Isolation of Chloroplasts from Marine Microalga Isochrysis galbana Parke for Their Lipid Composition Analysis     

ZHONG Ye  LI Yanrong  XU Jilin  CAO Jiayi  ZHOU Chengxu  YAN Xiaojun 《中国海洋大学学报(英文版)》2022,21(1):225-235

Marine microalga Isochrysis galbana is an important feed species with a high nutritional value.Different from other uni-cellular algae,its cell contains two chloroplasts which are the major sites for lipid synthesis.Here,we optimized a chloroplast isola-tion approach suitable for the isolation of I.galbana chloroplasts and determined the purity and integrity of the isolated chloroplasts through microscopic observations and enzyme activity assay.The chloroplast lipids were analyzed with a ultrahigh-performance li-quid chromatography-Q Exactive Orbitrap-mass spectrometry.This newly developed isolation approach is simple and reliable to isolate chloroplasts with high integrity and purity.The average yield of intact chloroplasts was 15.3%±0.1%.Glycolipids and acyl-glycerols were the main chloroplast lipids.Glycolipids accounted for 56.6%of chloroplast lipid.Digalactosyldiacylglycerol(DGDG),monogalactosyldiacylglycerol(MGDG)and sulfoquinovosyldiacylglycerol(SQDG)were the main glyceroglycolipids.The fatty acyl R1/R2 were mostly 18:4/16:1,18:3/16:1 and 18:4/18:5 in DGDGs,14:0/18:4,18:4/18:5,18:4/18:4 and 18:3/18:4 in MGDGs and 16:0/14:0,16:0/18:3,and 18:4/18:3 in SQDGs.In addition,diacylglycerol(DAG)was the most abundant acylglycerols;the content of 22:6/18:4-DAG was the highest.There was a little amount of glycosphingolipid(GSL)in chloroplast.Digalactosylmonoglyceride(DGMG),monogalactosylmonoglyceride(MGMG),sulfoquinovosylmonoacylglycerol(SQMG),monoglyceride(MAG),phospholi-pids(PLs),ceramide(Cer)and betaine lipids were nearly undetectable in chloroplast.The fatty acid proportions of DGDGs,MGDGs,SQDGs,DAGs,triglycerides(TAGs)and GSLs were either higher or lower than or similar to those of whole-cell.Collectively,our isolation approach is applicable to many aspects of chloroplast biology,and may offer a reference for the isolation of chloroplasts from other marine microalgae.  相似文献   

应用Cornell方法对重口裂腹鱼[Schizothorax (Racoma)davidi]幼鱼适宜常量营养物质需求量研究     

宁毅  冯健  何姝珍  孙晓峰  陈斌 《海洋与湖沼》2012,43(4):748-755

采用Cornell方法研究了初始重为(2.0±0.1)g的重口裂腹鱼幼鱼日粮中常量营养物质的适宜需求量。日粮中蛋白质、脂肪和碳水化合物梯度变化范围分别为33%—51%、3%—18%和24%—33%,试验周期为60d。试验结果表明,除日粮蛋白质33%组鱼成活率明显下降外(P<0.05),其它试验日粮中3种常量营养物质含量对重口裂腹鱼幼鱼成活率没有显著影响(P>0.05)。当日粮蛋白质水平≤45%时,其特定生长率随着日粮蛋白质水平上升而明显增加;当日粮蛋白质水平在33%—36%时,重口裂腹鱼的特定生长率显著降低(P<0.05)。除12%日粮脂肪组试验鱼外,9%日粮脂肪组试验鱼的特定生长率显著性高于其它脂肪水平日粮组(P<0.05),但当日粮中脂肪含量≥15%后,重口裂腹鱼鱼体脂肪含量显著上升(P<0.05),肝细胞脂肪变性明显。日粮中不同的碳水化合物水平对重口裂腹鱼特定生长率没有显著影响(P>0.05),但当日粮中碳水化合物为33%时,鱼体糖原含量显著增加(P<0.05)。试验表明,重口裂腹鱼幼鱼日粮蛋白质、脂肪和碳水化合物3种常量营养物质的适宜需求量分别为45%、9%和≤30%。日粮总能为19.0MJ/kg,蛋白质能量比(P/E)为23mg/J。  相似文献   

微藻总脂含量测定方法概述   总被引：1，自引：0，他引：1  

梁英  石伟杰  田传远 《中国海洋大学学报(自然科学版)》2012,42(5):35-40

概述微藻总脂含量测定方法的研究现状并比较各方法的优缺点:(1)有机溶剂提取法使用较普遍、易操作、成本低,缺点是所需样品量大、所得总脂含量较低、溶剂有毒易污染环境、操作耗时费力;(2)索氏提取法测定微藻总脂含量较准确,缺点是所需试剂和样品量大,提取时间长;(3)荧光染料测定法、傅里叶变换红外光谱法、磷酸香草醛法、铜试剂法、苏丹黑染色法以及核磁共振法所需样品量少、所用时间短,但测定的总脂含量均为相对值.其中,荧光染料测定法不需要提取油脂且灵敏度高,但其易受各种因子影响;傅里叶变换红外光谱法和核磁共振法对样品无损伤,无需或仅需很少的预处理,后者重复性很好;磷酸香草醛法较稳定;铜试剂法成本低,但对其报道极少;苏丹黑染色法操作简单,不需要对藻细胞破碎及化学抽提,但不适用于不均匀样品的测定;(4)超临界CO2提取法和离子液体提取法提取效率高,所得总脂含量较高,但测定成本较高.  相似文献   

Tissue-specific bioaccumulation and oxidative stress responses in juvenile Japanese flounder (Paralichthys olivaceus) exposed to mercury   总被引：1，自引：0，他引：1  

黄伟  曹亮  叶振江  林龙山  陈全震  窦硕增 《中国海洋湖沼学报》2012,30(4):569-579

To understand mercury (Hg) toxicity in marine fish, we measured Hg accumulation in juvenile Japanese flounder (Paralichthys olivaceus) and assessed the effects on growth and antioxidant responses. After Hg exposure (control, 5, 40, and 160 μg/L Hg) for 28 d, fish growth was significantly reduced. The accumulation of Hg in fish was dose-dependent and tissue-specific, with the maximum accumulation in kidney and liver, followed by gills, bone, and muscle. Different antioxidants responded differently to Hg exposure to cope with the induction of lipid peroxidation (LPO), which was also tissue-specific and dosedependent. As Hg concentration increased, superoxide dismutase (SOD) and catalase (CAT) activities increased significantly, whereas glutathione S-transferase (GST) activity and glutathione (GSH) levels decreased significantly in the gills. SOD and glutathione peroxidase (GPx) activities and the GSH level increased significantly in the liver. SOD activity and GSH levels increased significantly, but CAT activity decreased significantly with an increase in Hg concentration in the kidney. LPO was induced significantly by elevated Hg in the gills and kidney but was least affected in the liver. Therefore, oxidative stress biomarkers in gills were more sensitive than those in the liver and kidney to Hg exposure. Thus, the gills have potential as bioindicators for evaluating Hg toxicity in juvenile flounder.  相似文献   

Lipid and Fatty Acid Compositions of Cod(Gadus morhua),Haddock(Melanogrammus aeglefinus)and Halibut(Hippogiossus hippoglossus)     

ZENG Duan  MAI Kangsen  AI Qinghui  Joyce E.Milley  Santosh P.Lall 《中国海洋大学学报(英文版)》2010,9(4)

This study was conducted to compare lipid and fatty acid composition of cod,haddock and halibut.Three groups of cod(276 g±61 g),haddock(538 g±83 g)and halibut(3704 g±221 g)were maintained with commercial feeds mainly based on fish meal and marine fish oil for 12 weeks prior to sampling.The fatty acid compositions of muscle and liver were determined by GC/FID after derivatization of extracted lipids into fatty acid methyl esters(FAME).Lipids were also fractionated into neutral and polar lipids using Waters silica Sep-Pak(R).The phospholipid fraction was further separated by high-performance thin-layer chromatography(HPTLC)and the FAME profile was obtained.Results of the present study showed that cod and haddock were lean fish and their total muscle lipid contents were 0.8% and 0.7%,respectively,with phospholipid constituting 83.6% and 87.5% of the total muscle lipid,respectively.Halibut was a medium-fat fish and its muscle lipid content was 8%,with 84% of the total muscle lipid being neutral lipid.Total liver lipid contents of cod,haddock and halibut were 36.9%,67.2% and 30.7%,respectively,of which the neutral lipids accounted for the major fraction(88.1%-97.1%).Polyunsaturated fatty acids were the most abundant in cod and haddock muscle neutral lipid.Monounsaturated fatty acid level was the highest in halibut muscle neutral lipid.Fatty acid compositions of phospholipid were relatively constant.In summary,the liver of cod and haddock as lean fish was the main lipid reserve organ,and structural phospholipid is the major lipid form in flesh.However,as a medium-fat fish,halibut stored lipid in both their liver and muscle.  相似文献   

饲料脂肪水平对红鳍东方鲀幼鱼肝脏抗氧化酶活力及组织结构的影响     

孙阳  姜志强  李艳秋  毛明光  吴洪  宫雪 《广东海洋大学学报》2013,(3)

以脂肪质量分数分别为5.77%、7.71%、8.93%、11.89%、13.75%、16.42%的饲料饲喂红鳍东方鲀(Takifugurubripe)幼鱼56 d,测定其肝脏抗氧化酶活力和总抗氧化能力,并对其肝脏、肠及肌肉进行组织学观察,研究不同饲料脂肪水平对红鳍东方鲀幼鱼肝脏抗氧化酶活力及组织结构的影响。结果表明,5.77%组过氧化氢酶(CAT)活力与其他各组间有显著性差异(P<0.05)。总抗氧化能力(T-AOC)活力的最高值出现在8.93%组,且显著高于其他各组(P<0.05)。饲料脂肪水平大于8.93%的实验组间超氧化物歧化酶(SOD)活力差异不显著(P>0.05),且显著低于8.93%组;5.77%、7.71%、8.93%组呈增加趋势,且各组间差异显著(P<0.05)。谷胱甘肽过氧化物酶(GSH-PX)活力的峰值出现在13.75%组,且显著高于其他各组(P<0.05),总体呈先增加后降低的趋势。组织学观察表明,5.77%、7.71%、8.93%组,虽然肝脏也有脂肪空泡存在,但是肝脏、肠和肌肉组织结构完整,无明显病变现象;11.89%、13.75%、16.42%组的肝脏、肠和肌肉均有不同程度的病变现象,各组间差别明显。由此认为,脂肪水平为8.93%组,抗氧化酶活力最高,该脂肪水平下的饲料对红鳍东方鲀幼鱼肝脏代谢最有利,组织结构完整无病变。  相似文献   

30株海洋绿藻的总脂含量和脂肪酸组成   总被引：9，自引：2，他引：7  

曹春晖  孙世春  麦康森  梁英 《中国海洋大学学报(自然科学版)》2000,30(3):428-434

对 11属 (小球藻属、绿囊藻属、微绿球藻属、海绿球藻属、卵胞藻属、原球藻属、咸胞藻属、杜氏藻属、裂丝藻属、塔胞藻属和衣藻属 )的 30株海洋绿藻进行特定条件下的一次性培养 ,在指数生长末期收获 ,进行了总脂含量和脂肪酸组成的分析。 2 1株海洋绿藻的总脂含量超过干重的10 % ,达 11.6 1%～ 34.4 9% ,其它 9株在 4 .2 5%～ 9.4 8%之间 ,绿藻的 16碳和 18碳脂肪酸最为丰富 ,有着含量较高的 16∶ 0、16∶ (n- 3)、18∶ 2 (n- 6 )和 18∶ 3(n- 3)脂肪酸。两株小球藻 (C95,C97)的2 0∶ 5(n- 3)脂肪酸含量较高 ,分别为 2 0 .8%和 2 6 .1%。另一株小球藻 (C10 2 )和两株裂丝藻 (C19和C2 0 ) EPA含量居中 ,分别为 8.0 % ,6 .0 %和 8.1%。其它藻株一般只含有少量的 2 0∶ 5(n- 3)和 2 2∶ 6 (n- 3)或不含 2 2∶ 6 (n- 3)  相似文献   

基于脂质生物标志物评估河道型水库表层沉积物有机质来源     

宿苧仁  Emilie Jardé  尹月鹏  Marine Liotau  张雯 《湖泊科学》2024,36(1):137-148

水电开发导致河岸生境破碎化程度加剧和水库沉积物截留问题日益突出,而目前关于河道型水库有机质(OM)溯源及分布特征的研究较少,进而造成OM在水库复杂环境介质中迁移机理尚不清楚。本研究将脂质生物标志物分析方法应用于大渡河瀑布沟水库表层沉积物,经过气相色谱-质谱法对沉积物OM脂质组分进行了分析,将其量化并分为自然、成岩、热源和污水标志物4种来源。结果表明:OM平均含量坝前(10.7%±4.8%)>坝后(7.3%±0.9%)>库中(6.7%±1.8%),说明水库对沉积物OM的截留有显著影响。可提取脂质组分主要来源于自然标志物(45.9%±13.9%),碳偏好指数、陆源/水生比和陆生与水生植物输入比显示OM主要来源于陆生高等植物,占自然定量分子的70.5%±9.1%。此外,坝前汉源城镇下游D6和D7点位人为标志物(44.6%±25.1%)分子明显比自然标志物(24.2%±3.7%)含量高,并进一步通过比例研究强调了污水标志物的贡献率较为突出(31.6%±12.8%),这与该地住宅区生活污水处理设施覆盖率较低相关,表明OM污染源组成变化与水库滞留效应的耦合作用有关。  相似文献   

乳化碘油增强CT检测早期肝癌的实验研究(英)     

许乙凯  张嘉宁  刘杏元  乔东访 《CT理论与应用研究》1999,(2)

目的是探讨乳化碘油增强CT对早期肝癌的诊断作用。方法采用乳化碘油：由碘化油、卵磷脂、甘油等经超声乳化而成：用二乙基亚硝胺诱导法建立大鼠肝癌模型（n＝4）：观察静脉注射乳化碘油（0.4ml／只）后,正常大鼠（n＝4）,及大鼠肝癌病灶的强化情况,并与病理学检查进行对照分析。结果得到：乳化碘油能引起肝脾组织特异性显著强化：透视电镜证实肝细胞参与碘油乳粒的摄取：选取的4只大鼠肝癌模型中,共有14个瘤结,大小4、5±3.4mm;乳化碘油增强CT检出率85.7％（12／14）的瘤结,明显高于平扫时,病灶显示率（14．3％）。结论为乳化碘油是一种潜在的肝脾特异性造影剂,对早期肝癌的检出具有重要价值。  相似文献   

Evaluation of anthropogenic and biogenic inputs into the western Mediterranean using molecular markers     

Jordi Dachs  Josep M. Bayona  Joëlle Fillaux  Alain Saliot  Joan Albaigs 《Marine Chemistry》1999,65(3-4)

Aliphatic hydrocarbons (AHs), sterols (ST), and lipid classes were determined in suspended particles collected in the Catalan Sea, northwestern Mediterranean. Principal Component Analysis of the data set revealed a clustering of samples depending on the sources of organic matter, i.e., coastal influenced, open sea and frontal zone. Terrestrial inputs were recognized in particles collected in the surface waters, at the vicinities of river outflow (i.e., Rhône and Ebro), by a predominance of C₂₉ and C₃₁ n-alkanes, 24-ethylcholest-5-en-3β-ol (S12), and the anthropogenic 5β(H)-cholestan-3β-ol, coprostanol (S1). Phytoplanktonic molecular markers (n-C₁₇, 27-nor-24-methylcholesta-5,22E-dien-3β-ol, cholesta-5,22-dien-3β-ol and 24-methylcholest-5-en-3β-ol) were widespread but relatively more apparent in the open sea and frontal zones. A similar distribution was observed for lipid classes, with higher concentrations of phospholipids, and an enrichment in free fatty acids in the areas influenced by river discharges. Total sterol, the unresolved complex mixture of hydrocarbons and the pristane–phytane ratio were highest at a persistent frontal zone, possibly reflecting the combination of a higher primary productivity and fossil hydrocarbon absorption on to suspended particles. Moreover, vertical profiles exhibited a subsurface concentration maximum at 20–30 m water depth, concurrently with the chlorophyll.  相似文献