黄海沉积物多细胞趋磁原核生物的特性研究

多细胞趋磁原核生物（Multicellular magnetotactic prokaryotes，MMPs） 是一类由7～45 个含有磁小体的革兰氏阴性细胞聚集而成的球形或者椭圆形 的细胞聚集体，是研究生命起源与进化、细胞分化和生物矿化的模式生物， 目前仅在大西洋沿岸具有一定盐度的层化水体或沉积物中发现。 本文通过光学显微镜和电子显微镜研究了黄海沉积物MMPs 的超微结 构、运动特点和分裂方式等生物学特征，调查了MMPs 的生态分布特征，并 对其尝试培养。 根据形态差别，黄海沉积物的MMPs 可分为花瓣型MMPs（rosette-like MMPs）、菠萝型MMPs（pineapple-like MMPs）和松球型MMPs（pinecone-like MMPs）。花瓣型MMPs 是由23±4 个卵圆形的细胞螺旋形排列而成的球形聚 集体，直径为5.4±0.8 μm，鞭毛周生。细胞内外膜附近有子弹头形/和方形的 铁氧化物型磁小体。菠萝型MMPs 是由39±9 个方形细胞组成的大小为9.6±1.2 μm ×7.8±0.9 μm 的椭圆形聚集体，鞭毛周生。这类MMPs 由多环细胞组成的， 从椭圆体的赤道面向两极，细胞环的直径变小；在每一环内，细胞像书本似 并列相连；相邻两环的细胞为交错式相连。这种结构比花瓣型MMPs 的更为 紧密。菠萝型MMPs 的磁小体均为子弹头形铁氧化物，磁小体的排列与MMPs 的长轴近似平行。松球型MMPs 是由多个长条形的细胞围绕中心的一个凹陷 辐射排列而成的球形聚集体，直径在9.0～14.2 μm 之间。尼罗红和DAPI 染 色发现三种MMPs 均具有脂类颗粒，花瓣型MMPs 和菠萝型MMPs 在聚集体 的表面具有一层外膜，这说明MMPs 的细胞排列具有高度组织性，在一定程 度证明它属于多细胞生物。 花瓣型MMPs 和菠萝型MMPs 分裂时均保持多细胞形式，但花瓣型 MMPs 沿着聚集体的短轴分开，而菠萝型MMPs 沿着长轴分开。两种MMPs 具有MMPs 典型的逃逸运动，花瓣型MMPs 和菠萝型MMPs 的运动速度分别 为55±26 μm/s 和99±50 μm/s。 黄海花瓣型MMPs 的超微结构、运动方式和分裂特点与大西洋沿岸多个 地区发现的MMPs 相似，花瓣型MMPs 可能是MMPs 的优势类群。菠萝型 MMPs 从整体形态、细胞排列和分裂方式上与花瓣型MMPs 显著不同，是一 类新的MMPs。松球型MMPs 是一类尚未报道的MMPs。 对MMPs 的生态分布调查发现，花瓣型MMPs 广泛分布于砂质沉积物中， 最大丰度出现在氧化还原跃层（redoxcline）。菠萝型MMPs 多分布在砾石沉 积物的表层。两种MMPs 占据不同的生态位，暗示着两者可能具有不同的生 理代谢途径。 对MMPs 的培养发现，在实验室内MMPs 可存活8 个月，MMPs 丰度随 着时间变化出现周期性的变化，推测其繁殖周期可能是10～15 天。 本文为太平洋沿岸MMPs 的首次研究，支持MMPs 在全球广泛分布的观 点，并展示了MMPs 的形态多样性。

Differential selection on gene translation efficiency between the filamentous fungus Ashbya gossypii and yeasts

Background: The filamentous fungus Ashbya gossypii grows into a multicellular mycelium that is distinct from the unicellular morphology of its closely related yeast species. It has been proposed that genes important for cell cycle regulation play central

Fatty acid composition of some brown seaweeds (Phaeophyta) from the coast of Karachi

Ten species of filamentous and multicellular thalloid algae (Phaeophyta) belonging to six genera of the class Dictyophyceae were collected from different seawater habitats of Karachi, Pakistan at the northern boundary of the Arabian Sea during September 1997 and July 1998. They were extracted in chloroform: methanol, saponified, subjected to column chromatography (CC, TLC), esterified and analysed for fatty acid (FA) composition initially by gas-liquid-chromatography (GLC) and finally by gas chromatography-mass spectrometry (GC-MS). Algae of the classes Laminariophyceae and Fucophyceae (Phaeophyta) displayed only a few SCFAs, PUFAs and substituted FAs, no VLCFA, C22 UFA, CFA, DCFA and monoynoic FA, large amount of C16:0, very large quantity of C18:1, very small RCCL and FA-diversity, C18 UFAs up to four DBs, C20 UFAs up to three DBs only. They were characterized by the largest amount of C18:1, lowest degree of unsaturation of C20 UFAs, lack of C22 UFAs, the shortest RCCL and the smallest FA-diversity as compared to other phyla.

Fatty acid composition of more brown seaweeds (Phaeophyta) from the coast of Karachi

Six species of multicellular, thalloid, brown algae, belonging to five genera of the classes Laminariophyceae and Fucophyceae were collected from the coastal areas of Buleji near Karachi (Pakistan) during October 1997 and February 1998. They were extracted in chloroform: methanol, saponified, subjected to column chromatography (CC, TLC), esterified and analysed for fatty acid (FA) composition initially by gas-liquid-chromatography (GLC) and finally by gas chromatography-mass spectrometry (GC-MS). They displayed only a few SCFAs, PUFAs and substituted FAs, no VLCFA, C22 UFA, CFA, DCFA and monoynoic FA, large amount of CI6:0, very large quantity of C18:1, very small RCCL and FA-diversity, C18 UFAs up to four DBs, C20 UFAs up to three DBs only. They were characterized by the largest amount of C18:1, lowest degree of unsaturation of C20 UFAs, lack of C22 UFAs, the shortest RCCL and the smallest FA-diversity as compared to other phyla.

Characterizing the mechanics of cultured cell monolayers.

One-cell-thick monolayers are the simplest tissues in multicellular organisms, yet they fulfill critical roles in development and normal physiology. In early development, embryonic morphogenesis results largely from monolayer rearrangement and deformation due to internally generated forces. Later, monolayers act as physical barriers separating the internal environment from the exterior and must withstand externally applied forces. Though resisting and generating mechanical forces is an essential part of monolayer function, simple experimental methods to characterize monolayer mechanical properties are lacking. Here, we describe a system for tensile testing of freely suspended cultured monolayers that enables the examination of their mechanical behavior at multi-, uni-, and subcellular scales. Using this system, we provide measurements of monolayer elasticity and show that this is two orders of magnitude larger than the elasticity of their isolated cellular components. Monolayers could withstand more than a doubling in length before failing through rupture of intercellular junctions. Measurement of stress at fracture enabled a first estimation of the average force needed to separate cells within truly mature monolayers, approximately ninefold larger than measured in pairs of isolated cells. As in single cells, monolayer mechanical properties were strongly dependent on the integrity of the actin cytoskeleton, myosin, and intercellular adhesions interfacing adjacent cells. High magnification imaging revealed that keratin filaments became progressively stretched during extension, suggesting they participate in monolayer mechanics. This multiscale study of monolayer response to deformation enabled by our device provides the first quantitative investigation of the link between monolayer biology and mechanics.

Sound radiation from point-driven shell structures

A numerical study is presented showing the structural response and sound radiation from a range of thin shell structures excited by a point force: a baffled flat plate, a sphere, a family of spheroids and a family of closed circular cylinders. All the structures have the same material properties, thickness and total surface area so the asymptotic modal density is the same. Dramatic differences are shown in the total radiated sound power for the different shells. It was already known that the flat plate and the sphere behave very differently. These results show that the cylinders and, particularly, the spheroids show patterns that are not intermediate between the two but instead display new features: in certain frequency ranges the radiated sound power can be at least an order of magnitude greater than either the plate or the sphere. © 2013 Elsevier Ltd.

Margulis' theory on division of labour in cells revisited

Division of labour is a marked feature of multicellular organisms. Margulis proposed that the ancestors of metazoans had only one microtubule organizing center (MTOC), so they could not move and divide simultaneously. Selection for simultaneous movement and cell division had driven the division of labour between cells. However, no evidence or explanation for this assumption was provided. Why could the unicellular ancetors not have multiple MTOCs? The gain and loss of three possible strategies are discussed. It was found that the advantage of one or two MTOC per cell is environment-dependent. Unicellular organisms with only one MTOC per cell are favored only in resource-limited environments without strong predatory pressure. If division of labour occurring in a bicellular organism just makes simultaneous movement and cell division possible, the possibility of its fixation by natural selection is very low because a somatic cell performing the function of an MTOC is obviously wasting resources. Evolutionary biologists should search for other selective forces for division of labour in cells.

Primmorphs from archaeocytes-dominant cell population of the sponge Hymeniacidon perleve: Improved cell proliferation and spiculogenesis

Marine sponges (Porifera) possess an extraordinary diversity of bioactive metabolites for new drug discovery and development. In vitro cultivation of sponge cells in a bioreactor system is very attractive for the sustainable production of sponge-derived bioactive metabolites; however, it is still a challenging task. The recent establishment of sponge primmorphs, multicellular aggregates from dissociated mixed-cell population (MCP), has been widely acknowledged to hold great promise for cultivation in vitro. Here we present a new method to establish an in vitro sponge primmorph culture from archaeocyte-dominant cell population (ADCP) enriched by a Ficoll gradient, rather than a mixed-cell population (MCP). Our rationale is based upon the totipotency (the ability of a cell to differentiate into other cell types) of archaeocyte cells and the different biological functions of various sponge cell types. A sponge, Hymeniacidon perleve collected from the China Yellow Sea was used as a model system for this investigation. Distinct dynamics of primmorph formation were observed while significant increases in DNA synthesis, cell proliferation (up to threefold), and cell growth (up to fourfold) were achieved. Furthermore, a time-dependent spiculogenesis was clearly demonstrated in our longterm culture, indicating high metabolic activity of primmorphs from the ADCP. This new method represents an important step forward to advance sponge cell culture in vitro that may lead to commercial exploitation of sponge-derived drugs. (C) 2003 Wiley Periodicals, Inc.

The comparative estimation of phytoplanktonic, microphytobenthic and macrophytobenthic primary production in the oceans

The contributions of the planktonic unicellular algae [phytoplankton), the benthic unicellular algae [microphytobenthos) and the benthic multicellular algae (macrophytobenthos) to the primary production of the world ocean are evaluated, together with the respective limitations regarding data, concepts and methods. The use of “free-water” methods (e.g. in situ oxygen or CO2 budgets) is recommended in complement to the more specific measurements on enclosed organisms. For phytoplankton, a previous estimate of 30 . lo9 t C y-’ is retained as a minimal estimate. Earlier estimates of the world benthic production have been based on indirect calculations; revised estimates are suggested here which still lack precision but rely on the actual measurements available at present. Primary production of the micro- and macrobenthic algae amount to 50 and 375 g C m-? y-’ respectively as averages for the whole photic layer they can colonize, and total 2.9 . 10‘ t C y-’ for the world ocean. Thus, benthic algae contribute some 10% of the total marine primary production. On the continental shelf alone, the contributions of benthic and planktonib algae are commensurate and nearly equivalent.

Variations in morphology and PSII photosynthetic capabilities during the early development of tetraspores of Gracilaria vermiculophylla (Ohmi) Papenfuss (Gracilariales, Rhodophyta)

Background: Red algae are primitive photosynthetic eukaryotes, whose spores are ideal subjects for studies of photosynthesis and development. Although the development of red alga spores has received considerable research attention, few studies have focused on the detailed morphological and photosynthetic changes that occur during the early development of tetraspores of Gracilaria vermiculophylla (Ohmi) Papenfuss (Gracilariales, Rhodophyta). Herein, we documented these changes in this species of red algae. Results: In the tetraspores, we observed two types of division, cruciate and zonate, and both could develop into multicellular bodies (disks). During the first 84 hours, tetraspores divided several times, but the diameter of the disks changed very little; thereafter, the diameter increased significantly. Scanning electron microscopy observations and analysis of histological sections revealed that the natural shape of the disk remains tapered over time, and the erect frond grows from the central protrusion of the disk. Cultivation of tissue from excised disks demonstrated that the central protrusion of the disk is essential for initiation of the erect frond. Photosynthetic (i.e., PSII) activities were measured using chlorophyll fluorescence analysis. The results indicated that freshly released tetraspores retained limited PSII photosynthetic capabilities; when the tetraspores attached to a substrate, those capabilities increased significantly. In the disk, the PSII activity of both marginal and central cells was similar, although some degree of morphological polarity was present; the PSII photosynthetic capabilities in young germling exhibited an apico-basal gradient. Conclusions: Attachment of tetraspores to a substrate significantly enhanced their PSII photosynthetic capabilities, and triggered further development. The central protrusion of the disk is the growth point, may have transfer of nutritive material with the marginal cells. Within the young germling, the hetero-distribution of PSII photosynthetic capabilities might be due to the differences in cell functions.

Isolation and characterization of photosystem II of Porphyra yezoensis ueda

The thylakoid membranes were isolated and purified from gametophyte of Porphyrayezoensis Ueda (P yezoensis) by sucrose density gradient ultracentrifugation. After R yezoensis gametophyte thylakoid membranes were solubilized with SDS, the photosystem 11 (PSII) particles were isolated and purified. The activity of PSII particles was determined with DCIP (2,6-dichloroindophenol) photoreduction reaction. The composition of purified PSII particles was detected by SDS-PAGE. As a result, seven proteins including 55 kD protein, 47 kD protein, 43 kD protein, 33 kD protein, 31 kD protein, 29 kD protein, and 18 kD protein were found. Compared with PSII particles of higher plants and other algae, they were identified as D1/D2 complex, CP47, CP43, 33 kD protein, D1, D2 and cyt c-550 respectively. Besides, other three new proteins of 20 kD, 16 kD and 14 kD respectively were found. Among these extrinsic proteins, the 16 kD and 14 kD proteins had not been reported previously, and the 20 kD protein was found for the first time in multicellular red algae.

Study on early-stage development of conchospore in Porphyra yezoensis Ueda

Porphyra yezoensis Ueda (Rhodophyta) is a seaweed of economic importance with a typical dimorphic life cycle consisting of a leafy gametophyte and a filamentous sporophyte. Recently, it has been recognized as a model system for fundamental and applied studies in marine biological sciences. Conchospore, a major spore linking the two distinct multicellular phases in the life cycle, is most widely used in the breeding of P. yezoensis. In this paper, the early-stage development of conchospore, including the attachment and the cell wall formation, was studied with fluorescent reagents staining and Scanning Electron Microscopy detection. Results displayed: (I) the cell wall began to be generated after culturing for 4 h in the attached conchospores; (2) the initially released conchospores were plastids with some filmy, amorphous substance on the surface, and they attached to the fibers firmly via the actively secreted mucilaginous substances after their touch to the fibers; (3) cellulase and pectolase prohibited the attachment of conchospores in the different ways; and (4) only attached conchospores generated cell walls and developed normally, while the suspending ones could not. It indicated that the cellulose played crucial roles in the permanent attachment as the pectin did in the initial attachment. The conchospore attachment seemed to trigger the cell wall formation and the further development. Affects of light on the development of conchospores were also discussed. The results showed that high intensity (200 mu mol.m(-2).s(-1)) and long-wave (>= 580 nm) light facilitated the division rate of conchospores. (C) 2008 Elsevier B.V. All rights reserved.

Molecular cloning and expression of a novel Kazal-type serine proteinase inhibitor gene from Zhikong scallop Chlamys farreri, and the inhibitory activity of its recombinant domain

Serine proteinase inhibitors (SPIs) play important roles in host physiological and immunological processes in all multicellular organisms. A novel Kazal-type SPI gene was cloned from the Zhikong scallop Chlamys farreri (designated as CfKZSPI) by expressed sequence tag (EST) and rapid amplification of cDNA ends (RACE) approaches. The full-length cDNA of CfKZSPI was of 1788 nucleotides with a canonical polyadenylation signal sequence AATAAA and a polyA tail, and an open reading frame (ORF) encoding a polypeptide of 509 amino acids with a putative signal peptide of 22 amino acids. The deduced amino acid sequence of CfKZSPI contained 12 tandem Kazal domains with high similarity to other Kazal-type SPIs. The temporal expression of CfKZSPI in hemocytes after Vibrio anguillorum challenge was recorded by quantitative real-time RT-PCR. The relative mRNA expression level of CfKZSPI was up-regulated and reached 43.6-fold at 3 h post-challenge. After a decrease at 6 h, the expression Level increased again and reached 207.8-fold at 12 h post-challenge. The 12th Kazal domain of CfKZSPI was recombined into pET-32a(+) and expressed in Escherichia coli Rosetta-gami (DE3) to investigate its inhibitory activity. The purified recombinant protein (rCf KZSPI-1 2) showed significant inhibitory activity against trypsin but no activity against thrombin. When the molar ratio of inhibitor to trypsin reached 1:1, almost 90% of the enzyme activity could be inhibited, which suggested that one molecule of rCfKZSPI-12 was able to inhibit one molecule of trypsin. Kinetics analysis with Dixon plot showed that the inhibition constant (K-i) of rCfKZSPI-12 to trypsin was 173 nmol L-1. These results indicated that CfKZSPI was a novel Kazal-type SPI with significant inhibitory activity against trypsin, and was suspected to be involved in scallop immune response. (c) 2008 Elsevier Ltd. All rights reserved.

Early development of germlings of Sargassum thunbergii (Fucales, Phaeophyta) under laboratory conditions

Morphology and culture studies on germlings of Sargassum thunbergii (Mertens et Roth) Kuntze were carried out under controlled laboratory conditions. Growth characteristics of these germlings grown under different temperatures (from 10 to 25A degrees C), irradiances (from 9 to 88 mu mol photons m(-2) s(-1)), and under blue and white light conditions are described. The development of embryonic germlings follows the classic "8 nuclei 1 egg" type described for Sargassaceae. Fertilized eggs spent 5-6 h developing into multicellular germlings with abundant rhizoids after fertilization. Under conditions of 20A degrees C, 44 mu mol photons m(-2) s(-1) and photoperiod of 12 h, young germlings with one or two leaflets reached 2-3 mm in length after 8 weeks. Temperature variations (10, 15, 20, 25A degrees C) under 88 mu mol photons m(-2) s(-1) significantly influenced the growth rate within the first week, although this effect became less obvious after 8 weeks, especially at 15 and 20A degrees C. Variation in germling growth was highly significant under different irradiances (9, 18, 44, 88 mu mol photons m(-2) s(-1)) at 25A degrees C. Low temperature (10A degrees C) reduced germling growth. Growth of germlings cultured under blue light was lower than in white light. Optimal growth of these germlings occurred at 25A degrees C and 44 mu mol photons m(-2) s(-1).

Molecular cloning and responsive expression to injury stimulus of a defender against cell death 1 (DAD1) gene from bay scallops Argopecten irradians

Apoptosis is an active process of cell death, which is an integral part of growth and development in multicellular organisms. The defender against cell death 1 (DAD1), the regulatory protein to inhibit the apoptosis process, was first cloned from the bay scallop Argopecten irradians by randomly sequencing a whole tissue cDNA library and rapid amplification of cDNA end (RACE). The full-length cDNA of the A. irradians DAD1 was 607 bp, consist of a 5'-terminal untranslated region (UTR) of 63 bp, a 3'-terminal UTR of 205 bp with a canonical polyadenylation signal sequence AATAAA and a poly (A) tail, and an open reading frame of 339 bp. The deduced amino acid sequence of the A. irradians DAD1 showed 75.5% identity to Araneus ventricosus, 74.5% to Drosophila melanogaster, and 73.6% to Homo sapiens, Sus scrofa, Mesocricetus auratus, Rattus norvegicus and Mus musculus. Excluding the Saccharomyces cerevisiae DAD1 homologue, all animal DAD1 including A. irradians DAD1 homologue formed a subgroup and all plant DAD1 proteins formed another subgroup in the phylogenetic analysis. The A. irradians DAD1 was expressed in all examined tissues including adductor muscle, mantle, gills, digestive gland, gonad and hemolymph, suggesting that A. irradians DAD1 is expressed in most body tissues. Furthermore, the mRNA expression levels of A. irradians DAD1 gene of hemolymph were particularly high after injury, suggesting that the gene is responsive to injury stimuli.