海洋大型绿藻条浒苔与微藻三角褐指藻相生相克作用的研究

以种群密度为变量,采用实验生态学手段研究了大型海藻条浒苔与赤潮藻类三角褐指藻间的生长竞争关系,以及条浒苔水溶性抽提液对三角褐指藻生长的影响。结果表明,在无营养盐限制条件下,低密度条浒苔(0.3~0.7 g/L)均能抑制三角褐指藻(起始浓度104cell/ml)的生长,最大抑制率为74.5%;低起始浓度(102~103cell/ml)的三角褐指藻,对条浒苔具有促生长效应,而高起始浓度(104~105cell/ml)的三角褐指藻在一定程度上抑制条浒苔的生长。条浒苔水溶性抽提液(0.3~0.7 g/L)对三角褐指藻的生长皆表现出明显的抑制效应,平均抑制率为75.2%,抑制效果较条浒苔鲜组织更为明显;其最大抑制效应(84.8%)表现在接种后的第8天,三角褐指藻的生长抑制量随条浒苔水溶性抽提液浓度的增加而增大。表明条浒苔可能通过相生相克作用影响共培养体系中三角褐指藻的生长。     

Allelochemical ethyl 2-methyl acetoacetate (EMA) induces oxidative damage and antioxidant responses in Phaeodactylum tricornutum

Ethyl 2-methyl acetoacetate (EMA) is a novel allelochemical exhibiting inhibitory effects on the growth of marine unicellular alga Phaeodactylum tricornutum (P. tricornutum). Oxidative damage and antioxidant responses in P. tricornutum were investigated to elucidate the mechanism involved in EMA inhibition on algal growth. The increase in reactive oxygen species (ROS) levels and malondialdehyde (MDA) contents following exposure to EMA suggested that alga was suffered from oxidative stress and severely damaged. The decrease in cell activity and cellular inclusions suggested that cell growth was greatly inhibited. The activities of the antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione peroxide (GSH-PX) and glutathione S-transferase (GST) increased with the exposure concentration and decreased with the prolongation of exposure time. Cellular ascorbic acid (AsA) and reduced glutathione (GSH) systems were also involved in resisting oxidative stress of EMA by altering the composition of AsA and GSH pools. EMA exposure increased the contents of AsA, GSH, dehydroascorbate (DAsA) and glutathione (GSSG). However, the regeneration rate of AsA/DAsA did not change obviously between treatments and the control, while that of GSH/GSSG decreased significantly under 14 mmol/L EMA exposure on the 3rd day. These results showed that EMA-induced oxidative damage might be responsible for EMA inhibition on P. tricornutum growth and cellular antioxidant enzymes and non-enzymatic antioxidants were improved to counteract the oxidative stress.     

盐度对三角褐指藻生长及有机质积累的影响

以三角褐指藻为材料,探究盐度对该藻生长、总脂等有机质含量的影响,探讨有利于油脂生产的盐度条件。结果显示,盐度变化对三角褐指藻生物量和有机质积累均有显著影响。在盐度70‰的海水中培养的细胞,其比生长速率比正常海水盐度（35‰）培养组降低了22.6%。正常海水盐度下,生物量及总脂含量最高,分别达1.92g/L和29.98%,且总脂单位体积产量极显著（p＜0.01）地高于其他各组,藻油产量高达47.67mg•L-1•d-1。在盐度17‰时,多糖含量最高,达5.5%。在盐度70‰时,蛋白质含量最高,达17.82%。       

氮元素对三角褐指藻岩藻黄素和油脂合成关键酶基因表达与代谢合成的影响

为阐明氮元素对微藻次生代谢积累和调控的影响,以三角褐指藻为试验材料,研究不同氮浓度[896(CK)、448、112、28和0 μmol·L^-1]处理对三角褐指藻细胞生长、岩藻黄素含量、油脂含量以及叶绿素a含量的影响,并对岩藻黄素-叶绿素蛋白复合体基因(FCPb)和酰基-酰基载体蛋白去饱和酶基因(FAB2)的表达进行实时荧光定量PCR分析。结果表明,氮限制极显著抑制了三角褐指藻细胞的生长和岩藻黄素的合成,但促进了油脂的合成。当氮浓度为112 μmol·L^-1时,三角褐指藻岩藻黄素含量最低(0.084 mg·g^-1DW),而油脂含量最高,较CK提高了1.36倍。叶绿素a与岩藻黄素含量变化趋势一致。相关性分析表明,氮限制条件下,三角褐指藻岩藻黄素含量与油脂含量显著相关(R²=0.998 8)。实时荧光定量PCR分析表明,氮限制抑制了三角褐指藻中FCPb的表达,促进了FAB2的表达。综上,氮限制通过调控三角褐指藻岩藻黄素、油脂生物合成途径相关基因的表达影响了岩藻黄素和油脂的积累。本研究为进一步探究岩藻黄素与脂类物质代谢合成的关联性提供了一定的理论参考。     

三角褐指藻固定化培养的初步研究

用褐藻酸钙对三角褐指藻进行固定化培养实验,测定了不同胶粒大小,不同胶粒密度及不同接种量对该藻的影响,比较了自由生长和固定细胞的生长曲线,还进行了常温组和低温组固定化细胞的活化复苏实验,三角褐指藻在褐藻酸钙胶中仍具有呼吸和光合作用的能力,球径为２．５ｍｍ时,细胞生长快,每５０ｍｌ培养液中加入１５０个胶球时细胞生长量大,接种量不能低于１０＾５个细胞／ｍｌ,在冰箱（４℃）中可保存２１个月,本藻适合用固定     

Immune Status and Hepatic Antioxidant Capacity of Gilthead Seabream Sparus aurata Juveniles Fed Yeast and Microalga Derived β-glucans

This work aimed to evaluate the effects of dietary supplementation with β-glucans extracted from yeast (Saccharomyces cerevisiae) and microalga (Phaeodactylum tricornutum) on gene expression, oxidative stress biomarkers and plasma immune parameters in gilthead seabream (Sparus aurata) juveniles. A practical commercial diet was used as the control (CTRL), and three others based on CTRL were further supplemented with different β-glucan extracts. One was derived from S. cerevisiae (diet MG) and two different extracts of 21% and 37% P. tricornutum-derived β-glucans (defined as Phaeo21 and Phaeo37), to give a final 0.06% β-glucan dietary concentration. Quadruplicate groups of 95 gilthead seabream (initial body weight: 4.1 ± 0.1 g) were fed to satiation three times a day for 8 weeks in a pulse-feeding regimen, with experimental diets intercalated with the CTRL dietary treatment every 2 weeks. After 8 weeks of feeding, all groups showed equal growth performance and no changes were found in plasma innate immune status. Nonetheless, fish groups fed β-glucans supplemented diets showed an improved anti-oxidant status compared to those fed CTRL at both sampling points (i.e., 2 and 8 weeks). The intestinal gene expression analysis highlighted the immunomodulatory role of Phaeo37 diet after 8 weeks, inducing an immune tolerance effect in gilthead seabream intestine, and a general down-regulation of immune-related gene expression. In conclusion, the results suggest that the dietary pulse administration of a P. tricornutum 37% enriched-β-glucans extract might be used as a counter-measure in a context of gut inflammation, due to its immune-tolerant and anti-oxidative effects.     

温度影响三角褐指藻生长和脂肪酸组成的初步探讨

实验对指数生长期、指数生长末期、静止期和衰亡期的三角褐指藻脂肪酸进行研究,发现在指数生长末期二十碳五烯酸(Eicosapentaenoic acid,EPA)含量最高。实时定量PCR分析三角褐指藻Δ5脂肪酸去饱和酶基因在不同生长时期的表达变化水平差异显著,其中指数生长期表达量最高。为了探讨温度对微藻生长和脂肪酸组成的影响,作者在不同温度下对三角褐指藻进行了培养,并测定其生长和脂肪酸组成。结果表明,三角褐指藻的生长随着温度的升高到达指数生长末期所需时间缩短;不同温度下三角褐指藻的主要脂肪酸为C14∶0、C16∶0、C16∶lΔ9、Cl8∶lΔ9、C18∶2Δ9,12、C20∶5n3(EPA),总脂肪酸随着温度的升高依次呈现先升高后降低的趋势,在15℃时EPA占总脂肪酸的百分含量达到最高值25.71%。     

乙酰水杨酸对三角褐指藻岩藻黄质含量的影响及其分子机理研究

为了研究不同浓度乙酰水杨酸(ASA)对三角褐指藻(Phaeodactylum tricornutum Bohlin)细胞生长、岩藻黄质积累及岩藻黄质生物合成相关基因的表达水平的影响,通过分光光度计、高效液相色谱(HPLC)以及荧光定量PCR测定不同浓度ASA处理后的三角褐指藻的各项指标。结果表明,三角褐指藻的生长和岩藻黄质积累与ASA浓度密切相关。一定浓度的ASA抑制了三角褐指藻的细胞数量;岩藻黄质含量也随着ASA浓度的升高呈现先上升后下降的趋势,当ASA浓度为10 mg·L^-1时,岩藻黄质含量最高,达1.78 g·kg^-1 DW,为对照组的1.9倍。在不同浓度ASA诱导下,与岩藻黄质生物合成途径有关的PSY、PDS、ZDS、CRTISO、LCYB和ZEP基因表达均上调,当ASA浓度为10 mg·L^-1时,相关基因的表达量极显著高于对照组(P<0.01)。主成分分析(PCA)结果表明,PSY、PDS、ZDS、CRTISO、LCYB和ZEP基因与岩藻黄质积累具有相关性,其中,ZEP基因对岩藻黄质生物合成贡献率最高。综上,适当浓度的ASA不仅提高了岩藻黄质合成途径中相关基因的表达水平,而且促进了岩藻黄质的积累。本研究结果为进一步深入研究三角褐指藻中岩藻黄质合成的分子机制奠定了理论基础。