Role of phosphate solubilizing bacteria on rock phosphate solubility and growth of aerobic rice

Use of phosphate-solubilizing bacteria (PSB) as inoculants has concurrently increased phosphorous uptake in plants and improved yields in several crop species. The ability of PSB to improve growth of aerobic rice (Oryza sativa L.) through enhanced phosphorus (P) uptake from Christmas island rock phosphate (RP) was studied in glasshouse experiments. Two isolated PSB strains; Bacillus spp. PSB9 and PSB16, were evaluated with RP treatments at 0, 30 and 60 kg ha(-1). Surface sterilized seeds of aerobic rice were planted in plastic pots containing 3 kg soil and the effect of treatments incorporated at planting were observed over 60 days of growth. The isolated PSB strains (PSB9 and PSB16) solubilized significantly high amounts of P (20.05-24.08 mg kg(-1)) compared to non-inoculated (19-23.10 mg kg(-1)) treatments. Significantly higher P solubilization (24.08 mg kg(-1)) and plant P uptake (5.31 mg plant(-1)) was observed with the PSB16 strain at the highest P level of 60 kg ha(-1). The higher amounts of soluble P in the soil solution increased P uptake in plants and resulted in higher plant biomass (21.48 g plant(-1)). PSB strains also increased plant height (80 cm) and improved root morphology in aerobic rice. The results showed that inoculation of aerobic rice with PSB improved phosphate solubilizing activity of incorporated RP.     

Spatial distribution of bacteria associated with the marine sponge Tethya californiana

Microbial diversity and spatial distribution of the diversity within tissue of the marine sponge Tethya californiana was analyzed based on 16S rRNA gene sequences. One candidate division and nine bacterial phyla were detected, including members of all five subdivisions of Proteobacteria. Moreover, chloroplast-derived Stramenopiles- and Rhodophyta-affiliated 16S rRNA gene sequences were found and Stramenopiles represented the most abundant clones (30%) in the clone library. On the phylum-level, the microbial fingerprint of T. californiana showed a similar pattern as its Mediterranean relative T. aurantium. An interesting difference was that Cyanobacteria that were abundantly present in T. aurantium were not found in T. californiana, but that the latter sponges harbored phototrophic Stramenopiles instead. Surprisingly, the phototrophic microorganisms were evenly distributed over the inner and outer parts of the sponge tissue, which implies that they also reside in regions without direct light exposure. The other phyla were also present in both the outer cortex and the mesohyl of the sponges. These results were confirmed by analysis on the operational taxonomic unit level. This leads to the conclusion that from a qualitative point of view, spatial distribution of microorganisms in T. californiana tissue is quite homogeneous. Thirty-two percent of the operational taxonomic units shared less than 95% similarity with any other known sequence. This indicates that marine sponges are a rich source of previously undetected microbial life.     

Trace metal distribution in different chemical fractions of marine sediments along the eastern Aegean shelf

The major objective of this study was to carry out sequential chemical extraction for the partitioning of particulate trace metals in sediment samples, collected along the eastern Aegean shelf during cruises July‐August 1994, in the framework of a National Marine Measurement and Monitoring Programme for the Aegean Sea.

Five metals, Cd, Pb, Cu, Zn and Cr were examined in each of sediment samples. Three chemical fractions of the sediments were separated and concentrations of the trace metals were determined by AAS techniques. The three different leaches used were hydroxylamine hydrochloride‐acetic acid, hydrogen peroxide and nitric‐perchloric acids.

Metals were concentrated mainly in the fraction extracted by nitric‐perchloric acids. Lead in the first fraction were found in the sediments of Northern part of Aegean, where the concentration of organic material was high.

The total concentrations of Cd, Cu, Zn, Cr were higher in Izmir Bay than the other sampling points. The distribution of Pb concentrations was the highest in Edremit Bay and Izmir Bay.     

洞庭湖底泥中二英污染现状及水动力对其分布的影响?

2013年9月采集洞庭湖区三口四水入湖口,东、西、南洞庭湖湖区以及出湖口沉积物,采用同位素稀释高分辨气相色谱－高分辨质谱法测定了沉积物中的二英（ PCDD／Fs）．结果表明洞庭湖沉积物中二英的浓度范围为153—7144 pg·g－1 dw （干重）,小河嘴最低,虞公庙最高．对比国内外其他淡水湖泊河流二英浓度,洞庭湖污染程度相对较低．二英污染水平依次为洞庭湖湖区＞出湖口＞入湖口,湖区内污染水平依次为南洞庭湖＞东洞庭湖＞西洞庭湖．主要同类物为OCDD,贡献率范围为77％—97％．PCDD／Fs的污染水平比1995年下降1—2个数量级,但和2004年污染水平相当．沉积物中二英的含量与水的流速成反比．洞庭湖出口处PCDD／Fs浓度相比入湖口和湖区浓度处于中间水平,表明洞庭湖中的二英可能会随水流进入长江中下游．     

Activity of marine psychrophilic bacteria at elevated hydrostatic pressures and low temperatures

Uptake and metabolic turnover of a variety of radioactively labeled substrates by a number of bacterial isolates from several depths in offshore Atlantic waters were studied in laboratory experiments. The variable conditions were hydrostatic pressure (1 to 400 atm), temperature (1.50 to 15.0°C), concentration of substrate (2 to 500 g/ml), and time of incubation (4 h to 8 weeks). Elevated hydrostatic pressure and low temperature retarded the rate of metabolism (biosynthesis more than respiration) in all isolates tested, but considerably less in those of psychrophilic character.Contribution No. 2944 of the Woods Hole Oceanographic Institution.Supported by the National Foundation Grants GA-33405 and GA-29665.     

Microbial RNA and DNA synthesis in marine sediments

A technique for measuring rates of RNA and DNA synthesis in sedimentary microbial communities has been adapted from methods developed for marine and freshwater microplankton research. The procedure measures the uptake, incorporation and turnover of exogenous [2, ³H]-adenine by benthic microbial populations. With minor modification, it is applicable to a wide range of sediment types. Measurement of nucleic acid synthesis rates are reported from selected benthic marine environments, including coral reef sediments (Kaneohe Bay, Oahu, Hawaii), intertidal beach sands (Oahu and southern California) and California borderland basin sediment (San Pedro Basin), and comparisons are made to selected water-column microbial communities. Biomass-specific rates of nucleic acid synthesis in sediment microbial communities were comparable to those observed in water-column assemblages (i.e., 0.02 to 2.0 pmol deoxyadenine incorporated into DNA [ng ATP]^-1 h^-1 and 0.2 to 8.9 pmol adenine incorporated into RNA [ng ATP]^-1 h^-1). DNA synthesis rates were used to calculate carbon production estimates ranging from 2 g C cm^-3 h^-1 in San Pedro Basin sediment (880 m water depth) to 807 g C cm^-3 h^-1 in coral reef sediment from the Kaneohe Bay. Microbial community specific growth rate, (d^-1), estimated from DNA synthesis rates in surface sediments ranged from 0.1 in San Pedro Basin to 4.2 in Scripps Beach (La Jolla, California) intertidal sand.     

Respiration and physiological state in marine bacteria

The relationship between oxygen consumption (R) and respiratory electron-transport-system (ETS) activity was investigated in batch cultures of 5 species of marine bacteria, Vibrio adaptatus, V. anguillarum, a partially identified Vibrio sp. SA774, Serratia marinorubra, and Pseudomonas perfectomarinus. Although cellular levels of R and ETS varied widely among the species tested, the R:ETS ratios for growing or senescent populations were relatively constant among the species; these ratios were 5.02 in growth and 0.426 in senescence, with coefficients of variation of 29 and 20%, respectively. The lower senescent-phase R:ETS ratio was due to a depression of the respiration rates following growth termination. The regression log (R per cell) = 0.832 log (ETS per cell) + 0.727 for the growing populations was similar to that determined for marine zooplankton. The slight dependency of the R:ETS ratio on organism dry weight found for zooplankton was supported by our data. Planktonic respiration rates estimated from measured ETS-depth profiles in the eastern tropical North Pacific Ocean using the senescent-phase R:ETS ratio were similar to published oxygen consumption rates in the deep sea.Contribution No. M79-61 from the University of Washington's Department of Oceanography, and No. 79032 from the Bigelow Laboratory for Ocean Sciences.     

Arsenic metabolism in marine bacteria and yeast

Arsenic metabolism was studied for two marine microorganisms, a facultative anaerobic bacterium, Serratia marinorubra, and an obligately aerobic yeast, Rhodotorula rubra. Both were cultivated in media with (⁷⁴As) arsenate (As V), and the products of arsenate metabolism were determined qualitatively. Both the bacterium and the yeast produced arsenite (AS III) and methylarsonic acid [CH₃AsO(OH)₂]. In addition to the foregoing, only the yeast produced dimethylarsinic acid (CH₃)₂AsO(OH) and volatile alkylarsines. In contrast, the bacterium growing anaerobically with cobalamine as a cofactor did not synthesize gaseous forms of arsenic such as methylarsines. Neither organism synthesized arsoniumphospholipids such as those produced by marine phytoplankton or terrestrial fungi. The yeast did not accumulate arsenite, but instead transported some of it into the culture medium and methylated the remainder first to methylarsonic acid and then to dimethylarsinic acid. Finally, the latter compound was methylated further and volatile alkylarsines were formed. In contrast, the bacterium retained all products of arsenate metabolism intracellularly. Both the bacterium and the yeast, therefore, converted relatively toxic arsenate, the most abundant arsenic compound in seawater, to products that were presumably less toxic.     

Adenosine triphosphate and adenylate energy charge in marine sediments

The determination of adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP) concentrations in sediments from both Puget Sound and the Washington State continental shelf (USA) has been investigated. Major losses during the extraction procedure included sediment sorption, coprecipitation, and unreactivity of the soluble adenylates presumably through irreversible binding to solubilized sediment organics. Addition of phosphoric acid and the non-ionic detergent Triton X-100 to cold sulfuric acid significantly increased the extraction efficiency and sensitivity over that of sulfuric acid alone, although both methods yielded similar results when corrected for losses. Vertical profiles of ATP, total adenylates (A_T), and adenylate energy charge (EC) in sediments from both areas are presented. Profiles of A_T are similar in both areas, but EC values in the shelf sediments are significantly lower, which suggests a depressed physiological condition in the shelf population relative to that in Puget Sound.Contribution No. 513 from the University of Washington's College of Fisheries, and Contribution No. 1135 from the University of Washington's Department of Oceanography     

磷细菌筛选及其对苗期玉米生长的影响

从潮土、水稻土、砂姜黑土、石灰土上植物根际土壤和根中分离了86株磷细菌,通过NBRIP液体摇瓶培养3 d,培养液水溶磷质量浓度为4.2～387.3 mg.L^-1,水溶磷质量浓度与培养液pH呈显著负相关（r^2=0.621 6）。筛选出3株磷细菌进行玉米盆栽试验,结果表明,1株磷细菌处理的玉米干物质量和吸磷量与对照（处理4）相比无显著差异,2株磷细菌处理的玉米干物质量和吸磷量与对照相比有明显增加,干物质量增加了19.6%～37.5%,吸磷量增加了22.7%～40.2%,其中编号为HCW115解磷菌株的效果相当于施用无机磷（P）10 mg.kg^-1处理。     

Metal distribution in marine sediments of the Mar Piccolo in Taranto (Ionic Sea,southern Italy)

The spatial distribution of selected metals (Fe, Ni, Mn, Zn, Cu, Pb, Hg and Cd) and their partition in the different chemical forms (speciation) were determined in benthic sediments from Mar Piccolo, Taranto (Ionic Sea, southern Italy). The aim of this study was to assess the degree of sediment contamination and bioavailability of metals, very important for Italian mussel culture, in a semi-enclosed marine ecosystem. In sediments, grain size distribution, total organic carbon (TOC), and humic (HA) and fulvic (FA) acids contents were also determined. The compounds TOC, HA and FA, in fact, have a great influence on metal complexation. High concentrations of copper, lead, zinc and mercury were found, especially in sediments of the first inlet of Mar Piccolo. Statistical analysis of results shows significant correlation among Cu, Pb and Zn concentrations and the influence of organic matter on the sediment distribution of Cu, Pb and Zn. Metal speciation analysis, carried out with sequential extraction method, shows that Fe and Ni are present in sediments mainly as sulphides. Mn and Pb are present principally in an ion-exchangeable form or bound to carbonate and Cu is associated to organic matter. Metal pollution index (MPI), calculated as a weighed mean of the metal contamination factors in each station, shows that with regard to Cu, Ni, Pb and Zn, some stations investigated are unpolluted (MPI <1), whereas in other areas, pollution level is not very high. On the contrary, for mercury, the pollution level in the first inlet of the basin is very high.     

Light stimulation of phosphate uptake in marine phytoplankton

A phosphate uptake system responding positively to light would offer the algae a competitive advantage over bacteria, particularly when phosphorus is limiting. Effects of light on phosphate uptake in phytoplankton were investigated in view of conflicting literature reports, to test the hypothesis that both the P status and the light history of the population controlled the response. Maximum stimulation, to about 2.7x dark uptake, was observed at a Strait of Georgia, British Columbia, Canada station where phytoplankton was phosphorus sufficient but light limited. Stimulation was similar, about 1.8 x the dark rate, in the Northern Sargasso Sea, where phytoplankton was probably limited by phosphate but not by light. Minimal stimulation was observed in the Sheepscot Estuary, Maine, USA, where the population appeared to be neither phosphorus nor light limited.     

Sulfide and thiosulfate-oxidizing bacteria in anoxic marine basins

The occurrence of sulfide and thiosulfate-oxidizing bacteria was examined in waters and sediments of the Black Sea and the Caraco Trench. Isolates obtained by enrichment-culture techniques exhibited facultative autotrophic-growth characteristics and oxidized thiosulfate or sulfide. Acid was not formed during growth in mineral medium. Most of the isolates were facultatively anaerobic, using nitrate as the terminal hydrogen acceptor. Strictly aerobic bacteria were found only in oxygenated waters. Comparison of the metabolic capabilities of the isolates during growth in sulfide and thiosulfate media with chemical parameters measured in the Black Sea and Cariaco Trench waters suggests that the biological oxidation of thiosulfate is the rate-limiting step in the oxidative turnover of sulfur compounds in the basins. Evidence for the oxidation of sulfide in the anoxic waters is presented, and possible explanations for the occurrence of this process in the absence of suitable electron acceptors (O₂. NO₃) are discussed. A maximum dark assimilation of carbon dioxide was located just below the oxygen-sulfide interface in the Cariaco Trench. Problems of interpreting these data are considered.Contribution No. 2958 of the Woods Hole Occanographic Institution. Supported by the National Science Foundation Grants GA 29665 and GA 33405.     

Dehydrogenases as tools in the study of marine sediments

A method is outlined for the measurement of NADH₂-dependent dehydrogenase activity in whole samples as well as in fractions of carbonate sediments. The method is based on extraction of enzyme activity with phosphate buffer containing 0.12% Triton X-100, dialysis, and a photometric assay which permits initial velocity determinations of the enzyme reaction under controlled conditions of pH, temperature, and substrate concentration. The dehydrogenases extracted from carbonate sediment of a subtropical beach at Bermuda are characterized by low K _m values (0.03 to 0.07 mM NADH₂/1), pH maxima around 8.5, and temperature maxima between 35° and 40°C. Their vertical distribution in the sediment agrees with what is known about the distribution of biomass on beaches of this type, maximum activity occurring in the topmost centimetre. Approximately 80 to 90% of the total electron transport activity is contained in the grain fraction, the remainder in the interstitial fraction of a sediment sample. From this it can be inferred that the interstitial fauna contributes only little to the total energy budget of this type of sediment, which is dominated by the Aufwuchs on the sand grains. The electron transport activity (ETA) measured by means of the dehydrogenase assay is taken to represent the maximum transport capacity (ETC) of all the organisms living in the sand ecosystem. From published evidence it is inferred that oxygen consumption of sediment samples and of individual organisms measured in vivo usually accounts for 10 to 20% of the maximum electron transport capacity measured in vitro with saturating concentrations of substrates and with Triton X-100 as part of the enzyme assay. It is suggested further that the ratios of dehydrogenase activities in different fractions or zones of sediment, or of different organisms, may represent one of the best indicators, so far available, of energy relationships in marine sediments.Contribution No. 652 from the Bermuda Biological Station for Research.     

Heavy metal and nutrient contamination of marine sediments in Hong Kong

A monitoring programme was performed to use marine sediments as an indicator of marine contamination in Hong Kong. A total of 51 samples were collected from eight marine sites and analysed for pH, redox potential, salinity, total‐N, total‐P, total organic carbon, and total Cu, Zn Pb, Cr and Cd. Sites with high industrial or aquacultural activities contained high total organic carbon contents, total‐N and total‐P contents. Sediments from sites with high industrial activities also enriched with heavy metals. Metal speciation results indicate that Cu and Cr existed mainly in the organic fraction, Pb and Zn were distributed approximately equally in both the organic and residual fractions while Cd mainly occurred in the residual fraction. The Cu, Cr and Zn contents were highly correlated with total organic carbon contents. All samples were classified as non‐acid forming according to the net acid generation test and only samples from the industrial site released a high concentration of heavy metals under a complete oxidising condition. The present study reveals that organic and heavy metal pollution is serious in the marine sediments of Hong Kong, and industrial and aquacultural activities are probably the major sources of contamination.     

Pools of chlorophyll and live planktonic diatoms in aphotic marine sediments

Chlorophyll a and numbers of live pelagic diatoms were recorded from sediment depth profiles at 11 stations in the oligotrophic Øresund, Denmark, in late-June. Extraction efficiency of chlorophyll a analysed fluorometrically did not differ significantly between paired samples of frozen-thawed and fresh sediment. The depth profiles of chlorophyll a could be explained by a diagenetic model involving two different chlorophyll pools: one reactive pool declining exponentially with core depth, and one non-reactive pool, of about 1 µg Chl ml^-1 wet sediment, being constant with depth. The number of live diatoms, quantified by the dilution-extinction method, and expressed in terms of most probable number (MPN), declined from an average of about 300,000 g^-1 in the surface sediment to zero values at a depth of 13 cm. The number of live cells was significantly correlated with the sediment chlorophyll a, and the profiles of live cells as well as reactive chlorophyll followed the same exponential decline with core depth, suggesting that the main source of chlorophyll in the sediment was live pelagic diatoms. Taxonomic composition of diatoms in the sediment, dominated by the pelagic genera Chaetoceros, Thalassiosira and Skeletonema, matched the species composition in the water column 3 months earlier during the spring bloom. Regular recordings of the phytoplankton community in the water column showed that only these specific bloom species could be the source of the sediment content of diatoms and chlorophyll a. Further, the ratios between live cells and chlorophyll a were similar in the sediment and in the spring bloom. A conservative estimate of depth-integrated pools of diatoms in the sediment suggested that about 44% of the total phytoplankton biomass during the spring bloom was still present as live cells in the sediment after 3 months. This indicates that the spring bloom input to the sediment is not degraded immediately by the benthic fauna.     

The addition of FeOOH binds phosphate in organic matter-rich sediments

Due to climate change and anthropogenic nutrients’ runoff into freshwater or shallow lakes, eutrophication caused by phosphorus (P) can be seen in the frequent occurrence of cyanobacterial blooms and excessive growth of macrophytes. Subsequently, decomposition of cyanobacterial bloom biomass (CBB) and macrophytes leads to massive autochthonous organic matter (OM) and creates hypoxia in bodies of water. In this study, we investigated the effects of OM and iron on phosphorus release from lake sediments under anaerobic conditions. As with CBB, the addition of cellulose also enhanced P release from sediments during microcosm experiments, while total phosphorus (TP) concentration in the overlying water displayed an inverse relationship to cellulose amendment, with high TP concentration (0.41?±?0.07?mg?L^?1) observed in the treatment of less cellulose amendment (1?g of cellulose). In addition, P release from OM-rich sediments was effectively inhibited when amorphous FeOOH was added to the microcosms. P release was inhibited by 66–92% when the weight ratio between total Fe and total P in sediments varied from 18 to 60. Thus, iron treatment was useful to inhibit P release from OM-rich sediments, and could alleviate eutrophication problems.     

Diurnal variation of nitrogen cycling in coastal,marine sediments

A closed chamber technique was developed to determine the emission of microbially produced N₂O from an estuarine sediment. A diurnal variation was observed; maximum emissions of 0.4 to 4.0 mol N₂O–N m^-2 h^-1 were recorded at night whereas the rates were low or even negative, -0.4 to 0.4 mol N₂O–N m^-2 h^-1, during the day. The bacterial denitrification located in the uppermost centimeter was apparently the major source of the emitted N₂O. The diurnal emission pattern was thus inversely related to the O₂ availability at the sediment surface; in the dark, the lack of O₂ production by benthic photosynthesis allowed the denitrification to occur closer to the sediment-water interface and was likely to enhance the release of N₂O to the water. The daily averages for the emission were about 40 mol N₂O–N m^-2 d^-1 for three investigation periods in autumn (November), winter (February) and spring (April), whereas no significant emission was recorded in the NO ₃ ^- -depleted sediment in early summer (June). In this estuary, the N₂O emissions from the sediment were significant contributions to the overall release of N₂O to the atmosphere.     

Inorganic nitrogen metabolism in marine bacteria: Nitrate uptake and reduction in a marine pseudomonad

Growth experiments in batch cultures indicated that the uptake of nitrate by the marine pseudomonad PL1 was inhibited in the presence of ammonia provided that the ammonia concentration was higher than 1 mM. At ammonia concentrations of less than about 1 mM, however, both nitrate and ammonia were utilised simultaneously. The saturation constants for nitrate and ammonia uptake were both 2.6x10^-4 M, and similar to the Michaelis constants of nitrate reductase for nitrate (2.9x10^-4 M) and glutamine synthetase for ammonia (2x10^-4 M). Nitrate reductase activity linked to NADH was detected in chemostat-grown cultures with nitrate as nitrogen source, and in cultures containing limiting concentrations of nitrate and ammonia, ammonia or glutamate. Enzyme synthesis appeared to be repressed in cultures containing an excess of ammonia or glutamate. Chemostat cultures utilised ammonia or glutamate in preference to nitrate, while there was no marked preference between ammonia and glutamate.     

海洋沉积物中甾醇类物质的气相色谱/质谱方法分析

应用超声提取技术,结合硅胶-中性氧化铝柱层析净化分离,BSTFA+1%TMCS衍生,及气相色谱-质谱定性定量技术,建立了海洋表层沉积物中8种甾醇类化合物的定量分析方法.实验采用正交实验优化了提取过程中提取剂种类、试剂体积和超声时间,同时对比并优化了柱层析淋洗液的配比、用量以及衍生剂的用量.结果表明,50 mL二氯甲烷/甲醇(V/V,2∶1),超声40 min,超声3次,总甾醇的萃取率可达99.6%;3 g硅胶+2 g中性氧化铝层析,35 mL二氯甲烷/甲醇(V/V,9∶1)淋洗净化回收最佳;8种甾醇在0—848μg.L-1范围内有良好的线性关系;方法检测限为1.2—2.4 ng.g-1.在3种浓度水平0.05、0.1和1.0μg.g-1下,其平均回收率为76.2%—100.9%,相对标准偏差为1.0%—10.3%.应用本方法检测大连湾的3个沉积物样品,8种甾醇的含量在0.079—6.833μg.g-1范围内.本方法的灵敏度高、准确度好,适合用于沉积物样品中甾醇物质的检测要求.