基于海洋硅藻的生物燃油生产

微藻与陆生作物相比,具有生物量大、生长周期短、脂质含量高、不占用耕地等特点,已成为生物燃油原料的首选。微藻生物燃油的经济性生产,需要建立能保持单一藻种优势的开放型微藻生产系统。采用海洋硅藻——角毛藻(Chaetoceros)作为讨论对象,综述了目前基于海洋硅藻的生物燃油生产系统设计所应用的技术:在开放型系统中生产硅藻,从生长介质中收获硅藻,从收获的硅藻中提取油脂和脂肪酸,以及整体的经济性。     

秦岭火地塘林区硅藻植物的研究

【目的】研究秦岭火地塘林区硅藻植物的种类组成与生态特征。【方法】对采自秦岭火地塘林区8个样点的样品进行固定、酸处理后,在显微镜下对硅藻门植物进行观察、计数和拍照,研究林区硅藻的种类组成,并分析其生态类型及分布特点。【结果】共鉴定出硅藻门植物126个分类单位,包括91种29变种6变型,分别隶属于2纲6目10科29属,优势种包括Diatoma hiemale(Roth)Heib.、D.hiemale var.mesodon(Ehr.)Grun.、Cymbella ventricosa Agardh、Gomphonema lanceolatum Ag.、G.subclavatum(Grun.)Grun.、Cocconeis placentula Ehr.和C.placentulavar.euglypta(Ehr.)Grun.。火地塘林区大部分硅藻的生态类型为淡水性种类,并有较多的普生性种类及高山冷水性种类,其余种类较少。从硅藻的生活习性来看,49.2%的种类既可营浮游生活又可营附着生活,42.1%的种类为浮游硅藻种类;61.9%的硅藻为水生种类,31.0%的种类为水生或气生种类。【结论】探明了秦岭火地塘林区硅藻植物种类组成和生态特征。     

Improvement of Neutral Lipid and Polyunsaturated Fatty Acid Biosynthesis by Overexpressing a Type 2 Diacylglycerol Acyltransferase in Marine Diatom Phaeodactylum tricornutum

Microalgae have been emerging as an important source for the production of bioactive compounds. Marine diatoms can store high amounts of lipid and grow quite quickly. However, the genetic and biochemical characteristics of fatty acid biosynthesis in diatoms remain unclear. Glycerophospholipids are integral as structural and functional components of cellular membranes, as well as precursors of various lipid mediators. In addition, diacylglycerol acyltransferase (DGAT) is a key enzyme that catalyzes the last step of triacylglyceride (TAG) biosynthesis. However, a comprehensive sequence-structure and functional analysis of DGAT in diatoms is lacking. In this study, an isoform of diacylglycerol acyltransferase type 2 of the marine diatom Phaeodactylum tricornutum was characterized. Surprisingly, DGAT2 overexpression in P. tricornutum stimulated more oil bodies, and the neutral lipid content increased by 35%. The fatty acid composition showed a significant increase in the proportion of polyunsaturated fatty acids; in particular, EPA was increased by 76.2%. Moreover, the growth rate of transgenic microalgae remained similar, thereby maintaining a high biomass. Our results suggest that increased DGAT2 expression could alter fatty acid profile in the diatom, and the results thus represent a valuable strategy for polyunsaturated fatty acid production by genetic manipulation.     

Effect of an Introduced Phytoene Synthase Gene Expression on Carotenoid Biosynthesis in the Marine Diatom Phaeodactylum tricornutum

Carotenoids exert beneficial effects on human health through their excellent antioxidant activity. To increase carotenoid productivity in the marine Pennales Phaeodactylum tricornutum, we genetically engineered the phytoene synthase gene (psy) to improve expression because RNA-sequencing analysis has suggested that the expression level of psy is lower than other enzyme-encoding genes that are involved in the carotenoid biosynthetic pathway. We isolated psy from P. tricornutum, and this gene was fused with the enhanced green fluorescent protein gene to detect psy expression. After transformation using the microparticle bombardment technique, we obtained several P. tricornutum transformants and confirmed psy expression in their plastids. We investigated the amounts of PSY mRNA and carotenoids, such as fucoxanthin and β-carotene, at different growth phases. The introduction of psy increased the fucoxanthin content of a transformants by approximately 1.45-fold relative to the levels in the wild-type diatom. However, some transformants failed to show a significant increase in the carotenoid content relative to that of the wild-type diatom. We also found that the amount of PSY mRNA at log phase might contribute to the increase in carotenoids in the transformants at stationary phase.     

鲍采苗板上底栖硅藻种类变化的观察

底栖硅藻作为鲍幼体的前期饵料,其种类与质量对鲍人工育苗的成败起着至关重要的作用,其种类适宜且数量充足可以延长鲍幼体的附着变态率,延长在板时间,实现剥离大规格苗种,达到提高成活率的目的。据报道在底栖硅藻自然种群中,以小型的卵形藻(Cocconesp.)和舟形藻(Naviculasp.)的     

Using micro particle diets to replace diatoms for feeding postlarval abalone Haliotis discus discus (Reeve.)

Feeding artificial micro diets instead of diatoms may allow a more constant and controlled form of nutrition, and thus results in lower mortality rate and increased growth. A 4-week trial was conducted to determine the growth and survival of postlarval Haliotis discus discus (Reeve.) postlarvae fed three experimental micro particle diets, and naturally occurring diatoms. A completely randomized design was adopted with each diet being replicated three times. All diets varied in terms of particle size and gross composition. Survival rate was significantly lower ( P  < 0.05) in the diatom diet (19.5 ± 2.1%) when compared with the K-1 diet (38.8 ± 8.9%), the K-2 diet (43.7 ± 5.6%) and the MD-Q diet (44.2 ± 14.8%). The size of postlarvae fed the four diets was not significantly different ( P  > 0.05). There was also little difference in the daily growth rate. Based on the results of this work for the preliminary implementation of artificial food as a replacement diet for diatoms, it seems that postlarval H . discus discus readily consumes artificial micro particle diets and has a superior survival rate when compared to that of diatoms.     

赤水河流域秋季底栖硅藻群落结构特征及水质评价

为研究赤水河流域底栖硅藻群落特征及水质状况,2019年9月测定了赤水河流域36个代表性样点的水体理化指标,调查了底栖硅藻群落组成,并应用硅藻指数评价了赤水河流域的水质状况。结果表明：秋季赤水河流域上游的电导率和总磷、总氮浓度均高于下游;调查采集到底栖硅藻29属138种,其中扁圆卵形藻多孔变种(Cocconeis placentula var. euglypta)、变异直链藻（Melosira varians）、极细微曲壳藻（Achnanthes minutissima）和谷皮菱形藻（Nitzschia palea）等为秋季赤水河流域优势种;下游底栖硅藻群落稳定性高于中上游;CCA结果显示可溶活性磷（SRP）和总氮（TN）是影响赤水河流域底栖硅藻群落的主要环境因子。TDI和CEE硅藻指数评价结果表明,赤水河流域水质等级整体为“中等”,55.6%的样点水质等级为“中等”以上,8.3%的样点水质等级为“极差”。     

The concentrating method of benthic diatom affects the growth of juvenile sea cucumber (Apostichopus japonicus) and water quality

Concentrating methods generally affect the vitality of the microalgae and may alter its feeding and environmental effect. This article studied the concentrating methods (centrifugation and settlement) on primary productivity of the microalgae Cylindrotheca fusiformis. The effects of free settled C. fusiformis, centrifuged C. fusiformis and macroalgae Sargassum polycystum on growth of the sea cucumber, farming water and sediment quality under two light intensities (1500 lx and 500 lx) were studied by a six‐week rearing experiment. The results showed that the primary productivity of free settled C. fusiformis was 3–4 times higher than that of those centrifuged, indicating centrifugation inhibited the photosynthesis process. In the rearing experiment, feed, light intensities and their interactions significantly affected the specific growth rate (SGR) of the animals (P < 0.01). The animals reared in higher light intensity also had higher SGR, but only those fed free settled C. fusiformis reached a significant level (P < 0.05). The group also had the highest SGR (P < 0.05), suggesting that the photosynthesis ability of C. fusiformis directly contributed to the growth of the animals. During the experiment, the soluble inorganic nitrogen, reactive phosphorus in farming water, total nitrogen and total organic carbon in the sediment gradually increased in all treatments. The S. polycystum group had slightly higher level of nutrients in water and sediment at the end of the experiment for its high organic content. These results provide evidence that the harvesting methods of C. fusiformis directly affect its vitality, which consequentially alter the feeding and environmental effect, especially under higher light densities.     

武汉月湖沉积物中硅藻的垂直分布与水质变化

采用活塞取样器采集了月湖湖中央2处钻孔沉积物,分析了其中硅藻种类与密度的垂直变化,同时采用210Pb/137Cs推测沉积物的沉积速率.并进行了总有机质、生物硅含量分析,以了解月湖各个不同年代的水质变化.结果表明,2个柱状沉积物中硅藻的优势属为:小环藻属、直链藻属、舟形藻属、Cyclostephanos属.Z-1钻孔优势种为:梅尼小环藻、颗粒直链藻、Navicula porifera、Cyclostephanos tholifomis.Z-2钻孔中优势种为:梅尼小环藻、颗粒直链藻、舟形藻、Cyclostephanos invisitatus.就硅藻密度而言,总趋势是上层最多,平均7.14×105～58.65×105 ind·g-1,中层其次,平均0.08×105～13.98×105 ind·g-1,下层最少,平均0.24×105～0.64×105 ind·g-1.硅藻种类数变化虽然没有明显规律,但种类最少的样品出现在中层,210Pb/137Cs测年数据表明中层为19世纪90年代-20世纪70年代.相关分析表明:Z-1孔生物硅与沉积硅藻密度显著正相关(r=0.933,P=0.021<0.05),总有机质含量与种类数显著负相关(r=-0.972,P=-0.005<0.05);Z-2孔生物硅与沉积硅藻密度显著正相关(r=0.985,P=0.002<0.05),沉积硅藻密度与种类数显著负相关(r=-0.933,P=0.021<0.05).研究结果表明,月湖沉积物中硅藻种类与密度的垂直变化可以反映月湖的富营养演化历程:19世纪60年代-19世纪80年代,月湖水质较好;19世纪90年代-20世纪10年代,月湖开始由贫营养状态向中营养状态转变,为水体富营养化开始发生时间;20世纪20年代-20世纪40年代,月湖水体营养等级持续增加,已富营养化;20世纪50年代-20世纪70年代,周边人口数量急剧上升与污水的直接排放导致月湖水质持续恶化;20世纪80年代至月湖截污前这段时期使月湖最终沦为劣五类水体的湖泊.     

不同方法提取海洋硅藻粗多糖的比较

采用不同提取方法(水提法、酸提法、碱提法、超声辅助法、冻融辅助法等)从室外培养的海洋硅藻生物质中提取粗多糖。通过比较不同方法下的粗多糖提取率、多糖总糖、蛋白质及硫酸基含量筛选较优的方法。试验结果显示,水提法的粗多糖提取率最低,浓碱法粗多糖提取率最高。物理法(超声、冻融结合高温法)提取的粗多糖中总糖含量总体比化学试剂法(酸碱法)高,以冻融+90 ℃最高,稀酸法最低;硫酸基含量则以稀酸法最高,浓碱法最低。综合单位质量海洋硅藻生物质粗多糖提取率与单位质量粗多糖组成的结果可知,单位质量硅藻生物质提取的总糖量大小为:浓碱+冻融法>浓碱+冻融+90 ℃法>浓碱法>稀酸法>超声+冻融+90 ℃>稀碱法>超声+冻融>冻融+90 ℃;硫酸基质量以浓碱+冻融法最高,稀碱法最低。这些结果表明,利用浓碱+冻融法可从海洋硅藻中提取出较多多糖并使硫酸基处于较高水平,利于硅藻多糖后续的纯化与利用。进一步利用浓碱+冻融法提取4种海洋硅藻(角毛藻、菱形藻、茧形藻、海链藻)粗多糖,结果表明,单位质量生物质提取的粗多糖总糖含量以茧形藻最高(约24%),海链藻最低(约5%);硫酸基含量则以角毛藻最高(5.05%),茧形藻最低(0.51%)。