Phosphatase activity of extra-radical arbuscular mycorrhizal hyphae: A review

Phosphatase activity of arbuscular mycorrhizal (AM) fungi has attracted attention in three fairly distinct domains: intracellular enzymes with defined metabolic functions that have been studied in intraradical hyphae, histochemical staining of alkaline phosphatase as an indicator of fungal activity measured both intra- and extraradically, and extracellular activity related to mineralization of organic P (P_o) compounds that may enhance mycorrhizal utilization of an important nutrient pool in soil. This review focuses on the latter subjects with emphasis on extraradical mycelium (ERM), while it draws on selected data from the vast material available concerning phosphatases of other organisms. We conclude that histochemical staining of alkaline phosphatase is a sensitive and suitable method for monitoring the effect of adverse conditions encountered by ERM both as a symbiotically functional entity in soil, and in vitro without modifying interference of soil or other solid substrates. Furthermore, the quantitative importance of extracellular enzymes for P nutrition of AM plants is estimated to be insignificant. This is concluded from the low quantitative contribution extracellular hyphae of AM fungi give to the total phosphatase activity in soil, and from estimations of which processes that may be rate limiting in organic P mineralization. Maximum values for the former is in the order of a few percent. As for the latter, solubilization of P_o seems to be far more important than P_o hydrolysis for utilization of P_o by AM fungi and plants, as both endogenous soil phosphatase activity and phosphatases of other soil organisms are ubiquitous and abundant. Our discussion of mycorrhizal phosphatases supports the view that extracellular phosphatases of roots and micro-organisms are to a large extent released incidentally into soil, and that the source has limited benefit from its activity. This revised version was published online in June 2006 with corrections to the Cover Date.     

Hydrolytic enzyme activities of extracted humic substances during the vermicomposting of a lignocellulosic olive waste

Humic substances and three hydrolytic enzymes (beta-glucosidase, phosphatase and urease) were extracted by neutral sodium pyrophosphate from an olive waste (dry olive cake), alone or mixed with municipal biosolids, during a nine month vermicomposting process. Easily degradable compounds decreased during the vermicomposting process because of microbial consumption. When municipal biosolids were added to dry olive cake, microbial activity increased and the amounts of compounds extracted by pyrophosphate were three times lower than olive cake alone. In both instances, beta-glucosidase, phosphatase and urease activities of the organic extracts either increased or remained the same after a nine month period of vermicomposting, thus suggesting that the humus enzyme complexes resisted microbial and earthworm attack. It is known that humus immobilised enzymes also remain active in soil environments, reactivating the nutrient cycles in soil. The use as amendments of vermicomposted olive cake, alone or when mixed with biosolids, could be a good alternative to reactivate the C, P and N-cycles in degraded soils for regeneration purposes.     

Ancient DNA reveals traces of Iberian Neolithic and Bronze Age lineages in modern Iberian horses

Multiple geographical regions have been proposed for the domestication of Equus caballus . It has been suggested, based on zooarchaeological and genetic analyses that wild horses from the Iberian Peninsula were involved in the process, and the overrepresentation of mitochondrial D1 cluster in modern Iberian horses supports this suggestion. To test this hypothesis, we analysed mitochondrial DNA from 22 ancient Iberian horse remains belonging to the Neolithic, the Bronze Age and the Middle Ages, against previously published sequences. Only the medieval Iberian sequence appeared in the D1 group. Neolithic and Bronze Age sequences grouped in other clusters, one of which (Lusitano group C) is exclusively represented by modern horses of Iberian origin. Moreover, Bronze Age Iberian sequences displayed the lowest nucleotide diversity values when compared with modern horses, ancient wild horses and other ancient domesticates using nonparametric bootstrapping analyses. We conclude that the excessive clustering of Bronze Age horses in the Lusitano group C, the observed nucleotide diversity and the local continuity from wild Neolithic Iberian to modern Iberian horses, could be explained by the use of local wild mares during an early Iberian domestication or restocking event, whereas the D1 group probably was introduced into Iberia in later historical times.     

汞对土壤酶活性的影响

利用室内模拟方法,研究了重金属Hg对不同土样脲酶、转化酶和中性磷酸酶活性的影响.结果表明,Hg可显著地抑制土壤脲酶和转化酶的活性,但不同土样Hg对两种酶活性的抑制程度有很大差别.HgCl₂浓度与两种酶活性之间的关系均可用对数方程很好地描述(P<0.05).4个土样的脲酶ED₅₀(生态剂量)分别为87.99、5.47、24.05和19.88 mg·kg^-1;转化酶的ED₅₀分别为76.68、727.49、236.52和316.59 mg·kg^-1.脲酶对Hg污染比转化酶敏感;有机质对土壤酶活性有一定的保护作用.除连续2年施用大量有机肥的草甸棕壤土样中Hg对中性磷酸酶有显著的激活作用外(P<0.05),其它土样无显著变化,表明中性磷酸酶活性对Hg污染反应不敏感.     

碳添加下黑钙土胞内、胞外脲酶活性变化及其机制

土壤脲酶作为能够催化尿素水解的最重要酶类,对草地生态系统氮素供应具有重要作用。目前探讨不同碳添加对草地土壤胞外脲酶影响的研究报道相对较多,但碳添加对土壤胞内脲酶的影响,以及胞内和胞外脲酶对碳添加的响应是否一致等尚需深入研究。本研究依托额尔古纳森林草原过渡带生态系统研究站开展的碳添加野外试验平台(以葡萄糖为碳源),选取无碳添加(C₀)、250(C₂₅₀)和500(C₅₀₀) kg C·hm^-2·a^-1处理为供试对象,探讨碳添加下黑钙土胞内、胞外脲酶活性响应及其与土壤性质的关系。结果表明: 碳添加显著提高了土壤胞内脲酶活性,增加了土壤胞内脲酶活性占总脲酶活性的比例,但对土壤胞外脲酶活性没有显著影响。土壤胞内脲酶活性与微生物生物量具有显著正相关关系,表明胞内脲酶活性增加主要是由微生物生物量增加引起的。结构方程模型(SEM)分析表明,碳添加通过影响土壤微生物生物量间接提高了土壤胞内脲酶活性。     

华西雨屏区不同退耕模式细根(包括草根)分解过程中土壤酶动态

采用原状土芯(intact core)法, 探讨了四川洪雅柳江退耕模式——光皮桦(Betula luminifera)与扁穗牛鞭草(Hemarthria compressa)复合模式(HN)、扁穗牛鞭草草地模式(NC)、柳杉(Cryptameria fortunei)人工林模式(LS)、光皮桦人工林模式(H)细根(包括草根)分解过程中土壤酶动态。结果表明: 1) HN下的土壤脲酶、蔗糖酶、酸性磷酸酶活性较大, LS下的土壤脲酶、酸性磷酸酶活性最小, 显著低于其他模式(p < 0.05)。2) HN、NC和LS下的土壤脲酶与细根(包括草根)分解速率显著相关, HN的蔗糖酶、NC的酸性磷酸酶、LS的多酚氧化酶活性与细根(包括草根)分解速率也呈显著正相关关系(p < 0.05)。3) 除H外, 土壤脲酶活性与细根C/N、纤维素绝对含量呈显著负相关关系(p < 0.05); 除NC外, 多酚氧化酶活性与细根纤维素绝对含量呈显著负相关关系。4)土壤脲酶活性与需氧固氮细菌或与真菌数量显著相关, HN下的土壤蔗糖酶活性与细菌和纤维素分解菌数量呈正相关关系, H与NC下的土壤酸性磷酸酶还分别与细菌和纤维素分解菌数量呈正相关关系(p < 0.05)。以上结果显示: 由光皮桦与扁穗牛鞭草不同生活型植物构成的复合模式有利于土壤酶活性的提高; 土壤脲酶活性高低能够反映这几种退耕模式细根(包括草根)分解速率的快慢, 细根(包括草根)的C/N是影响土壤脲酶活性的一个重要因素; 土壤酶活性与土壤真菌、需氧固氮细菌、纤维分解菌及细菌数量有关。     

Activity of acid and alkaline phosphatases (intracellular and extracellular) in rhyzosphere fungi from Arachis hypogaea (Papiloneaceae)

The potential role of the fungi, isolated from the peanut rhizosphere, in the production of extracellular and intracellular acid and alkaline phosphatase, was evaluated in vitro. Acid and alkaline extracellular phosphatases showed the highest activities, and the Penicillium species were the most efficient in their production. The correlation analysis showed that extracellular acid and intracellular acid phosphatase produced by Aspergillus niger A. terreus, Penicillium sp. y P. brevicompactum were negatively correlated; while the extracellular and intracellular phosphatase activities, were positively correlated. The extracellular acid phosphatase activities produced in vitro by majority of fungi assayed, were not correlated with the acid phosphatase activity present in the peanut soil rhizosphere. Nevertheless, the extracellular alkaline phosphatase activities produced in vitro, were negatively correlated with the extracellular alkaline phosphatase activities present in the rhizosphere. The ability of phosphatase production by fungi isolated from peanut rhizosphere suggests they have great potential to contribute to the P mineralization in this zone.     

Assessing temporal and geographic contacts across the Adriatic Sea through the analysis of genome-wide data from Southern Italy

Southern Italy was characterised by a complex prehistory that started with different Palaeolithic cultures, later followed by the Neolithization and the demic dispersal from the Pontic-Caspian Steppe during the Bronze Age. Archaeological and historical evidences point to a link between Southern Italians and the Balkans still present in modern times. To shed light on these dynamics, we analysed around 700 South Mediterranean genomes combined with informative ancient DNAs. Our findings revealed high affinities of South-Eastern Italians with modern Eastern Peloponnesians, and a closer affinity of ancient Greek genomes with those from specific regions of South Italy than modern Greek genomes. The higher similarity could be associated with a Bronze Age component ultimately originating from the Caucasus with high Iranian and Anatolian Neolithic ancestries. Furthermore, extremely differentiated allele frequencies among Northern and Southern Italy revealed putatively adapted SNPs in genes involved in alcohol metabolism, nevi features and immunological traits.     

Microbiological indicators of soil quality and degradation following conversion of native forests to continuous croplands

Deforestation resulting from forest conversion to agricultural land use is an important issue worldwide. This phenomenon is known to influence the activity and size of soil microbial community due to changes in environmental conditions with subsequent losses of soil organic matter (SOM) and soil quality degradation. The objective of this study was to investigate the relationship between soil organic carbon (SOC) losses and enzyme activities following land use conversion from native forests to continuous croplands. The amount of soil microbial biomass carbon (SMBC) and the activity of five soil enzymes (i.e., urease, invertase, alkaline phosphatase, acid phosphatase and arylsulfatase) were measured in croplands derived from forests and adjacent natural forests all on similar soil type at Gorgan site located in Northeast Iran. The content of SMBC decreased (47–83%) with deforestation at both soil sampling depths (0–20 and 20–40 cm). With the exception of phosphatases, the absolute activities of soil enzymes (activity on a soil mass basis) tended to decrease significantly (15–35%) with continuous cultivation. However, the specific enzyme activities expressed either per unit of SOC or SMBC tended to increase (about 1.5–5.5 times) with conversion of forestlands to croplands. The significant positive correlation between enzyme activity per SMBC and C turnover rate may imply that a faster C cycle and loss due to deforestation is related to a greater enzymatic activity by a smaller size of microbial biomass in cropland soils. In brief, the specific activities of soil enzymes could be used to reveal SOM losses and soil degradation in natural forest ecosystems, and to identify changes in soil quality and fertility following deforestation. Changes or improvements in soil management such as cessation of cultivation or implementing agricultural practices that stop or minimize soil disturbance are most likely needed to stop further soil degradation, restore soil quality and rebuild SOC stocks to offset CO₂ emissions in these ecosystems.     

Differentiating Intracellular from Extracellular Alkaline Phosphatase Activity in Soil by Sonication

Differentiating intracellular from extracellular enzyme activity is important in soil enzymology, but not easy. Here, we report on an adjusted sonication method for the separation of intracellular from extracellular phosphatase activity in soil. Under optimal sonication conditions [soil:water ratio  =  1/8 (w/v) and power density  =  15 watt ml^-1], the activity of alkaline phosphomonoesterase (phosphatase) in a Haplic Cambisol soil increased with sonication time in two distinct steps. A first plateau of enzyme activity was reached between 60 and 100 s, and a second higher plateau after 300 s. We also found that sonication for 100 s under optimal conditions activated most (about 80%) of the alkaline phosphatase that was added to an autoclaved soil, while total bacteria number was not affected. Sonication for 300 s reduced the total bacteria number by three orders of magnitude but had no further effects on enzyme activity. Our results indicate that the first plateau of alkaline phosphatase activity was derived from extracellular enzymes attached to soil particles, and the second plateau to the combination of extracellular and intracellular enzymes after cell lysis. We conclude that our adjusted sonication method may be an alternative to the currently used physiological and chloroform-fumigation methods for differentiating intracellular from extracellular phosphatase activity in soil. Further testing is needed to find out whether this holds for other soil types.     

线粒体全基因组揭示嫩江流域史前人群遗传结构的动态变化

嫩江流域是中国东北地区古代先民的重要栖息地之一。自新石器时代开始这里的先民一直以渔猎经济为主要生活方式,直到新石器时代晚期至早期青铜时代才开始兼营畜牧业和少量的种植业。嫩江流域青铜时代的生业模式的转变是否伴随着外来人群的融合与替代一直是考古研究的热点。为了探讨嫩江流域新石器时代与青铜铁器时代人群的构成是否改变,我们对嫩江流域新石器时代至青铜铁器时代的24个个体进行了线粒体全基因组分析。分析结果表明：嫩江流域青铜铁器时代人群与新石器时代人群具有一定遗传连续性的同时,晚期人群与西辽河地区古代人群有着更近的遗传联系,表明西辽河地区古代居民对嫩江流域青铜铁器时代人群具有部分遗传贡献。结合考古学文化、古气候学数据以及语言学证据,我们推测距今4000-3000年间,西辽河地区古代居民曾迁入到嫩江流域,并留下遗传印记。     

Nitrogen inputs accelerate phosphorus cycling rates across a wide variety of terrestrial ecosystems

? Biologically essential elements--especially nitrogen (N) and phosphorus (P)--constrain plant growth and microbial functioning; however, human activities are drastically altering the magnitude and pattern of such nutrient limitations on land. Here we examine interactions between N and P cycles of P mineralizing enzyme activities (phosphatase enzymes) across a wide variety of terrestrial biomes. ? We synthesized results from 34 separate studies and used meta-analysis to evaluate phosphatase activity with N, P, or N×P fertilization. ? Our results show that N fertilization enhances phosphatase activity, from the tropics to the extra-tropics, both on plant roots and in bulk soils. By contrast, P fertilization strongly suppresses rates of phosphatase activity. ? These results imply that phosphatase enzymes are strongly responsive to changes in local nutrient cycle conditions. We also show that plant phosphatases respond more strongly to fertilization than soil phosphatases. The tight coupling between N and P provides a mechanism for recent observations of N and P co-limitation on land. Moreover, our results suggest that terrestrial plants and microbes can allocate excess N to phosphatase enzymes, thus delaying the onset of single P limitation to plant productivity as can occur via human modifications to the global N cycle.     

Genetic variation in prehistoric Sardinia

We sampled teeth from 53 ancient Sardinian (Nuragic) individuals who lived in the Late Bronze Age and Iron Age, between 3,430 and 2,700 years ago. After eliminating the samples that, in preliminary biochemical tests, did not show a high probability to yield reproducible results, we obtained 23 sequences of the mitochondrial DNA control region, which were associated to haplogroups by comparison with a dataset of modern sequences. The Nuragic samples show a remarkably low genetic diversity, comparable to that observed in ancient Iberians, but much lower than among the Etruscans. Most of these sequences have exact matches in two modern Sardinian populations, supporting a clear genealogical continuity from the Late Bronze Age up to current times. The Nuragic populations appear to be part of a large and geographically unstructured cluster of modern European populations, thus making it difficult to infer their evolutionary relationships. However, the low levels of genetic diversity, both within and among ancient samples, as opposed to the sharp differences among modern Sardinian samples, support the hypothesis of the expansion of a small group of maternally related individuals, and of comparatively recent differentiation of the Sardinian gene pools. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

New approaches to establish optimum moisture content for compostable materials

This paper reports on the persistence of total and immobilised enzyme activities (urease and phosphatase) in a soil amended with organic wastes containing high levels of total-urease and phosphatase activity. Fresh organic materials showed the highest values for both total-enzymatic activities. The addition of organic waste to soil increased both total-enzymatic activities in the soil, which, after 360 days, showed values above those of the control. Immobilised enzymes were also higher in the fresh wastes than in the soil with compost, while the specific enzymatic activity levels (enzymatic activity per unit of carbon) were similar. The immobilised urease activity was greater in the amended soil than in the control. At the beginning of the incubation period, the immobilised urease activity was significantly higher in the soil amended with fresh organic wastes than with compost. However, this activity decreased with incubation, whilst the compost-immobilised urease activity increased with time. The effect of organic amendment on immobilised phosphatase activity was similar to that shown by immobilised urease but less pronounced. The persistence of both enzymes was significantly higher in the soil amended with compost than in that amended with fresh materials.     

Phosphomonoesterase hydrolysis of polyphosphoinositides in rat kidney: Properties and subcellular localization of the enzyme system.

Tthe properties of diphosphoinositide and triphosphoinositide phosphatases from rat kidney homogenate were studied in an assay system in which non-specific phosphatase activity was eliminated. The enzymes were not completely metal-ion dependent and were activated by Mg2+. The detergent sodium deoxycholate, Triton X-100 and Cutscum inhibited the reaction; cetyltrimethylammonium bromide only activated when added with the subtrates and in the presence Mg2+. Both enzymes had a pH optimum of 7.5. Ca2+ and Li+ both activated triphosphoinositide phosphatase, but Ca2+ inhibited and L+ had little effect on diphosphoinositide phosphatase. Cyclic AMP had no effect on either enzyme. The enzymes were three times more active in kidney cortex than in the medulla. On subcellular fractionation of kidney-cortex homogenates by differential and density-gradient centrifugation, the distribution of the enzymes resembled that of thiamin pyrophosphatase (assayed in the absence of ATP), suggesting localization in the Golgi complex. However, the distribution differed from that of the liver Golgimarker galactosyltransferase. Activities of both diphosphoinositide and triphosphoinositide phosphatases and thiamin pyrophosphatase were low in purified brush-border fragments. Further experiments indicate that at least part of the phosphatase activity is soluble.     

Localization of alkaline phosphatase in cells of Bacillus intermedius

Alkaline phosphatase, an enzyme secreted by Bacillus intermedius S3-19 cells to the medium, was also detected in the cell wall, membrane, and cytoplasm. The relative content of alkaline phosphatase in these cell compartments depended on the culture age and cultivation medium. The vegetative growth of B. intermedius on 0.3% lactate was characterized by increased activity of extracellular and membrane-bound phosphatases. The increase in lactate concentration to 3% did not affect the activity of membrane-bound phosphatase but led to a decrease in the activity of the extracellular enzyme. Na2HPO4 at a concentration of 0.01% diminished the activity of membrane-bound and extracellular phosphatases. CoCl2 at a concentration of 0.1 mM released membrane-bound phosphatase into the medium. By the onset of sporulation, phosphatase was predominantly localized in the medium and in the cell wall. As is evident from zymograms, the multiple molecular forms of phosphatase varied depending on its cellular localization and growth phase.     

Phosphate regulation of phosphatases in submerged cultures of Claviceps purpurea 129 producing clavine alkaloids

Summary Claviceps purpurea strain 129 was cultivated under submerged conditions in a sucrose-citrate medium containing high (36.8 mM) or low (1.84 mM) KH₂PO₄ concentrations. The permeabilized cells and culture supernatants contained alkaline and acid phosphatases. In the medium containing a high phosphate concentration, the synthesis of extracellular phosphatases was repressed, but that of cellular phosphatases was not. Extracellular phosphatases, especially alkaline phosphatases, were derepressed by transferring the mycelium into a phosphate-free medium. This derepression was inhibited by cycloheximide. In the presence of cycloheximide, the activities of the cellular phosphatases decreased markedly, indicating turnover of these enzymes. The cellular acid phosphatase was inhibited by phosphate (0.025 M–0.1 M) and NaF (0.01 M) while the cellular alkaline phosphatase was only inhibited by phosphate. Both cellular and extracellular alkaline phosphatases were more sensitive to repression by phosphate than the acid phosphatases. The alkaloid synthesizing enzymes were: a) present in mycelia grown in high levels of phosphate and b) activated by decreasing the intracellular phosphate level.     

ACCUMULATION OF WATER SOLUBLE PHOSPHORUS AND HYDROLYSIS OF POLYPHOSPHATES BY CLADOPHORA GLOMERATA (CHLOROPHYCEAE)1,2

Concentrations of hot-water extractable phosphorus from most samples of Cladophora glomerata (L.) Kütz. were relatively high (0.06–0.68%) and correlated closely with total dissolved P in ambient Lake Michigan water. Cladophora was able to hydrolyze polyphosphates by enzymes found in intracellular, extracellular and cell wall fractions. The intracellular phosphatase activity is pH dependent with the optimal hydrolysis rate at pH 7.8. Secretion of phosphatases is affected by pH, with maximum rate at 7, but affected little by light intensity. Magnesium is the most effective metallic cofactor required for maximal rates of intracellular phosphatase activities.     

模拟氮沉降增加条件下土壤团聚体对酶活性的影响

氮沉降增加改变了森林土壤生态系统物质输入,影响土壤生物及酶活性,而土壤团聚体内相对稳定的微域生境可能减弱或延缓土壤生物和酶对氮沉降增加的响应强度。以广东省东莞大岭山森林公园荷木人工林为研究对象,用模拟N沉降方法,分析了2011年12月到2012年11月一年内氮沉降增加条件下表层混合土壤和土壤团聚体内脲酶、蔗糖酶和酸性磷酸酶活性的变化及影响因素,旨在理解氮沉降增加条件下土壤团聚体对酶活性的影响。结果表明:氮沉降增加对表层混合土壤中脲酶和蔗糖酶的抑制作用不显著,而酸性磷酸酶受氮沉降显著影响,表现为低氮(50 kg N hm-2a-1)促进,高氮(300 kg N hm-2a-1)抑制的规律。表层土壤团聚体内脲酶活性随氮沉降增加而降低,N300处理显著低于对照;蔗糖酶和酸性磷酸酶活性随氮沉降增加先降低后增加,N100处理最低,分别比其他处理降低了6.46%—25.53%和42.33%—68.25%。试验区内各粒径土壤团聚体内酶活性高于混合土壤,且随团聚体粒径增加酶活性均为先增加后降低。不同粒径土壤团聚体的3种酶活性均以2—5 mm最高,但脲酶、酸性磷酸酶在各团聚体粒径间差异不显著,蔗糖酶活性2—5 mm显著高于5—8 mm。土壤酶相对活性指数和相对活性综合指数结果显示,超过85%的团聚体粒径内的相对酶活性指数大于1,而土壤酶相对活性综合指数均大于1。以上结果表明,氮沉降增加条件下土壤团聚体对其团聚体内的土壤酶活性有隔离保护作用,但其隔离保护效果与酶的种类和土壤团聚体粒径有关。     

Differential Distribution of Metals and Enzymes in Quanzhou Bay Estuarine Wetland Soils under Three Mangrove Species

This study aimed to investigate the effects of three dominant mangrove species (Avicennia marina, Aegiceras corniculatum, and Kandelia obovata) on the distribution of the metal elements Fe, Mn, Cu, and Zn and the enzymes urease, phosphatase, polyphenol oxidase, and catalase. Concentrations of the metallic elements and enzymatic activities were quantified in soils under the three mangrove species in the Quanzhou Bay estuarine wetland, a typical coastal wetland in China. Results showed that A. corniculatum promoted aggregation of Mn, urease, and phosphatase, while K. obovata contributed to accumulation of Cu, urease, and phosphatase. Furthermore, A. marina induced activation of Zn and accumulation of urease, phosphatase, and catalase. The characteristic enzyme and metal distributions induced by the different mangrove species are likely to result from different planting times, root systems, and soil pH and salinity. Moreover, the three mangrove species were found to influence the diversity of elemental and enzymatic stoichiometry through differences in the soil microenvironment, which can promote biodiversity in wetland ecosystems. These findings provide a useful guideline to develop strategies for restoration of estuarine wetlands.