应用原子力显微镜研究盐度对棕囊藻生长的影响

研究不同盐度梯度下棕囊藻的生长,实验设计了10个盐度梯度,结果表明:在盐度8~32范围内,棕囊藻的生长正常,最佳生长盐度为16,当盐度低于8或高于32时,藻的生长明显受到抑制,细胞数量增长缓慢,最大细胞数远低于16盐度组。研究发现:棕囊藻在盐度为0的环境下仍能缓慢生长,说明海洋中的棕囊藻赤潮在内陆的淡水湖也可能暴发,用原子力显微镜研究比较盐度为0及36的棕囊藻细胞表面形态,发现两者差异较显著,前者藻细胞的直径大于后者,且后者藻细胞表面形成了明显的塌陷。     

球形棕囊藻(Phaeocystis globosa Scherffel)赤潮成因分析

对1997年冬季和1999年夏季发生于广东省饶平枯林湾海域的棕囊藻赤进行了现场调查和监测，结果表明，棕囊藻赤潮的爆发是海洋环境变化和棕囊藻自身特性共同决定的，长期集约化养殖引起枯林湾海域的N、P、Fe、Mn浓度较高，特别是Fe浓度明显高于广东沿海的其它海区，为赤潮的爆发提供了物质基础，棕囊藻增殖过程中对DIN（可溶性无机氮）的需求量较高，特别对NO3^－-N有明显的主动吸收，与DIP（可溶性无机氮）相比，海水DIN浓度恢复速率较慢。可能成为限制棕囊藻赤潮的规模和持续时间的关键因子，海水温度的异常增高及降雨量偏少，与棕囊藻赤潮的发生紧密相关。     

二氯异氰脲酸钠和二氯异氰脲酸对棕囊藻细胞去除的研究

研究了二氯异氰脲酸钠和三氯异氰脲酸对球形棕囊藻的灭杀和控制作用,及其投药时间对除藻效果的影响,并比较2种药剂的除藻效果,初步探讨了除藻机理.结果表明,这2种除藻剂可以缓释次氯酸,提高其稳定性,有较长的药效期.当有效氯浓度达到4.5mg/L时,2种药剂均能有效地控制和灭杀球形棕囊藻,具有高效、低毒的优点.有效氯浓度为5.0mg/L,藻细胞去除率在24h可达到90%以上.在等于和大于5.0mg/L有效氯浓度时,三氯异氰脲酸的除藻效果优于二氯异氰脲酸钠,通过对2种药剂除藻率的t检验进一步证实了这一结论.     

溶藻活性物质对棕囊藻溶藻及其脂肪酸影响的模拟

为了探讨芽孢杆菌B1分泌的胞外溶藻活性物质对球形棕囊藻的溶藻特性和藻毒素物质脂肪酸的影响,比较了模拟自然水体中叶绿素a、p H、溶解氧DO、高锰酸盐指数和营养元素N、P浓度在溶藻前后的变化,并利用GC-MS检测了球形棕囊藻脂肪酸的成分和含量.用体积比1∶100的芽孢杆菌B1胞外活性物质处理模拟水体14 d,发现水体中叶绿素a、p H值和高锰酸盐指数随处理时间的增加而降低,DO和N、P浓度随处理时间的增加而增加.在第14 d时,处理组水体中p H值由8.50降低到7.51,叶绿素a降低82.3%(P0.05),DO增加29.5%(P0.05),高锰酸盐指数降低55.2%(P0.01).NH+4-N、NO-2-N、NO-3-N和PO3-4-P浓度分别增加了0.46、1.50、6.24和1.30倍.投加活性物质处理14 d后,球形棕囊藻藻毒素中的主要3种脂肪酸C18:2、C16:0和C18:1分别降低了100%、97.7%和85.4%(P0.01),总脂肪酸含量降低83.4%(P0.01).结果表明,芽孢杆菌B1胞外溶藻活性物质在模拟自然水体中能有效抑制球形棕囊藻的生长,并降低藻毒素脂肪酸的含量.研究结果为芽孢杆菌B1胞外活性物质的生态安全性应用提供理论基础.     

细菌B1胞外活性物质对球形棕囊藻的溶藻机制初探

为探讨溶藻细菌B1对球形棕囊藻的溶藻作用机理及起作用的胞外活性物质的类别,研究了从珠海香洲码头赤潮海水中分离获得的溶藻细菌BI的无菌滤液对球形棕囊藻生长过程中的叶绿素a含量、丙二醛含量(MDA)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性的影响,并对溶藻细菌B1分泌的胞外活性物质进行了初步分析.当100 mL藻液中加入1 mL的无菌滤液,除藻率达到了83％.B1无菌滤液使藻中丙二醛的含量显著上升,48 h达到最大值153％,球形棕囊藻的SOD、CAT活性在处理开始后保持下降趋势,分别在96 h、48 h达到最低值27％、33％.研究表明B1无菌滤液(不大于1 mL)对棕囊藻的溶藻效果与加入的量成正比,且主要通过降低机体中保护酶的活性和加剧膜脂质过氧化的程度等来抑制球形棕囊藻的生长;细菌B1分泌的胞外活性物质中可能含有芳香醛类且包括载有磷基团的有机物,但不含蛋白质.     

球形棕囊藻与红树林细菌Flavobacterium sp.相互关系的研究

从深圳福田红树林中分离出一株细菌,对球形棕囊藻有溶藻活性,通过形态鉴定为假单胞菌属细菌。将球形棕囊藻、藻过滤液和提取的溶血毒素与细菌共培养,通过荧光显微计数及平板计数研究活细菌数和可培养细菌数,结果表明:当细菌直接与棕囊藻共培养时,细菌很快进入VBNC状态,细菌能够促进棕囊藻溶血毒素的积累。而将细菌置于透析袋中与棕囊藻一起共培养时,棕囊藻对细菌的VBNC状态影响不大,细菌对棕囊藻溶血毒素的积累无影响。对数期、稳定期棕囊藻培养过滤液和溶血毒素均可使细菌很快进入VBNC状态,溶血毒素可能在球形棕囊藻在被牧食及抑制其他竞争者中(化感作用)扮演着重要的角色。     

芽孢杆菌 B1胞外活性物质对球形棕囊藻的溶藻特性研究

从珠海香洲码头赤潮海水中分离获得1株对球形棕囊藻有显著溶藻效果的芽孢杆菌B1,研究了B1对棕囊藻的溶藻作用方式,溶藻过程中藻细胞结构变化,并采用透析、乙醇沉淀、有机溶剂萃取、酸碱及热稳定性分析等方法探讨了溶藻活性物质的性质.结果表明,B1无菌滤液对棕囊藻有较强的溶藻效应,除藻率达94.9%,B1通过分泌活性物质间接对球形棕囊藻的生长产生抑制;藻培养液中加入B1无菌滤液16 h后,藻细胞发生团聚,细胞壁失去完整性,56 h后藻细胞破碎,胞内物质溶出;相对分子质量<3 500的分泌物是溶藻过程中起主要作用的活性物质,具有较强极性和热稳定性,在121℃加热20 min后,仍然有良好的溶藻能力,除藻率达92.6%,活性物质在pH 9.0左右溶藻能力较强,在乙醇中不发生沉淀反应,由此推测该活性物质为含有酸性或碱性基团的非生物活性分子,不属于蛋白质、核酸、多糖等物质.     

Phytoremediation of Cadmium-Contaminated Soils by Rorippa globosa Using Two-Phase Planting (5 pp)

Background Phytoextraction of contaminated soils by heavy metals can provide a great promise of commercial development. Although there are more than 400 species of hyperaccumulators found in the world, phytoremediation technology is rarely applied in field practice for remedying contaminated soils, partially due to low biomass and long growth duration for most of discovered hyperaccumulating plants. In order to enhance the metal-removing efficiency in a year, the two-phase planting countermeasure of phytoextraction by harvesting anthesis biomass was investigated on the basis of the newly found Cd-hyperaccumulator Rorippa globosa (Turcz.) Thell. with 107.0 and 150.1 mg/kg of the Cd accumulation in stems and leaves, respectively, when soil Cd added was concentrated to 25.0 mg/kg. Methods The field pot-culture experiment was used to observe the distribution property of R. globosa aboveground biomass and to examine characteristics of accumulating Cd by the plant at different growth stages. The concentration of Cd in plants and soils was determined using atomic absorption spectrophotometry (AAS). Results and Discussion The results indicated that the total dry stem and leaf biomass of R. globosa harvested at the flowering phase was up to 92.3% of that at its full maturity and the concentration of Cd in stems and leaves harvested at the flowering phase was up to 73.8% and 87.7% of that at the mature phase, respectively. The Cd-removing ratio by shoots of R. globosa harvested at the flowering phase was up to 71.4% of that at the mature phase. It was also found, by observing the growth duration of R. globosa, that the frostless period at the experiment site was twice as long as the growth time from the seedling-transplanted phase to the flowering phase of the hyperaccumulator. Conclusion R. globosa could be transplanted into contaminated soils twice in one year by harvesting the hyperaccumulator at its flowering phase based on climatic conditions of the site and traits of the plant growth. In this sense, the extraction efficiency of Cd in shoots of R. globosa increased 42.8% compared to that of at its single maturity when the plant was transplanted into contaminated soils after it had been harvested at its flowering phase and the plant accumulated Cd from soil at the same extraction ratio at its second flowering phase. Thus, the method of anthesis biomass regulation by the two-phase planting is very significant to increase the Cd-removing efficiency by phytoremediation used in practice over the course of a year. Recommendation and Outlook As for some hyperaccumulators that the growth duration from the seedling-transplanted phase to the flowering phase are short and the concentrations of heavy metals accumulated in their shoots at the flowering phase are high, the efficiency of phytoremediation can greatly be improved using the method of the two-phase planting.     

尿素对中国近海3种典型赤潮藻生长的影响

采用实验室一次性培养的方法,研究了尿素对中国近海3种赤潮藻---球形棕囊藻(Phaeocystis globosa)、锥状斯氏藻(Scrippsiellatrochoidea)和中肋骨条藻(Skeletonema costatum)生长的影响.结果表明,这3种赤潮藻均能利用有机氮源尿素生长,且表现出明显的浓度效应:当尿素浓度介于0~8.82μmol.L-1(以N计,下同)时,3种赤潮藻的比生长速率均较低,细胞密度也较低,与对照组相比无显著差异;当尿素浓度大于8.82μmol.L-1时,这3种赤潮藻表现出明显的生长效应.然而,3种赤潮藻适宜生长的尿素浓度并不一致,当尿素浓度分别为882、8820和8820μmol.L-1时,球形棕囊藻、锥状斯氏藻和中肋骨条藻分别达到最大比生长率0.76、0.54和0.66d-1.将3种赤潮藻在不同尿素浓度下的最大比生长速率用Monod方程拟合,得出球形棕囊藻、锥状斯氏藻和中肋骨条藻在以尿素为唯一氮源条件下的最大比生长速率和半饱和常数分别为0.73、0.47、0.63d-1和12.36、21.83和20.34μmo.lL-1.由此可见,这3种藻能够利用有机氮源尿素,无疑扩展了其氮营养来源,在无机氮源缺乏时,具有竞争优势.相比较而言,球形棕囊藻对尿素的亲和力最高,在低浓度尿素条件下,球形棕囊藻具有较高的竞争力.     

有机磷农药草甘膦异丙胺盐对球形棕囊藻的刺激效应

设置0、0．001、0．01、0．1、l和10mg·L-16个质量浓度梯度,测定了草甘膦异丙胺盐（Glyphosate—isopropylammonium）农药对球形棕囊藻（Phaeocystis globosa）生长、叶绿素a含量、SOD和CAT活性的影响,探讨了有机磷农药对球形棕囊藻的毒物刺激效应。结果表明,低浓度的草甘膦异丙胺盐对球形棕囊藻的生长呈现出刺激效应。在实验所设定的6个质量浓度梯度范围内,0．001mg·L0和0．01mg·L-1质量浓度处理下的刺激效应最为显著。该两个质量浓度处理下,球形棕囊藻的平均相对增长率比对照分别提高了1．27％和0．45％,叶绿素a含量分别比对照提高了9．2％和2．5％,SOD活性提高了5．9％和7．2％,CAT活性提高了93％和155％;而当草甘膦异丙胺盐质量浓度在0．1mg·L-1以上时,球形棕囊藻的生长率、叶绿素a含量、SOD活性均显著低于对照。草甘膦异丙胺盐在0．001mg·L-1和0．01mg·L-1范围内对球形棕囊藻均呈现刺激效应,而高于0．1mg·L-1时球形棕囊藻各生理指标表现出显著抑制效应。研究结果为了解有机磷农药对海洋微藻生长的影响特征,揭示有机磷农药对微藻产生低促高抑的规律和为深入探索赤潮暴发的成因提供依据。