形成条斑紫菜离体组织再生苗的几种方式

紫菜属的不少种类的叶状体通过形成单孢子直接产生叶状体苗,也可以以其他方式直接成苗、一些报道表明,由紫菜叶状体营养细胞直接成苗在不少种类中存在。尤其是离体营养组织块,与离体细胞或原生质体相比,容易获得而且出苗量大,在生产上有潜在的利用价值。作者以条斑紫菜为材料开展的研究取得了以下结果。１材料与方法样品１～６号为中国科学院实验海洋生物学开放研究室海藻种质种苗组保种并在１９９７年９月育成的６个品系的苗网,在大连旅星海珍品公司海区养成后,１１月３０日自栽培网上采下,７号样品取自江苏海头养殖海区紫菜网。８…     

坛紫菜体细胞的几种发育途径

以坛紫菜(Porphyra haitaneusis)叶状体为材料,利用单细胞克隆技术,研究记录坛紫菜体细胞的发育途径。本文选择介绍了坛紫菜体细胞3种途径8种发育方式:1)多数体细胞的叶状体途径:A通过极性分裂直接形成单细胞苗;B通过叶状分裂成苗;C先形成细胞团后形成畸形苗;2)来自雌、雄叶片未成熟区域体细胞的性细胞途径:D发育形成精子囊并放散精子;E单细胞发育形成类似果孢子囊结构再放散果孢子,果孢子萌发成丝状体;3)来自雄叶片体细胞的丝状体途径:F单细胞发育至多细胞团,放散后萌发成丝状体;G经过不断分裂形成愈伤组织,直接形成丝状体;H分裂成2细胞后,一个细胞侧面突起发育成丝状体。     

温度对皱紫菜(Pyropia crispata)壳孢子萌发及叶状体形态建成的影响

近几年,紫菜"高温烂菜"现象频发,为开发耐高温紫菜栽培种质,本文研究了4种温度(15、20、25、28℃)对皱紫菜壳孢子萌发及叶状体形态建成的影响,并观察了藻体的繁殖特性。结果显示,在试验设定的培养条件下,适宜皱紫菜壳孢子萌发生长温度为20—25℃,最适温度为20℃。皱紫菜壳孢子幼苗萌发初期细胞分裂为横分裂,叶状体细胞呈直线排列,生长至7—10个细胞后,细胞开始出现纵分裂。15、20℃温度组藻体多为不规则的半圆形裂片。25、28℃温度组藻体多为细长型叶片。有性生殖是皱紫菜主要的繁殖方式,但15—20℃培养组的部分藻体能够少量形成并放散类似无性生殖的单孢子并萌发形成叶状体。     

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.     

条斑紫菜单细胞固体培养的初步研究

成熟条斑紫菜叶状体酶解后获得单细胞,分别培养于添加了不同浓度N,P营养盐的0．5％和10％的琼胶培养基上,观察细胞的分化发育情况。结果显示：(1)营养细胞大部分先发育为细胞团,细胞团放散孢子,再由孢子萌发形成小苗。其余营养细胞极性分裂形成正常细胞苗或带假根的形状不规则的片状体。孢子茁和营养细胞直接形成的幼苗生长到一定大小均可放散单孢子,单孢予能够正常萌发形成单孢子苗。(2)在0．5％的琼胶培养基上或较高浓度的营养盐中,细胞发育快。添加5mg／L NO3-N和0．5mg／L H2PO4—P营养盐不能满足细胞生长发育的需要,培育20d绝大部分仍为单细胞,不能形成正常幼苗。     

不同培养条件对坛紫菜叶状体体细胞发育分化影响的研究

本文以野生坛紫菜(Porphyra hatanensis)叶状体为材料,通过海螺酶酶解获得叶状体单离体细胞,探讨了细胞密度、光照强度对坛紫菜叶状体体细胞发育分化的影响.研究结果表明:细胞密度对坛紫菜叶状体单离体细胞的分裂、出苗具有明显的影响.其中,细胞培养3～5d时,分裂率随着细胞密度的增大而降低;5d时,1×103个/孔组的分裂速度显著大于其余各组的(p＜0.05);12～19d时,随着细胞密度的增大,体细胞出苗率相应减小.光照强度对坛紫菜叶状体单离体细胞的分裂、出苗也具有显著影响.细胞培养5d时,分裂率随着光照强度的增大而增大;5-7d时,体细胞分裂速度随着光照强度增大而增大;9～19d培养期内,出苗率随着光照强度的增大而增大.本实验结果为坛紫菜转基因育种等研究提供了可资借鉴的基础资料.     

坛紫菜耐高温品系选育及经济性状的初步研究

将人工杂交选育的坛紫菜新品系Z-26,Z-61,Z-81,Z-17的子3代(F₃)与野生型对照组进行遗传性状的比较,旨在筛选耐高温品系。实验结果表明:(1)在高温29℃下培养40 d,人工选育的4个新品系丝状体均能正常生长发育,其中Z-26丝状体生长较快,Z-61在29℃下较易成熟,95%的营养藻丝发育成孢子囊枝,Z-17只有30%形成孢子囊枝;(2)Z-26和Z-61品系的叶状体在30℃下培养10 d,虽然藻体变白,但没有死亡,此时将其移至21℃下恢复培养3~4 d,藻体逐渐恢复为原有的色泽;(3)Z-81和Z-17的叶状体在29℃下培养7~10 d,虽然藻体发白但尚未死亡,移至21℃下培养基本可恢复正常,但在30℃下培养10 d,藻体腐烂不可恢复;(4)野生型对照组不耐高温,藻体在27℃下培养5 d就腐烂死亡,人工选育的4个品系耐受高温的幅度比野生型对照组高1~3℃;(5)4个品系叶状体在26~28℃温度下培养10 d后,其主要的藻胆蛋白和叶绿素a的含量与21℃培养下的含量差异不显著,说明短时间的高温培养并不影响其品质。     

60Co-γ射线辐照坛紫菜叶状体及单克隆培养的研究

本文报道了坛紫菜叶状体经60Coγ射线辐照后,用酶解方法获得离体细胞,观察了体细胞的分裂分化及进行单克隆培养叶状体.结果显示:体细胞发育和分化的趋势分为正常叶状体、畸形叶状体、细胞团、丝状体和根丝细胞5种方式;通过单克隆的培养,初步筛选出1个抗逆性较强、生长速度较快的新品系D,该品系1m以上长度的叶状体平均生长速度为9.4cm/d,快的可达11.5cm/d.对新品系的子一代叶状体再度酶解和观察,获得纯系的丝状体.     

条斑紫菜冷冻前不同处理对细胞活性的影响

将新鲜幼嫩条斑紫菜叶状体在不同温度和通风状态下,干燥处 理后冻存于-20℃冰箱中。30d后酶解成单细胞,检查其活性和再生能力。结果表明,条斑紫 菜含水量为81.8%,在含水35%时冻藏可保持最大的活性。适宜的含水量范围是30%～40%。冷 冻前含水量高于40%时,复苏后细胞外观虽然正常,但酶解后存活率低,发育迟缓;冷冻前 含水量低于30%时,复苏后在叶状体上可见成片细胞死亡,色素弥散,形成红色斑块。酶解 后细胞死亡率高。阴干处理期间,温度的变化(10℃～25℃)对冷冻后的紫菜细胞活力没有明 显影响。     

坛紫菜叶状体营养细胞与生殖细胞叶绿素荧光特性比较

利用叶绿素荧光技术对坛紫菜(Porphyra haitanensis)叶状体营养细胞和生殖细胞的光能利用特性进行了比较。结果表明:坛紫菜叶状体营养细胞和雌性生殖细胞的实际量子效率(ΔF/Fm′)差异不明显,但显著高于雄性生殖细胞;快速光曲线测定表明雌、雄藻体营养细胞的最大相对电子传递速率(rETRmax)相近,但显著高于雄性生殖细胞;不同生殖细胞半饱和光强(Ik)无显著差异;在生长光强下,营养细胞和雌性生殖细胞rETR、光化学猝灭(qP)和非光化学猝灭(NPQ)差异不明显,而雄性生殖细胞rETR、qP等荧光参数均显著低于营养细胞和雌性生殖细胞。本文表明坛紫菜叶状体营养细胞和雌性生殖细胞具有较高的光能利用能力,能够将吸收的光能多数用于电子传递,而雄性生殖细胞对光能的利用能力较低。     

Microbial Biomass and Organic Nutrients in the Deep-Sea Sediments of the Central Indian Ocean Basin

In order to assess the impact of deep-sea mining on the in situ benthic life, we measured the microbial standing stock and concentration of organic nutrients in the deep-sea sediments of the Central Indian Ocean Basin in the Indian pioneer area. Sediments were collected using box core and grab samples during September 1996. The total bacterial numbers ranged from 10 10 -10 11 cells per g -1 dry weight sediment. There was a marginal decrease in the number of bacteria from surface to 30 cm depth, though the subsurface section registered a higher number than did the surface. The highest numbers were encountered at depths of 4-8 cm. The retrievable number of bacteria were two orders less in comparison with the direct total counts of bacteria. An almost homogeneous distribution of bacteria, total organic carbon, living biomass, and lipids throughout the depth of cores indicates active microbial and benthic processes in the deep sea sediments. On the other hand, a uniform distribution of total counts of bacteria, carbohydrates, and total organic carbon in all the cores indicates their stable nature and suggests that they can serve as useful parameters for long-term monitoring of the area after the benthic disturbance. Further studies on temporal variability in this region would not only verify the observed norms of distribution of these variables but would also help to understand restabilization processes after the simulated benthic disturbance.     

Assessing the Validity of Seismo-Acoustic Predictor Equations for Obtaining Seabed and Sub-Surface Sediment Physical Properties

The interdependence between the seismo-acoustic properties of a marine sediment and its geotechnical/physical parameters has been known for many years, and it has been postulated that this should allow the extraction of geotechnical information from seismic data. Though in the literature many correlations have been published for the surficial layer, there is a lack of information for greater sediment depths. In this article, a desktop study on a synthetic seafloor model illustrates how the application of published near-surface prediction equations to subsurface sediments (up to several tens of meters burial depth) can lead to spurious predictions. To test this further, acoustic and geotechnical properties were measured on a number of sediment core samples, some of which were subjected to loading in acoustically-equipped consolidation cells (oedometers) to simulate greater burial depth conditions. For low effective pressures (representing small burial depths extending to around 10 meters subsurface), the general applicability of established relationships was confirmed: the prediction of porosity, bulk density, and mean grain size from acoustic velocity and impedance appears generally possible for the investigated sedimentary environments. As effective pressure increases through, the observed relationships deviate more and more from the established ones for the near-surface area. For the samples tested in this study, in some instances increasing pressure even resulted in decreasing velocities. There are several possible explanations for this abnormal behavior, including the presence of gas, overconsolidation, or bimodal grain size distribution. The results indicate that an appropriate depth correction must be introduced into the published prediction equations in order to obtain reliable estimates of physical sediment properties for greater subsurface depths.     

Geoacoustic Inversion via Genetic Algorithm and Its Application to Manganese Sediment Identification

An acoustic inversion method using a wide-band signal and two near field receivers is proposed and applied to multiple layered seabed models including a manganese sediment. The inversion problem can be formulated into a probabilistic model comprised of signals, a forward model, and additive noise. The forward model simulates wide-band signals, such as chirp signals, and is chosen to be the source-waveletconvolution plane wave modeling method. The wavelet matching technique, using weighted least-squares fitting, estimates the sediment sound-speed and thickness on which determination of the possible numerical ranges for a priori uniform distribution is based. The genetic algorithm is applied to a global optimization problem to find a maximum a posteriori solution for determined a priori search space. Here the object function is defined by an L 2 norm of the difference between measured and modeled signals. Not only the marginal pdf but also its statistics are calculated by numerical evaluation of integrals using the samples selected during importance sampling process of the genetic algorithm.     

海上大直径钢管桩打桩过程中桩周土强度弱化研究

海上大直径钢管桩打桩过程中,桩周土体受到强烈扰动而发生强度弱化,掌握桩周土体强度弱化规律对于准确预测打桩过程、保证工程安全具有重要意义。为研究土体强度弱化规律,开展了环剪试验模拟打桩对桩周土体的扰动,测试土体强度随剪切速率的变化规律,建立了描述土体强度弱化规律的拟合公式,引入到打桩分析软件中。研究结果表明：土体的强度折减程度不仅与土体本身的性质有关还受到土体的埋深和剪切速率的影响,埋深越深土体强度折减程度越低,剪切速率越高土体强度折减越高,在打桩分析中可采用这里推荐的线性折减方法来模拟不同深度处土体强度的折减规律。     

Interannual Sea Level Variations and Annual Tides in the Northwestern Pacific

Specific properties of the interannual sea level variations and annual tides in the Northwestern Pacific were studied. Several tide stations were monitored. The monthly mean sea level for the year of 1995 was analyzed at each tide station. A seismic event in 1995, some tectonic activity around the subject area, and the Kuroshio (the oceanic western boundary current) may possibly contaminate results which would have occurred from the astronomical annual tide alone.     

Flow Cytometric Analysis of Coastal Lagoon Bacterioplankton and Picophytoplankton: Fixation and Storage Effects

The effect of different fixation and storage protocols on the flow cytometric (FCM) simultaneous analysis of bacterioplankton and phytoplankton in coastal seawater samples (Mediterranean coastal lagoons) was investigated. FCM measurements (cell number, fluorescence and scatter characteristics) were obtained through DAPI staining. Three fixatives [glutaraldehyde (GA), formaldehyde (FA) and paraformaldehyde (PFA)] and two storage (3 months duration) methods (5 °C and −196 °C) were tested. Two dominant populations were detected in studied samples: bacteria and eukaryotic picophytoplankton. Adding fixatives (2% final concentration) appears necessary to obtain FCM exhaustive counts of all the bacteria and phytoplanktonic cells. This was related to the permeation effect of fixatives which allowed a better DAPI staining of the cells. Maximum fluorescence, i.e. optimal staining of the cells was obtained with FA or PFA, and significant lower fluorescences with GA. Fixed samples stored at 5 °C induced rapid cell loss. Only storage in liquid nitrogen of samples fixed with FA or PFA, allows mid-term (≥4 months) preservation of bacteria or picophytoplankton cell numbers, and limited evolution of DAPI-induced fluorescence and scatter characteristics.     

INFORMATION FOR AUTHORS

<正>Acta Oceanologica Sinica(AOS)is a comprehensive academic journal edited by the Editorial Committee of Acta Oceanologica Sinica and is designed to provide a forum for important research papers of the marine scientific community which reflect the information on a worldwide basis.The journal publishes scholarly papers on marine science and technology,including physics,chemistry,biology,     

Deep-Sea Sediment Compression Curves: Some Controlling Factors, Spurious Overconsolidation, Predictions, and Geophysical Reproduction

Oedometer tests have been carried out on 70 undisturbed surficial clays (at approximately 250 mm below the mudline), mostly collected by free-fall corers from sites widely scattered throughout the deep-sea North Atlantic. Acoustic measurements were also made, initially on contiguous samples and ultimately on the same sample using a geophysically instrumented oedometer which also collected electrical resistivity data. Apart from those quiescent areas below the carbonate compensation depth, such as north of the West Indies where very fine clays exist, most of the samples are silty clays whose geotechnical-geophysical properties are dependent on the type of clay minerals present (and their ability to take in moisture), the sand-size fraction, and the quantity of carbonate present. Thus the pure clays have high compressibilities which decrease on the addition of coarse particles, while the converse is true for the acoustic parameters, these increasing with the sand fraction. Using the notion of the intrinsic compression line for all samples, and comparison to it of the measured compression curves, it is clear that, contrary to some previously held ideas, most deep-sea clays are normally consolidated; the addition of carbonate has the effect of creating an open, stronger sediment skeleton. Interestingly, where information is available, the variation with depth of a sample's acoustic velocity follows the void ratio pressure relationship of the compression curve. This allows the construction of an in-situ sediment compression curve using the in-situ geophysical observations.