MIXOTROPHIC ALGAE CONSTRAIN THE LOSS OF ORGANIC CARBON BY EXUDATION1

Algae of various taxonomic groups are capable of assimilating dissolved organic carbon (DOC) from their environments (mixotrophy). Recently, we reported that, with increasing biomass of mixotrophs, heterotrophic bacteria did not increase. We hypothesized that algal uptake of external DOC may outweigh their release of DOC by exudation (H1). Here, we addressed an alternative hypothesis that algae did not assimilate external DOC but constrained the release of DOC (H2). In chemostat experiments, we cultured the mixotrophic Chlamydomonas acidophila Negoro together with heterotrophic bacteria. As external substrates, we used glucose, which was potentially available for both bacteria and algae, or fructose, which was available only for bacteria. We increased the biomass of algae by the stepwise addition of phosphorus. Bacterial biomass did not increase in experiments using glucose or when fructose was offered, suggesting that mechanisms other than algal mixotrophy (H1) kept concentrations of bacteria low. Measured exudation rates (percent extracellular release, PER) of mixotrophic algae (Cd. acidophila, Chlorella protothecoides W. Krüger) were very low and ranged between 1.0% and 3.5% at low and moderately high phosphorus concentrations. In contrast, an obligately phototrophic alga (Chlamydomonas segnis H. Ettl) showed higher exudation rates, particularly under phosphorus limitation (70%). The results support H2. If mixotrophy is considered as a mechanism to recycle organic exudates from near the cell surface, this would explain why algae retained mixotrophic capabilities although they cannot compete with bacteria for external organic carbon.     

AUTOTROPHIC AND HETEROTROPHIC NUTRITIONAL BUDGET OF SALT MARSH EPIPHYTIC ALGAE1

This paper addresses the nutrition and relative importance of primary production to heterotrophy of select unicellular green algae and diatoms from the same assemblage. Algal growth responses to nutrient additions varied widely. Such responses included: inhibition and enhancement of growth by amino acid additions: nonstimulation or inhibition by most sugars 10 mM concentration; glucose stimulation of 6 and inhibition of 3 species; fructose inhibition of 6- and 3- fold stimulation of one species; stimulation of most species by a vitamin mixture, some natural products, metabolite mixtures, etc. Photoassimilation of glucose and aculeate in 8 of 12 species occurred. Nine of 12 species took up only a fraction of the total carbon fixed as organic substrate. Enhancement of photosynthesis by glucose and inhibition by acetate was common. The data suggest that attached littoral and shoal marine algal assemblages may play mixed trophic roles lower levels of the detrital food web.     

污水净化湿地模拟系统中细菌和藻类的生态分布研究

以湿地土壤－微生物模拟系统为基础,研究了细菌和藻类在不同处理单元中的生态分布和种类变化。随着系统中微生物的净化作用,污水中污染物浓度下降,污染负荷降低,细菌和藻类的种类和数量也发生相应的变化。系统中,藻类种类由耐污种类向喜清种类过渡,显示了藻类是良好的水质评价和监测的指示生物。细菌总量降低,第４单元中的细菌总量比第１单元要低２个数量级。Ｍａｒｇａｌｅｆ多样性指数逐渐升高,第１单元为０．３８,第２单元为０．６０,第３单元为０．６９,第４单元为０．８７。系统中的反硝化率在第２单元处最高,在第４单元处最低,垂直分布以２ｃｍ深度处最高。     

UREASE GENE SEQUENCES FROM ALGAE AND HETEROTROPHIC BACTERIA IN AXENIC AND NONAXENIC PHYTOPLANKTON CULTURES1

While urea has long been recognized as an important form of nitrogen in planktonic ecosystems, very little is known about how many or which phytoplankton and bacteria can use urea as a nitrogen source. We developed a method, targeting the gene encoding urease, for the direct detection and identification of ureolytic organisms and tested it on seven axenic phytoplankton cultures (three diatoms, two prymnesiophytes, a eustigmatophyte, and a pelagophyte) and on three nonaxenic Aureococcus anophagefferens Hargraves et Sieburth cultures (CCMP1784 and two CCMP1708 cultures from different laboratories). The urease amplicon sequences from axenic phytoplankton cultures were consistent with genomic data in the three species for which both were available. Seven of 12 phytoplankton species have one or more introns in the amplified region of their urease gene(s). The 63 urease amplicons that were cloned and sequenced from nonaxenic A. anophagefferens cultures grouped into 17 distinct sequence types. Eleven types were related to α‐Proteobacteria, including three types likely belonging to the genus Roseovarius. Four types were related to γ‐Proteobacteria, including two likely belonging to the genus Marinobacter, and two types were related to β‐Proteobacteria. Terminal restriction fragment length polymorphism (TRFLP) analyses suggested that the sequenced amplicons represented approximately half of the diversity of bacterial urease genes present in the nonaxenic cultures. While many of the bacterial urease sequence types were apparently lab‐ or culture‐specific, others were found in all three nonaxenic cultures, suggesting the possibility of specific relationships between these bacteria and A. anophagefferens.     

凤眼莲根区异养细菌的群落特征与异养活性的研究

本文研究了凤眼莲根区(根际与根面)异养细菌的群落特征与异养活性。从根区分离出24株优势菌,经鉴定有10个属。其中以气单胞菌属、微球菌属、假单胞菌属、土壤杆菌属和芽孢杆菌属为主要菌属。细菌总数:根际>根面>水体。异养活性以氧化葡萄糖的速率计算。在30℃和pH7.0的条件下。根区菌群都显示较高的活性,且按下列顺序增强:老根区菌群>幼根区菌群>枯根区菌群;而根面菌群活性均大于根际菌群。但在10℃和pH5.0、8.6时,其活性明显地受到抑制。     

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.     

PlANT-ASSOCIATED BACTERIA AS TOOLS FOR THE PHYTOREMEDIATION OF OILY NITROGEN-POOR SOILS

The rhizospheres and phyllospheres of peas, beans, tomatoes, and squash raised in a desert sand soil mixed with 0.5% crude oil were rich in oil-utilizing bacteria and accommodated large numbers of free-living diazotrophic bacteria, with potential for hydrocarbon utilization. According to their 16S rRNA-sequences, the cultivable oil-utilizing bacteria were affiliated with the following genera, arranged in decreasing frequency: Bacillus, Ochrobactrum, Enterobacter, Rhodococcus, Arthrobacter, Pontola, Nocardia, and Pseudoxanthomonas. Diazotrophic isolates were affiliated with Rhizobium, Bacillus, Rhodococcus, Leifsonia, Cellulosimicrobium, Stenotrophomonas, Kocuria, Arthrobacter, and Brevibacillus. The crude oil–utilizing and diazotrophic isolates grew, with varying growth intensities, on individual aliphatic (C₈ to C₄₀) and aromatic hydrocarbons, as sole sources of carbon and energy. Quantitative gas liquid chromatographic measurements showed that representative bacterial isolates eliminated pure n-hexadecane, n-decosane, phenanthrene, and crude oil from the surrounding liquid media. Cultivation of oily sand–soil samples with any of the four tested crops led to enhanced oil degradation in that soil, as compared with the degradation in uncultivated oily sand–soil samples.     

AN UNUSUAL METHOD FOR CULTURE OF UNICELLULAR MARINE ALGAE1

A method for the axenic culture of marine microscopic algae that utilizes various types of paper as solid substrates for growth is described. Investigations into the growth of 39 algal, strains on various paper substrates were conducted. Applications of the method to bioassay are discussed.     

EVIDENCE A PHOTOPROTECTIVE FOR SECONDARY CAROTENOIDS OF SNOW ALGAE1

Snow algae occupy a unique habitat in high altitude and polar environments. These algae are often subject to extremes in nutrient availability, acidity, solar irradiance, desiccation, and ambient temperature. This report documents the accumulation of secondary carotenoids by snow algae in response to the availability of nitrogenous nutrients. Unusually large accumulations of astaxanthin esters in extra-chloroplastic lipid globules produce the characteristics red pigmentation typical of some snow algae (e.g. Chlamydomonas nivalis (Bauer) Wille). Consequently these compounds greatly reduce the amount of light available for absorption by the light-harvesting pigment-protein complexes, thus potentially limiting photoinhibition and photodamage caused by intense solar radiation. The esterification of astaxnthin with fatty acids represents a possible mechanism by which this chromophore can be concentrated within cytoplasmic globules to maximize its photoprotective efficiency.     

AN IMPROVED METHOD OF CELL ENUMERATION FOR FILAMENTOUS ALGAE AND BACTERIA1

A new simple method for estimating the number of individual cells per milliliter in suspensions of filamentous microorganisms is described. This 2-part procedure utilizes a standard microscopic counting chamber and is independent of filament length or individual cell size. A statistical analysis of the method is also presented.     

N-METHYL-N'-NITRO-N-NITROSOGUANIDINE AS A MUTAGEN FOR BLUE-GREEN ALGAE: EVIDENCE FOR REPAIR1

Survival and mutagenesis have been examined in the marine coccoid blue-green alga, Agmenellum quadruplicatum, after treatment with the chemical mutagen, N-methyl-N'-nitro-N-nitrosoguanidine (NTG). When cells were immediately returned to growth conditions following NTG treatment, the survival level was consistently low and essentially independent of the treatment conditions. The types of mutants found paralleled those previously described in the freshwater coccoid, Anacystis nidulans. If, however, cells were kept under very low light intensity, a nongrowth condition, following NTG treatment, viable cell recovery was dramatically increased. This “dim light” repair in A. quadruplicatum has characteristics similar to those reported for the dark repair systems of bacteria and yeast.     

MEASUREMENT OF ACETYLCHOLINE TURNOVER WITH GLUCOSE USED AS PRECURSOR: EVIDENCE FOR COMPARTMENTATION OF GLUCOSE METABOLISM IN BRAIN

The turnover of acetylcholine in whole mouse brain in vivo has been determined using [U-¹⁴C]glucose as a precursor of the acetyl moiety. The standard requirements for the measurement of turnover were met: the injection did not change the concentrations of precursor or product, the amount of radioactivity in the brain was proportional to the amount injected, and the relationship between the specific activity of glucose and that of acetylcholine was typical of a precursor and a product. The value for acetylcholine turnover was 64 pmol/min per mg protein, approx 6.4 nmol/min per g brain. Treatment with amobarbital (0.16 mmol/kg) decreased the incorporation of glucose into acetylcholine by 73 × 7%, and treatment with atropine increased it by 18 × 6%. These values agree with those using choline as a precursor, supporting the validity of the values for turnover obtained with either labelled precursor. The specific activity of acetylcholine was higher than that of pyruvate at all times in mouse brain in vivo and in rat brain slices in vitro. These observations demonstrate compartmentation of glucose metabolism with respect to acetylcholine synthesis in the brain. They agree with observations by others of compartmentation of acetyl metabolism. They provide an explanation for the close linkage which has been observed between carbohydrate catabolism and acetylcholine synthesis in the CNS.     

INTERCEPTION OF ATMOSPHERIC FIXED NITROGEN AS AN ADAPTIVE ADVANTAGE OF SCUM FORMATION IN BLUE-GREEN ALGAE1

Scum formation by blue-green algae has been interpreted as a nonadaptive failure of the buoyancy control mechanism and, alternatively, as an adaptive mechanism to reduce inorganic carbon limitation. Data from Lake Valencia, Venezuela, are used to illustrate the principle that scum formers intercept significant amounts of fixed N deposited from the atmosphere at the water surface. Interception of fixed N at the surface can confer a significant selective advantage for populations that are strongly N limited, as in Lake Valencia. This additional adaptive advantage to scum formation may be alternative lo or complementary to the possible relief of carbon limitation.     

COLONIZATION AND SUCCESSION OF ALGAE AND SOIL-ALGAL INTERACTIONS ASSOCIATED WITH DISTURBED AREAS1

The number and abundance of algal species were not significantly different between non-topsoiled areas and top-soiled areas. Successional trends were evident at both sites, since the number of species and their abundance increased over time. Soil-algal interactions were determined by regression analysis. A conceptual model was developed that illustrated the influence of soil chemical properties on algal abundance. Important factors affecting the soil algae were sodium, calcium, potassium, magnesium and certain trace elements. The possible influence of these factors on algal growth is discussed.     

SPECIFIC ASSOCIATIONS OF THE BLUEGREEN ALGAE ANABAENA AND APHANIZOMENON WITH BACTERIA IN FRESHWATER BLOOMS1

Scanning electron micrographs and autoradiographs of Anabaena circinalis Rabenh. and Aphanizomenon flos-aquae (L.) Ralfs in samples from fresh-water communities show that bacteria are attached specifically at the polar region of heterocysts of these known N₂ fixers. This algal-bacterial association occurs most frequently during bloom conditions. The possible roles of this association in maintaining nuisance bloom conditions are discussed.     

DIRECT ENUMERATION OF BACTERIA FROM MACROALGAE BY EPIFLUORESCENCE MICROSCOPY AS APPLIED TO THE FLESHY RED ALGAE KAPPAPHYCUS ALVAREZII AND GRACILARIA SPP. (RHODOPHYTA)1

This report describes a simplified method for direct counting of total bacteria associated with the fleshy red algae Kappaphycus alvarezii (Doty) Doty and Gracilaria spp. A Nuclepore® polycarbonate membrane (0.2–μm pore size) fitted to a vacuum filtration apparatus was used to filter algal tissue homogenate after serial dilution and staining with the fluorochrome 4′,6-diamidino-2-phenylindole. Using epifluorescence microscopy, it is possible to count bacteria without preseparating them from the algae. The technique requires homogenized algal tissue diluted with 0.2-μm-filtered, autoclaved seawater to a level appropriate for counting. Dilution reduces the amount of autofluorescent algal debris, which may interfere with Counting. The membrane filtration method yielded a bacterial count two orders of magnitude higher than that of the conventional agarspread plate technique. This method offers a more accurate approach to counting the total number of bacteria on macroalgae.     

用中度嗜热菌氧化预处理含砷难浸金精矿回收金的研究

用中度嗜热菌MP30菌株氧化预处理含砷难浸金精矿回收金。该菌株在45～48℃,pH1.7～2.2和混合营养条件下,在金精矿上获得最佳生长和氧化效果。毒砂是金精矿中主要含砷矿物,砷的最终脱除取决于包裹毒砂的黄铁矿完全氧化。砷脱除后可直接氰化提金,不必再行火法冶炼。金的提取量与载金矿物(主要是黄铁矿和毒砂)氧化程度相关。强化氧化条件下,MP30能使载金硫化物矿物完全氧化分解,使金充分暴露,继而氰化提金,其提取率可达90％～98％。用这一工艺从含砷难浸金精矿中回收金是可行的。     

ESTUARINE HETEROTROPHIC CRYPTOPERIDINIOPSOIDS (DINOPHYCEAE): LIFE CYCLE AND CULTURE STUDIES1

Cryptoperidiniopsoids are an unclassified group of delicately thecate heterotrophic dinoflagellates known to be common in eastern U.S. estuarine waters. Over the past 10 years cryptoperidiniopsoids were isolated from different geographical regions and cultured with cryptophyte algal prey. In the seven clonal isolates examined, reproduction was strongly linked to the availability of prey cells. The dinoflagellates phagocytized the contents of prey cells through a tube‐like peduncle, similarly as close relatives of Pfiesteria spp. and several other heterotrophic species. Cell division occurred while encysted, most commonly yielding two biflagellated offspring. Abundant fusing gametes, phagotrophic planozygotes, and cysts with a pronounced nuclear cyclosis characterized persistent sexuality. Cysts with nuclear cyclosis produced two flagellated offspring cells. The resistance of reproductive cysts to antimicrobial treatments was examined, and a simple high‐yield technique was developed for population synchronization while ridding the dinoflagellates of most contaminating vacuolar prey DNA and external contaminants. The DNA content and population DNA profiles of synchronously excysted cryptoperidiniopsoids from different isolates were measured using flow cytometry and were related to the life history of these and other dinoflagellates. Cryptophyte‐fed cultures with versus without extracellular bacteria were compared, and bacteria apparently promoted cryptoperidiniopsoid feeding and growth. Externally bacteria‐free dinoflagellates were cultured in media enriched with dissolved organic nutrients, and nutritional benefit may have occurred in some treatments. The potential for mixotrophic nutrition from maintenance of cryptophyte chloroplasts was examined using flow cytometrically sorted cells, but evidence of kleptoplastidy was not found in these isolates under the conditions imposed.     

PROTOCOLS FOR THE REMOVAL OF BACTERIA FROM FRESHWATER BENTHIC DIATOM CULTURES1

In this study, we describe different combinations of physical separation and antibiotic treatment to remove associated bacteria from freshwater diatoms. Diatoms were purified either from natural epilithic biofilms or from unialgal cultures. We determined that for most strains, different purification procedures have to be combined individually. In a new approach, we show that for some diatom strains, the substitution of associated aquatic bacteria by an antibiotic‐sensitive Escherichia coli strain and subsequent treatment with antibiotics may be a successful strategy to obtain axenic diatom cultures. Axenic diatom cultures are essential to study the physiology and biochemistry of individual strains as well as their responses to environmental changes without interference of accompanying bacteria.