The effect of afforestation with Scots pine (Pinus silvestris L.) of sandy post-arable soils on their selected properties. II. Reaction, carbon, nitrogen and phosphorus

Despite the extensive literature on the effect on soil properties of afforestation of former arable land, we still lack full understanding of whether the changes proceed in the same direction and at the same rate, and of how long is required to achieve a state of soil equilibrium typical of a natural forest ecosystem. Therefore, as part of a study comparing post-arable sandy soils (Dystric Arenosols) afforested with Scots pine (Pinus silvestris L.) with arable soils and soils of continuous coniferous forests, the range and direction of changes in pH, organic carbon (C_org), total nitrogen (N_tot), ammonium (N-NH₄) and nitrates (N-NO₃) in soil solution, total (P_tot) and available (P_av) phosphorus were determined. The studies were carried out in south-east Poland (51°30′-51°37′N, 22°20′-22°35′E). Ten paired sites of afforested soils (five with 14- to 17-year-old stands and five with 32- to 36-year-old stands) with adjacent cultivated fields, and five sites of continuous forest with present stands of ca. 130–150 years old were selected. Soil samples were taken from the whole thickness of master horizons and, in the case of the A horizon of the afforested soils, from three layers: 0–5 (A_0–5), 5–10 (A_5–10) and 10–20 cm (A_10–20). The cultivated soils in the Ap horizon showed higher pH (by ca. 1.0 unit), lower C_org and C:N, similar N_tot, lower N-NH₄, higher N-NO₃, higher P_tot and P_av contents compared with the Ah horizon of continuous forest soils. The results indicated decreased soil pH in the former plough layer of the afforested soils, with the greatest decrease observed in the 0–5 cm layer. In these soils, the C_org content was considerably higher in the A_0–5 layer, but lower in the two deeper layers and in the whole A horizon (0–20 cm) compared with the Ap horizon of the arable soils. The results indicate that the C_org content, after an initial phase of decline, again achieved a level characteristic of arable soils. The N_tot content in all layers of the A horizon of the afforested soils was lower than in the Ap horizon of the arable soils, and showed a reduction with stand age, especially in deeper layers. The C:N ratios in the mineral topsoil increased with stand age. N-NH₄ content increased and N-NO₃ decreased after afforestation. The P_tot and P_av contents in all layers and in the whole A horizon of the afforested soils, on stands of both ages, was lower than in the Ap of the cultivated soils. From the results, it could be concluded that, after more than 30 years of tree growth, the soils of the A horizon were still more similar to arable than to continuous forest soils with respect to C_org, P_tot and P_av. With respect to pH, N-NH₄ and N-NO₃, especially in the 0–5 cm layer, they were more similar to continuous forest soils than to cultivated soils, but with respect to N_tot and C:N ratio they were somewhere in between.     

Soil changes in different age classes of Norway spruce (Picea abies (L.) Karst.) on afforested farmland

The aim of this study was to test the hypothesis that afforestation changes the content and distribution of soil organic carbon, nutrients and pH in the A-horizon of land previously used in agriculture, and that such soil changes depend on stand development. The investigation was evaluated as a completely randomised design with three treatments representing different age classes of trees: 20 years (Y20), 40 years (Y40) and 55 years (Y55). Eighteen trial plots, six per treatment, were established in plantations of Picea abies (L.) Karst. on soils of similar texture and mineralogy. Tree volume was 220 m³ ha^-1 in Y20, 400 in Y40 and 440 m³ ha^-1 in Y55.Concentrations of carbon (C) and nitrogen (N) were significantly higher in the uppermost part of the soil in the older stands Y40 and Y55 than in Y20. The total amount of organic C in the litter layer plus the top 15 cm of the soil differed between age classes, with Y40 and Y55 having the largest amounts. A reference layer (15–20 cm) was used in calculating the amount of soil C that had accumulated in the horizon since afforestation, being about 10 tonnes ha^-1 of C in Y20 and 19 tonnes ha^-1 in Y40 and Y55.Cation exchange capacity (CEC) and base saturation (BS) was higher in the older stands. Carbon contents and CEC were strongly correlated. In Y40 and Y55, pH was significantly lower than in Y20 in the lower part of the soil horizon. There was a general decrease with depth of C, N, CEC, K⁺ and Mg²⁺ in the soil horizon. BS, Ca²⁺, Na⁺ and pH showed a somewhat different pattern of distribution, with deceasing values in the upper part of the soil horizon and increasing values in the lower part of the soil horizon.Abbreviations BD Bulk density - CEC cation exchange capacity - BS base saturation - Ca²⁺ calcium ion - Mg²⁺ magnesium ion - K⁺ potassium ion - Na⁺ sodium ion - C carbon - Ca _a accumulated carbon content - C _t total carbon content - N nitrogen - Y20 age class 20 years - Y40 age class 40 years - Y55 age class 55 years     

Effect of an epoxy derivative of 2′5′-trioligoadenylate on human neuroblastoma cells

We studied the effect of an epoxy derivative of dephosphorylated 2′,5′-trioligoadenylate (5′,5′ApApAepoxy) resistive to the action of cellular phosphodiesterase on cells of human neuroblastoma IMR 32 cultured in vitro. Twenty-two hours after the addition of 5·10⁻⁶ M 2′,5′ApApAepoxy to the culture medium, the number of cells decreased by 20% (P < 0.05), while the content of protein in these cells increased, on average, by 52% (P < 0.01), as compared with the control. The activities of Na⁺,K⁺-and Ca²⁺, Mg²⁺-ATPases in a microsomal fraction obtained from cells cultured in the presence of 2′, 5′ ApApAepoxy decreased by 50% (P < 0.001) as compared with those in the control cells. Our data indicate that 2′,5′ApApAepoxy possess antiproliferative activity. According to our findings, the antiproliferative effect of 2′,5′ ApApAepoxy can, to a great extent, be explained by the fact that this oligoadenylate derivative significantly modulates the activities of Na⁺,K⁺-and Ca²⁺,Mg²⁺-ATPases. Neirofiziologiya/Neurophysiology, Vol. 38, No. 2, pp. 97–102, March–April, 2006.     

Changes in soil properties after afforestation of former intensively managed soils with oak and Norway spruce

In many European countries, surplus agricultural production and ecological problems due to intensive soil cultivation have increased the interest in afforestation of arable soils. Many environmental consequences which might rise from this alternative land-use are only known from forest establishment on less intensively managed or marginal soils. The present study deals with changes in soil properties following afforestation of nutrient-rich arable soils. A chronosequence study was carried out comprising seven Norway spruce (Picea abies (Karst.) L.) and seven oak (Quercus robur L.) stands established from 1969 to 1997 on former horticultural and agricultural soils in the vicinity of Copenhagen, Denmark. For comparison, a permanent pasture and a ca. 200-year-old mixed deciduous forest were included. This paper reports on changes in pH values, base saturation (BS_eff), exchangeable calcium, soil N pools (N_min contents), and C/N ratios in the Ap-horizon (0–25 cm) and the accumulated forest floor. The results suggest that afforestation slowly modifies soil properties of former arable soils. Land-use history seems to influence soil properties more than the selected tree species. An effect of tree species was only found in the forest floor parameters. Soil acidification was the most apparent change along the chronosequence in terms of a pH decrease from 6 to 4 in the upper 5 cm soil. Forest floor pH varied only slightly around 5. Nitrogen storage in the Ap-horizon remained almost constant at 5.5 Mg N ha^–1. This was less than in the mineral soil of the ca. 200-year-old forest. In the permanent pasture, N storage was somewhat higher in 0–15 cm depth than in afforested stands of comparable age. Nitrogen storage in the forest floor of the 0–30-year-old stands increased in connection with the build-up of forest floor mass. The increase was approximately five times greater under spruce than oak. Mineral soil C/N ratios ranged from 10 to 15 in all stands and tended to increase in older stands only in 0–5 cm depth. Forest floor C/N ratios were higher in spruce stands (26.4) as compared to oak stands (22.7). All stands except the youngest within a single tree species had comparable C/N ratios.     

Patterns and mechanisms of soil acidification in the conversion of grasslands to forests

Grassland to forest conversions currently affect some of the world's most productive regions and have the potential to modify many soil properties. We used afforestation of native temperate humid grassland in the Pampas with eucalypts as an experimental system to 1) isolate forest and grassland imprints on soil acidity and base cation cycling and 2) evaluate the mechanisms of soil acidification. We characterized soil changes with afforestation using ten paired stands of native grasslands and Eucalyptus plantations (10–100 years of age). Compared to grasslands, afforested stands had lower soil pH (4.6 vs.5.6, p < 0.0001) and 40% lower exchangeable Ca (p < 0.001) in the top 20 cm of the soil. At three afforested stands where we further characterized soil changes to one meter depth, soil became increasingly acidic from 5 to 35 cm depth but more alkaline below 60 cm compared to adjacent grasslands, with few differences observed between 35 and 60 cm. These changes corresponded with gains of exchangeable acidity and Na in intermediate and deeper soil layers. Inferred ecosystem cation balances (biomass + forest floor + first meter of mineral soil) revealed substantial vertical redistributions of Ca and Mn and a tripling of Na pools within the mineral soil after afforestation. Soil exchangeable acidity increased 0.5–1.2 kmol_c.Ha^–1.yr^–1 across afforested stands, although no aboveground acidic inputs were detected in wet + dry deposition, throughfall and forest floor leachates. Our results suggest that cation cycling and redistribution by trees, rather than cation leaching by organic acids or enhanced carbonic acid production in the soil, is the dominant mechanism of acidification in this system. The magnitude of soil changes that we observed within half a century of tree establishment in the Pampas emphasizes the rapid influence of vegetation on soil formation and suggests that massive afforestation of grasslands for carbon sequestration could have important consequences for soil fertility and base cation cycles.     

137Cs and 40K isotopes in forest and wasteland soils in a selected region of eastern Poland 20 years after the Chernobyl accident

The vertical ¹³⁷Cs profile of forest and wasteland soils was analyzed in the south of the Podlasie Lowland area (Eastern Poland) about 20 years after the Chernobyl accident. In addition, the concentration of ⁴⁰K in soils of the investigated area was measured. Below the litter layer (mean thickness 3 cm), the soil samples were collected up to a depth of 12 cm and then divided into three layers: 0–3, 3–7, 7–12 cm. The behavior of ¹³⁷Cs and ⁴⁰K isotopes in soils was analyzed depending on the depth from which the soil samples were collected, as well as on the content of organic carbon, pH of soil and its granulometric composition. It was established that the density of ¹³⁷Cs in the litter layer equals 2.17 kBq m⁻²; it is the highest in layer 0–3 cm where it equals 3.44 kBq m⁻², and it decreases with the depth to the value of 0.76 kBq m⁻² in layer 7–12 cm. No similar pattern was observed in wasteland soils. The concentrations of ⁴⁰K in forest and wasteland soils did not change significantly with depth.     

Effect of Sesbania,Azolla and rice straw incorporation on the kinetics of NH4, K,Fe, Mn,Zn and P in some flooded rice soils

In a greenhouse study, with and without rice plants, of five flooded Philippine rice soils whose organic C (OC) content varied from 0.5 to 3.6%, incorporation ofSesbania rostrata, Azolla microphylla and rice straw affected the kinetics of soil solution NH ₄ ⁺ −N, K⁺, Fe²⁺, Mn²⁺, Zn²⁺, and P. Sesbania and Azolla increased NH ₄ ⁺ −N concentration above the control treatment, whereas rice straw depressed it. In all soils Azolla released less NH ₄ ⁺ −N than Sesbania. The apparent net N release depended on the soil and ranged from 44–81% for Sesbania and 27–52% for Azolla. These effects persisted throughout the growth of IR36. Soil solution and exchangeable NH ₄ ⁺ −N increased initially but levelled off between 30 to 80 days and between 20 to 40 days after flooding (DF), respectively. With rice, soil solution NH ₄ ⁺ −N concentration, reached a peak at 15–40 DF and declined to very low levels (<4mg L⁻¹). In the 3 soils of low OC content nitrogen derived from green manure ranged from 34–53% and the apparent revovery of added green manure N varied from 29–67%. Almost all N released from both Azolla and Sesbania were recovered in the rice plant in all soils except Concepcion with only 77%. The concentration of K⁺, Fe²⁺, Mn²⁺ and P in the soil solution were higher with rice straw than Sesbania and Azolla in all soils except Hanggan which showed no change in Fe²⁺ and Mn²⁺ but increased K⁺ and P. In general, rice straw, Sesbania and Azolla decreased Zn²⁺ concentration in all soils.     

Effects of land use on 15N natural abundance of soils in Ethiopian highlands

We used the natural abundance of ¹⁵N in soils in forests, pastures and cultivated lands in the Menagesha and Wendo-Genet areas of Ethiopia to make inferences about the N cycles in these ecosystems. Since we have described the history of these sites based on variations in ¹³C natural abundance, patterns of δ¹⁵N and δ¹³C values were compared to determine if shifts of ¹⁵N correlate with shifts of vegetation. At Menagesha, a > 500-yr-old planted forest, we found δ¹⁵N values from −8.8 to +3.5‰ in litter, from −3.5 to +4.5‰ in 0–10 cm soil layer, and from −1.5 to +6.8‰ at >20 cm soil depth. The low δ¹⁵N in litter and surface mineral soils suggests that a closed N cycle has operated for a long time. At this site, the low δ¹³C of the surface horizon and the high δ¹³C of the lower soil horizons is clear evidence of a long phase of C₄ grass dominance or cultivation of C₄ crops before the establishment of the forest >500 years ago. In contrast, at Wendo-Genet, high δ¹³C of soils reveals that most of the land has been uncovered by forests until recently. Soil δ¹⁵N was high throughout (3.4–9.8‰), and there were no major differences between forested, cultivated and pasture soils in δ¹⁵N values of surface mineral soils. The high δ¹⁵N values suggest that open N cycles operate in the Wendo-Genet area. From the points of view of soil fertility management, it is interesting that tall forest ecosystems with relatively closed N cycling could be established on the fairly steep slopes at Menagesha after a long period of grass vegetation cover or cultivation. This revised version was published online in June 2006 with corrections to the Cover Date.     

Kinetic and thermodynamic properties of wall bound invertase in heat tolerant and susceptible cultivars of wheat

We have identified two types of invertases, one bound ionically and the other covalently to the particulate fraction in grains of heat tolerant C 306 and heat susceptible WH 542 cultivars of wheat (Triticum aestivum L.). The cell walls contained a high level of invertase activity, of which 79.2–72.8% was extractable by 2 M NaCl and 14.9–21.1% by 0.5% EDTA in C 306 and WH 542, respectively. The NaCl-released invertase constituted the predominant fraction. Using 5–100 mM sucrose and pH range of 4.0–7.0, the apparent Michaelis constant (K _m, enzyme substrate affinity measure) of enzyme ranged from 5.73 to 16.06 mM for C 306 and from 6.08 to 19.86 mM for WH 542. The V _max (maximum catalytic rate) values at these pH were higher in C 306 (0.63–11.04 μg sucrose hydrolysed min⁻¹) than WH 542 (0.51–8.73 μg sucrose hydrolysed min⁻¹). By employing photo-oxidation and by studying the effect of pH on K _m and V _max, the involvement of histidine and α-carboxyl groups at the active site of the enzyme was indicated. The two cultivars also showed differential response in terms of thermodynamic properties of the enzyme i.e. energy of activation (E _a), enthalpy change (ΔH) and entropy change (ΔS). NaCl-released invertase showed differential response to metal ions in two cultivars suggesting their distinctive nature. Mn²⁺, Cu²⁺, Hg²⁺, Mg²⁺, Zn²⁺ and Cd²⁺ were strong inhibitors in WH 542 as compared to C 306 while K⁺, Ca²⁺ were stimulators in both the cultivars. Overall the results suggest that genetic differences exist in wall bound invertase properties of wheat grains as evident in its altered kinetic behaviour.     

Transgenic aequorin monitors cytosolic calcium transients in soybean cells challenged with β-glucan or chitin elicitors

Transgenic soybean (Glycine max L.) cells expressing aequorin were used to monitor changes in cytosolic Ca²⁺ concentrations in response to treatment with fungal elicitors. After an apparent lag phase of about 60 s, both chitin fragments and β-glucan elicitors caused a rapid increase in cytosolic Ca²⁺ concentration, which peaked within 2–2.5 min of treatment. The Ca²⁺ concentration then decreased and reached the basal level after about 5 min in the case of the treatment with chitin fragments, while a second rise in the Ca²⁺ concentration with a maximum occurring after about 7–8 min was observed in the case of β-glucan treatment. Calibration of the signals showed that the elicitors enhanced the cytosolic Ca²⁺ concentration from resting concentrations as low as 0.1 lM to highest levels of about 2 lM. Dose-response experiments showed that the concentration of elicitors giving a Ca²⁺ response at the 50% level was 0.4 nM for the chitin fragment and 28 lM and 72 lM, respectively, for a synthetic hepta-β-glucoside and a fungal β-glucan fraction. The β-glucan- or N,N′,N′′,N′′′-tetraacetyl chitotetratose (CH4)-induced Ca²⁺ signals were inhibited by both the Ca²⁺ chelator 1,2-bis-(2-aminophenoxy) ethane-N,N,N′,N′-tetraacetic acid (BAPTA) and by the Ca²⁺-channel inhibitor La³⁺. Neomycin, whose target in plant cells has not yet been clearly identified, reduced predominantly the expression of the second peak of the biphasic Ca²⁺ curve following β-glucan treatment. Bacterial cyclic β-glucans known to suppress β-glucan-induced phytoalexin production were also found to function as a suppressor for the Ca²⁺ response that was elicited by the fungal β-glucans. The results clearly show that the increase in the cytosolic Ca²⁺ concentration is an early and rapid event in the elicitor-sensing mechanism of soybean cells, and is probably connected with the subsequent activation of defence responses. Received: 23 July 1998 / Accepted: 16 October 1998     

Metal–bipyridine complexes in DNA backbones and effects on thermal stability

Modified oligonucleotides are showing potential for multiple applications, including drug design, nanoscale building blocks, and biosensors. In an effort to expand the functionality available to DNA, we have placed chelating ligands directly into the backbone of DNA. Between one and three nucleosides were replaced with 2,2′-bipyridine phosphates in 23-mer duplexes of DNA. An array of metal ions were added (Fe²⁺, Co²⁺, Ni²⁺, Cu²⁺, Zn²⁺, and Pt²⁺) and the influences on duplex stability were examined by melting temperature studies. Titrations and UV–vis absorption spectroscopy were used to provide insights into the nature of the metal complexes formed. We found that Ni²⁺ binding to 2,2′-bipyridine typically provided the greatest increase in duplex stability relative to the other metal ions examined. For example, addition of Ni²⁺ to one 2,2′-bipyridine–DNA duplex increased the melting temperature by 13 °C, from 65.0 ± 0.3 to 78.4 ± 0.9 °C. These studies show that metal ions and backbone ligands can be used to regulate DNA structure and stability.     

Two modes of sulfite oxidation in the extremely thermophilic and acidophilic archaeon Acidianus ambivalens

Sulfite-oxidizing enzyme activities were analyzed in cell-free extracts of aerobically grown cells of Acidianus ambivalens, an extremely thermophilic and chemolithoautotrophic archaeon. In the membrane and cytoplasmic fractions, two distinct enzyme activities were found. In the membrane fraction, a sulfite:acceptor oxidoreductase activity was found [530 mU (mg protein)^–1; apparent K _m for sulfite, 3.6 mM]. In the cytoplasmic fraction the following enzyme activities were found and are indicative of an oxidative adenylylsulfate pathway: adenylylsulfate reductase [138 mU (mg protein)^–1], adenylylsulfate:phosphate adenyltransferase [“ADP sulfurylase”; 86 mU (mg protein)^–1], adenylate kinase [650 mU (mg protein)^–1], and rhodanese [thiosulfate sulfur transferase, 9.2 mU (mg protein)^–1]. In addition, 5′,5′′′-P¹,P⁴-di(adenosine-5′) tetraphosphate (Ap₄A) synthase and Ap₄A pyrophosphohydrolase activities were detected. Received: 17 August 1998 / Accepted: 29 April 1999     

Indicators for nitrogen status and leaching in subtropical forest ecosystems,South China

The deposition of nitrogen (N) is high in subtropical forest in South China and it is expected to increase further in the coming decades. To assess effects of increasing deposition on N cycling, we investigated the current N status of two selected 40–45-year-old masson pine-dominated Chinese subtropical forest stands at Tieshanping (TSP, near Chongqing City) and Caijiatang (CJT in Shaoshan, Hunan province), and explored the applicability of several indicators for N status and leaching, suggested for temperate and boreal forest ecosystems. Current atmospheric N deposition to the systems is from 25 to 49 kg ha⁻¹ year⁻¹. The concentration of total N in the upper 15 cm of the soil is from as low as 0.05% in the B₂ horizon to as high as 0.53% in the O/A horizon. The concentration of organic carbon (C) varies from 0.74 (B₂) to 9.54% (O/A). Pools of N in the upper 15 cm of the soils range from 1460 to 2290 kg N ha⁻¹, where 25–55% of the N pool is in the O/A horizon (upper 3 cm of the soil). Due to a lack of a well-developed continuous O horizon (forest floor), the C/N ratio of this layer cannot be used as an indicator for the N status, as is commonly done in temperate and boreal forests. The net N mineralization rate (mg N g⁻¹ C year⁻¹) in individual horizons correlates significantly with the C/N ratio, which is from as high as 18.2 in the O/A horizon to as low as 11.2 in the B₂ horizon. The N₂O emission flux from soil is significantly correlated with the KCl extractable NH₄⁺–N in the O/A horizon and with the net nitrification in the upper 15 cm of the soil. However, the spatial and temporal variation of the N₂O emission rate is high and rates are small and often difficult to detect in the field. The soil flux density of mineral N, defined as the sum of the throughfall N input rate and the rate of in situ net N mineralization in the upper 15 cm of the soil, i.e., the combination of deposition input and the N status of the system, explains the NO₃⁻ leaching potential at 30 cm soil depth best. The seasonality of stream water N concentration at TSP and CJT is climatic and hydrologically controlled, with highest values commonly occurring in the wet growing season and lowest in the dry dormant season. This is different from temperate forest ecosystems, where N saturation is indicated by elevated NO₃⁻ leaching in stream water during summer.     

Effects of deforestation and cultivation on soil CEC and contents of exchangeable bases: A case study in Simlipal National Park,India

Deforestation in the tropics seems to be a serious problem probably because of the reduction in soil CEC and the consequent losses of nutrients from the soils. Here, changes in these parameters as influenced by deforestation as well as vegetative cover were studied; statistical methods were applied to interpret the results. Cultivation causes a significant reduction in CEC, total content of the exchangeable bases and exchangeable Ca2+ and Mg2+ levels compared to the adjoining unmanaged forest land. Levels of exchangeable K⁺ and Na⁺, however, do not change significantly. Evergreen forest soils have the highest levels of CEC, total exchangeable bases, exchangeable Ca²⁺ and K⁺. Deciduous forest, grassland and cultivated soils have statistically similar contents of exchangeable Ca²⁺, Mg²⁺, K⁺ and Na⁺. Exchangeable Mg²⁺, however, is not affected by vegetative cover. Soil CEC shows fairly good correlation with the organic carbon content only in evergreen forest soils. In others, organic carbon apparently does not influence CEC significantly. All soils show excellent correlation between their CEC and total exchangeable bases. It is concluded that for regeneration of weathered tropical soils, an evergreen cover provides the most effective means; deciduous vegetation or grass cover do not seem promising.     

Effect of dephosphorylated 2′,5′-trioligoadenylate on the entry of sodium ions into human neuroblastoma cells

Using a radioisotope technique, we studied the effect of dephosphorylated 2′,5′-trioligoadenylate (2′,5′ ApApA) on the entry of sodium ions into cultured human neuroblastoma cells (IMR 32 strain). Short-term (nearly 1 h) action of 2′,5′ ApApA did not influence the entry of sodium ions through voltage-operated sodium channels in the absence of neurotoxins modulating the characteristics of these channels (toxin of a scorpion, Leiurus quinquestriatus, + veratrine). At the same time, 2′,5′ ApApA weakened in a dose-dependent manner the entry of sodium ions through sodium channels opened upon the action of the above neurotoxins. In cells cultured for 22 h in a medium containing 5 · 10⁻⁶ M 2′,5′ ApApA, the entry of sodium ions in the absence of neurotoxins was 25% greater, while in the presence of neurotoxins it was 24% smaller than that in the control cells. Tetrodotoxin (TTX, 4 · 10⁻⁷ M) blocked completely sodium entry through sodium channels in cells cultured in the absence of 2′,5′ ApApA, while in cells cultured in the presence of this adenylate TTX decreased the entry by 64%. It is hypothesized that long-lasting action of 2′,5′ ApApA results in the appearance of voltage-operated TTX-insensitive sodium channels in the plasma membrane of IMR 32 cells. Our data show that 2′,5′ ApApA is capable of modulating to a considerable extent the functioning of mechanisms controlling transport of sodium ions in cells of human neuroblastoma cells of the IMR 32 strain. Neirofiziologiya/Neurophysiology, Vol. 40, No. 1, pp. 3–8, January–February, 2008.     

Purification and properties of extracellular nuclease from tobacco pollen

Proteins diffusing from tobacco pollen grains exhibit different phosphohydrolytic activities. Molecular sieving produces nuclease fractionation into forms I, II and III with apparent molecular masses ≥ 60 × 10³, 32.9 × 10³ and 24.6 × 10³, respectively, and separation of principal forms II and III from phosphatase and major part of 5′- and 3′-nucleotidase activities. These forms did not differ in the mode of substrate attack and were combined for further enzyme characterization. The preparation had 3′-nueleotidase activity even after further purification by DEAE-cellulose chromatography. The enzyme is an endonuclease with preference for single stranded molecules. The endolytical cleavage of native DNA occurs simultaneously in both strands and generates limit products of about 58 pairs of nucleotides. DNA duplex polymers are also cleaved by a terminally-directed, exonuclease-like process. The products of DNA degradation are oligonucleotides and 5′-mononucleotides. In the presence of NaCl, both endolytical and exonucleaselike activities on bihelical DNA are inhibited and the proportion of mono-to oligonucleotides produced increases. The enzyme can rapidly convert superhelical plasmid DNA to a nicked open circular form, and then to a unit-length linear molecule. On the basis of these properties and of those found earlier (sugar-unspecificity, acidic pH optimum, activation by Zn²⁺ ions), the extracellular nuclease of tobacco pollen can be classified as plant nuclease I (EC 3.1.30.x).     

Benzothiazole bipyridine complexes of ruthenium(II) with cytotoxic activity

A series of benzothiazole-substituted trisbipyridine ruthenium(II) analogues {[Ru(bpy)₂(4,5′-bbtb)]²⁺, [Ru(bpy)₂(5,5′-bbtb)]²⁺ and [Ru(bpy)₂(5-mbtb)]²⁺ [bpy is 2,2′-bipyridine, bbtb is bis(benzothiazol-2-yl)-2,2′-bipyridine, 5-mbtb is 5-(benzothiazol-2-yl),5′-methyl-2,2′-bipyridine]} have been prepared and compared with the complex [Ru(bpy)₂(4,4′-bbtb)]²⁺ reported previously. From the UV–vis spectral studies, substitution at the 5-position of the bpy causes the ligand-centred transitions to occur at considerably lower energy than for those with the functionality at the 4-position, while at the same time causing the emission to be effectively quenched. However, substitution at the 4-position causes the metal-to-ligand charge transfer to occur at lower energies. Fluorescent intercalator displacement studies indicate that the doubly substituted complexes displace ethidium bromide from a range of oligonucleotides, with the greater preference shown for bulge and hairpin sequences by the Λ enantiomer. Since the complexes only show small variation in the UV–vis spectra on the introduction of calf thymus DNA and a small increase in fluorescence they do not appear to be intercalators, but appear to associate within one of the grooves. All of the reported bisbenzothiazole complexes show reasonable cytotoxicity against a range of human cancer cell lines. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Kinetics of formation and stability of {Pt(dien)}2+ complexes with octamer and 14-mer DNA oligonucleotides containing a GG sequence

 Reaction of [Pt(dien)Cl]⁺ (1) with the 14-mer oligonucleotide 5′-d(ATACATGGTACATA) (I) gave rise to two major species which corresponded to the 5′-G and 3′-G platinated monofunctional adducts, and a minor amount of the bis-platinated adduct formed during the later stages of the reaction. The reaction of (1) with the related octamer 5′-d(ATACATGG) (II) was also investigated. Kinetic data obtained by HPLC showed that the 5′-G and 3′-G bases of the 14-mer oligonucleotide were platinated at similar rates: the second-order rate constant is 53×10^–2 M^–1 s^–1 at 298 K in 0.1 M NaClO₄. However, the platination rate of 5′-G of the octamer (II) (k=69×10^–2 M^–1 s^–1) was enhanced by a factor of three compared to the rate of platination at 3′-G (k=22×10^–2 M^–1 s^–1). All the adducts were separated by HPLC and characterized by NMR spectroscopy, enzymatic digestion and MALDI-TOF mass spectrometry. ¹H and ¹⁵N NMR shifts suggest that there are distinct conformational differences between 14-mer duplexes platinated at 5′-G (I5′ _ds) and 3′–G (I3′ _ds). Molecular mechanics modelling indicates that rotation around the Pt-N7 bond is more restricted in the case of the 5′-G adduct than in that of the 3′-G adduct. The binding of {Pt(dien)}²⁺ to 5′-GN7 and 3′-GN7 in the monofunctional adducts of (I) was shown to be reversible upon the addition of high concentrations of chloride ions. Received: 3 July 1998 / Accepted: 10 November 1998     

Microbial communities of the stratified soda Lake Doroninskoe (Transbaikal region)

The physicochemical properties, species composition, and vertical distribution of microorganisms in the water column, shoreline microbial mat, and small shoreline mud volcanoes of the stratified soda Lake Doroninskoe were investigated in September 2007. The lake is located in the Transbaikal region, in the permafrost zone (51°25′N; 112°28′E). The maximal depth of the contemporary lake is about 6 m, the pH value of the water is 9.72, and the water mineralization in the near-bottom horizon is 32.3 g l⁻¹. In summer, the surface oxygen-containing horizon of the water column becomes demineralized to 26.5 g l⁻¹; at a depth of 3.5–4.0 m, an abrupt transition occurs to the aerobic zone containing hydrosulfide (up to 12.56 g l⁻¹). Hydrosulfide was also detected in trace quantities in the upper water horizons. The density stratification of the water column usually ensures stable anaerobic conditions until the freezing period (November and December). The primary production of oxygenic phototrophs reached 176–230 μg l⁻¹. High rates of dark CO₂ assimilation (61–240 μg l⁻¹) were detected in the chemocline. Within this zone, an alkaliphilic species of sulfur-oxidizing bacteria of the genus Thioalkalivibrio was detected (10⁴ cells ml⁻¹). Lithoheterotrophic bacteria Halomonas spp., as well as bacteriochlorophyll a-containing aerobic anoxygenic phototrophic bacteria (AAP) Roseinatronobacter sp. capable of thiosulfate oxidation, were isolated from samples collected from the aerobic zone (0–3 m). The water transparency in September was extremely low; therefore, no visible clusters of anoxygenic phototrophic bacteria (APBs) were detected at the boundary of the hydrosulfide layer. However, purple sulfur bacteria which, according to the results of the 16S rRNA gene analysis, belong to the species Thioalkalicoccus limnaeus, Ectothiorhodospira variabilis, “Ect. magna,” and Ect. shaposhnikovii, were isolated from samples of deep silt sediments. Ect. variabilis and Ect. shaposhnikovii were the major APB species in the shoreline algo-bacterial mat. The halotolerant bacterium Ect. shaposhnikovii, purple nonsulfur bacteria of the genus Rhodobacter, and AAP of Roseococcus sp. were isolated from the samples collected from mud volcanoes. All these species are alkaliphiles, moderate halophiles, or halotolerant microorganisms.     

Organic matter properties in soils afforested with Pinus radiata

Aims

Afforestation causes important alterations in SOM content and composition that affect the soil functions and C balance. The aim of this study was to identify the mechanisms that determine the changes in SOM composition following afforestation of grasslands.

Methods

The study included 4 chronosequences and 5 paired plots comprising pastures and land afforested with Pinus radiata. The SOM was characterized by ¹³C CP-MAS NMR spectroscopy and differential scanning calorimetry.

Results

During the first 10–20 year after afforestation, the changes in SOM content varied from slight gains to large losses (>40 %). The analyses revealed that even SOM compounds considered resistant to decomposition were degraded during this time. The SOM gains, observed 20 year after stand establishment, were favoured by the higher recalcitrance of pine litter and possibly by soil acidification. The concentrations of most SOM compounds, particularly the stable compounds, were higher at the end of the rotation. The low degree of protection, along with the favourable climatic conditions, may also explain the rapid decomposition of SOM, including resistant compounds, in these soils. DSC analysis complemented the information about SOM composition provided by other techniques.

Conclusions

The accumulation of stable SOM compounds at the end of the rotation suggests a longer soil C turnover in these afforested soils, which may alleviate the gradual loss of SOC in intensively managed forest soils.