富油微藻富碳培养下油脂积累及固碳研究

微藻固碳是一种新型节能减排技术,具有长期可持续发展的潜力。本文对两株富油微藻（球等鞭金藻和微拟球藻）进行了富碳培养下生长特性及中性脂积累特性的研究。两株富油微藻的最佳培养条件为10%CO₂浓度和f培养基。本研究对两株富油微藻的最大生物量产率、总脂含量、最大油脂产率、微藻的C含量和CO₂固定率进行了测定。球等鞭金藻的各参数指标分别为：142.42±4.58g/（m²·d）,39.95%±0.77%,84.47±1.56g/（m²·d）,45.98%±1.75%和33.74±1.65g/（m²·d）。微拟球藻的各参数指标分别为：149.92±1.80g/（m²·d）,37.91%±0.58%,89.90±1.98g/（m²·d）,46.88%±2.01%和34.08±1.32g/（m²·d）。实验结果显示,两株海洋微藻均属于高固碳优良藻株,适合应用于微藻烟气减排技术开发,具备用于海洋生物质能耦合CO₂减排开发的潜力。     

温度对四种产油微藻生长和油脂特性的影响

本文采用气相质谱联用技术对三株海水微藻—三角褐指藻(Phaeodactylum tricornutum,CCMM2012)、小球藻(Chlorella vulgaris,CCMM4004)、微拟球藻(Nannochloropsis sp.,CCMM6004),和一株淡水微藻—栅藻(Scenedesmus quadricauda,CCMM4002)在不同温度下的生长及油脂变化规律进行研究。结果表明:三角褐指藻、小球藻和栅藻在20℃下的油脂产率最大,分别为2.94、5.79和5.83mg/L/d,它们在20℃的培养条件下比较适合应用于生物能源开发;而微拟球藻在25℃下油脂产率最大,达6.45mg/L/d,因此此温度下更适合用于生物能源开发。三角褐指藻在15℃条件下DHA和EPA含量最多,为16.47%,而微拟球藻在25℃下DHA和EPA最多,为22.74%;因此三角褐指藻和微拟球藻在15℃和25℃下均比较适合应用于多不饱和脂肪酸的开发,而小球藻和栅藻的EPA和DHA含量在三个温度下普遍较少,不适合用于多不饱和脂肪酸的开发。     

采用加速溶剂萃取法提高微藻油脂萃取量的优化条件研究

设计4因子5水平的正交实验方案[采用正交表L25（5＾6）],通过确定加速溶剂萃取法（ASE）处理的最优化条件,包括萃取溶剂、萃取温度、萃取时间和萃取循环次数,达到快速萃取微藻油脂并且萃取量高的效果．结果表明,除循环次数外,其他萃取条件因子均能显著影响油脂萃取量（P〈0．035）．在萃取溶剂为无水乙醇或丙酮、温度为175℃、萃取时间为16min、循环次数为3次的最优处理条件下,微藻油脂萃取量最高（19．20％～21．36％）．与常规的索氏萃取法相比,萃取量可提高39．08％～47．09％．采用ASE法在最优化条件下萃取微藻油脂可以达到良好效果．     

苏丹黑B染色处理细胞技术在快速检测微藻油脂含量中的应用

探讨了采用苏丹黑B染料染色处理微藻细胞技术估测微藻油脂含量的可行性.结果表明,微藻细胞经苏丹黑B染色处理后在645 nm光波长处有最大吸收峰值A645,微藻油脂含量与经苏丹黑B染色处理后微藻细胞的吸光值A645呈明显的相关性.建立了考察的7种微藻油脂含量与吸光值A645的线性回归方程,线性关系显著（p〈0.01）.基于藻湿重,微藻油脂含量的线性范围为1.79~39.63 mg/g,估算的相对偏差〈10%.因而,采用苏丹黑B染色处理微藻细胞技术适宜用来检测微藻的油脂含量.     

富油金藻与绿藻的耐光适应性比较

以金藻(Isochrysis sp.)、球等鞭金藻(Isochrysis galbana Parke)、微拟球藻(Nannochloropsis sp.)、东营小球藻(Chlorella sp.)、小球藻(Chlorella capsulata Guillard)等五株富油微藻为实验材料,通过对比研究强弱光驯化后其叶绿素荧光和快速光曲线(RLC)的变化,比较了金藻与绿藻的光适应性。结果发现:弱光驯化后,五株微藻有效量子产量(ΦPSII)随着光照强度(PAR)的上升均呈下降趋势,但是其下降速率在不同藻株之间存在差异,其中以球等鞭金藻下降速率最小,小球藻下降速率最大;两株金藻的最大电子传递速率(rETRm)及半饱和光强(IK)均高于三株绿藻,但是初始斜率(α)小于绿藻;强光驯化后,五株富油微藻的ΦPSII及α均明显下降,但两株金藻下降幅度均小于三株绿藻,而金藻的rETRm及IK始终高于绿藻。同样,不同藻株的快速光曲线下降斜率(β)也都呈下降趋势,但金藻β的下降幅度大于绿藻。上述结果表明:绿藻在弱光下有较强的光吸收能力,但其光利用能力及热耗散能力相对较差,在强光下易发生光抑制;强光驯化后,金藻比绿藻有更好的强光可塑性,属于耐光性强的潜力产油微藻。     

融合微藻Tetraselmis sp.-1兼养培养条件下的动力学研究

利用统计分析系统(Statistic Analysis System,SAS),对融合微藻Tetraselmis sp.-1兼养培养进行研究,建立了兼养条件下的细胞生长、底物消耗和产物生成3种动力学模型.经过数据验证,3种模型均能较好的反映微藻培养过程.     

缺氮条件对栅藻油脂积累与光合作用的影响

栅通过油脂分析、光合放氧、叶绿素荧光等手段,研究了缺氮处理对栅藻(Scenedesmus dimorphus)油脂含量、油脂组分以及光合作用影响。结果显示,总脂含量由22.4%±0.6%提高到36.3%±0.7%,其中甘油三酯含量由4.6%±1.2%提高到68.3%±2.5%而磷脂含量由92.8%±1.6%降低到26.8%±2.0%。缺氮处理1d后,叶绿素含量明显下降,呼吸速率明显升高。缺氮处理初始阶段,栅藻光系统Ⅱ最大光能转化效率(Fv/Fm)和非光化学猝灭系数(NPQ)维持稳定,4 d后Fv/Fm显著降低而NPQ显著升高。上述变化可能是光能过剩引起的防御反应,也可能是自身代谢模式发生转变的结果而与光能耗散无关。     

三种海洋浮游微藻的分子鉴定及培养条件研究

采用分子生物学手段结合形态特征对海洋中生活着的浮游单细胞海藻进行准确无误的分类鉴定,正在成为一种普遍接受的方式。这样做的前提是首先需要从海水中分离得到纯藻株, 掌握它们的生活条件, 获得生物量, 并进而完成鉴定。作者对一株裸甲藻类似种和两株新分离的赤潮单细胞海藻进行了分离、培养和鉴定。核糖体18S RNA 基因(18S rDNA)和转录单元内间隔区(ITS)序列分析结果显示这三株藻分别为共生甲藻(Symbiodinium sp.)、锥状斯氏藻(Scrippsiella trochoidea)和赤潮异弯藻(Heterosigma akashiwo)。作者以广泛使用的光密度值A₆₈₀作为衡量藻体生物量变化的指标, 研究了这三种藻在不同培养条件下的生长。结果表明, 在批次培养中, 共生甲藻、锥状斯氏藻和赤潮异弯藻的生物量都在22℃达到最大, 分别为9.28×10⁵个/mL、7.9×10⁴个/mL 和4.75×10⁵个/mL; 强光和长时间光照促进生长, 但锥状斯氏藻在强光下更早进入衰退期; 三种藻都表现出较强耐受盐度变化的能力,在盐度为15～36 的培养液中生长良好; 营养实验表明, 使用882 μmol/L 的NO₃^- 和36.2 μmol/L 的PO₄^3-能够维持三种海藻的快速生长。     

不同摄食水平条件下鮻的碳收支研究

在水温20℃和盐度30的条件下研究摄食水平对鮻碳收支的影响。摄食水平分为饥饿（不投喂）、1、2、3、4（指日摄食量占鱼体重百分比）和自由摄食共6组，实验持续21d。结果表明：摄食水平对碳消化率没有影响。碳转化率、净生长率与摄食水平之间呈显著正相关关系，在自由摄食状态下分别达到9．29％和10．41％。非粪便流失碳随摄食水平增加而显著降低。粪便碳占摄入碳比例变化不大，与摄食水平间无规律性关系。在自由摄食状态下，大部分摄入碳量均通过非粪便途径流失，其中大部分是呼吸过程消耗的。余下的部分中，粪便流失量和鱼体保存量各占一半。鱼体每增长1g体重伴有979．56mg碳流失。在自由摄食状态下鮻碳收支方程式为：100摄入碳=9．29生长碳＋9．69粪便碳＋79．94非粪便流失碳。     

铜在海洋食物链中的积累与传递规律的研究

在实验室条件下,研究铜在海水及以扁藻、轮虫、对虾等生物组成的食物链中的积累与传递规律。结果表明,各种海洋生物都能直接从海水中吸收铜,铜可沿着海水→扁藻→轮虫→仔虾传递。文章还探讨了本食物链各环节积累铜的主要途径以及影响生物个体铜的累积量的诸因素。     

Dark respiration in the light and in darkness of three marine macroalgal species grown under ambient and elevated CO2 concentrations

Dark respiration (non-photorespiratory mitochondrial respiration),which occurs both in the light and in darkness,is vital for growth and survival of algae and plays a critical role in modulating the carbon balance of them.In the present study,we have investigated dark respiration in the light (R L) and in darkness (R D) in three marine macroalgal species,Hizikia fusiformis (phaeophyta),Gracilaria lemaneiformis (Rhodophyta) and Ulva lactuca (Chlorophyta),cultured at 20℃ using aeration with two CO 2 conditions:current ambient (CO 2 concentration about 380 μl/L) and elevated CO 2 (approximately 720 μl/L) air.R L was estimated by using the Kok method,whereas R D was determined as the rate of O 2 influx at zero light.The results showed that both R L and R D were unchanged for the elevated CO 2-grown algae relative to ambient CO 2 concentration for all the algal species tested.However,R L was significantly lower than R D across all the algal species and growth CO 2 treatments,demonstrating that daytime respiration was partly depressed by the light.The percentage of inhibition of respiration by light was similar between ambient and elevated CO 2 grown algae.The ratio of respiration to photosynthesis,which tended to decrease when estimated using R L instead of R D,was not altered for the elevated relative to ambient CO 2 concentration.The results suggest that R L,rather than R D,is a more accurate estimate of nonphotorespiratory carbon loss in marine macroalgae during the daytime.It would not be anticipated that elevated atmospheric CO 2 would exert a substantial influence on respiratory flux either in the light or in darkness in these particular marine macroalgal species.     

CO2加富培养对鼠尾藻克制赤潮异弯藻效果的影响

采用实验生态学的方法,研究了大型海藻鼠尾藻(Sargassum thunbergii)对赤潮微藻中肋骨条藻(Skeletonema costatum)的克生效应及其对 CO2加富的响应变化.结果表明,正常培养的鼠尾藻新鲜组织、干粉末和水溶性抽提液能够导致中肋骨条藻的细胞密度显著降低(P<0.05), CO2加富培养的鼠尾藻新鲜组织、干粉末和水溶性抽提液能够更加明显地抑制中肋骨条藻种群生长(P<0.01).实验结果证实鼠尾藻对中肋骨条藻具有克生效应,而 CO2加富培养引起了鼠尾藻对中肋骨条藻克生效应的变化,使其对中肋骨条藻生长的抑制作用显著加强.正常培养的鼠尾藻培养水过滤液能够显著抑制中肋骨条藻的生长(P<0.05),而 CO2加富培养的鼠尾藻培养水过滤液对中肋骨条藻的生长没有表现出明显的影响(P>0.05),因此推测 CO2加富培养可能改变了鼠尾藻对中肋骨条藻克生作用的方式,作用方式由分泌克生物质转变为细胞直接接触转递.     

Distributions and air-sea fluxes of CO2 in the summer Bering Sea

The 3rd Chinese National Arctic Research Expedition(CHINARE–Arctic III) was carried out from July to September in 2008. The partial pressure of CO2(pCO2) in the atmosphere and in surface seawater were determined in the Bering Sea during July 11–27, 2008, and a large number of seawater samples were taken for total alkalinity(TA) and total dissolved inorganic carbon(DIC) analysis. The distributions of CO2 parameters in the Bering Sea and their controlling factors were discussed. The pCO2 values in surface seawater presented a drastic variation from 148 to 563 μatm(1 μatm = 1.013 25×10-1 Pa). The lowest pCO2 values were observed near the Bering Sea shelf break while the highest pCO2 existed at the western Bering Strait. The Bering Sea generally acts as a net sink for atmospheric CO2 in summer. The air-sea CO2 fluxes in the Bering Sea shelf, slope, and basin were estimated at-9.4,-16.3, and-5.1 mmol/(m2·d), respectively. The annual uptake of CO2 was about 34 Tg C in the Bering Sea.     

夏季东海西部表层海水中的pCO2及海-气界面通量

根据2001年夏季长江口及东海西部海域表层海水pCO     

大气CO2浓度升高对繁茂膜海绵滤食海水细菌能力的影响

通过模拟大气CO_2浓度升高,研究其对海绵滤食细菌功能的影响。在模拟大气CO_2浓度升高生态系统中,探究了大气CO_2浓度387、500、750和1 000mmol/mol环境下繁茂膜海绵(Hymeniacidon perlevis)滤食灭菌海水中大肠杆菌(Escherichia coli AS 1.1017)和灿烂弧菌(Vibrio splendidus)的能力。结果表明:在24 h实验期间,模拟大气CO_2从目前约387mmol/mol升高至500mmol/mol,繁茂膜海绵滤食大肠杆菌和灿烂弧菌效率提高了。当模拟大气CO_2 750mmol/mol时,繁茂膜海绵滤食海水中大肠杆菌和灿烂弧菌功能都下降了,说明繁茂膜海绵已经受到大气较高浓度CO_2损害。模拟大气CO_2为1 000mmol/mol时,繁茂膜海绵基本丧失了滤食海水中大肠杆菌和灿烂弧菌的功能。上述结果可为了解大气CO_2浓度对近岸海洋生态系统的影响提供科学依据。     

海浪对北太平洋海-气二氧化碳通量的影响

利用4种海-气界面气体传输速率公式对比研究了北太平洋气体传输速率及其CO₂通量的季节变化特征。与单纯依赖风速的算法相比, 考虑波浪影响的气体传输速率和CO₂通量在空间分布和季节变化上具有明显差异。在低纬度地区(0°～30°N), 波浪参数使气体传输速率下降, 海洋对大气CO₂的吸收减少, 而在30°N以北范围内则出现新的气体传输速率高值区, 海洋对大气的吸收增加。进一步研究了黑潮延伸体区域的气候态月平均气体传输速率和CO₂通量。结果表明, 该区域气体传输速率和CO₂通量最大值分别出现于冬季和春季, 引入波浪参数后, 虽然该区域气体传输速率和CO₂通量平均值没有明显差异, 但季节变化强度显著增强。     

High-resolution measurement of Southern Ocean CO2 and O2/Ar by membrane inlet mass spectrometry

This paper evaluates the simultaneous measurement of dissolved gases (CO₂ and O₂/Ar ratios) by membrane inlet mass spectrometry (MIMS) along the 180° meridian in the Southern Ocean. The calibration of pCO₂ measurements by MIMS is reported for the first time using two independent methods of temperature correction. Multiple calibrations and method comparison exercises conducted in the Southern Ocean between New Zealand and the Ross Sea showed that the MIMS method provides pCO₂ measurements that are consistent with those obtained by standard techniques (i.e. headspace equilibrator equipped with a Li–Cor NDIR analyser). The overall MIMS accuracy compared to Li–Cor measurements was 0.8 μatm. The O₂/Ar ratio measurements were calibrated with air-equilibrated seawater standards stored at constant temperature (0 ± 1 °C). The reproducibility of the O₂/Ar standards was better than 0.07% during the 9 days of transect between New Zealand and the Ross Sea.The high frequency, real-time measurements of dissolved gases with MIMS revealed significant small-scale heterogeneity in the distribution of pCO₂ and biologically-induced O₂ supersaturation (ΔO₂/Ar). North of 65°S several prominent thermal fronts influenced CO₂ concentrations, with biological factors also contributing to local variability. In contrast, the spatial variation of pCO₂ in the Ross Sea gyre was almost entirely attributed to the biological utilization of CO_2, with only small temperature effects. This high productivity region showed a strong inverse relationship between pCO₂ and biologically-induced O₂ disequilibria (r² = 0.93). The daily sea air CO₂ flux ranged from − 0.2 mmol/m² in the Northern Sub-Antarctic Front to − 6.4 mmol/m² on the Ross Sea shelves where the maximum CO₂ influx reached values up to − 13.9 mmol/m². This suggests that the Southern Ocean water (south of 58°S) acts as a seasonal sink for atmospheric CO₂ at the time of our field study.     

2008年夏季加拿大海盆净二氧化碳吸收量评估

The third Chinese National Arctic Research Expedition(CHINARE) was conducted in the summer of 2008.During the survey,the surface seawater partial pressure of CO_2(pCO_2) was measured,and sea water samples were collected for CO_2 measurement in the Canada Basin.The distribution of pCO_2 in the Canada Basin was determined,the influencing factors were addressed,and the air-sea CO_2 flux in the Canada Basin was evaluated.The Canada Basin was divided into three regions:the ice-free zone(south of 77°N),the partially ice-covered zone(77°–80°N),and the heavily ice-covered zone(north of 80°N).In the ice-free zone,pCO_2 was high(320 to 368μatm,1 μatm=0.101 325 Pa),primarily due to rapid equilibration with atmospheric CO_2 over a short time.In the partially ice-covered zone,the surface pCO_2 was relatively low(250 to 270 μatm) due to ice-edge blooms and icemelt water dilution.In the heavily ice-covered zone,the seawater pCO_2 varied between 270 and 300 μatm due to biological CO_2 removal,the transportation of low pCO_2 water northward,and heavy ice cover.The surface seawater pCO_2 during the survey was undersaturated with respect to the atmosphere in the Canada Basin,and it was a net sink for atmospheric CO_2.The summertime net CO_2 uptake of the ice-free zone,the partially ice-covered zone and the heavily ice-covered zone was(4.14±1.08),(1.79±0.19),and(0.57±0.03) Tg/a(calculated by carbon,1Tg=10~(12) g),respectively.Overall,the net CO_2 sink of the Canada Basin in the summer of 2008 was(6.5±1.3) Tg/a,which accounted for 4%–10% of the Arctic Ocean CO_2 sink.     

基于蛋白组学分析杜氏盐藻对不同浓度CO2的响应

用气体混合仪设置不同的CO_2浓度处理组,测定了杜氏盐藻(Dunaliella salina)细胞密度、碳酸酐酶活性、甘油含量等生理指标,结合蛋白质组学分析方法,比较了不同CO_2浓度下细胞内主要代谢路径蛋白表达的差异。结果表明:在一定范围内,随着CO_2浓度的升高,杜氏盐藻的生理活性及光合活性提高;而CO_2浓度过高对盐藻生长呈抑制作用; 3%CO_2浓度最适于本实验杜氏盐藻藻株的生长。随着CO_2浓度的升高,胞外碳酸酐酶活性下降。低浓度的CO_2有利于β-胡萝卜素的积累,且光系统Ⅱ(PSⅡ)光合活性在CO_2浓度0.03%～3%范围内上升, CO_2浓度达9%时降低,与光合作用相关蛋白的表达趋势接近。上述结果说明杜氏盐藻可能通过调节光合作用中叶绿素等捕光色素的合成及相关蛋白的表达笼统,以响应CO_2浓度的变化;而过高浓度的CO_2可对细胞产生氧化损害,引起热激蛋白和超氧化物歧化酶等蛋白含量的上调以应对氧化胁迫。