太湖不同解有机磷菌株胞外碱性磷酸酶活性对蓝藻碎屑的响应

浮游细菌胞外碱性磷酸酶在湖泊磷循环过程中具有关键作用, 但其与生物可利用性磷和颗粒态有机质之间的关系尚未得到充分的研究。研究以从太湖沉积物分离的3 株解有机磷细菌(OPB)和东湖水中OPB菌群为实验对象, 以蓝藻干物质为颗粒态有机质, 系统分析了模拟条件下OPB 的数量、不同大小颗粒所表现的胞外碱性磷酸酶活性(APA)与不同形态磷的浓度及其相互关系。从总体上讲, APA 与OPB 数量显著正相关, 且明显反比于溶解反应性磷(SRP)浓度, 在模拟实验过程中, 不同菌株(群)所对应的SRP 浓度均显著不同, 故其解磷能力各异。从绝对活性及其在总活性中所占的比例来看, 各处理间小颗粒(0.22—3.0 μm)APA亦明显不同, 而大颗粒(>3.0 μm)APA和溶解态(8 CFU/mL), SRP 浓度逐渐增加。实验后期(20—33 d)细菌数量大幅度减少, 且伴随SRP 和溶解态有机磷浓度的显著升高。因此, 在缺磷且富含颗粒态有机质的条件下, OPB 将产生胞外碱性磷酸酶, 分解有机磷, 进而满足其大量生长的需要, 同时有效改变溶解态磷的形态与生物可利用性。       

Dynamics of water-extractable phosphorus during the degradation of Microcystis aeruginosa by four bacteria species

We carried out a laboratory experiment for studying the dynamics of water-extractable phosphorus release during the decomposition of dead Microcystis aeruginosa driven by the four bacteria strains. Our results showed that water-extractable phosphorus had significant correlation to alkaline phosphatase but without obvious correlation to acid phosphatase, suggesting that alkaline phosphatase was the primary enzyme decomposing organic phosphorus. Also, water-extractable phosphorus exhibited an obvious correlation to pH, indicating that the alkaline environment contributed to the release of water-extractable phosphorus. The quantity of bacteria hardly correlated to the release of water-extractable phosphorus for all species. This could result from decrease of bacterial quantity after consumption of substrate in closed system. Obvious variation in water-extractable phosphorus was observed across the strains in two water treatments. Sporolactobacillus sp. and Bacillus macerans had the strongest ability for decomposing organic phosphorus among five strain treatments in the lake water. In the distilled water, Bacillus sp. had the strongest ability to degrade organic phosphorus. In both water treatments, mixed species exhibited intermediate ability for decomposing organic phosphorus. It was true that water quality strongly affected the release of phosphorus by regulating phosphatase activity.     

Diversity and biogeography of bacterial assemblages in surface sediments across the San Pedro Basin, Southern California Borderlands

Sediment bacteria play important roles in the biogeochemistry of ocean sediments; however, factors influencing assemblage composition have not been extensively studied. We examined extractable sediment bacterial abundance, the composition of bacterial assemblages using a high-throughput molecular fingerprinting approach, and several sediment biogeochemical parameters (organic matter content and alkaline phosphatase activity), along a 35 km transect from Point Fermin, Southern California, to Santa Catalina Island, across the approximately 900-m-deep San Pedro Basin. Automated rRNA intergenic spacer analysis (ARISA) demonstrated that in two spatially isolated shallow (approximately < 60 m, on opposite sides of the channel) sediment environments, assemblages were more similar to each other than to deeper communities. Distinct communities existed in deeper and shallower sediments, and stations within the deep basin over 2 km apart contained remarkably similar assemblage fingerprints. The relative contribution to total amplified DNA fluorescence of operational taxonomic units (OTUs) was significantly correlated to that of other OTUs in few comparisons (2.7% of total), i.e. few bacterial types were found together or apart consistently. The relative proportions within assemblages of only a few OTU were significantly correlated to measured physicochemical parameters (organic matter content and wet/dry weight ratio of sediments) or enzyme (alkaline phosphatase) activities. A low percentage of shared OTU between shallow and deep sediments, and the presence of similar, but spatially isolated assemblages suggests that bacterial OTU may be widely dispersed over scales of a few kilometres, but that environmental conditions select for particular assemblages.     

Diel changes in the alkaline phosphatase activity of bacteria and phytoplankton in the hypereutrophic Villerest reservoir (Roanne,France)

The diel changes of the size fractioned alkaline phosphatase activity (APA) were studied in relation to several abiotic and biotic factors in Villerest reservoir (located on the Loire river, near the city of Roanne, France), bihourly during two days in July 1992. The APA measured in this work exceeded considerably those reported in the literature, suggesting that dissolved mineral phosphorus was not available to microorganisms. At 1 m, the APA was primarily due to bacteria which actively assimilated organic P compounds released by photosynthetic algal metabolism. At 5, 10 and 20 m, the APA was predominantly algal. The high concentrations in SRP (soluble reactive phosphorus) would indicate that orthophosphates were not bioavailable. The reverse (i. e availability to phytoplankton) would have resulted in undetectable levels of P-PO _inf4 ^sup3– due to the massive proliferation of algae in Villerest reservoir.     

Ecological significance of phosphomonoesters and phosphomonoesterase activity in a small Mediterranean river and its estuary

In 1998, the concentration of phosphomonoesters, a biologically available fraction of the dissolved organic phosphorus pool, was measured along Palmones River (a small Mediterranean river in Southern Spain) and its estuary. Due to the extremely low phosphomonoester concentration in the river (usually under detection limits by analytical procedures), a method using chromatographic cartridges to concentrate this compound was used. Phosphomonoester concentration was usually in the nanomolar range, although values up to 6 m were measured in a sampling station near an effluent of domestic sewage and a cellulose pulp plant. Concentrations were always lower than the dissolved reactive phosphorus (mostly orthophosphate). Phosphomonoesters represented a variable percentage of the dissolved organic phosphorus, from negligible percentages up to 56%. Besides, a highly significant correlation between these two phosphorus fractions was found.In addition, phosphomonoesterase (alkaline phosphatase) activity was measured in the water and macrophytobenthos (both algae and aquatic phanerogams) along the river and the estuary. In water, most of the phosphatase activity upstream was regarded as soluble. This was confirmed by the difficulty of performing kinetic studies in the unfiltered samples. On the contrary, the phosphomonoesterase activity in the estuary was attributed to different size fractions, suggesting that phosphomonoesters are used actively as a phosphate source by bacteria and phytoplankton, with relative contributions depending on the sampling period.     

Longitudinal and Vertical Trends of Bacterial Limitation by Phosphorus and Carbon in the Mediterranean Sea

The effect of phosphate (P), nitrate (N), and organic carbon (C, glucose) enrichment on heterotrophic bacterial production was examined along two longitudinal transects covering the whole Mediterranean Sea during June and September 1999. During these cruises, integrated bacterial production ranged from 11 to 349 mgC m(-2) d(-1) for the 0-150 m layer. P was found to stimulate bacterial production (BP) in 13 out of 18 experiments, in the eastern and in the western Mediterranean Sea. Organic carbon stimulation of bacterial production was observed at two stations in the Alboran Sea, where the highest bacterial production was recorded (216 and 349 mg C m(-2) d(-1)) and in the Sicily Strait. Maximum rates of alkaline phosphatase (AP) increased from the Alboran to the Levantine Sea whereas AP turnover time decreased. Moreover, alkaline phosphatase activity was not systematically reduced following additions of P. In cases of P limitation, however, the alkaline phosphatase activity to bacterial production ratio was severely reduced in the P and NPC enrichments. Generally, the addition of the limiting factor--whether P or C--had a synchronous stimulating effect on bacterial production and ectoaminopeptidase activity and induced a decline in the amino acid respiration percentage. At two selected stations in the eastern and northwestern Mediterranean, response to enrichment was tested on vertical profiles. Bacteria shifted from P to C limitation at a depth where soluble reactive phosphorus was still undetectable, but corresponding to a strong increase in alkaline phosphatase turnover time. Our results showed that values of AP turnover time lower than 100 h corresponded to situations of P limitation of bacterial production.     

Phosphatases; origin,characteristics and function in lakes

Phosphatases catalyze the liberation of orthophosphate from organic phosphorus compounds. The total phosphatase activity in lake water results from a mixture of phosphatases localized on the cell surfaces of algae and bacteria and from dissolved enzymes supplied by autolysis or excretion from algae, bacteria and zooplankton. External lake water phosphatases usually have pH optima in the alkaline region. Acid phosphatases generally seem to be active in the internal cell metabolism. The synthesis of external alkaline phosphatases is often repressed at high phosphate concentrations and derepressed at low phosphate concentrations. Phosphatase activity has therefore been used as a phosphorus deficiency indicator in algae and in natural plankton populations. The possibilities for this interpretation of phosphatase activity in lake water are limited, however, and this is discussed. The in situ hydrolysis capacity, i.e. the rate by which orthophosphate is released from natural substrates, is unknown. However, we advocate that this process is important and that the rate of substrate supply, rather than phosphatase activity, limits the enzymatic phosphate regeneration.     

Viral dynamics in two trophically different areas in the Central Adriatic Sea

To understand the activity of marine viruses, experiments on viral production, viral decay and the percentage of lytic and lysogenic bacterial cells among the total number of bacterial cells were carried out seasonally at two stations in the Adriatic Sea with different trophic conditions. Additionally, we are providing an insight on the enrichment with dissolved and particulate organic matter by viral lysis in the studied area. Viral production was higher at the coastal station than at the open-sea station. Viral decay rates were also higher at the coastal sea station than at the open-sea station, and accounted for approximately 40% of viral production at both investigated stations. The percentage of lysogenic infection was lower than that of lytical infection, which indicates the prevalence of the lytic cycle at both stations. Viruses had a significant influence on bacterial mortality through high daily removal of the bacterial standing stock at the coastal and open-sea station. The viruses contributed to the restoration of dissolved organic carbon, nitrogen and phosphorus in the microbial loop by lysing the bacterial cells at the studied stations. All the above suggest that viruses are important in the microbial food web and an important factor in the control of bacterial populations within the study area.     

Nutrient Utilization Strategies of Algae and Bacteria after the Termination of Nutrient Amendment with Different Phosphorus Dosage: A Mesocosm Case

The impacts of nutrient amendment termination on the growth strategies of algae and bacteria were conducted in experimentally designed mesocosm in which two different phosphorus (P) dosages were treated. The algal community composition did not change greatly in Group A (low phosphorus) and Group B (high phosphorus). In Group A, the secretion of bacterial alkaline phosphatase (AP) after nutrient termination stimulated bacterial phosphorus acquisition, which caused the decrease in algal phosphorus levels, in terms of the increase of bacterial abundance and bacterial production, as well as the decrease in chlorophyll a and particulate organic carbon. The algal collapse resulted in dissolved organic carbon secretion, further fuelling bacterial growth. In Group B, excess phosphorus input urged algae to store phosphorus as poly-phosphate. When phosphorus input ceased, in order to maintain their used high phosphorus demand, algae strengthened to gain phosphorus through the hydrolysis of dissolved organic phosphorus in water column and ploy-phosphate inside the cells by AP, evidenced by high algal alkaline phosphatase activity, algal growth continuation, and bacterial growth decline. These facts indicated that phosphorus content should reduce to a lower level than expected, so that algal bloom can be effectively controlled in eutrophic water bodies.     

Alkaline phosphatase activity and cellular phosphorus as an index of the phosphorus status of phytoplankton in Minnesota lakes

SUMMARY. Total alkaline phosphatase activity is inversely related to cellular phosphorus in the particulate organic matter of waters above and below the thermocline in nine Minnesota lakes. Diagnostic criteria for assessing surplus and limiting levels of phosphorus are proposed. When applied to seasonal changes in phosphorus status in Lake Josephine, they demonstrate severe phosphorus starvation in late summer and autumn.     

秸秆和生草覆盖对桃园土壤养分含量、微生物数量及土壤酶活性的影响

以秸秆(覆盖重量分别为小麦(Triticum aestivum)秸3.25 kg·m^-2、玉米(Zea mays)秸1.97 kg·m^-2、禾本科杂草3.67 kg·m^-2)和生草(白三叶草(Trifolium repens)、高羊茅(Festuca arundincea)和紫花苜蓿(Medicago sativa), 播种量均为50 kg·hm^-2)为覆盖材料, 以不覆盖为对照, 研究了不同覆盖材料对桃园土壤微生物数量和酶活性的影响, 及其与土壤养分的关系。结果表明, 与对照相比, 除覆盖生草根际和非根际土壤全磷和速效磷含量差异均不显著外, 其他处理根际和非根际土壤碱解氮、速效钾、全氮、全钾和有机质含量差异均达到显著水平; 所有处理根际和非根际土壤氨化细菌、真菌和放线菌数量、土壤含水率和pH值、土壤脲酶和磷酸酶活性差异均达到显著水平。白三叶草处理的根际和非根际土壤碱解氮、速效钾、全氮、全钾、有机质含量, 土壤氨化细菌和真菌数量, 土壤脲酶和磷酸酶活性的平均升幅均最高, 分别为99%、270%、267%、117%、272%、158%、141%、156%和64%。氨化细菌、真菌、放线菌、脲酶和磷酸酶分别与土壤碱解氮、速效钾(放线菌和磷酸酶除外)、全氮、全钾和有机质呈显著或极显著的正相关。通径分析表明, 在3种土壤微生物和2种酶对养分含量的影响中, 脲酶是影响土壤碱解氮、速效钾、全氮、全钾和有机质的主要因子。     

Isolation and Characterization of Organic Phosphorus-Mineralizing Bacteria in Sediment of a Chinese Large Shallow Eutrophic Lake (Lake Taihu)

The organic phosphorus mineralizing bacteria (OPB) play an important role in phosphorus cycling in lake sediment, to which less attention has been paid. Monthly sediment samples in 2009 ending in October, together with the samples from different seasons (May, July, October, and December) in 2008, were collected at 6 sites in a Chinese large shallow eutrophic lake (Lake Taihu). The sediment OPB numbers ranged from 2.2 × 10⁶ to 1.79 × 10⁸ cells g^?1 (dry weight) at different sampling sites and in different seasons, with the average being 3.88 × 10⁷ cells g^?1 (dry weight). Its number was highest at the most polluted site and peaked in spring and summer, which can be explained by the enrichment of organic matter in sediment. Furthermore, there existed a significant positive relationship between the OPB numbers and alkaline phosphatase activities in the sediment. The 6 OPB strains isolated from the sediment were distinct in terms of their colony morphology on the yolk agar, biochemical characteristics and phosphorus release abilities. According to the 16S rDNA sequences, these OPB belong to Bacillus cereus, Stenotrophomonas maltophilia, Stenotrophomonas sp., Bacillus cereus, Xanthomonas sp., Pseudomonas sp. They were distinguished from the OPB species recorded in a Chinese small shallow eutrophic lake whose sediment organic content was remarkably higher. Taken together, organic matter shaped the OPB community not only quantitatively but also qualitatively, which in concert facilitated the enzymatic hydrolysis of organic phosphorus in lake sediment.     

祁连山东段高原鼢鼠洞道土壤微生物和土壤酶

高原鼢鼠繁杂的洞道系统对高寒草甸土壤的理化性质有着重要影响。本研究采集高原鼢鼠活动洞道土壤样本,并以无鼢鼠扰动的同深度土层土壤作为对照。对比分析了2个处理土壤微生物（细菌、真菌、放线菌）、土壤酶（脲酶、蔗糖酶、碱性磷酸酶）活性以及土壤全氮、有机质、速效磷含量的差异。结果表明：高原鼢鼠洞道土壤速效磷和全氮的含量均高于对照,其中春季和秋季的速效磷达到显著水平（P<0.05）;秋季无鼢鼠扰动的土壤有机质和真菌的含量显著高于洞道土壤（P<0.05）;3种酶活性各处理间无显著差异（P>0.05）;对照区春季土壤的细菌和秋季真菌的含量显著高于洞道土壤（P<0.05）;洞道和对照区土壤的细菌、放线菌、速效磷、全氮和有机质的含量均在夏季达到最大值,各种指标其它月份差异不显著（P>0.05）。基于以上结果,本研究发现高原鼢鼠活动洞道土壤微生物和土壤酶与对照区土壤无显著差异（P>0.05）。     

模拟践踏和降水对高寒草甸土壤养分和酶活性的影响

为明晰牦牛和藏羊践踏对高寒草甸的分异影响,通过2年模拟践踏和降水双因子控制试验,研究了践踏和降水对高寒草甸土壤养分和酶活性的影响。研究结果表明,践踏处理提高了0—20 cm土层土壤速效氮和速效钾含量,降低了0—20 cm全磷、脲酶和0—10 cm速效磷、碱性磷酸酶和有机质含量,且适度践踏促进了全氮的矿化。随降水强度的增加,0—30 cm土层土壤全氮和0—20 cm全磷和脲酶活性呈单峰曲线的变化态势,在平水下达到峰值;降水显著降低了0—30 cm土层土壤速效氮、磷、钾和0—10 cm土层土壤全钾含量,对土壤有机质含量无显著影响(P0.05)。同一放牧强度下,藏羊践踏区的土壤养分和酶活性优于牦牛践踏区,但差异不显著(P0.05)。综合可得,家畜的践踏作用促进了土壤氮和钾的矿化,抑制了磷的累积且加速了表层土壤有机质的耗竭,降低了土壤脲酶和碱性磷酸酶活性;适度降水提高了土壤全氮、全磷含量及酶活性,降水过多则相反。适度的家畜践踏与降水相耦合下草地土壤的养分循环和酶活性要优于重度践踏和不践踏小区。在对草地的适度放牧利用前提下,应注重土壤含水量和放牧畜种对草地的影响。草地干旱或土壤含水量过高时,应适当减少放牧畜种中牦牛比例增加藏羊比例,以期使草地得到健康可持续发展。     

Experimental evidence for interactions between bacterial peptidase and alkaline phosphatase activity in the Baltic Sea

From the observed pattern of aminopeptidase and alkaline phosphatase activities in the Baltic Sea, the question arose whether there is an interaction between the activities of both enzymes. In experiments with 0.8 m filtered seawater, the effects of commercial alkaline phosphatase on bacterial aminopeptidase, the effects of commercial peptidase on bacterial alkaline phosphatase activity (APA), and the effects of proteins, carbohydrates and inorganic nutrients on the activities of both enzymes were investigated.Addition of commercial alkaline phosphatase stimulated bacterial aminopeptidase activity and, similarly, the addition of commercial peptidase increased the APA in bacteria. The proteins, albumin and casein, stimulated aminopeptidase activity and APA simultaneously. Experiments using ammonium and glucose suggested that stimulation of APA by peptidase could be mediated by nitrogen and carbon availability. There were also some indications that stimulation of aminopeptidase activity by alkaline phosphatase functioned by catalysing phosphate release from organic phosphorus compounds.     

Differential strain response in alkaline phosphatase activity to available phosphorus in Microcoleus autumnalis

Toxic, benthic cyanobacterial proliferations have increased in frequency and severity globally and can have negative impacts on aquatic ecosystems, recreation and human health. Microcoleus autumnalis has been associated with numerous animal fatalities and is causing increasing concern. It tends to grow in systems with moderate dissolved inorganic nitrogen and very low dissolved reactive phosphorus. Acquisition of nutrients, particularly phosphorus, from organic sources may explain how M. autumnalis can reach the high biomass in these relatively nutrient deplete environments. In the present study the effect of phosphorus concentration and source on alkaline phosphatase activity was investigated in toxic and non-toxic M. autumnalis strains. Toxic strains exhibited significantly higher alkaline phosphatase activity than non-toxic strains (p < 0.05), and alkaline phosphatase activity increased in all strains under phosphorus-depleted conditions (p < 0.05). Alkaline phosphatase activity was also present in environmental M. autumnalis mats, though at lower levels than in laboratory experiments. The presence of alkaline phosphatase activity indicates that the acquisition of phosphorus from organic phosphorus sources may contribute to the ability of M. autumnalis to grow in systems with low dissolved reactive phosphorus.     

Mineralization of dissolved organic phosphorus from a shallow eutrophic lake

Release of soluble reactive phosphorus (SRP) from dissolved organic phosphorus (DOP), concentrated by reverse osmosis of water samples from Lough Neagh Northern Ireland, was measured in the presence of enzymes and cultures of lake water bacteria in a basal liquid medium adjusted to the pH of lake water (7.6). No hydrolysis of unfractionated DOP was observed in the presence of alkaline phosphatase but a combination of alkaline phosphatase and phosphodiesterase mineralized 14% of DOP in a 30 day incubation period at 15 °C. A similar amount of mineralization was attained by phytase. Phytase induced the same degree of mineralization in a range of DOP fractions varying from MW > 100 000 to c. 500. A mixed culture of lake water bacteria mineralized 12% of unfractionated DOP. Single cultures of lake water bacteria displayed low mineralizing activity (mean of 49 cultures = 5% DOP hydrolysed). Results indicate that DOP from Lough Neagh in the above molecular weight range is predominantly recalcitrant to bacterial mineralization under natural lake conditions.     

Apatite genesis: A biologically induced or biologically controlled mineral formation process?

Apatite formation from organic matter (ribonucleic acid) and calcium carbonate (cuttlebone) requires intervention of microorganisms. We have attempted to characterize this mineral formation process by locating the alkaline phosphatase and the crystals formed. Alkaline phosphatase, which is important for the liberation of the necessary components, was localized in the periplasmic space of Providencia rettgeri in the same manner as in Escherichia coli. Accordingly, the release of inorganic phosphate and the formation of apatite may occur at this site. However, electron microscope observations revealed the presence of extracellular apatite; moreover, apatite particles that were formed with or without bacteria (with alkaline phosphatase from hydrolyzed ribonucleic acid as phosphorus source) were closely similar in size and appearance. The formation of apatite can thus be qualified as biologically induced mineralization. Nevertheless, a bacterial cell can also act as a nucleator for apatite crystallization, but this would appear exceptional.     

Effects of Viruses on Nutrient Turnover and Growth Efficiency of Noninfected Marine Bacterioplankton

The effects of virus infection and lysis of a marine Vibrio sp. on C, N, and P turnover and the growth efficiency of noninfected bacterioplankton were studied in a series of dilution cultures. The cultures were enriched with various sources of organic matter and N and P. The growth of the Vibrio host and the growth of the natural bacterioplankton were measured by immunofluorescence and 4(prm1),6-diamidino-2-phenylindole staining methods, respectively. Lysis products resulting from infection of the Vibrio sp. caused an increase in metabolic activity and cell production by the noninfected bacterioplankton. In P-limited cultures, the addition of viruses increased the uptake of dissolved organic carbon by 72% and the potential alkaline phosphatase activity by 89% compared with control cultures without viruses. Our data suggest that input of available phosphorus through virus-induced Vibrio lysates occurred, which caused an increase in the bacterial nutrient uptake. The growth efficiency of noninfected bacteria was reduced in the presence of viruses compared with the control without viruses (growth efficiencies, 0.08 (plusmn) 0.03 and 0.24 (plusmn) 0.02, respectively). We suggest that the decrease in growth efficiency may be explained by an increase in bacterial energy demand associated with extracellular degradation of polymeric organic nitrogen and phosphorus in cell lysates.     

Phosphatase activity and its rôle in the mineralization of organic phosphorus in coastal sea water

Alkaline phosphatase activity in sea-water samples taken from Tokyo, Sagami, and Suruga Bays in Japan was measured by a sensitive fluorometric method. There is a relationship between the phosphatase activity and bacterial biomasses in these three bays. The phosphatase-producing bacteria accounted for 40, 46, and 41% of heterotrophic bacteria in Tokyo, Sagami and Suruga Bays, respectively. Significant amounts of phosphatase-hydrolysable organic phosphorus were found in the euphotic zones of these bays and this organic phosphorus fraction accounted for 19 and 50% of organic phosphorus in Tokyo and Sagami Bays, respectively. Phosphatase-hydrolysable organic phosphorus was decomposed completely in the euphotic zone suggesting that this organic fraction is re-cycled in the primary production of the bays. Decomposition of natural organic phosphorus by enzymes was followed by measuring the release of inorganic orthophosphate from samples saturated with chloroform. Release of inorganic phosphorus proceeds rapidly for 2 or 3 days followed by a slow release. Enzymes present in the samples contributed to the decomposition of 50% of the organic phosphorus and the phosphatase enzyme was responsible for the decomposition of one-third of the hydrolysable organic phosphorus in the samples. Potential phosphatase activity in the samples was found to be indicative of the extent and rate of decomposition of organic phosphorus in coastal waters.