小珊瑚藻对赤潮异弯藻的克生效应及其 对UV-B辐射增强的响应

通过共培养方法,以细胞密度为主要测定指标,研究了小珊瑚藻对赤潮异弯藻的克生效应及其对UV-B辐射增强的响应.结果表明：小珊瑚藻的新鲜组织和水溶性抽提液对赤潮异弯藻的生长表现出显著的抑制作用(P＜0.05),说明小珊瑚藻对赤潮异弯藻具有克生效应;而小珊瑚藻干粉末和培养水过滤液对赤潮异弯藻生长则无明显影响(P>0.05).将小珊瑚藻用不同剂量的UV-B辐射预处理后,再与赤潮异弯藻共培养,除培养水过滤液外,其新鲜组织、干粉末和水溶性抽提液对赤潮异弯藻生长的作用效果均有所改变,高剂量(3.0 J·m^-2)UV-B辐射使小珊瑚藻对赤潮异弯藻的抑制作用减弱(P＜0.05),而低剂量(0.9 J·m^-2) UV-B辐射则使抑制作用增强(P＜0.05).     

缘管浒苔对赤潮异弯藻的克生效应

利用共存培养系统研究了缘管浒苔(E nterom orpha linza)新鲜组织和干粉末对赤潮异弯藻(H eterosigm a akash iw o)生长的克生效应。共存实验结果表明,缘管浒苔新鲜组织和干粉末对赤潮异弯藻生长均有强烈的克生效应。同时,研究了缘管浒苔培养水过滤液对赤潮异弯藻的克生作用。赤潮异弯藻在缘管浒苔培养水过滤液的半连续添加方式下生长受到明显的克制作用,但在一次性培养方式下生长未受到明显的抑制作用,说明克生物质的连续分泌是有效克制赤潮异弯藻生长的关键;煮沸的大藻培养水过滤液对微藻的生长无抑制作用,表明克生物质在高温下不稳定和易分解。     

不同起始密度对3种赤潮微藻种间竞争的影响

通过共培养的方法,研究了不同起始密度对赤潮异弯藻和中肋骨条藻、塔玛亚历山大藻和赤潮异弯藻、塔玛亚历山大藻和中肋骨条藻之间种间竞争的影响。结果表明1赤潮异弯藻对中肋骨条藻的生长有一定的抑制作用,随着初始接种时赤潮异弯藻细胞密度的提高,这种抑制作用愈加明显。2塔玛亚历山大藻和中肋骨条藻与塔玛亚历山大藻和赤潮异弯藻之间的种间竞争结果相类似,即起始密度比例为A∶S(H)=1∶4时,中肋骨条藻和赤潮异弯藻分别在竞争中占优势;当起始密度比例为A∶S(H)=1∶1和A∶S(H)=4∶1,塔玛亚历山大藻在竞争中占优势。     

赤潮水体中胶体物质对赤潮异弯藻(Heterosigma akashiwo)和中肋骨条藻(Skeletonema costatum)生长的影响

2004年2月9～10日在胶州湾发生的柔弱几内亚藻(Guinardia delicatula)赤潮期间分别在3个典型赤潮站位采集表层水样,利用过滤(0.7μm)和切向超滤(1000 Daltons)分离赤潮暴发水体中的胶体物质,分析了不同滤液及截留液的有机碳含量,并将赤潮异弯藻 (Heterosigma akashiwo)和中肋骨条藻(Skeletonema costatum)在<1000 Daltons的超滤液及具有不同含量胶体物质(1000 Daltons～0.7μm)的截留液中进行培养.结果发现在发生柔弱几内亚藻赤潮期间水体中的溶解有机碳浓度会增加,特别是胶体有机碳浓度会成倍增长;在不同胶体含量海水中培养的赤潮异弯藻的最大生长率μmax和最大生物量Bf与对照组相比会随着胶体含量的增加而显著增大,而中肋骨条藻的最大生长率μmax和最大生物量Bf与对照组相比会随着胶体含量增加而显著降低,表明柔弱几内亚藻赤潮水体中的胶体物质对赤潮异弯藻的生长起促进作用,而对中肋骨条藻的生长起抑制作用.这说明柔弱几内亚藻在增殖过程中所产生的胶体物质具有他感作用,会影响其它微藻的生长,从而可能对柔弱几内亚藻赤潮的形成起重要作用.     

UV-B辐射增强对三种赤潮微藻DNA的伤害效应

运用生态毒理学和生物化学方法研究了UV-B辐射增强对赤潮异弯藻、亚历山大藻和中肋骨条藻DNA的伤害作用.结果表明,3种赤潮微藻的生长状况对UV-B辐射增强的敏感性不同;对UV-B辐射增强的敏感性由高到低依次是赤潮异弯藻、亚历山大藻和中肋骨条藻.随着UV-B辐射剂量的增加,3种赤潮微藻的DNA损伤程度提高,而且赤潮异弯藻DNA的损伤程度明显高于亚历山大藻和中肋骨条藻,亚历山大藻DNA的伤害程度又远远高于中肋骨条藻.UV-B辐射处理解除后,损伤DNA可明显恢复.赤潮异湾藻和亚历山大藻恢复培养6d,损伤DNA可明显恢复(P＜0.05);而中肋骨条藻恢复培养3d,损伤DNA可明显恢复(P＜0.05),说明3种赤潮微藻的DNA损伤水平不适合作为指示UV-B辐射增强的生物学指标.     

大米草对赤潮藻的抑制作用及其抑藻物质的分离鉴定

以赤潮异弯藻和海洋原甲藻为研究对象,研究了海洋滩涂植物大米草对两种赤潮微藻的抑制作用及抑藻物质.结果表明：大米草对赤潮异弯藻的影响表现为高浓度抑制生长,低浓度则有一定的促进作用,大米草新鲜组织、干粉末和提取物浓度分别为4.8、0.8和0.5 mg·ml^-1以上时对赤潮异弯藻有致死作用;大米草对海洋原甲藻则表现出明显的抑制作用,且大米草新鲜组织、干粉末和提取物浓度分别为9.6、1.6和1.25 mg·ml^-1以上时对海洋原甲藻有致死作用.对大米草提取物进行抑藻活性物质的分离纯化,从中分离鉴定了2个具有抑藻活性的黄酮糖苷类化合物：异鼠李素-3-O-槐二糖-7-O-鼠李糖苷和丁香亭-3-O-半乳糖苷.     

小珊瑚藻对赤潮异弯藻的化感效应

研究了不同浓度的小珊瑚藻组织4种不同极性有机溶剂(甲醇、丙酮、乙醚、氯仿)提取物对赤潮异弯藻的生长抑制作用.结果表明：小珊瑚藻组织甲醇提取物对赤潮异弯藻的生长抑制活性最强,并且在较高浓度下能使赤潮异弯藻完全死亡,其他3种有机溶剂提取物对赤潮异弯藻的生长无明显影响.表明小珊瑚藻组织中含有的对赤潮异弯藻有抑制作用的活性物质具有较高的极性.对小珊瑚藻的甲醇提取物进行液液萃取,将其分离为石油醚相、乙酸乙酯相、正丁醇相和蒸馏水相, 并对赤潮异弯藻进行生物活性检测.结果发现石油醚相和乙酸乙酯相具有较强的杀藻活性,表明脂肪酸可能是小珊瑚藻组织内抑制赤潮异弯藻生长的化感物质的重要组成成分之一.     

浒苔干粉末提取物对东海原甲藻和中肋骨条藻的克生作用

研究不同溶剂的浒苔干粉末组织提取液对两种赤潮藻—东海原甲藻和中肋骨条藻生长的克生效应。结果表明, 浒苔提取物中确实含有可以影响赤潮藻类生长的克生物质,克生作用具有较明显的浓度效应,添加浓度低时可能会表现为一定的促进生长的作用,添加浓度较高时表现为抑制作用,添加浓度越大,抑制作用越强,即“低促高抑”的特点,这与浒苔提取物对其他微藻的作用相似。其中,蒸馏水提取物对这两种赤潮藻的克生作用小于有机溶剂提取物,有机溶剂中,甲醇和乙酸乙酯提取物对这两种赤潮藻的克生效果最好,正己烷相对较差。根据相似相溶原理,可以初步推断最有效的克生物质应为具有相对较高的极性的有机物。两种赤潮藻对克生物质的敏感程度不同。东海原甲藻对克生物质的敏感性高于中肋骨条藻。蒸馏水、甲醇、乙酸乙酯、正己烷的浒苔干粉末提取物影响东海原甲藻生长的致死作用阈值浓度分别为5.00 g/L、0.50 g/L、0.50 g/L、0.63 g/L（相当于浒苔新鲜藻体浓度为22.00 g-wet/L、2.20 g-wet/L、2.20 g-wet/L、2.75 g-wet/L）;影响中肋骨条藻生长的致死作用阈值浓度则分别为20.00 g/L、1.25 g/L、1.25 g/L、2.50 g/L（相当于新鲜藻体浓度为88.00 g-wet/L、5.50 g-wet/L、5.50 g-wet/L、11.00 g-wet/L）。     

3种海洋赤潮微藻蛋白质和核酸合成对UV-B辐射增强的响应

应用同位素标志法, 研究了赤潮异弯藻 (Heterosigmaakashiwo) 、亚历山大藻 (Alexandriumtamarense ) 和中肋骨条藻 (Skeletonemacostatum) 核酸和蛋白质合成对紫外线B波段 (UV_B, 2 80~ 32 0nm) 辐射增强的响应变化。结果表明 :1) 按照由高到低的顺序, 3种海洋赤潮微藻对UV_B辐射增强的敏感性依次是赤潮异弯藻、亚历山大藻和中肋骨条藻。 2 ) UV_B辐射增强抑制赤潮异弯藻的生长和脱氧核糖核酸 (DNA) 的合成, 而低剂量的UV_B辐射对中肋骨条藻和亚历山大藻的生长与DNA的合成表现出刺激作用, 高剂量则表现出抑制作用。 3) 随着UV_B辐射的增强, 3种海洋赤潮微藻核糖核酸 (RNA) 和蛋白质的合成速度下降, 其中赤潮异弯藻合成速度的下降幅度明显大于中肋骨条藻和亚历山大藻, 表明赤潮异弯藻RNA和蛋白质的合成对UV_B辐射增强的敏感性高于中肋骨条藻和亚历山大藻。     

有机磷农药草甘膦异丙胺盐对赤潮异弯藻的毒物干扰效应

以我国典型赤潮藻赤潮异弯藻(Heterosigma akashiwo)为研究对象,设置了6个浓度梯度(0 mg·L^-1、0.001mg·L^-1、0.01 mg·L^-1、0.1 mg·L^-1、1 mg·L^-1和10 mg·L^-1)的草甘膦异丙胺盐处理,研究了草甘膦异丙胺盐暴露对赤潮异弯藻的生长、叶绿素a含量和可溶性蛋白含量等指标的影响.结果表明,草甘膦异丙胺盐对赤潮异弯藻具有明显的毒性效应,10 mg·L^-1浓度处理下,赤潮异弯藻细胞大量死亡,藻细胞密度以及叶绿素a、可溶性蛋白的含量显著降低(p<0.05);当草甘膦异丙胺盐浓度在0.001～1 mg·L^-1范围内,在培养的第3 d草甘膦异丙胺盐能够显著促进赤潮异弯藻的细胞密度增加,叶绿素a含量也明显高于对照组(p<0.05),表现出毒物刺激效应;在暴露实验的中后期(第7 d、第9 d和第11 d),赤潮异弯藻的各生长指标均与对照无显著差异,可能是随着培养时间的延长,农药的降解、生物体对农药的适应、进入细胞的农药减少等原因,藻细胞生理状态逐渐恢复到正常水平.     

Allelopathic effects of Ulva pertusa , Corallina pilulifera and Sargassum thunbergii on the growth of the dinoflagellates Heterosigma akashiwo and Alexandrium tamarense

The allelopathic effects of fresh tissue, dry powder and aqueous extracts of three macroalgae, Ulva pertusa, Corallina pilulifera and Sargassum thunbergii, on the growth of the dinoflagellates Heterosigma akashiwo and Alexandrium tamarense were evaluated using coexistence culture systems in which concentrations of the three macroalga were varied. The results of the coexistence assay showed that the growth of the two microalgae was strongly inhibited by using fresh tissue, dry powder and aqueous extracts of the three macroalga; the allelochemicals were lethal to H. akashiwo at relatively higher concentrations of the three macroalga. The macroalgae showing the most allelopathic effect on H. akashiwo and A. tamarense using fresh tissue were U. pertusa and S. thunbergii, using dry powder were S. thunbergii and U. pertusa, and using aqueous extracts were U. pertusa and C. pilulifera. We also examined the potential allelopathic effect on the two microalgae of culture filtrate of the three macroalga; culture medium filtrate initially exhibited no inhibitory effects when first added but inhibitory effects became apparent under semi-continuous addition, which suggested that continuous release of small quantities of rapidly degradable allelochemicals from the fresh macroalgal tissue were essential to effectively inhibit the growth of the two microalgae.     

Comparative studies on the allelopathic effects of two different strains of Ulva pertusa on Heterosigma akashiwo and Alexandrium tamarense

Allelopathic effects of fresh tissue and dry powder of a nonsexual and a sexual strain of the macroalga Ulva pertusa on the growth of the microalgae Heterosigma akashiwo and Alexandium tamarense were evaluated using long-term coexistence culture systems in which several concentrations of macroalga fresh tissue and dry powder were used. The effects of macroalga culture medium filtrate on the two HAB algae were also investigated. Moreover, isolation co-culture systems were built to confirm the existence of allelochemicals and preclude the growth inhibition by direct contact. Short-term algicidal effect assays of macroalgae on H. akashiwo were carried out to measure the rate of algal cell lysis. The results of the coexistence assays showed that the growth of H. akashiwo and A. tamarense was strongly inhibited by fresh tissue and by dry powder of both strains of U. pertusa. The allelochemicals were lethal to H. akashiwo at relatively higher concentrations. The macroalga culture medium filtrate exhibited no apparent growth inhibitory effect on the two HAB algae under initial or semicontinuous filtrate addition, which suggested that continuous release of small quantities of rapidly degradable allelochemicals from the fresh tissue of both strains of U. pertusa was essential to effectively inhibit the growth of H. akashiwo and A. tamarense.     

Effects of two species of macroalgae—Ulva pertusa and Gracilaria lemaneiformis—on growth of Heterosigma akashiwo (Raphidophyceae)

The effects of fresh thalli, culture filtrate, water-soluble extract and dry powder of two species of macroalgae, Ulva pertusa (Chlorophyta) and Gracilaria lemaneiformis (Rhodophyta), on the growth of a bloom-forming microalga, Heterosigma akashiwo, were studied in co-culture under controlled laboratory conditions. Both fresh thalli and culture filtrate of U. pertusa and G. lemaneiformis, particularly in the form of fresh thalli, significantly inhibited microalgal growth; indeed, the microalga was completely killed during the course of the experiment. A clear concentration-dependent relationship was observed between the initial concentration of fresh thalli (either U. pertusa or G. lemaneiformis) and its inhibitory effect on H. akashiwo. Simultaneous nutrient assays showed that nitrate and phosphate were almost exhausted in G. lemaneiformis fresh thalli co-culture but remained well above nutrient limitation for microalgal growth in U. pertusa co-culture, in which the microalgal cells were completely killed. However, daily f/2 medium repletion would obviously alleviate the growth inhibition in G. lemaneiformis co-culture. Since the present study was carried out under controlled conditions, fluctuations in environmental factors (i.e., light, temperature, carbon limitation, bacterial presence and pH) were limited during the experiment. We thus concluded that allelopathy was the most likely explanation for microalgal growth inhibition in U. pertusa co-culture, while the combined roles of allelopathy and nutrient limitation were responsible for growth inhibition in G. lemaneiformis co-culture. Similarly, macroalgal water-soluble extracts and dry powders affected the co-cultured H. akashiwo greatly, with more obvious effects observed in water-soluble extract co-cultures. A dose-dependent relationship was also observed over the course of the experiment. It can be concluded that macroalgal thalli contain some bioactive compounds. The results of the present study suggest that U. pertusa and G. lemaneiformis, especially in the form of fresh thalli, effectively inhibit the growth of H. akashiwo and could thus be potential candidates for use in the control and mitigation of H. akashiwo blooms.     

Comparative Studies on the Allelopathic Effects of Ulva pertusa Kjellml, Corllina pilulifera Postl et Ruprl, and Sargassum thunbergii mertl O. Kuntze on Skeletonema costatum （Grev.）Cleve

In the present study, we evaluated the allelopathic effects of three macroalgae, namely Ulva pertusa Kjellml, Corallina pilufifera Postl et Ruprl, and Sargassum thunbergii Mertl O. Kuntze, on the growth of the microalga Skeletonema costaturn （Grev.） Creve using culture systems in which the algae coexisted. The effects of the macroalgal culture medium filtrate on S. costatum were also investigated. Moreover, isolated co-culture systems were built to confirm the existence of allelochemicals and preclude growth inhibition by direct contact. The coexistence assay data demonstrated that the growth of S. costaturn was strongly inhibited when fresh tissues, dry powder and aqueous extracts were used; the allelochemicals were lethal to S. costatum at relatively higher concentrations. The effects of the macroalgal culture medium filtrate on the microalga showed both species specificity and complexity. The inhibitory effect of fresh macroalgal tissue and culture medium filtrate on the microalga was due to the alleochemicals released by the macroalgae. The results of the present study show that the allelopathic effects of macroalgae on the microalga are complex. The present study could shed light onto the basis of the interaction between macro- and microalgae.     

凤眼莲浸出液对东海原甲藻生长的抑制作用

研究了凤眼莲浸出液对赤潮藻东海原甲藻生长的影响。浸出液浓度以单位体积溶液中所含凤眼莲鲜重表示。结果显示,在浸出液浓度大于2g·L^-1时,活体凤眼莲浸出液和干凤眼莲浸出液对东海原甲藻的生长都有明显的抑制作用。在浸出液浓度达到8g·L^-1时,活体凤眼莲浸出液和干凤眼莲浸出液对东海原甲藻都具有杀灭作用,东海原甲藻全部死亡出现的时间分别为培养后的第4t和第5t。经121℃高温处理20min后的活体凤眼莲浸出液对东海原甲藻的抑制作用仍然存在,抑制作用并没有显著减弱。研究结果表明凤眼莲的抑藻物质具有用来控制赤潮生物东海原甲藻的潜在价值。     

东海原甲藻与中肋骨条藻的种间竞争特征

对东海原甲藻和中肋骨条藻按照起始Chl-a比1∶5、1∶1和5∶1进行了f/2条件下的共培养实验,以探讨这两种藻的种间竞争特征。实验结果表明在共培养体系中,中肋骨条藻完全占优势,而东海原甲藻的生长受到明显的抑制。应用Lotka-Volterra种间竞争模型对共培养实验进行模拟的结果表明,东海原甲藻与中肋骨条藻的种间竞争结果与初始密度配比无关,中肋骨条藻总会竞争胜过东海原甲藻。为了探讨他感作用对东海原甲藻和中肋骨条藻种间竞争的影响,采用了中肋骨条藻的无藻细胞滤液来进行培养实验。实验结果显示,中肋骨条藻滤液对东海原甲藻及其本身的生长均无明显影响,这表明他感作用并非中肋骨条藻获得优势的主要竞争方式。