EXCRETION OF GLYCOLATE BY A SPECIES OF CHLORELLA (CHLOROPHYCEAE)1

The claim that Chlorella sp. (CCAP 211/8p), sometimes referred to as C. fusca, Shihira and Krauss, does not excrete glycolate has been reexamined. Chlorella sp. grown on 5% CO₂in air, excreted glycolate when incubated in light in 10 mM bicarbonate. Excretion ceased 30–60 min after transfer of the cells to air and no excretion could be detected with air-grown cells or with cells grown on 5% CO₂in media buffered at pH 8.0. Incubation with 10 mM isonicotinyl hydrazide, a glycolate pathway inhibitor, caused excretion in air-grown cells and stimulated excretion in CO₂-grown cells indicating that both the rate of glycolate synthesis and metabolism is higher in CO₂grown cells than in air-grown cells. Enhanced glycolate synthesis and excretion in CO₂-grown cells is correlated with law photosynthetic rate in 10 mM bicarbonate, and the photosynthetic rate of these cells doubles over a period of 2–2.5 h after initial transfer from high CO₂to bicarbonate. This correlation of photosynthetic induction with cessation of glycolate excretion is similar to that reported in a bluegreen alga and thought to occur in other green algae. These results indicate that glycolate excretion and its regulation in this species of Chlorella is not different from that in other algae.     

STIMULATION OF GROWTH IN SCENEDESMUS OBLIQUUS (CHLOROPHYCEAE) BY HUMIC ACIDS UNDER IRON LIMITED CONDITIONS1,2

Stimulatory affects of humic acids of molecular weight 30,000 or greater on iron-starved Scenedesmus obliquus (Turp.) Kütz. in association with bacteria were studied by growth and Fe uptake experiments. Humic acids stimulated growth of Fe-starved cells by decreasing the lag phase and extending the growth phase. Humic acids stimulated increased algal growth in medium containing EDTA as well as in medium containing no EDTA, indicating humic acids are not stimulating algal growth under Fe limiting conditions by creating a soluble Fe pool. Humic acids decreased Fe availability to Fe-starved S. obliquus. Iron bound to humic acids is unavailable for uptake by Fe-starved cells indicating growth stimulation is not due to chelation effects alone. Stimulation of growth is not a membrane phenomenon as humic acids show the same stimulatory effect when in contact with cells or separated by dialysis membrane. Humic acids also stimulate growth in the dark, with and without aeration, indicating use as a heterotrophic substrate. A photoheterotrophic mechanism is indicated by increased algal growth caused by illuminating cultures, containing humic acids but excluding CO₂.     

STEROL,FATTY ACID,AND PIGMENT CHARACTERISTICS OF UTEX 2341, A MARINE EUSTIGMATOPHYTE IDENTIFIED PREVIOUSLY AS CHLORELLA MINUTISSIMA (CHLOROPHYCEAE)1

An examination of the sterols of UTEX 2341, a small (ca. 2μm), nonmotile unicellular marine alga identified as Chlorella minutissima Fott et Novakova, yielded results inconsistent with any of 35 Chlorella strains analyzed previously. UTEX 2341 contained cholesterol as the principal sterol, with 24-methylenecholesterol, fucosterol, and isofucosterol also present; these are not dominant sterols in any other Chlorella species. Presence of eicosapentaenoic acid in UTEX 2341 also contrasted with fatty acids of Chlorella strains analyzed previously. Pigment analysis of UTEX 2341 revealed that it contained chlorophyll a, but not chlorophylls b or c; violaxanthin was the only major xanthophyll pigment. Both lipid and pigment compositions suggest that UTEX 2341 is not a member of the genus Chlorella but, rather, belongs in the Eustigmatophyceae; it may be Nannochloropsis sp. Cells with possible extracellular structures were present at an appreciable percentage of the stationary-phase population studied; centrifuging removed or collapsed these structures. The high cholesterol and polyunsaturated fatty acid contents of UTEX 2341 make it attractive as a potential aquaculturefeed, provided it is, or can be made, digestible.     

THE UPTAKE OF MANNITOL AND SORBITOL BY A SPECIES OF CHLORELLA (CHLOROPHYCEAE)1

Chlorella saccharophila (Krüger) Nadson takes up mannitol and sorbitol in the light and the dark. The rate of uptake is concentration dependent. is not affected by pH in the range pH 6.0 to 8.0 and ii not stimulated by light. Uptake is inhibited by the respiration inhibitor sodium azide (10^-2 M) but not by 3-(3,4-dichlorophenyl)-1,1-di-methyl urea (10^-6 M), an inhibitor of photosynthesis. Sorbitol. but not mannitol, stimulates the rate of dark respiration but both support the heterotrophic growth of the alga. Both compounds permeate the cells of C. miniata. and two strains of C. pyrenoidosa but do not support the heterotrophic growth of these algae. The cells of C. vulgaris are impermeable to both compounds.     

EFFECTS OF LOWERING TEMPERATURE DURING CULTURE ON THE PRODUCTION OF POLYUNSATURATED FATTY ACIDS IN THE MARINE DIATOM PHAEODACTYLUM TRICORNUTUM (BACILLARIOPHYCEAE)1

The composition of fatty acids and contents of eicosapentaenoic acid (EPA) and polyunsaturated fatty acids (PUFAs) of the economically important marine diatom, Phaeodactylum tricornutum (Bohlin), were investigated to see whether reducing the culture temperature enhances the production of EPA and PUFAs. The contents of EPA and PUFAs of P. tricornutum were found to be higher at lower temperature when cultured at 10, 15, 20, or 25°C. When the cells grown at 25°C were shifted to 20, 15, or 10°C, the contents per dry mass of PUFAs and EPA increased to the maximal values in 48, 24, and 12 h, respectively. The highest yields of PUFAs and EPA per unit dry mass (per unit volume of culture) were 4.9% and 2.6% (12.4 and 6.6 mg·L^?1), respectively, when temperature was shifted from 25 to 10°C for 12 h, both being raised by 120% compared with the control. The representative fatty acids in the total fatty acids, when temperature was lowered from 25 to 10°C, decreased proportionally by about 30% in C_16:0 and 20% in C_16:1(n?7) but increased about 85% in EPA. It was concluded that lowering culture temperature of P. tricornutum could significantly raise the yields of EPA and PUFAs.     

INHIBITION OF GROWTH OF ULVA EXPANSA (CHLOROPHYTA) BY ULTRAVIOLET-B RADIATION1

We examined the effect of ultraviolet-B radiation (UV-B, 290–320 nm) on the growth rate of the intertidal marine alga Ulva expansa (Setch.) S. & G. (Chlorophyta). Segments of thallus collected from a natural population were grown in outdoor seawater tanks. Combinations of UV-B-opaque screens, UV-B-transparent screens, and UV-B lamps were used to investigate the effects of solar UV-B and solar plus supplemental UV-B on the growth of these segments. Growth was measured by changes in segment surface area, damp weight, and dry weight. Growth rates of segments were inhibited under both solar UV-B and solar plus supplemental UV-B treatments. Growth rates were also inhibited by high levels of photosynthetically active radiation, independent of UV-B fluence. These results indicate that increases in UV-B resulting from further ozone depletion will have a negative impact on the growth of this alga.     

CO2 ACCUMULATION BY CHLORELLA SACCHAROPHILA (CHLOROPHYCEAE) AT LOW EXTERNAL pH: EVIDENCE FOR ACTIVE TRANSPORT OF INORGANIC CARBON AT THE CHLOROPLAST ENVELOPE1

The CO₂ uptake characteristics of the acid-tolerant alga Chlorella saccharophila (Krüger) Nadson have been examined. These, together with direct estimations of internal inorganic C concentrations, indicate that air-adapted cells of this alga possess a CO₂ accumulation mechanism similar to that reported in other members of the Chlorophyceae and in Cyanophyceae. This accumulation mechanism operates at external pH values of 4 and less. Under these conditions CO₂ must be the carbon species crossing the plasmlemma, and since preliminary evidence suggests that the transport mechanism is active primary uniport of HCO₃^–, it is proposed that the site of the CO₂ accumulation mechanism is the chloroplast envelope.     

珍稀水生动物白暨豚油中脂肪酸的GC／MS分析

白暨豚是一种珍稀水生动物.本文报道了运用 Dexsil-300GC 毛细管柱色质联用仪,对白(?)豚油中脂肪酸进行了分析鉴定,并检测到了22种脂肪酸,其中主要脂肪酸是十四碳烯酸、十六碳烯酸和十八碳烯酸.     

MODULATION OF FATTY ACIDS IN THE MEMBRANES OF ANACYSTIS NIDULANS (CYANOBACTERIA): INCORPORATION OF ODD-NUMBERED CARBON FATTY ACIDS1

Anacystis nidulans Richt., a unicellular cyanobacterium, can incorporate exogenously supplied fatty acids, including odd-numbered carbon fatty acid (OFAs), into the acylglycerols of cell membranes. Data are presented for the uptake of undecanoic acid (11:0) into cells of A. nidulans, the subsequent elongation up to C17, and incorporation of OFA into the four major membrane acylglycerols. The incorporation of OFAs was followed by desaturation of part of the saturated fatty acid to monoenoic fatty acid. Positional analyses of the double bonds of these manoenoic fatty acids suggest that there is one desaturase that inserts a Δ⁹ bond in both odd- and even-numbered fatty acids of varying chain length. Our data also suggest that there is no positional specificity for chain length on the glycerol backbone by the acyltransferases.     

ISOLATION AND CHARACTERIZATION OF CHLORELLA SOROKINIANA GXNN01 (CHLOROPHYTA) WITH THE PROPERTIES OF HETEROTROPHIC AND MICROAEROBIC GROWTH1

Heterotrophic and anaerobic microalgae are of significance in both basic research and industrial application. A microalga strain was isolated from a wastewater treatment pond and identified as Chlorella sorokiniana Shihira et W. R. Krauss GXNN01 in terms of morphology, physiology, and phylogeny. The strain grows rapidly in heterotrophic or mixotrophic conditions with addition of various carbon sources, and even in anaerobic conditions. The maximum growth rate reached 0.28 d^?1 when using d,l ‐malate as the carbon source, and the protein content of the microalgae was 75.32% in cell dry weight. The strain was shown to be capable of (1) utilizing d,l ‐malate only with light, (2) inhibiting photosynthesis in mixotrophic growth, and (3) growing in anaerobic conditions with regular photosynthesis and producing oxygen internally. This study demonstrates the influence of oxygen (aerobic vs. anaerobic) and metabolic regime (autotrophy, mixotrophy, heterotrophy) on the physiological state of the cell.     

EXOGENOUS STEROIDS AND GROWTH OF NEOSPONGIOCOCCUM SP. (CHLOROPHYTA)1

Of the various types of steroids found in nature, only sterols (steroids whose molecules possess an 8- to 10-carbon atom hydrocarbon side chain at position 17 of the perhydrocyclopentanophenanthrene ring) are known to be common constituents of algae. Little is known of the effects of steroids in the environment upon the growth and survival of algae. This paper investigates the growth of the green alga Neospongiococcum sp. in medium containing steroids. Bile acids are not inhibitory, even at a concentration of 100 ppm. Some sterols inhibit growth when present in the medium at a concentration of 100 ppm, but not at 10 ppm. Testosterone and β-estradiol, which have no carbon atom side chain at ring position 17, inhibit growth at a concentration of only 10 ppm. Steroids whose molecules possess a 2-carbon atom side chain at ring position 17 and a keto group at the α-carbon of this side chain, such as pregnenolone, inhibit growth at a concentration of as little as 1 ppm. Respiration is also inhibited by pregnenolone at this level .     

麦红吸浆虫幼虫脂肪酸变化的分析测定(英文)

采用气相色谱法测定了不同时期麦红吸浆虫Sitodiplosismosellana (Gehin)幼虫脂肪酸的变化。结果表明 :幼虫脂肪酸组分有 1 0～ 1 6种 ,其中主要以棕榈酸 (C1 6:0 )、油酸 (C1 8:1 )和亚油酸 (C1 8:2 ) 3种组分为主 ,占全部脂肪酸总量的 95 %以上 ;其次是硬脂酸 (C1 8:0 )含量较高 ,约占 2 %～ 3 .5 % ;其它组分脂肪酸含量很少 ,均在 1 %以下。从 5月中下旬麦红吸浆虫幼虫离穗入土进入越夏至翌年 4月化蛹羽化前 ,脂肪酸组分表现了由少到多的变化趋势。越夏、越冬幼虫及越冬后未进入活动状态的圆茧其脂肪酸组分中不含花生酸 (C2 0 :0 ) ,而且饱和脂肪酸含量明显少于越冬后进入活动状态的幼虫和麦穗幼虫 ;而越冬后进入活动状态的幼虫和麦穗幼虫脂肪酸组分中含有一定量的花生酸 ,不饱和脂肪酸含量明显少于越夏、越冬幼虫及越冬后未进入活动状态的圆茧 ,故花生酸的有无及饱和脂肪酸与不饱和脂肪酸含量的比例变化可作为判断麦红吸浆虫幼虫活动状态 ,特别是滞育不同深度的生化指标之一     

MOBILIZATION OF β-1,3-GLUCAN AND BIOSYNTHESIS OF AMINO ACIDS INDUCED BY NH4+ ADDITION TO N-LIMITED CELLS OF THE MARINE DIATOM SKELETONEMA COSTATUM (BACILLARIOPHYCEAE)

Mobilization of the reserve β-1,3-glucan (chrysolaminaran) in N-limited cells of the marine diatom Skeletonema costatum (Grev.) Cleve (Bacillariophyceae) was investigated. The diatom was grown in pH-regulated batch cultures with a 14:10-h light:dark cycle until N depletion. In a pulse-chase experiment, the cells were first incubated in high light (200 μmol photons·m ^{− 2}·s ^{− 1}) with ¹⁴C-bicarbonate until dissolved inorganic carbon was exhausted. Unlabeled bicarbonate (1 mM) was then added, and the cells were incubated in the dark and subsequently in low light (20 μmol photons·m ^{− 2}·s ^{− 1}) with additions of 40 μM NH₄ ⁺ . In the ¹⁴C pulse phase with high light and N depletion, β-1,3-glucan accumulated and accounted for 85% of incorporated ¹⁴C. In the subsequent ¹⁴C chase phases, added NH₄ ⁺ was assimilated at an N-specific rate of 0.11 h ^{− 1} in both the dark and low light, and in both cases it caused a significant mobilization of β-1,3-glucan (dark, 26%; low light, 19%). Biochemical fractionation of organic ¹⁴C showed that free amino acids were most rapidly labeled in the early stage of NH₄ ⁺ assimilation, whereas proteins and polysaccharides were labeled more rapidly after 1.2 h. Analysis of the cellular free amino acids strongly indicated that de novo biosynthesis was occurring, with a Gln:Glu ratio increasing from 0.4 to 10 within 1.2 h. After the NH₄ ⁺ was exhausted, the cellular pools of glucan and amino acids became constant or slowly decreased. In another experiment, N-limited cells were first incubated in high light until dissolved inorganic carbon was exhausted and were further incubated in high light with 150 μM NH₄ ⁺ under inorganic carbon limitation. Added NH₄ ⁺ was assimilated at an N-specific rate of 0.023 h ^{− 1}, and cellular β-1,3-glucan decreased by 15% within 6 h. Hence, β-1,3-glucan was mobilized during NH₄ ⁺ assimilation, even though inorganic carbon was modifying the metabolic rates. The results provide new evidence of β-1,3-glucan supplying essential precursors for biosynthesis of amino acids and other components in S. costatum in both the dark and subsaturating light and even saturating light under inorganic carbon limitation.     

CHANGES OF FATTY ACIDS IN LARVAE OF THE WHEAT BLOSSOM MIDGE,SITODIPLOSIS MOSELLANA (GEHIN) (DIPTERA: CECIDOMYIIDAE)*

Abstract The changes of fatty acids in larvae of the wheat blossom midge, Sitodiplosis mosellana (Gehin) at different periods were examined by gas choromatography. There were 10–16 kinds of fatty acids, of which the predominant ingredients were palmitic (C_16:0), oleic (C_18:1) and linoleic (C_18:2) acids which were more than 95% in total fatty acids, stearic acid (C_18:0) about 2%‐3.5% and any of the others was less than 1%. The fatty acid compositions increased from mid‐May, when larvae of the wheat blossom midge left the wheat‐ears and fallen on the ground, to April of next year before pupating and emerging. No arachidic acid (C_20.0) was discovered in over‐summering, over‐wintering as well as inactive over‐wintered larvae. The content of saturated fatty acids in over‐summering, overwintering as well as inactive over‐wintered larvae were less than those of in active over‐wintered larvae and wheat‐ear larvae. Therefore, changes of the arachidic acid and the proportions of saturated fatty acids/unsaturated fatty acids could be used as one of the biochemical criteria to determine the active state and the degree of diapause in larvae of the wheat blossom midge.     

INFLUENCE OF GROWTH STATUS AND NUTRIENTS ON EXTRACELLULAR POLYSACCHARIDE SYNTHESIS BY THE SOIL ALGA CHLAMYDOMONAS MEXICANA (CHLOROPHYCEAE)1

Increased levels of nitrogen in liquid growth medium bring about increased growth and a delay in extracellular polysaccharide production by Chlamydomonas mexicana Lewin on a per-cell basis. Addition of nitrogen to stationary phase cultures causes renewed growth and a temporary lag in polysaccharide synthesis until growth again ceases. Removal of nitrogen terminates growth, causing an immediate increase in polysaccharide synthesis. Phosphate-starved cells show a response similar to nitrogen-starved cells, indicating that the beginning of stationary phase and not nitrogen depletion causes the stimulation in extracellular polysaccharide synthesis. As similar results are assumed to occur on soil, the significance of this response is discussed.     

MODULATION OF VERY-LONG CHAIN (C28) HIGHLY UNSATURATED FATTY ACIDS IN PROROCENTRUM MININUM (DINOPHYCEAE) BY SELENIUM

Sargassum is one of the most species-rich genera in the brown algae with over 400 described species worldwide. The bulk of these species occurs in Pacific-Indian ocean waters with only a small portion found on the Atlantic side of the Isthmus of Panama. Sargassum also has one of the most subdivided and complex taxonomic systems used within the algae. Systematic distinctions within the genus are further complicated by high rates of phenotypic variability in several key morphological characters. Molecular analyses in such systems should allow testing of systematic concepts while providing insights into speciation and evolutionary patterns. Global molecular phylogenetic analyses using both conserved and variable regions of the Rubisco operon ( rbc L and rbc L-IGS-rbcS) were performed with species from the Gulf of Mexico, Caribbean, and Pacific basin. Results confirm earlier analyses based on rbc L-IGS- rbc S from Pacific species at the subgeneric and sectional level while providing additional insights into the systematics and phylogenetics on a global scale. For example, species east of the Isthmus of Panama form a distinct well-resolved clade within the tropical subgenus. This result in sharp contrast to traditional systematic treatments but provides a window into the evolutionary history of this genus in the Pacific and Atlantic Ocean basins and a possible means to time speciation events.     

DIVERSITY IN THE MECHANISM OF CARBON DIOXIDE FIXATION IN DUNALIELLA TERTIOLECTA (CHLOROPHYCEAE)1

The mechanism of photosynthetic carbon dioxide fixation in the green flagellate Dunaliella tertiolecta Butcher varies during growth in batch culture. Evidence for this change comes from three sources: i) algae from the stationary phase incorporated a greater proportion of the fixed carbon into amino arids and protein than did cells from the mid-exponential phase; ii) the activity of phosphoenolpyruvate carboxylase relative to that of ribulose-1, 5-di-phosphate carboxylase increased with age in batch culture; and, iii) cells from the stationary phase appeared to utilize the bicarbonate ion as the substrate for photosynthesis, whereas those from mid-exponential phase appeared to utilize fire carbon dioxide. These data suggest that a change of photosynthetic mechanism can occur within a single species of alga, depending on its physiological state.     

INVOLVEMENT OF INDOLE‐3‐ACETIC ACID PRODUCED BY THE GROWTH‐PROMOTING BACTERIUM AZOSPIRILLUM SPP. IN PROMOTING GROWTH OF CHLORELLA VULGARIS1

Involvement of indole‐3‐acetic acid (IAA), produced by the microalgae‐growth‐promoting bacteria Azospirillum brasilens and A. lipoferum, in promoting growth of the microalga Chlorella vulgaris Beij. was studied. Four wildtype strains of Azospirillum and their IAA‐deficient mutants were co‐immobilized with C. vulgaris in alginate beads. Cultures were grown in synthetic growth medium supplemented with tryptophan. Growth promotion of microalgae and production of exogenous IAA by Azospirillum spp. were monitored. All wildtype Azospirillum spp. produced significant but varying amounts of IAA, while their mutant forms produced significantly less. The results demonstrated a significant growth promotion in Chlorella cultures when immobilized with the four wildtype strains of Azospirillum, while very low or no enhanced growth was induced by the four IAA‐deficient mutants, compared to when C. vulgaris is immobilized alone. A complementation experiment, where an IAA‐attenuated mutant (A. brasilense SpM7918) was supplemented with IAA produced by its parental wildtype strain (A. brasilense Sp6), restored growth promotion in the microalgae‐mutant culture.     

DEGRADATION OF ADENINE BY PROTOTHECA ZOPFII (CHLOROPHYTA)1

The purines 6-amino-2-hydroxypurine and 6-amino-8-hydroxypurine, not normally associated with purine degradation in algae, were isolated by high-pressure liquid chromatography from cell extracts of Prototheca zopfii Krüger grown with adenine as the sole nitrogen source .