紫菜丝状体DNA的提取

紫菜(Porphyra spp.)是一种大型红藻,它的孢子体世代是丝状体.丝状体适于在常温条件下(20℃左右)长期保存和培养,是筛选丝状体优良栽培品系,利用光生物反应器进行紫菜育苗的理想材料.紫菜细胞含有大量胞外多糖,主要以硫酸基多糖和羧基多糖形式存在,它们比陆生植物的中性多糖更易溶于水,与DNA产生共沉淀,从而影响限制性内切酶的消化和PCR扩增.另外,紫菜细胞DNA含量较少,DNA酶含量却很高,使紫菜DNA提取的得率不足[1].     

应用响应曲面法优化萱藻丝状体单室孢子囊发育条件

为优化萱藻丝状体单室孢子囊发育条件,提高孢子囊比例,研究以萱藻丝状体为材料,在单因素试验的基础上,应用响应曲面法对丝状体单室孢子囊的发育条件进行优化。结果显示,诱导萱藻丝状体单室孢子囊发育的最佳条件为:温度17.67°C,光照强度30.32μmol/(m²·s),外加氮浓度37.37 mg/L,外加磷浓度10.63 mg/L。在该条件下诱导培养20 d,丝状体孢子囊比例为48.92%±5.58%,约为对照组的1.41倍。试验结果与理论预测值的相对误差小于5%,模型可靠。为评价优化培养后萱藻丝状体孢子囊的发育情况,对丝状体进行了阴干刺激(17°C,2h),结果显示,优化培养后的丝状体游孢子放散量(3.33×10⁶ ind./g FW)约为对照组(1.67×10⁶ ind./g FW)的1.99倍。研究表明,在萱藻丝状体单室孢子囊诱导过程中,对诱导环境条件进行有目的性的调控,不仅可以提高孢子囊比例,还可以促进孢子囊发育和游孢子放散的同步性。     

基于第二代测序技术的条斑紫菜(Pyropia yezoensis)贝壳丝状体附生菌群研究

条斑紫菜(Pyropia yezoensis)贝壳丝状体黄斑病给育苗产业造成危害,但目前对健康状态下的菌群结构及其发病后变化仍缺乏认识。本研究基于第二代测序技术和16S r RNA序列分析,对健康和患病丝状体的细菌组成与变化进行比较。测序共产生1.6Gb配对末端序列,获得了第一个高通量的条斑紫菜贝壳丝状体细菌群落数据集。通过数据分析共识别出7,833种可操作分类单元(operational taxonomic units,OTUs),比对确定为18个细菌门。相对于健康样本,黄斑病丝状体样本的微生物多样性显著增加。主坐标分析和非加权组平均法(unweighted pair group method using arithmetic average,UPGMA)聚类均将健康和黄斑病丝状体分成不同的组,且健康组样本间的距离明显小于黄斑病组。上述结果说明,健康贝壳丝状体保持着相似的微生物群落结构,而患病后的丝状体则发生了不同的变化;健康样本和黄斑病样本组间差异显著的细菌属有39个,其中29个在健康样本中的相对丰度明显高于病患组样本。本研究从菌群生态角度分析了健康贝壳丝状体微生物菌群结构、发病后菌群的变化及其对丝状体生长的潜在作用,为丝状体病害的检测和预防提供有价值的信息。     

包埋脱水法超低温保存萱藻(Scytosiphon lomentaria)丝状体的研究

以本实验室保存的萱藻(Scytosiphon lomentaria)丝状体为材料,用包埋脱水法超低温保存萱藻丝状体,研究了蔗糖预培养浓度和时间、胶球含水量、化冻温度以及胶球恢复时间对萱藻丝状体存活率和冻存后生长发育能力的影响。结果显示:(1)萱藻丝状体胶球在0.4mol/L的蔗糖溶液中预培养6h能获得最高的存活率;(2)萱藻丝状体胶球的最佳含水量较低,为15%左右,当含水量高于27%时,冻存后的存活率为0;(3)40°C为最适的化冻温度;(4)化冻后,将萱藻丝状体胶球在黑暗条件下恢复18h能显著提高存活率;(5)在最佳条件下,萱藻丝状体经冻存后的存活率最高可达54.79%,恢复后的丝状体与冻存前丝状体并无区别,且经诱导后能够产生正常的孢子囊,放散的孢子可以发育成叶状体。     

颗石藻Pleurochrysis carterae形成休眠体的环境诱因分析

利用实验生理生态学方法对Pleurochrysis carterae种群增殖过程中出现丝状体的环境诱因进行了研究,从营养胁迫、钙化抑制胁迫、低温胁迫以及外源α-酮戊二酸的影响等几个方面,探讨了环境因子对P.carterae细胞休眠的诱导作用。结果表明,氮营养盐水平在P.carterae形成丝状体过程中有重要的作用,氮缺乏和氮限制会明显诱导丝状体的早期大量形成,即细胞快速进入休眠期。磷缺乏和磷限制对颗石藻丝状体形成的短期诱导作用不显著。正常钙化的颗石藻在缺钙环境中钙壳逐渐脱落、细胞裸露、没有种群增殖、不形成丝状体。低温胁迫下,颗石藻细胞不分裂,但细胞维持游泳的钙化单细胞状态,不形成丝状体。外源α-酮戊二酸对颗石藻形成丝状体具有浓度相关的效应,小于1.2mmol/L的浓度抑制丝状体的早期发生;1.2mmol/L及以上浓度的α-酮戊二酸能诱导丝状体早期发生。     

北欧东部古生代台地生物成因碳酸盐岩演化

通过追踪分布在北极刮北乌拉尔内超过1000km长的东北欧古生代台地生物成因碳酸盐岩,发现它们在前乌托尔前渊,伯朝拉陆槽和Timan均有发育。生物成因碳酸盐岩建造可分为5类：1）后生动物骨架;2）后生动物-钙质微生物骨架;3）钙质微生物骨架;4）非钙质微生物骨架,和5）生物粘结岩。从古生代初至末期,本区以生物礁建造为主的生物控制作用逐渐由强变弱。     

海洋微生物Paenisporosarcina quisquiliarum对Fe的生物矿化过程

为了解海洋微生物的生物矿化作用在大洋铁锰结核形成过程中的意义,选取芽孢八叠球菌作为实验菌株,通过实验室模拟中性有氧环境下的生物矿化实验,测定了Fe生物矿化过程中Fe离子浓度与价态的变化、细菌表面形态与矿物成分的变化。反应过程中,通过与死菌对照组和空白对照组的对比研究,有菌实验组的总Fe浓度、Fe²⁺浓度和Fe³⁺浓度下降最快,尤其是在前24 h。TEM照片显示细菌表面有明显的矿物颗粒生成,对这些矿物颗粒进行EDS分析测定,推测为Fe的氧化物或氢氧化物。实验结果表明,以芽孢八叠球菌为代表的海洋微生物在中性有氧条件下,通过主被动相结合的过程,对Fe产生了明显的矿化作用,揭示了微生物矿化作用对大洋铁锰结核形成的贡献。     

光温对紫菜丝状体生长发育的影响

紫菜的丝状体阶段比叶状体阶段更容易培养和保存,为了加深对丝状体阶段的认识,在相当长的一段时间内,对紫菜的研究热点集中在丝状体阶段,其中大量的工作涉及丝状体不同阶段对光温条件的需求上。1　光对紫菜丝状体生长发育的影响由于植物生活的地域环境和季节不同,导致了植物受光量的不同,植物对这种环境的长期适应结果使其对不同光强、光周期和光质产生不同的生理反应。所以,植物的生长发育与所处环境的光因子的变化密切相关。紫菜是一种潮间带藻类,其丝状体阶段却生活在潮下带;紫菜在全世界的分布很广,不同种类生活的地理区域(如纬度、水深…     

坛紫菜单性叶状体细胞的发育研究

研究采用雌雄营养组织块和营养细胞进行培养,以了解坛紫菜单性营养细胞的生长发育情况.结果表明,坛紫菜雌性营养细胞无论在组织水平还是在细胞水平均以产生红色果胞或果孢子细胞为主,有少量细胞发育成精子囊,并放散精子.雄性营养组织和营养细胞以产生精子为主,但不能再分化成丝状体;产生的少量果孢子细胞能发育成丝状体.因此,在室内培养条件下,坛紫菜单性叶状体都能产生两性细胞,它们的差异只是不同性细胞所占的优势不同,并讨论了单性叶状体的产生.     

大洋铁锰结核的微生物成矿过程及其研究进展

深海铁锰结核作为世界上潜在的巨大金属宝库已成为当今开发海底矿藏的热点,因而深入了解铁锰结核成矿过程成为其开发利用的先决条件。研究发现多金属铁锰结核中的铁锰矿物不仅仅是由单纯的物理作用形成的,同时也包含了海洋生物驱动的生物矿化的过程。本文介绍了运用分子生物学、矿物学和地球化学等多学科的研究方法对大洋中铁锰的生物成矿过程和成矿特征的研究。深海铁锰结核的生长速率缓慢且其生长演化伴随着微生物群落的活动,因此结核的生长过程同时也记录着不同时期微生物群落结构的变化并生成了大量的微生物化石。在铁和锰的生物矿化过程中,细菌可以通过酶促反应氧化Fe（Ⅱ）和Mn（Ⅱ）,同时可能伴随生物能量的生成,此外微生物还可以通过非酶促反应的方式促进Fe和Mn的富集沉淀。这些研究表明生物矿化作用在大洋铁锰结核成矿过程中有巨大贡献,对大洋铁锰结核的生物成因过程提供更加全面准确的理解,从而为今后进一步充实大洋铁锰结核的生物矿化理论及其开发利用提供依据。     

A study of the microbial mineralization in submarine black smoker chimneys from the Okinawa Trough

Large amounts of microfossil records discovered in the seafloor black smoker chimney are reported from the Okinawa Trough. They are well preserved and can be divided into four types of filamentous microfossils. It suggests that the fossils may be derived from sulfur or iron oxidation chemolithotrophic prokaryotes and fungi. Based on the comparison studies of the microbial mineralization processes, two steps of biomineralization were hypothesized:(1) biology controlled mineralization; and (2) biology induced mineralization. At the early stage of the mineralization, the biology controlling mineralization is dominating; at the later stage, the biology inducing mineralization is the main mechanism. The composition of the fluids and the species of the microbes will determine the types of the minerals formed.     

冲绳海槽中、南部热液活动区金属硫化物的地球化学特征

对新近(2014年)采自冲绳海槽南部Yonaguni Knoll IV热液活动区和中部伊平屋热液活动区的金属硫化物样品分别进行了X射线荧光光谱(XRF)和等离子体质谱(ICP-MS)分析,旨在通过对比分析两个热液活动区金属硫化物的元素地球化学特征,探讨两个不同热液活动区成矿元素的富集规律、热液成矿的作用过程及其区域差异。研究结果表明:在冲绳海槽中部和南部热液区分布有两种金属硫化物,分别为富Fe-Zn型硫化物和Ba-Zn-Pb型硫化物(+硫酸盐);相较于大洋中脊和其它弧后盆地的热液硫化物,研究区硫化物中Pb的含量明显较高,而Cu和Fe的含量相对较低,这与冲绳海槽存在较厚的沉积盖层有关;无论是常量元素、微量元素、还是稀土元素(REE),其含量和相关的特征值都变化较大,表明研究区热液硫化物形成于不同的热液成矿阶段;富Fe-Zn型硫化物是在高温热液活动阶段形成的,而Ba-Zn-Pb型硫化物(+硫酸盐)则形成于热液喷出海底与海水混合的过程中,成矿温度较低;即使在同一热液活动区,成矿环境也有较大变化,主要表现在成矿温度的波动上;研究区热液硫化物中REE的配分模式特征表明硫化物中REE的物源相同,但经历的过程(REE的迁移和富集)有所不同,反映了热液硫化物形成于不同的热液成矿阶段;热液成矿作用存在区域性差异,海槽南部的成矿温度较中部低,成矿物质来源方面,南部热液区硫化物的Pb含量较中部高,表明其成矿物质可能部分来源于沉积物。     

Hydrothermal Metalliferous Sediments and Waters Off the Lesser Antilles

Considerable metal enrichments have been found in hydrothermal fluids and metalliferous sediments off the central Lesser Antilles Volcanic Arc. The elements variably enriched in the fluids as a result of the hydrothermal activity are Fe, Mn, As, Si, B, Li and in the sediments, Fe, P, Mo, As, Sb, Hg, Cu and Pb. Variations in the concentrations of these elements in both fluids and sediments along the arc result from a number of factors, the most important of which is the stage that each island's volcano has reached in its eruptive cycle. Although hydrothermal mineralization on the sea floor off the islands is only low grade, phase separation in the hydrothermal fluids at depth could be leading to higher grade stockwork mineralization below the vent fields or the discharge of metal rich brines on the lower flanks of the volcanic islands.     

Hydrothermal Metalliferous Sediments and Waters Off the Lesser Antilles

Abstract

Considerable metal enrichments have been found in hydrothermal fluids and metalliferous sediments off the central Lesser Antilles Volcanic Arc. The elements variably enriched in the fluids as a result of the hydrothermal activity are Fe, Mn, As, Si, B, Li and in the sediments, Fe, P, Mo, As, Sb, Hg, Cu and Pb. Variations in the concentrations of these elements in both fluids and sediments along the arc result from a number of factors, the most important of which is the stage that each island's volcano has reached in its eruptive cycle. Although hydrothermal mineralization on the sea floor off the islands is only low grade, phase separation in the hydrothermal fluids at depth could be leading to higher grade stockwork mineralization below the vent fields or the discharge of metal rich brines on the lower flanks of the volcanic islands.     

A new method of evaluating the mineralization of particulate and dissolved photoassimilated organic matter

A new method of evaluating the rate of mineralization of photoassimilated organic matter is described. This method enables us to compare the rate of direct mineralization of particulate organic carbon (POC) to CO₂ with the rate of solubilization of photoassimilated organic carbon followed by the mineralization of the resultant dissolved organic carbon (DOC) under the same conditions. The direct mineralization of photoassimilated carbon from POC to CO₂ is a more significant process compared with the mineralization of extracellular released organic carbon. The first-order rate coefficients range from 0.132 to 0.434 day^–1 for direct mineralization and 0.034 to 0.189 day^–1 for solubilization.     

The effect of salinity on the microbial mineralization of two polycyclic aromatic hydrocarbons in estuarine sediments

Sediments from the lower Hudson River estuary and two other coastal environments were examined experimentally for their ability to mineralize (convert to CO₂) the polycyclic aromatic hydrocarbons (PAHs) naphthalene and anthracene over a range of salinities. Routine assays employed 1:1 (vol fresh sed:vol water) sediment slurrys in order to overcome natural variability in mineralization rates among replicates. Mineralization rates were stimulated by about 2·5 fold, compared to unslurried controls, while the coefficient of variation fell from 13% to 3·5%.Rates of naphthalene mineralization in surface sediments from along the mainstem of the Hudson River (salinities from 2 to 27%) ranged from 0·011 to 1·5 nmol cm⁻³ day⁻¹ (pool turnover [T_n] from 60 to 2040 days) with no discernible trends along the estuarine gradient. For two stations examined experimentally (mile point 5, salinity 23%; mile point 26, salinity 5%), microbial assemblages appeared acclimated to broad salinity variations as alter rates of mineralization compared to controls.Sediments from two upstream marshes of the Hudson (mile points 36 and 45) showed rates of naphthalene mineralization from 0·007 to 0·15 nmol cm⁻³ day⁻¹ (T_n from 14 to 368 days), while sediments from a third marsh in freshwater (mile point 76) had high rates (66 nmol cm⁻³ day⁻¹; T_n 40 days). For the two upstream marsh stations which rarely experienced salt intrusion, there was a substantial decrease in mineralization of naphthalene and anthracene with increasing salinity.Consistently high rates of naphthalene mineralization (780 to 1600 nmol cm⁻³ day⁻¹; T_n 5 to 6 days) were observed in petroleum contaminated sediments from Port Jefferson Harbor (PJH) on the north shore of Long Island. PJH has a relatively constant salinity regime (about 27%) and imposed decreases in salinity effected decreases in rates of naphthalene and anthracene mineralization. Lowest rates of naphthalene mineralization (0·003 to 0·004 nmol cm⁻³ day⁻¹; T_n from 714 days to 833 days) were found in sediments from two stations in the relatively pristine Carmans River estuary on the south shore of Long Island.The ability of increases or decreases in salinity to affect the rate of model PAH mineralization appeared to be dependent on the natural variation in the salinity regime from which a sample was obtained. Data from all the environments studied indicated a strong positive correlation between PAH concentration and the rates of mineralization of naphthalene. Rates of PAH mineralization in all environments examined appear to be primarily controlled by the extent of pollutant loading and not by natural variations in the salinity regime.     

超慢速扩张环境下超镁铁质岩系统的热液硫化物成矿机理以及启示

西南印度洋63.5°E热液区是在超慢速扩张洋脊发现的首个超镁铁质岩热液系统。对取自该区的热液硫化物样品进行了系统的矿物学和地球化学分析,矿物学分析结果表明:该热液区硫化物为富Fe型高温硫化物,且经历了较深程度的氧化蚀变,大量中间态的Fe氧化物充填在硫化物矿物间的孔隙及内部解理中;这些硫化物相以白铁矿为主,其次是等轴古巴矿和少量铜蓝,缺乏黄铁矿、闪锌矿。据推断,该区的热液成矿作用分为4个阶段:低温白铁矿阶段→高温等轴古巴矿阶段→自形白铁矿阶段→后期海底风化阶段（少量铜蓝以及大量的Fe的羟氧化物）。与之相对应,地球化学分析结果表明这些硫化物的Fe含量较高（31.57%~44.59%）,Cu含量次之（0.16%~7.24%）,而Zn含量普遍较低（0.01%~0.11%）;微量元素较为富集Co（328×10-6~2 400×10-6）和Mn（48.5×10-6~1 730×10-6）。该区硫化物中较高含量的Fe、Co与超镁铁质岩热液系统相似,明显高于镁铁质岩热液系统。独特的热液硫化物矿物学特征和元素组成可能与该区普遍出露的地幔岩、橄榄岩蛇纹石化作用以及拆离断层的广泛发育的环境有关。     

Mafic-hosted seafloor sulfide mineralization at the margin of a non-transform discontinuity on the southern mid-Atlantic ridge

Abstract

Mineralogical, geochemical and S-isotopic studies were carried out on seafloor massive sulfides (SMS) and hydrothermally altered rocks from the mafic-hosted TaiJi field (including TaiJi-1 and TaiJi-2 sites), which is located on the margin of a nontransform discontinuity (NTD) on the Southern Mid-Atlantic Ridge (SMAR). The main hydrothermal precipitates of TaiJi-1 were massive sulfides, while TaiJi-2 produced a large amount of semi-massive sulfides. Significant rock alteration is an important feature of this field, and large amounts of clay minerals (chlorite) occurred in the semi-massive sulfides. Geochemically, notable negative correlations were identified between rock forming elements (Mg) and major hydrothermal metal elements such as Zn?+?Cu and Co in semisulfides. Such mineralogical and geochemical characteristics, together with low Co/Ni ratio (similar to the surrounding rocks) of the semi-massive sulfides and the unique REE features of some altered rocks, tell us that the prominent mineralization mechanism for these semi-massive sulfides was probably related to rock mineralization. Thus, TaiJi-2 appears to differ significantly from the TaiJi-1 in terms of sulfide mineralization mechanism. Moreover, the TaiJi sulfides are remarkable for large δ³⁴S variation (7.2–15.3%). We suggest that seawater corrosion after sulfides precipitation were responsible for ³⁴S enrichment in sulfides. Furthermore, our work sheds some light on the comparison between TaiJi and other NTD-related fields along the Mid-Atlantic Ridge. We argue that rock mineralization may represent an important mineralization type in NTDs.     

Biomineralization of organic matter in cobalt-rich crusts from the Marcus–Wake Seamounts of the western Pacific Ocean

Organic matter in cobalt-rich crust(CRC) from the Marcus–Wake Seamounts of the western Pacific Ocean, Sample CM1D03, has been analyzed to understand the source, geochemistry and mineralization of organic matter, and the mineralization environment. Biomarkers, including n-alkanes, isoprenoids, terpanes and sterols, have been detected in various layers of the CRC sample, using gas chromatography(GC) and gas chromatography–mass spectrometry(GC–MS). The content of organic carbon(OC) and its stable isotope(δ13C), and the combined features of the biomarkers show that the mineralized organic matter in CM1D03 CRC was mainly derived from microorganisms and lower plankton(e.g., bacteria and algae, respectively) from marine surface water, with some terrestrial higher plant components. The ratio of chloroform bitumen "A": OC was high in the CRC, between 10.51 and 20.66, showing significant migration characteristics of n-alkanes. Four mineralization categories of organic matter were recognized based on GC chromatograms of n-alkane molecules:(1) primitive type(bacteria and algae), which is characterized by moderately mature of n-alkanes preserving the original characteristics of the organic matter from microorganisms and lower plankton;(2) microbial degradation type, which is characterized by low contents of n-alkanes and rising baseline in the chromatogram, with the "bulge" being the products of organic matter by biodegradation;(3) organic matter migration type, which is characterized by low carbon number of n-alkanes with n C18 as the main peak carbon, without odd even predominance, and low concentrations of isoprenoids and hydrocarbons with high carbon number; and(4) organic matter hydrothermal type, which is characterized by relatively low concentration of small molecular weight n-alkanes, pristane, and phytane, accompanied by higher concentration of n-alkanes with carbon number greater than n C18. This study shows that biomarkers can record controlling factors of mineralization and their variation.     

The influence of sediment resuspension on the degradation of phenanthrene in flow-through microcosms

The effect of sediment resuspension on the mineralization of phenanthrene was examined in microcosms and sediment slurries. In computer-controlled, flow-through microcosms, 14C-phenanthrene-amended sediments were resuspended into overlying oxic water at frequencies of 12, 4, 1, 0.25 and 0 d(-1). In slurry bottle experiments 14C-phenanthrene-amended sediments were continuously resuspended under oxic (excess air headspace) and anoxic (N2 headspace) conditions and mineralization was measured at periods from 2 h to 7 days. Our main findings were: (1) mineralization rate constants from the microcosms ranged from 0.001 to 0.01 d(-1) and increased with frequency of resuspension, (2) these rates fell between those measured in oxic and anoxic slurries and were predicted within a factor of 2.5 by a model in which mineralization depended on the degree of oxygen exposure, and (3) the phenanthrene-degrading bacterial community was more active in resuspended sediments incubated in the microcosms than in sediments which were not resuspended, or which were stored under refrigeration. We conclude from these experiments that the effects of sediment resuspension on phenanthrene degradation are consistent with a primary role of average oxygen exposure, and also an alteration in the PAH-degrading activity of microbial populations.