DEOXYRIBONUCLEIC ACID CONTENT OF THE THREE CELL TYPES OF PHAEODACTYLUM TRICORNUTUM BOHLIN1

There is no significant difference in the DNA content of the oval, fusiform, and triradiate cell types of Phaeodactylum tricornutum, suggesting that they do not represent an alternation of haploid and diploid generations. The triradiate cells have a shorter generation time than the oval or fusiform cells.     

CALCIUM DEFICIENCY CAN INDUCE THE TRANSITION FROM OVAL TO FUSIFORM CELLS IN CULTURES OF PHAEODACTYLUM TRICORNUTUM BOHLIN1,2

The diatom Phaeodactylum tricornutum Bohlin is known to be pleomorphic. Previously it was considered that growth on medium solidified with agar produced ovate cells, whereas growth in liquid medium gave rise to fusiform cells. We have shown that ovate cells can be propagated in liquid medium if the calcium content of the medium is below 15 mg/liter. If the concentration of calcium is raised, fusiform cells develop.     

ULTRASTRUCTURE OF A CHAIN-FORMING DIATOM PHAEODACTYLUM TRICORNUTUM1

The ultrastructure of a chain-forming clone of the polymorphic diatom Phaeodactylum tricornutum Bohlin has been studied by scanning and transmission electron microscopy. Both fusiform and tri-radiate cells are capable of forming chains. The cells, lacking any silica shell, are attached to each other at the central region of the theca, leaving the arms free. Neither homogenization nor sonication completely disrupts the chains. The attachment is due to fusion of the cell wall in the central region of the cell during cell wall deposition. This fusion results from failure of the cytoplasmic cleavage furrow to separate the plasma membranes of the two daughter cells sufficiently so that a single wall is deposited instead of two separate walls. Possible explanations for this are discussed.     

THE METABOLISM OF GUANINE BY THE DIATOM PHAEODACYLUM TRICONUTUM BOHLIN1

Guanine taken up by intact cells of Phaeodactylum tricornutum Bohlin was rapidly converted to allantoin which accumulated in the cells; the earleir view that the compound which accumulated was a methylahypoxanthine is shown to be erroneous. In contrast, cells of P. tricornutum, after premeabilisation with toluene, converted guanine only to xanthine, the reaction presumably being catalysed by guanine diaminase. Freshly harvested N-replete cells contained substantial guanine deaminase activity (ca. 200 nmol (10⁸ cells h)^?1); this activity doubled during 5 hours of N-deprivation. During the same period, the ability to take up guanine, which was initially low, increased by about 25x.     

THE POLYMORPHIC DIATOM PHAEODACTYLUM TRICORNUTUM: ULTRASTRUCTURE OF ITS MORPHOTYPES1,2

The ultrastructure of the oval, fusiform and triradiate morphotypes of Phaeodactylum tricornutum Bohlin is described. The organization and structure of the cytoplasmic organelles is similar in all three morphotypes, except that the vacuoles occupy the extra volume created by the arms of the fusiform and triradiate cells. The frustule in fusiform and triradiate cells is organic; in the oval type it may be organic or one of the valves may have a silica frustule surrounded by an organic wall. In all cells, the organic cell wall has up to 10 silica bands (13 nm wide) embedded in its surface in the girdle region, lacks girdle bands, and has an outer corrugated cell wall layer, except in the girdle region. Cell division, organic wall formation and silica deposition are described in detail. Four types of oval cells are also described. The relation to other diatoms is discussed.     

FIRST INSIGHTS INTO IMPROVEMENT OF EICOSAPENTAENOIC ACID CONTENT IN PHAEODACTYLUM TRICORNUTUM (BACILLARIOPHYCEAE) BY INDUCED MUTAGENESIS1

A strain improvement program was initiated based on mutagenesis with the goal of commercial production of eicosapentaenoic acid (EPA)from EPA-overproducing microalgal strains. Two rounds of mutation and selection were conducted using Phaeodactylum tricornutum Bohlin UTEX #640 as the parent strain. After the first round of mutagenesis, a putative mutant (provisionally labeled 114) was obtained. The EPA content (% of dry weight) of this mutant strain was 37% higher than that of the wild type. 114 was further mutated and another putative mutant (provisionally called II242) was isolated, the EPA content of which was 44% higher than that of the wild type. When cultured with aeration in 1-L flasks, EPA content of the wild type and putative mutants 114 and II242 was, 17.3 mg · g^?1, 31.5mg · g^?1, and 38.6 mg · g^?1 dry biomass, respectively. EPA productivity was 3.48 mg · L^?1· d^?1 4.01 mg · L^?1· d^?1, and 4.98 mg · L^?1· d^?1 respectively. These figures compare favorably with many other promising EPA-producing microorganisms and suggest that the use of a single methodology such as mutation and selection is a way to improve the polyunsaturated fatty acid content of microalgae and other microorganisms.     

CHANGES IN CELL COMPOSITION AND LIPID METABOLISM MEDIATED BY SODIUM AND NITROGEN AVAILABILITY IN THE MARINE DIATOM PHAEODACTYLUM TRICORNUTUM (BACILLARIOPHYCEAE)1

The effects of nitrogen starvation in the presence or absence of sodium in the culture medium were monitored in batch cultures of the marine diatom Phaeodactylum tricornutum Bohlin. During nitrogen starvation in the presence of sodium, cell nitrogen and chlorophyll a decreased, mainly as a consequence of continued cell division. These decreases were accompanied by decreases in the rates of photosynthesis and respiration. There was no change in either cell volume or carbohydrate, but both carbon and lipid increased. During nitrogen starvation in the absence of sodium, cell division ceased. Cell nitrogen and chlorophyll a remained constant, and respiration did not decrease, but the changes in the photosynthetic rate and the lipid content per cell were similar to cultures that were nitrogen-starved in the presence of sodium. The carbon-to-nitrogen ratio increased in both cultures. Nitrogen, in the form of nitrate, and sodium were resupplied to cultures that had been preconditioned in nitrogen- and sodium-deficient medium for 5 d. Control cultures to which neither nitrate or sodium were added remained in a static state with respect to cell number, volume, and carbohydrate but showed slight increases in lipid. Cells in cultures to which 10 mM nitrate alone was added showed a similar response to cultures where no additions were made. Cells in cultures to which 50 mM sodium alone was added divided for 2 d, with concomitant small decreases in all measured constituents. Cell division resumed in cultures to which both sodium and nitrate were added. The lipid content fell dramatically in these cells and was correlated to metabolic oxidation via measured increases in the activity of the glyoxylate cycle enzyme, isocitrate lyase. We conclude that lipids are stored as a function of decreased growth rate and are metabolized to a small extent when cell division resumes. However, much higher rates of metabolism occur if cell division resumes in the presence of a nitrogen source.     

酵母全细胞苯丙氨酸鲜氨酶（PAL）活性的紫外分光光度测定法

本文报告以L-苯丙氨酸 (L-phe) 为底物,酵母全细胞作酶源,酶促生成产物反式-肉桂酸 (t-Ca)测定苯丙氨解氨酸 (PAP,EC_(4.3.1.5) 活性的紫外分光光度法。测定程序包括标准物质t-Ca的加样试验,绝对回收率试验,线性回归分析的整套定量分析研宄步骤,建立了一套经过修改的Kalghatgi和Subba Rao(1975) PAL 活性测定法。此法具有良好的准确度和精密度,已经用于评价具有PAL活性的酵母菌株在液体培养物中细胞生长和PAL活性形成的时间过程研究。     

CHANGES IN ABSORBANCE SPECTRUM OF THE DIATOM PHAEODACTYLUM TRICORNUTUM UPON MODIFICATION OF PROTEIN STRUCTURE1,2

Changes in the absorbance spectrum, of the diatom Phaeodactylum tricornutum which are brought about by heat, urea, stilt, and formaldehyde have been studied. The changes brought about by all agents are similar. Each converts cell suspensions from orange-brown to yellow-green. This color change results from an increase in absorbance in the blue and a decrease in absorbance in the green. All agents, except salt, also bring about an increase in absorbance in the red at wavelengths below that of the red peak of chlorophyll, and a decrease in absorbance at longer wavelengths. On heating cells, the absorbance change in the green occurs more rapidly than the changes in the blue and red. The converse is true when cells are treated with 4 M NaCl. Heat or formaldehyde treatments chemically modify chlorophyll pigments which are present in untreated cells. The change in absorbance in the green is probably due to changes in the molecular environment of fucoxanthin which, results from protein modification. The changes in the blue and red are at least partly due to chemical changes in chlorophylls, but may also be due, in part, to protein modification.     

DEMONSTRATION OF NET INFLUX OF FREE AMINO ACIDS IN PHAEODACTYLUM TRICORNUTUM USING HIGH PERFORMANCE LIQUID CHROMATOGRAPHY1

Influx of glycine from dilute solution in the medium into Phaeodactylum tricornutum (Bohlin) was studied using radiochemical techniques. Comparison of these data with simultaneous measurements of the disappearance of primary amines from the medium by fluorometry indicates that influx of ¹⁴C-labeled glycine accurately reflects net entry of substrate. High performance liquid chromatography (HPLC) was employed to demonstrate net removal of fourteen different amino acids from dilute solution by P. tricornutum. HPLC was also used to demonstrate net removal of free amino acids naturally present in sea water.     

剪切应力对毛细血管内皮细胞代谢的影响

建立的平行平板流动腔装置适用于研究血管内皮细胞代谢对剪切流场的响应。将培养的人胚肾小球血管单层内皮细胞置于剪应力分别为５×１０－５Ｎ／ｃｍ２,１×１０－４Ｎ／ｃｍ２和１．５×１０－４Ｎ／ｃｍ２的定常层流中剪切２５小时,样品中的内皮素分泌量用放射免疫法测定。结果表明,剪应力水平对内皮细胞内皮素的代谢活动有显著影响。与静态培养对照,低水平的剪应力（５×１０－５Ｎ／ｃｍ２、１×１０－４Ｎ／ｃｍ２）促进内皮素的分泌,而较高水平的剪应力（１．５×１０－４Ｎ／ｃｍ２）抑制内皮素的分泌;剪应力对内皮素累积含量的影响比之分泌速率更大     

THE REFRACTIVE INDICES OF WHOLE CELLS

Refractive indices of intact sporangiophores of Phycomyces were computed from measurements of focal length and radius of curvature of the cells. For the six cells studied, effective values of n were obtained ranging from 1.35 to 1.40. The average effective n was 1.38. Senn''s determination of refractive indices of other plants cells gave much higher values: n = 1.37 to 1.52. The precision of the method and possible sources of this discrepancy are discussed.     

CHRONIC EFFECTS OF ULTRAVIOLET-B RADIATION ON GROWTH AND CELL VOLUME OF PHAEODACTYLUM TRICORNUTUM (BACILLARIOPHYCEAE)1

Cultures of the temperate estuarine diatom, Phaeodactylum tricornutum Bohlin (NEPCC Clone 31), were grown under ambient intensities of ultraviolet-A radiation (UVAR), photosynthetically active radiation (PAR), and various intensities of ultraviolet-B radiation (UVBR; 290–320 nm). Growth rates and cell volumes were monitored for 36 d. UVBR decreased growth rates and increased cell volumes. Sensitivity of growth to UVBR increased with time. Growth rates of P. tricornutum decreased with increasing ratios of UVBR:UVAR + PAR.     

PHOTOSYNTHESIS IN THE DEEP BLUE SEA: PHOTOPHYSIOLOGICAL RESPONSES OF PHAEODACTYLUM TRICORNUTUM TO DYNAMIC AND STATIC IRRADIANCES

The kinetics of xanthophyll-cycle pigment switching and fluorescence quenching dynamics in the marine diatom, Phaeodactylum tricornutum were determined in the context of dynamic and static growth light. Cultures were grown in a modified photobioreactor capable of producing dynamic light fields which exhibited attenuation characteristics similar to that of water; these cultures were pre-acclimated to high and low, static and dynamic, growth-light regimes for at least three days, and then examined under high, static and dynamic light. Pigment pools varied markedly. The two static light cultures had pigment complements that were very similar to “traditional” high and low-light static cultures. The dynamic-light grown cultures had pigment complements, which were very similar to each other but different from the static-grown cultures. The maximum xanthophyll-cycle pigment de-epoxidation state attainable under saturating light was equal for all four treatments. Induction of fluorescence quenching was significantly faster in the static-grown cultures, while xanthophyll-cycle de-epoxidation rates did not show as much variation. Minimum irradiances for xanthophyll-cycle induction were correlated to average growth irradiance. Taken as a whole, the results from this work suggest that dynamic light-grown phytoplankton have a unique photosynthetic functionality that is different from static light-grown phytoplankton. The significance of these observations in the context of realistic light fields, and the photosynthetic response capabilities of algae grown under them will be discussed.