度有关.对细胞密度为6×107—7×109个·l-1的铜绿微囊藻,FDA染色效率可达94%以上.对细胞密度为4×107—5×108个·l-1的水华鱼腥藻,FDA染色效率可达91%以上,但细胞密度增大到5×109个·l-1时,由于藻丝体易卷曲在一起,FDA染色率下降到67%.对死亡细胞,PI染色率基本都可达到100%.因此,用FDA-PI检测活细胞和死亡细胞混合的细胞悬液,可根据细胞所发出的不同荧光而判断细胞活性.     

作物根系对干旱胁迫逆境的适应性研究进展

主要就作物根系的形态性状、根系提水作用、生理代谢、根系细胞壁蛋白及其生长性能与干旱胁迫间的关系作了综述;指出应加强对干旱逆境下根系发育及根系生长性状变化上的遗传机理研究。     

Variability of spectral absorption efficiency within living cells of Pyrocystis lunula (Dinophyta)

This study examines variability in the spectral absorption efficiency in various parts of living Pyrocystis lunula cells. changes in spectral absorption efficiency measured within cells are attributed to light-induced chloroplast migration as well as reorganization of cellular material during the process of asexual reproduction. During the dark cycle, major pigment peaks were well resolved in those spectra measured in the distal cytoplasmic strands where chloroplasts were concentrated. In contrast, the absorption efficiencies measured in the granular central area that did not contain chloroplasts decreased gradually from the blue to the red portions of the spectrum and are similar to those published for detrital particles. When chloroplasts migrated toward the center of the cell in response to light, absorption efficiency curves for the granular central area were flatter than the curves measured in cytoplasmic strands containing chloroplasts. This was due to the combined absorption properties of the central area and the chloroplasts. Absorption efficiency spectra were also flattened in aplanospores within the parent vegetative cells because of the concentration of cellular material into smaller areas. These findings suggest that shapes of spectral absorption curves measured for the major phytoplankton groups cannot be assumed to remain constant over time. Furthermore, changes in cell structure may account for some of the reported diel changes in beam attenuation and stimulated fluorescence in natural waters.     

松科植物对干旱胁迫的反应

水分亏缺是制约树木生长的重要环境因子，植物通过形态、生理以及分子水平来适应水分亏缺．渗透调节使植物在低水势下维持正常生理活动，是植物忍耐水分亏缺的重要生理机制．干旱胁迫下，植物形态结构变化有利于水分吸收和传导，从而提高水分利用效率；同时，生物量向根部的分配增加，叶面积/边材面积比发生变化，这种生物量分配转移提高了根和茎向叶片输水能力，从而防止气穴现象；干旱胁迫容易引起光能过剩，过剩的光能会对光合器官产生潜在的危害．依赖于叶黄素循环的热耗散是光保护的主要途径；同时酶促及非酶促系统也是防止光合器官破坏的重要途径；脱落酸作为一种激素逆境信号，活化了与抗旱诱导有关的基因．本文从形态变化、渗透调节、气穴现象、光合作用、水分利用效率、脱落酸以及分子机理等方面阐述了松科植物对干旱胁迫的响应，并对耐旱指标的筛选进行了讨论，干旱胁迫下，各耐旱机理相互制约，需要联合各个方面的因素来考虑整个植物对干旱的反应．参99。     

五氯酚对鱼鳃丝组织结构及电解质元素的影响

利用场发射扫描电镜观察了罗非鱼在0．05mg L^-1和0．10mg L^-1五氯酚(PCP)中暴露3d，7d和10d后鳃丝上皮组织结构的变化，以及扫描电镜／X-射线能谱联用仪定量分析鳃丝中主要电解质元素Na，Ca和Cl的变化．与CK比较，随着暴露时问的延长和暴露浓度的增加，泌氯细胞出现的密度增加，部分泌氯细胞个体的面积也增大，扁平上皮细胞的微脊碎片数量增加，排列散乱，细胞受损现象明显；鳃丝中Na，Ca和Cl元素的含量总体上呈下降的趋势，在一定程度上说明与Ca^2 、Na^ 和Cl^-运转有关的酶的活性受到抑制．图2参14     

Replacement and aging of chloroplasts inStrombidium capitatum (Ciliophora: Oligotrichida)

The planktonic ciliateStrombidium capitatum (Leegaard, 1915) Kahl, 1932 retains functional chloroplasts derived from ingested algal cells. Chloroplast replacement and aging were experimentally investigated in cultured ciliates provided with a cryptophyte (Pyrenomonas salina), a prymnesiophyte (Isochrysis galbana) and a prasinophyte (Pyramimonas sp.) as sources of plastids. All three algae were ingested and chloroplasts from all were retained by the ciliate. Within 15 min of exposure to the cryptophyte, this alga was taken up by the ciliates. Initially, most of the cryptophyte chloroplasts were in intact algal cells in ciliate vacuoles. By 2 h, cryptophyte plastids were commonly found free in the ciliate cytoplasm. WhenS. capitatum was switched from a diet containing cryptophytes to a non-cryptophyte diet, most cryptophyte chloroplasts were diluted out of the ciliates by cell division and/or replaced by non-cryptophyte chloroplasts within 9 h. When the ciliates are not provided with algae, they decrease in size and number. However, the starving ciliate cells contain some chloroplasts for as long as they live (40 h or more). Under these conditions, cryptophyte chloroplasts persist longer than the other chloroplast types. Our observations suggest that chloroplast retention times inS. capitatum depend on the type of chloroplast as well as the availability of phytoplankton containing suitable new chloroplasts, and probably also on the physiological states of the ingested algae and the ciliates. It is interesting that we were not able to grow this ciliate when we provided it only with prey that lacked chloroplasts.Contribution No. 7241 from Woods Hole Oceanographic Institution. This research was supported by NSF grants OCE-8600765 and OCE-8709961 to D.K.S     

水体中氧化铁鞘细菌的分离鉴定及保藏

采用试管静止富集培养及生态模拟富集培养，结合平板划线、稀释涂布、平板滴加法，成功地从150份水样中分离到94株鞘细菌．利用Winogradsky液体试管法进行产铁氧化酶鞘细菌的快速初筛，从中筛选出24株产铁氧化酶能力较高的菌株，再经摇瓶复筛，获得产酶活性最高的菌株FC9901．采用多相鉴定方法对菌株FC9901细胞形态、培养特征、生理生化反应及各种碳源利用情况进行了研究，根据《伯杰氏细菌鉴定手册》第9版，把该菌株鉴定为第十四群鞘细菌类(sheathed bacteria)球衣菌属(Sphaerotilus)浮游球衣菌(Sphaerotilus natans)．对鞘细菌几种保藏方法进行比较研究，结果表明蒸馏水保藏法保藏时间长，是一种经济简便有效的保藏方法．图3表7参9     

Species and speciation in poikilothermal animals

On the basis of current scientific information related to intraspecific and interspecific variability of eco-physiological properties at the individual (organismic) and subindividual (tissues, cells, and molecules) levels and his own experimental data, the author comes to the conclusion that eco-physiological adaptation is one of the major processes determining speciation. During sequential multi-step adaptation at the most sensitive and plastic levels of organisation (whole organisms and their labile cells and proteins) to changed abiotic (physico-chemical) environmental factors, the latter directly affect the conservative proteins and polynucleotides that maintain the species' genetic information. Transition from phenotypical to genotypical changes may be achieved if the organisms have attained the limit of their phenotypical plasticity, via selection at cellular and molecular levels. The high rate of cellular mytotic division and a promiment rise in the speed of interrelated DNA-RNA protein synthesis under sudden and profound environmental changes may account for the high rates of speciation indicative of cataclysmic climatic changes. Evolutionary morphological changes are considered to follow physiological changes. The adaptive nature of morphological peculiarities in higher taxonomic categories (type, class, order, family; more seldom genus) may be accounted for by the selection of phenotypes possessing morphological peculiarities best suited to the ecological niches of the species concerned. These peculiarities (although often neutral to physiological changes that have determined the speciation) are transferred from species to species, manifesting both historical relationship and morphological unity. The suggested scheme of speciation is in accord with the conceptions of the intermittent character of speciation, which can proceed only if extreme, directed, climatic changes cover a larger portion of the species' area. As the organisms' organisation becomes more complex during phylogenesis, intraspecific variability decreases; this trend manifests itself in the progressive stabilization of body shape and other morphological properties, and in the increase of eco-physiological stability of cells and proteins.     

玉米铝（Al）胁迫研究初报

通过试验和分析，研究Ａｌ胁迫下的玉米生物性状和主要生理特性，明确活性铝浓度与玉米根系特征及地上部农艺性状之间存在极显著或显著的反比相关；活性铝的存在，降低玉米茎叶Ｎ、Ｐ、Ｋ等养分含量，提高叶片脯氨酸累积量；脯氨酸含量可作为玉米耐Ａｌ性的生理指标．     

一株产生物絮凝剂的Bacillus sp.的分离与鉴定

从重庆城南污水处理厂的活性污泥中分离筛选到一株絮凝剂产生菌,革兰氏染色呈阳性,好氧,有芽孢,可运动,短杆;分离菌的基因组DNA(G +C)mol%含量测定为5 0 .6 % ,16SrRNA序列分析表明,分离筛选出的絮凝剂产生菌的16SrRNA基因序列与Bacillus菌有高度同源性,从而确定絮凝剂产生菌RL - 2归于Bacillusbrevis菌属。絮凝实验表明:在有CaCl2 存在时用Bacillussp .RL - 2生物絮凝剂处理土壤悬液时用量少(10 0mL样品只需2mL絮凝剂) ,絮体形成快,矾花大,泥少;体系温度对活性影响很小,且在酸性与碱性条件下,絮凝活性均很高