响应面法对微拟球藻产微藻蛋白的发酵条件优化

以稳定期微藻蛋白浓度为评价指标,利用响应面设计对微拟球藻(Nannochloropsis gaditana)的分批发酵条件进行优化。在单因素试验的基础上,选取温度、p H、搅拌速度及通气量为影响因子,采用四因素三水平的Box-Benhnken中心组合法设计试验。结果表明:微拟球藻的最佳发酵条件为温度30℃、p H 6.9、搅拌速度340 r/min以及通气量0.65 vvm,在此优化条件下得到微藻蛋白浓度为6.18 g/L,与模型预测值基本相符,较优化前提高了9.18%。     

五种淡水微藻的适宜培养温度和光照强度

从淡水中分离得到绿球藻(Chlorococcum sp.)SHOU-F3、纤维藻(Ankistrodesmus sp.)SHOU-F33、小球藻(Chlorella sp.)SHOU-F46、空星藻(Coelastrum sphaericum)SHOU-F10和栅藻(Scenedesmus sp.)SHOU-FX,分别在光照培养箱中研究了温度、光照强度对5种微藻增殖的影响,并分析了5种微藻的细胞组成。结果表明:绿球藻SHOU-F3、纤维藻SHOU-F33、小球藻SHOU-F46、空星藻SHOU-F10和栅藻SHOU-FX的最适生长温度分别为29.8、23.5、31.4、34.4和24.7℃;最适光照强度分别为16、119、42、82和106μmol·m-2·s-1;在适宜培养条件下,绿球藻SHOU-F3的色素、蛋白以及总糖的百分含量最高,纤维藻SHOU-F33的脂肪百分含量最高。     

光生物反应器中螺旋藻培养条件的优化

利用正交实验对搅拌式光生物反应器中钝顶螺旋藻(Spirulina platensis Geitl)的培养条件即搅拌速度、通气量和光照强度进行优化.实验结果表明:当培养温度为30℃时,通过正交实验所获得的最佳培养条件为搅拌转速120 r·min-1,通气量80 L·h-1,光照强度5000lx.在最佳培养条件下,收获时螺旋藻的干重为1.922 g·L-1.根据回归模型得到相应的优化条件为:光照强度5000lx,通气量150L·h-1,搅拌转速111.70r·min-1,收获量(干重)的预测值为2.293 g·L-1.另外,10%的接种量有利于螺旋藻的生长.     

基于尼罗红染色分析金藻总脂动态积累简

通过尼罗红荧光染色对一株富油微藻金藻Isochrysis sp.CCMM5001建立和完善了一种方便快捷且能准确定量金藻油脂含量的方法,利用该方法探索了不同培养条件对金藻生长和总脂积累的影响。结果表明:经尼罗红染色后,金藻的单细胞荧光强度与其总脂含量呈良好的线性关系;金藻最适生长的氮浓度、光照强度和温度分别为1323μmol/L、148.0μmol/(m2·s)、25℃;最适总脂积累的氮浓度、光照强度和温度分别为441μmol/L、92.5μmol/(m2·s)、15℃;优化培养条件并采用两阶段培养法后总脂含量和油脂产率都有大幅提高,可分别高达63.3%和22 mg/(L·d)。     

环境因子对小球藻(Chlorella sp.XQ-20044)光合作用的影响

小球藻(Chlorella sp.XQ-20044)是一株具有应用潜力的产油微藻,本文利用测定净光合放氧速率的方法研究了光照强度、温度、pH值和盐度对其光合作用的影响。研究结果表明:小球藻适宜的光照强度为500~1200μmol·m-2·s-1,光补偿点约30μmol·m-2·s-1,光饱和点在600μmol·m-2·s-1附近;光合作用适宜的温度范围为30~42.5℃,最适温度为40℃;适宜的pH值范围7.0~10.0,最适pH值为8.0;适宜盐度范围0.1~0.3 mol/L,最适盐度为0.2 mol/L。从光合作用特性来看,小球藻能适应较强的光照强度、较高的温度、偏碱性和较高的盐度环境,其中可耐受较高盐度的特性,有助于预防敌害生物的污染,对于实现规模培养,特别是利用开放系统进行规模培养较为有利。     

单生卵囊藻(Oocystis solitaria)和月牙藻(Selenastrumsp.)的培养条件及其细胞组成

从海滨的淡水池沼中分离得到单生卵囊藻(Oocystis solitaria)和月牙藻(Selenastrum sp.),分别研究了温度、光照强度、盐度对2种微藻生长繁殖的影响,分析了2种微藻的细胞组成、脂肪酸组成及盐度对2种微藻脂肪积累的影响.结果表明:单生卵囊藻和月牙藻的适宜生长温度分别为35.9℃～40.5℃和29.7℃～32.8℃;单生卵囊藻和月牙藻的适宜光照强度分别为46～70μmol · m-2·s-1和17 ～54 μmol·m-2·s-1;单生卵囊藻在淡水培养液中生长最好,月牙藻在盐度为2的半成水培养液中生长最好;在适宜培养条件下,单生卵囊藻细胞蛋白、总糖和总脂肪分别为27.61％、22.00％和3.84％;月牙藻细胞的蛋白、总糖和总脂肪分别为28.06％、21.99％和12.53％;单生卵囊藻的脂肪酸组成中含有丰富的18∶3n3;月牙藻的脂肪酸组成中含有DHA;盐度影响2种微藻的总脂肪含量;单生卵囊藻和月牙藻分别在盐度4和10的半咸水培养液中细胞总脂含量最高.     

利用叶绿素荧光监测微藻的生长及生理状态

衣藻与扁藻是生产生物柴油的优势藻种,监测其生长和生理状况是利用它们生产生物柴油的关键环节。本研究通过叶绿素荧光对这两种微藻的细胞密度进行监测并利用多激发波长调制叶绿素荧光仪(MultiColor-PAM)检测其在不同生长条件下的生理状况,获得本研究中两种藻的适宜的培养温度/光照强度:衣藻为28℃/80μmol/m~2·s~(-1),扁藻为28℃/100μmol/m2·s~(-1),利用叶绿素荧光可以便捷准确的监测微藻的生长及生理状态,为微藻培养及其代谢产物积累的研究提供切实可行的无损伤的检测方法。     

缺氮和高光对集球藻光合生理与油脂积累的影响

微藻是生产生物柴油理想的生物资源,对产油微藻的生理生化特性的研究具有重要意义。本文以一种耐碱性产油微藻集球藻为研究对象,将其置于两种光强水平(100和600μmol photons·m~(-2)·s~(-1))下培养,探讨缺氮对集球藻光合生理及油脂积累的影响。结果表明:缺氮培养下,集球藻光系统II的最大光化学效率F_v/F_m下降,电子从QA-到QB的电子传递受阻,光合作用受到抑制,生长表现缓慢,蛋白质含量降低,而油脂含量增加;在高光的耦合作用下,其光合作用进一步受到抑制,蛋白质含量下降,但油脂含量与低光缺氮条件下相比,没有显著变化。研究表明,缺氮使集球藻碳同化进入油脂合成代谢,增加其油脂含量,而光强对油脂积累的诱导效应不明显。     

松墨天牛对共生真菌和病原真菌的差异性免疫响应

在中国,入侵生物松材线虫进行传播的主要媒介昆虫松墨天牛(Monochamus alternatus)可与一种中国本地蓝变真菌Sporothrix sp.1建立体外共生关系.然而在天牛和真菌共生互作过程中,松墨天牛对蓝变菌S.sp.1的免疫响应却未见报道.因此,本文通过比较体外共生真菌S.sp.1和昆虫病原真菌球孢白僵菌(Beauveria bassiana)感染松墨天牛蛹所产生的免疫反应,发现天牛对两种菌的免疫应答方式具有显著性差异.将真菌注射到刚化蛹的天牛体内,S.sp.1感染的天牛不会死亡,B.bassiana感染的天牛则全部死亡.转录组测序结果显示,B.bassiana感染48 h后的松墨天牛的差异表达基因数量是S.sp.1感染的2倍.值得注意的是,在体外共生真菌感染的松墨天牛免疫系统中起主导作用的是Toll和IMD信号通路;而在昆虫病原真菌感染松墨天牛后,只有Toll信号通路上调表达.此外,体外共生真菌S.sp.1在感染天牛48 h后可通过下调天牛的Toll通路和IMD通路来产生免疫逃避.本研究提供了松墨天牛转录组全测序数据,为后续松墨天牛和其相关真菌互作研究提供了基础.     

内生真菌对草坪植物病原真菌抑制作用的比较

分别从野生牧草羽茅(Achnatherum sibiricum (L.) Keng)、栽培种高羊茅(Festuca arundinacea Schreb.)(品种Millennium)、栽培种黑麦草(Lolium perenne L.) (品种Justus)中分离出内生真菌Neotyphodium sp.、N. coenophialum和N. lolli,通过体外培养法比较了这3种内生真菌对草坪植物病原真菌的抑制作用.结果表明,从羽茅中分离的内生真菌Neotyphodium sp.在两菌相交前对所有供试的病原真菌都有一定的抑制作用,其中对枝孢霉属(Cladosporium sp.)、弯孢霉属(Curvularia sp.)和拟茎点属(Phomopsis sp.)病原真菌的抑制效果尤为显著,对峙培养3d后的抑菌率分别达70.1%、52.3%和30.9%,营养竞争作用、重寄生作用是其主要的拮抗机制;从高羊茅中分离的内生真菌N. coenophialum对枝孢霉属病原真菌存在一定的抑制作用;而从黑麦草中分离的内生真菌N. lolli与病原真菌对峙培养时,病原真菌菌落慢慢侵占整个营养空间,内生真菌停止生长并逐渐褐变死亡.体外培养结果说明Neotyphodium sp.对供试病原真菌的拮抗效果优于N. coenophialum和N. lolli,由此推测Neotyphodium sp.与宿主植物羽茅的共生可能有利于宿主植物抵抗病原真菌的侵扰.     

Efficiency of sunlight utilization: tubular versus flat photobioreactors

The light saturation effect imposes a serious limitation on the efficiency with which solar energy can be utilized in outdoor algal cultures. One solution proposed to reduce the intensity of incident solar radiation and overcome the light saturation effect is "spatial dilution of light" (i.e., distribution of the impinging photon flux on a greater photosynthetic surface area), but consistent experimental data supporting a significant positive influence of spatial light dilution on the productivity and the photosynthetic efficiency of outdoor algal cultures have never been reported. We used a coiled tubular reactor and compared a near-horizontal straight tubular reactor and a near-horizontal flat panel in outdoor cultivation of the cyanobacterium Arthrospira (Spirulina) platensis under defined operating conditions for optimum productivity. The photosynthetic efficiency achieved in the tubular systems was significantly higher because their curved surface "diluted" the impinging solar radiation and thus reduced the light saturation effect. This interpretation was supported by the results of experiments carried out in the laboratory under continuous artificial illumination using both a flat and a curved chamber reactor. The study also showed that, when the effect of light saturation is eliminated or reduced, productivity and solar irradiance are linearly correlated even at very high diurnal irradiance values, and supported findings that outdoor algal cultures are light-limited even during bright summer days. It was also observed that, besides improving the photosynthetic efficiency of the culture, spatial dilution of light also leads to higher growth rates and lowers the cellular content of accessory pigments; that is, it reduces mutual shading in the culture. The inadequacy of using volumetric productivity as the sole criterion for comparing reactors of different surface-to-volume ratio and of the areal productivity for evaluating the performance of elevated photobioreactors operated outdoors is stressed; it is furthermore suggested that the photosynthetic efficiency achieved by the culture also be calculated to provide a suitable parameter for comparison of different algal cultivation systems operated under similar climatic conditions. Copyright 1998 John Wiley & Sons, Inc.     

Influence of physiological factors on the lysis effect of Cytophaga on the red microalga Rhodella reticulata

The influence of different factors on the lysis of the red microalga, Rhodella reticulata, by Cytophaga sp. LR2 was studied. The pathogenic bacterial strain was more resistant than the alga to the physiological parameters studied, which assured long-term survival of bacteria in algal cultures. Cytophaga sp. LR2 infected R. reticulata at temperatures between 15 and 30 degrees C, in the illuminated as well as the non-illuminated cultures, at pH values between 5.0 and 9.0, and in the presence of NaCl and CaCl2 in the culture medium. SEM showed a different morphology of the bacteria in algal cultures from those of axenic cultures of Cytophaga. Observations of specific associations between algal and bacterial cells revealed that the role of the slime extrusions on the bacterial surface was attachment of Cytophaga to algal cells, and that their clumping leads to rapid lysis.     

温度、光照、盐度及pH对旋链角毛藻生长的影响

以旋链角毛藻(Chaetoceros curvisetus)为实验材料,采用f/2培养基,设置了2个盐度梯度(25和30),4个温度梯度(15℃、20℃、25℃和30℃),3个光照梯度(29.3μE·m^-2·s^-1、78.12μE·m^-2·s^-1和126.95μE·m^-2·s^-1),3个pH梯度(7.0、7.5和8.3)的处理,并计算了不同培养条件下藻细胞在指数生长期的比生长率,比较研究了温度、光照、盐度及pH等环境因子对旋链角毛藻生长的影响。进一步进行盐度、光照的两因素正交实验,并检验两者的交互作用。其中盐度设25、30两个水平,光照强度设29.3μE·m^-2·s^-1、78.12μE·m^-2·s^-1和126.95μE·m^-2·s^-1三个水平。实验结果表明,温度为20℃,光照为78.12μE·m^-2·s^-1,盐度为25,pH在8.3时是旋链角毛藻的最佳生长条件,此时最大比生长率和生物量达到最高,温度对藻类生长的影响比盐度,光照,pH的要明显得多,实验中其他处理均会抑制其生长。     

Modeling the effects of light and temperature on algae growth: State of the art and critical assessment for productivity prediction during outdoor cultivation

The ability to model algal productivity under transient conditions of light intensity and temperature is critical for assessing the profitability and sustainability of full-scale algae cultivation outdoors. However, a review of over 40 modeling approaches reveals that most of the models hitherto described in the literature have not been validated under conditions relevant to outdoor cultivation. With respect to light intensity, we therefore categorized and assessed these models based on their theoretical ability to account for the light gradients and short light cycles experienced in well-mixed dense outdoor cultures. Type I models were defined as models predicting the rate of photosynthesis of the entire culture as a function of the incident or average light intensity reaching the culture. Type II models were defined as models computing productivity as the sum of local productivities within the cultivation broth (based on the light intensity locally experienced by individual cells) without consideration of short light cycles. Type III models were then defined as models considering the impacts of both light gradients and short light cycles. Whereas Type I models are easy to implement, they are theoretically not applicable to outdoor systems outside the range of experimental conditions used for their development. By contrast, Type III models offer significant refinement but the complexity of the inputs needed currently restricts their practical application. We therefore propose that Type II models currently offer the best compromise between accuracy and practicability for full scale engineering application. With respect to temperature, we defined as “coupled” and “uncoupled” models the approaches which account and do not account for the potential interdependence of light and temperature on the rate of photosynthesis, respectively. Due to the high number of coefficients of coupled models and the associated risk of overfitting, the recommended approach is uncoupled models. Most of models do not include the modeling of endogenous respiration and the modeling of light and temperature acclimation in spite of their potential effect on productivity.     

环境因子对小球藻(Chlorella sp. XQ-20044)光合作用的影响

小球藻（Chlorella sp.XQ-20044）是一株具有应用潜力的产油微藻,本文利用测定净光合放氧速率的方法研究了光照强度、温度、pH值和盐度对其光合作用的影响。研究结果表明：小球藻适宜的光照强度为500~1200 μmol·m^-2·s^-1,光补偿点约30 μmol·m^-2·s^-1,光饱和点在600 μmol·m^-2·s^-1附近;光合作用适宜的温度范围为30~42.5℃,最适温度为40℃;适宜的pH值范围7.0~10.0,最适pH值为8.0;适宜盐度范围0.1~0.3 mol/L,最适盐度为0.2 mol/L。从光合作用特性来看,小球藻能适应较强的光照强度、较高的温度、偏碱性和较高的盐度环境,其中可耐受较高盐度的特性,有助于预防敌害生物的污染,对于实现规模培养,特别是利用开放系统进行规模培养较为有利。     

Responses of the coastal bacterial community to viral infection of the algae Phaeocystis globosa

The release of organic material upon algal cell lyses has a key role in structuring bacterial communities and affects the cycling of biolimiting elements in the marine environment. Here we show that already before cell lysis the leakage or excretion of organic matter by infected yet intact algal cells shaped North Sea bacterial community composition and enhanced bacterial substrate assimilation. Infected algal cultures of Phaeocystis globosa grown in coastal North Sea water contained gamma- and alphaproteobacterial phylotypes that were distinct from those in the non-infected control cultures 5 h after infection. The gammaproteobacterial population at this time mainly consisted of Alteromonas sp. cells that were attached to the infected but still intact host cells. Nano-scale secondary-ion mass spectrometry (nanoSIMS) showed ∼20% transfer of organic matter derived from the infected ¹³C- and ¹⁵N-labelled P. globosa cells to Alteromonas sp. cells. Subsequent, viral lysis of P. globosa resulted in the formation of aggregates that were densely colonised by bacteria. Aggregate dissolution was observed after 2 days, which we attribute to bacteriophage-induced lysis of the attached bacteria. Isotope mass spectrometry analysis showed that 40% of the particulate ¹³C-organic carbon from the infected P. globosa culture was remineralized to dissolved inorganic carbon after 7 days. These findings reveal a novel role of viruses in the leakage or excretion of algal biomass upon infection, which provides an additional ecological niche for specific bacterial populations and potentially redirects carbon availability.     

Interactions between the unicellular red alga Rhodella reticulata (Rhodophyta) and contaminated bacteria

AIMS: To define the role of the bacterial strains LR1 and LR3 in the Rhodella cell destruction caused by Cytophaga sp.LR2. METHODS AND RESULTS: The bacteria were obtained from algal culture with destruction. They were isolated in pure culture and tested for biochemical activities using Polymicrotest. The ability of bacteria to degrade and utilize the algal polysaccharide was investigated. The bacteria were grown in a media containing Rhodella polysaccharide as a sole carbon source. The level of the reducing sugars in the culture media was determined. Scanning electron microscopy (SEM) was used to define the location of bacteria in extensively and intensively cultivated Rhodella reticulata previously infected by Cytophaga sp. LR2. CONCLUSIONS: The lysis of Rhodella reticulata cells is due to the joint action of the three bacterial strains with the former pathogen Cytophaga sp. LR2 playing the main role. The accumulation of the polysaccharide and the excreted metabolites of the strains LR1 and LR3 stimulated the development of Cytophaga sp. LR2. The adaptation of the strain to particular conditions of alga cultivation and the utilization of polysaccharide as a sole carbon source supported its stable growth in alga suspension and destruction of Rhodella cells. SIGNIFICANCE AND IMPACT OF THE STUDY: The predominance of Cytophaga sp. LR2 over the two other contaminants and the lysis of Rhodella reticulata cells resulted from the ability of the bacterium to attach to the algal polysaccharide sheath. The formation of slime and extrusions facilitated the phenomenon of bacterial adhesion to the algal surface as well as the formation of colonial alga - bacterial spherules. The sedimentation of these aggregates decreased the ability of the algal strain to photosynthesize, led to the lysis of the cells and finally caused the death of Rhodella.     

几种主要环境因子对布朗葡萄藻(Botryococcus braunii)光合作用的影响

利用测定净光合放氧速率的方法研究了光照强度、温度、pH值、盐度对布朗葡萄藻Botryococcus braunii UTEX 572和B.braunii UTEX 2441两个品系的光合作用的影响。B.braunii UTEX 572的适宜光照强度范围400～1600μmol·m^-2·s^-1,光饱和点在800μmol·m^-2·s^-1附近;适宜温度范围25～35℃,最适温度30℃;适宜pH范围5.0～8.0,最适pH7.0;适宜盐度范围0～0.2mol/L,最适盐度0.1mol/L。B.braunii UTEX 2441的适宜光照强度范围400～1600μmol·m^-2·s^-1,光饱和点在400μmol·m^-2·s^-1附近;适宜温度范围25～35℃,最适温度30℃;适宜pH范围5.0～8.0,最适pH7.0;对盐度的适应范围较小,盐度升高,光合放氧速率明显下降。两个布朗葡萄藻净光合放氧速率随光照强度、温度、pH值和盐度变化的规律,表明布朗葡萄藻的基本生理生态学特征:适应于较强的光照强度、较高的温度、中性偏酸的环境和较低的盐度。对布朗葡萄藻基本生理生态学特征的了解,为培养条件的优化提供了依据。2个布朗葡萄藻品系对光强、温度、pH值和盐度变化的反应有所不同:与B.braunii UTEX 2441相比,B.braunii UTEX 572具有更高的光饱和点,适应更高的温度,对pH值变化有更宽的适应范围,适当提高盐度对其光合作用有促进作用,表明B.braunii UTEX 572在快速生长繁殖方面具有更大的潜力,这一研究结果为筛选适合于大量培养的优良藻种提供了依据。     

内生真菌枝孢属Cladosporium sp.对丹参生长和丹酚酸含量的影响

本文介绍从药用植物丹参内生真菌中筛选新获得的一株对丹参生长和丹酚酸含量具有显著促进作用的菌株,Cladosporium sp. SM58。运用菌根生物技术,于离体、盆栽和田栽条件下,分别将丹参组培苗或幼苗接种SM58固体菌种后,观察测定菌株SM58对丹参组培苗生长的影响,以及丹参幼苗接菌培植6个月后菌株SM58对丹参丹酚酸有效成分含量和生物量的影响;同时进行显微观察,对菌株SM58侵入田栽丹参根组织的状况进行分析。结果表明,SM58菌株不仅对丹参组培苗的株高、根长和生物量均有显著促生长作用（P<0.01）,而且对盆栽和田栽丹参的丹酚酸有效成分含量和根干重的提高也均有显著效果（P<0.01）。与对照相比较,接菌组田栽丹参的根干重、总酚酸含量、丹酚酸A的含量分别提高68%、47%、11%。显微观察显示SM58菌丝主要通过侵染丹参的根表皮细胞和皮层细胞而对丹参产生影响。本研究表明,SM58菌株能对丹参品质和产量产生有益影响,是一株具有很好研究价值和开发应用的活性菌。