Ultrastructure of Single Cells, Callus-like and Monospore-like Cells in Porphyra yezoensis Ueda on Semi-solid Culture Medium

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Ultrastructure of Single Cells ,Callus—like and Monos—pore—like Cells in Porphyra yezoensis Ueda on Semi—slid Culture Medium

It had been demonstrated that individual or protoplasts isolated from Porphyra thallus by enzyme could develop into normal leafy thalli in the same way as monosproes,and that isolated cells develop in different way in liquid and on semi-solid media,The authors observed the ultrastructure of isolated vegetative cells cultured on semi-solid media and compared them with those of monospores and isolated cells cultured in liquid media,The results showed that subcellular struce-tures were quite different among cells in different conditions.In their development,isolated cells on semi-solid media did not show the characteristic subcellular eature of monospore formation ,such as production of fibrous vesicles,Callus-like cells formed on semi-solid media underwent a distinctive modification in cellular organization.They developed characteristic cell inclusions and a special 2-layer cell covering,Golgi bodies,ER ,starch grains,mitochondria,Vaculoes were not commonly found in them.     

THE ULTRASTRUCTURE OF SEPARATED AND CULTURED CELL OF PORPHYRA YEZOENSIS

There are many reports that cells (protoplasts) separated from the thallus of Porphyra by enzyme can develop to normal leafy thalli in the same way as monopores. But there are few investigations on the subcellular structure of the isolated vegetative cell for ccmparison with the subcellular structure of monospores. To clarify whether the separated and cultured cells undergo the same or similar ultrastructure changes during culture and germination as monospores undergo in their formation and germination, we observed their ultrastructure, ccxnpared them with those of the monospore and found that the ultrastructure of separated and cultured cells did not have the characteristic feature as that of monospore formarion, such as production of small and large fibrous vesicles, but was acccxnpanied by vacuolation andstarch mobilization like that in monospore germination. The paper also discusses the relations between monospores and separated and cultured cells.     

The ultrastructure of separated and cultured cell of Porphyra yezoensis

There are many reports that cells (protoplasts) separated from the thallus ofPorphyra by enzyme can develop to normal leafy thalli in the same way as monospores. But there are few investigations on the subcellular structure of the isolated vegetative cell for comparison with the subcellular structure of monospores. To clarify whether the separated and cultured cells undergo the same or similar ultrastructure changes during culture and germination as monospores undergo in their formation and germination, we observed their ultrastructure, compared them with those of the monospore and found that the ultrastructure of separated and cultured cells did not have the characteristic feature as that of monospore formation, such as production of small and large fibrous vesicles, but was accompanied by vacuolation and starch mobilization like that in monospore germination. The paper also discusses the relations between monospores and separated and cultured cells. Contribution No. 3875 from the Institute of Oceanology, Chinese Academy of Sciences. Project 39770593 supported by NSFC; Project 96-C01-05-01 of the 9th Five-Year Plan supported by Science and Technology Commision of China.     

Study on monospore-like cells fromPorphyra haitanensis

Porphyra haitanensis andP. yezoensis are two mainPorphyra species cultured in China. Their life histories are slightly different. So far we have not observed thatP. haitanensis naturally produces monospores developing into thalli.P. yezoensis produces monospores which directly germinate into young thalli used in cultivation (Zeng, et al., 1985). Some somatic cells inP. yezoensis develop in vitro into monospore-like cells which later grow into young thalli (Lu, 1983). Studies on whether or not somatic cells inP. haitanensis can produce in vitro monospore-like cells that later grow into young thalli is important for understanding its life history and for culturing new varieties.     

The influence of Fe^2＋ on growth and development of cells enzymatically isolated from Porphyra yezoensis blades

Fe^2＋ acted as an accessorial factor for many cellular enzymatic reactions is very important for seaweed growth and development, but the Fe^2＋ requirement in nori had not been seen. Porphyra yezoensis cells were separated enzymatically and cultured in a series of sterilized seawater media containing various concentrations of Fe^2＋. The growth development and cell were investigated in this work. Through this experiment, two biologically-meant concentration scales were found, one is low concentrations, 12.1-102.1μg/L, 10-100 times than that in seawater, favoring the development of isolated cells of Porphyra and the other was high concentrations, more than 10mg/L inhibiting the cell growth, leading to the deformity and shrinkage of the cells. At the concentration of 50 mg/L, the cells stopped growing and died eventually.     

米曲霉两步活化法对黄芩苷的生物转化及转化产物的纯化与鉴定

利用米曲霉把黄芩苷转化为黄芩素。转化条件为:液体发酵,pH6,30℃,利用综合马铃薯培养基与玉米芯培养基两步发酵。菌种采用两步活化法,将经综合马铃薯培养基培养48 h的米曲霉加入至玉米芯培养基培养48 h后,再加入底物黄芩苷继续培养3 d。玉米芯发酵液加甲醇混匀后过滤,通过高效液相色谱检测,结果表明米曲霉已经将黄芩苷转化为黄芩素,转化率为40.72%。同时用乙醚萃取法从玉米芯发酵液中萃取到黄色固体提取物,经过高效液相色谱与标准品对照鉴定,该提取物为黄芩素。     

Effects of Basic Fibroblast Growth Factor and Insulin-like Growth Factor on Cultured Cartilage Cells from Skate Raja porasa

Effects of basic fibroblast growth factor (bFGF) and insulin-like growth factor Ⅱ(IGF-Ⅱ) on cartilage cells from proboscis of skate, Raja porasa Glinther, were investigated in this study. The cartilage cells were cultured in 20% FBS-supplemented MEM medium at 24℃. Twelve hours after culture initiation, the cartilage cells were treated with bFGF and IGF-Ⅱ at different concentration combinations. It was found that 20 ng/ml of bFGF or 80 ng/ml of IGF-Ⅱ was enough to have obvious stimulating effect on the growth and division of skate cartilage cells. Test of bFGF and IGF-Ⅱ together, revealed that 20 ng/ml of bFGF and 80 ng/ml of IGF-Ⅱ together had the best stimulating effect on the growth and division of skate cartilage cells. The cartilage cells cultured could form a monolayer at day 7.     

INDUCTION OF ECTODERMAL CELLS FROM VEGETAL- ENDODERMAL BLASTOMERES OF AMPHIOXUS(BRANCHIOSTOMA BELCHERI TSINGTAUNESE) EMBRYOS BY THE CALCIUM IONOPHORE A23187

INTRODUCnONAInPhioxusl6-celleInbryosconsistoftwotiersofblastomeres,eighboInalblas-tomeres,A8,andeightvegetalblastomeres,V8(Fig.l).Tungetal.(l958)shOWedthaV8isolatedatthel6-cellstage,whenculndexplan,gaVeriseInainlytointestineandoccasionalltonotochoal,muscleandneUraltube,butneverformedepidends-bendng-ly,thedeveloPmentalfateofV8blastomeresap~tobedetendnedatthel6-cellStag.HOWeVer,thesamauthors(l961)shoWedIaterthatifvegetal-mostblastomersatthe32-cellstage,thedescendansofV8,wereisolateda…     

Enhancing the culturability of bacteria from the gastrointestinal tract of farmed adult turbot Scophthalmus maximus

Eighteen agar media were tested for the culture of gut-associated bacteria from farmed adult turbot(Scophthalmus maximus),including 16 agar media with or without 1% gastrointestinal(GI)supernatant,or with 2% or 4% GI supernatant.A total of 1 711 colonies were analyzed and 24 operational taxonomic units(OTUs)were identifi ed.The greatest bacterial diversity was isolated on Zobell 2216E/Zobell 2216E+ agar media,whereas MRS/MRS+ agar media produced a low diversity of colonies.Agar media with GI supernatant(1%,2%,or 4%)showed increased diversity and yielded different profi les of OTUs from the corresponding original media,suggesting that GI supernatant provides substances that enhance the culture effi ciency of bacteria from the turbot GI tract.The large majority of the colonies(82%)were γ-Proteobacteria,whereas 15.6% and 2.4% of colonies were Firmicutes and Actinobacteria,respectively.At the genus level,49.4% of all colonies were assigned to Vibrio.Other potential pathogens,including Pseudomonas,Photobacterium,and Enterobacter,and potential probiotics,including Bacillus,Paenibacillus,and Pseudomonas,were also isolated on agar media.Most cultured bacteria belonged to species that were fi rst described in the turbot GI tract.The impact of these species on turbot physiology and health should be investigated further.     

勒氏笛鲷外周血细胞显微结构的观察

报道了勒氏笛鲷外周血细胞的显微结构,血涂片经过Wright染色液染色,可区分出红血细胞、血栓细胞、嗜中性粒细胞、单核细胞、淋巴细胞等五种血细胞,没有发现嗜碱性粒细胞、嗜酸性粒细胞.白血细胞中,血栓细胞体积最小,单核细胞体积最大;血栓细胞数目最多,淋巴细胞和单核细胞数目最少;淋巴细胞有大小两种类型大淋巴细胞和小淋巴细胞.此外,在外周血液中还观察到少量未成熟的红细胞和嗜中性粒细胞,偶而可见正在分裂的红细胞,提示红细胞也可在外周血直接分裂.     

3D numerical simulation in acoustic-elastic coupled media with staggered-grid finite-difference method

Acoustic-elastic coupled media is often encountered in most marine explorations, and accurate simulation of acoustic-elastic coupled media is of great significance. At present, the study of acoustic-elastic coupled media still assumes that the solid of the acoustic-elastic coupled media is isotropic, but this assumption is not in accordance with the actual situation. In this paper, we derive the solid media of acoustic-elastic coupled media from isotropic media to anisotropic media, and propose an acoustic-elastic coupled medium based ontransverse isotropic media with vertical symmetric axes(VTI) to improve the accuracy of forward modeling. Based on the relationship between the Thomsen parameter and the coefficient matrix of the anisotropic elastic wave equation, we transform the Thomson parameter into a velocity model with anisotropic properties. We use a staggered grid finite difference method to simulate the propagation of a wavefield in a three-dimensional acoustic-elastic coupled media. We obtain the snapshots of the wave field when the solid of the acoustic-elastic coupled media is an isotropic medium and a VTI media. When the solid of the acoustic-elastic coupled media is considered VTI media, we can observe the qP wave and qS wave that cannot be observed in the isotropic medium from the wave field snapshot. We can also find that the seismic records obtained by the method we use are more realistic. The algorithm proposed in this paper is of great significance for high-precision ocean numerical simulation.     

Induction of ectodermal cells from vegetal-endodermal blastomeres of amphioxus (Branchiostoma belcheri tsingtaunese) embryos by the calcium ionophore A23187

Timing of vegetal-endodermal cell determination in amphioxus embryos remains uncertain. We tentatively tested effects of A23187, the calcium ionophore, on the development of vegetal blastomeres isolated at the 16-cell stage. It was found that when vegetal blastomeres committed to endoderm were treated with A23187 prior to gastrulation, they were transformed into ectodermal cells as evidenced by the cell morphology and function characteristic of epidermis. However, the developmental fate of the same blastomeres untreated or treated with DMSO at the same stage or of those treated with A23187 after gastrulation remained unchanged. Thus, vegetal-endodermal cells in amphioxus embryos are not irreversibly determined before the gastrula stage, and artificial increase in intracelluar Ca²⁺ concentration can induce transdetermination of the predetermined endodermal cells into ectodermal cells. Project 39470091 supported by NSFC and partly supported by Shandong Natural Science Foundation, Grant No.93D0140.     

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山地气候梯变效应与特色气候资源的开发利用

从空气中的氧、二氧化碳、水汽、固体粒子含量以及大气压力、接受的太阳辐射、光照时数、所获得热量、降水量等因素介绍了山地气候梯变效应的具体表现,分析了山地气候梯度效应对生态环境及人类活动的影响。从山地气候梯变效应的实际状况出发,通过分析山地气候资源的特点,指出人类应遵循生态系统、自然保护、景观和谐、因地制宜等原则,挖掘山地气候旅游资源、多样化气候资源和气候"小生境"等气候资源,突显其特色,发展异于平地的农业生产、旅游活动等,将有助于山地特色气候资源的充分利用。     

LIGHT AND ELECTRON MICROSCOPE OBSERVATIONS ON NEPHROSELMIS GAOAE SP.NOV.(PRASINOPHYCEAE)

Nephroselmis gaoae sp.nov.is described on the basis of light and electron microscope observations ofcultured material originally collected and isolated from seawater of Jiaozhou Bay,Qingdao,China.Theperiplasts on the cell body and flagella are covered by five types of scales,two types on the flagella andthree on the body.Among these,the morphology and the number of spines of large stellate body scalesdiffer remarkably from those of previously described species of Nephroselmis.Apart from these,the unusualfine structure of the eyespot(stigma)is very characteristic.As in the other species of Nephroselmis,theeyespot lies immediately under the two-membraned chloroplast envelope;unlike the others,however,it isnot composed of a number of osmiophilic globules,but consists of about 14 curved rod-shapedosmiophilic bodies arranged loosely and randomly.This feature distinguishes the present new species notonly from the other species of Nephroselmis but also from the other motile algal species,the eyespots struc-ture     

Cultural characteristics of chromium resistant unicellular cyanobacteria isolated from local environment in Pakistan

Many unicellular cyanobacteria were isolated from different places： fields, ponds, polluted water, and soils from Muredkey and Kasur tannery areas, near Lahore, Pakistan. Different media like BG 11 medium, Bold Basal medium, Chu＇s # 10 medium and Gotham＇s medium, in standard forms and with slight variations of ingredients, and different pH, temperature and light regimes were checked for the optimum growth of the isolates. The isolation pro- cedure was repeated with different concentrations of chromium to select the resistant strains. These selected strains grew on chromium of the range 100-200 μg/ml in BG 11 medium. Cyanobacteria were maintained in solid and liquid media with/without shaking. Cyanobacterial strains were collected from natural habitats that were accompanied by a diversified group of organisms including bacteria, protozoan, and rotifers etc. In order to eliminate these agents termed as contaminants, we used several methods including phenol treatment, use of antibiotic and careful manual picking of unicellular cyanobacteria. Resistance of these strains against different heavy metals （ZnSO4, MnSO4, NiSO4, COCl2, Pb（NO3）3, CuSO4, HgCl2, AgNO3 and CdCl2） and antibiotics （erythromycin, streptomycin, kanamycin, chloramphenicol, neomycin） was evaluated. Optimum temperature was 30℃ with variable pH for the reduction of Cr^6＋ in to Cr^3＋ in majority of strains.     

STUDY ON THE RELATIONSHIP BETWEEN SPECIATION OF HEAVY METALS AND THEIR ECOTOXICITY——I. TOXICITY OF Cu, Cd , Pb AND Zn IN SEAWATER TO THREE MARINE ALGAE IN THE PRESENCE OF DIFFERENT COMPLEXATION AGENTS

Heavy metal is a main pollutant in the marine ecosystem . so study on the effect of heavy metal on phytoplankton is important . Algae (Chaetoceros sp . , Dunaliella sp . ., Dicrateria zhanjiangenis Hu . var . sp .) were laboratory cultured to observe the effect of heavy metals on their growth . The effect of different metal ion concentration , the detoxication effect of complexation agents and the growth of algae in different media and different nutrition levels were studied to evaluate the effect of metal speciation . It is proved that trace amount of heavy metals can stimulate the growth of algae cells but that high concentration is lethal . The sequence of toxicity is Cd2+>Zn2+>Pb2+ . In ordinary nutrition conditions , the detoxication sequence of complexation agents to Chaetoceros sp . is EDTA > sodium salicylate > sodium oxalate > sodium citrate > sulfanilic acid > O-phenanthroline . This is in good conformity with the stability constant sequence of these agents with copper and good evidence that t     

Genetic transformation of <Emphasis Type="Italic">Nannochloropsis oculata</Emphasis> with a bacterial phleomycin resistance gene as dominant selective marker

The gene ble from Streptoalloteichus hindustanus is widely used as a selective antibiotic marker. It can control the phleomycin resistance, and significantly increase the tolerance of hosts to zeocin. The unicellular marine microalga Nannochloropsis oculata is extremely sensitive to zeocin. We selected ble as the selective marker for the genetic transformation of N. oculata. After the algal cells at a density of 2×10⁷ cells mL^?1 was digested with 4% hemicellulase and 2% driselase for 1 h, the protoplasts accounted for 90% of the total. The ble was placed at the downstream of promoter HSP70A-RUBS2 isolated from Chlamydomonas reinhardtii, yielding a recombinant expression construct pMS188. The construct was transferred into the protoplasts through electroporation (1 kV, 15 μS). The transformed protoplasts were cultured in fresh f/2 liquid medium, and selected on solid f/2 medium supplemented with 500 ng mL^?1 zeocin. The PCR result proved that ble existed in the transformants. Three transformants had been cultured for at least 5 generations without losing ble. Southern blotting analysis showed that the ble has been integrated into the genome of N. oculata. The ble will serve as a new dominant selective marker in genetic engineering N. oculata.