Actin cytoskeleton modulates calcium signaling during maturation of starfish oocytes

Before successful fertilization can occur, oocytes must undergo meiotic maturation. In starfish, this can be achieved in vitro by applying 1-methyladenine (1-MA). The immediate response to 1-MA is the fast Ca²⁺ release in the cell cortex. Here, we show that this Ca²⁺ wave always initiates in the vegetal hemisphere and propagates through the cortex, which is the space immediately under the plasma membrane. We have observed that alteration of the cortical actin cytoskeleton by latrunculin-A and jasplakinolide can potently affect the Ca²⁺ waves triggered by 1-MA. This indicates that the cortical actin cytoskeleton modulates Ca²⁺ release during meiotic maturation. The Ca²⁺ wave was inhibited by the classical antagonists of the InsP₃-linked Ca²⁺ signaling pathway, U73122 and heparin. To our surprise, however, these two inhibitors induced remarkable actin hyper-polymerization in the cell cortex, suggesting that their inhibitory effect on Ca²⁺ release may be attributed to the perturbation of the cortical actin cytoskeleton. In post-meiotic eggs, U73122 and jasplakinolide blocked the elevation of the vitelline layer by uncaged InsP₃, despite the massive release of Ca²⁺, implying that exocytosis of the cortical granules requires not only a Ca²⁺ rise, but also regulation of the cortical actin cytoskeleton. Our results suggest that the cortical actin cytoskeleton of starfish oocytes plays critical roles both in generating Ca²⁺ signals and in regulating cortical granule exocytosis.     

改变施肥管理后不同肥力稻田土壤CO2排放特征

利用一个长达30a水稻土长期定位试验,在保证原有定位试验继续正常开展的前提下,将原化肥处理改施有机肥,原有机肥处理改施化肥或者增施有机肥。通过观测田间试验2012—2013年双季稻轮作周期内不同肥力水平稻田土壤施肥管理改变后的土体CO2排放通量(FCO2),研究不同后续施肥管理对不同肥力红壤性水稻土CO2排放的影响。结果表明:变更施肥能明显改变CO2排放动态变化,其中长期施用有机肥处理改施化肥后其FCO2明显减小,长期施用化肥或有机肥处理增施有机肥后其FCO2显著增大。有机肥和土壤有机碳均可促进土体CO2排放,有机肥处理有机物料碳添加量与CO2-C年排放量呈极显著的正相关关系(r=0.9015**,n=21),单施化肥处理土壤有机碳含量与土体CO2-C年排放量符合线性方程:y=10.962x-68.86(R2=0.7507,n=9,P0.01)。长期施用有机肥土壤改施化肥会导致其有机碳矿化损失,土壤有机碳含量越高,矿化损失量越多,最终其有机碳水平将与长期施用化肥的土壤有机碳平衡值一致;长期施用化肥或有机肥土壤改施或增施有机肥可促进土壤有机碳积累,外源添加碳越多,土壤积累碳越多;相同有机肥施用量下土壤有机碳含量越高,有机物料表观分解率越大,积累于土壤中的有机碳越少,不同有机碳水平土壤在相同有机肥管理下其有机碳最终会达到相同的平衡值。在有机碳水平较低(20.46 g/kg)红壤稻田上增施有机肥是提升已培肥水稻土有机碳含量的可持续发展措施,而在有机碳水平较高(14.45 g/kg)红壤稻田上应避免改施化肥。总之,在有机碳含量较高或者较低的中国南方红壤性水稻土上,持续的有机肥施用是保持或者提高其有机碳水平的必要措施。     

Structure and stability of aerobic granules cultivated under different shear force in sequencing batch reactors

The cultivation of stable aerobic granules as well as granular structure and stability in sequencing batch reactors under different shear force were investigated in this study. Four column sequencing batch reactors (R1–R4) were operated under various shear force, in terms of superficial upflow air velocity of 0.8, 1.6, 2.4, and 3.2 cm s⁻¹, respectively. Aerobic granules were formed in all reactors in the experiment. It was found that the magnitude of shear force has an important impact on the granule stability. At shear force of 2.4 and 3.2 cm s⁻¹, granules can maintain a robust structure and have the potential of long-term operation. Granules developed in low shear force (R1, 0.8 cm s⁻¹ and R2, 1.6 cm s⁻¹) deteriorated to large-sized filamentous granules with irregular shape, loose structure and resulted in poor performance and operation instability. Granules cultivated under high shear force (R3, 2.4 cm s⁻¹ and R4, 3.2 cm s⁻¹) stabilized to clear outer morphology, dense and compact structure, and with good performance in 120 days operation. Fractal dimension (D_f) represents the internal structure of granules and can be used as an important indicator to describe the structure and stability of granules. Due to the combined effects of shear force and growth force, the mature granules developed in R3 and R4 also displayed certain differences in granular structure and characteristics.     

Ponderosa Pine Responses to Elevated CO2and Nitrogen Fertilization

The effects of elevated CO₂ (ambient, +175, and +350 μl l⁻¹) and nitrogen fertilization (0, 100, and 200 kg N ha⁻¹ yr⁻¹ as ammonium sulfate) on C and N accumulations in biomass and soils planted with ponderosa pine (Pinus ponderosa Laws) over a 6-year study period are reported. Both nitrogen fertilization and elevated CO₂ caused increases in C and N contents of vegetation over the study period. The pattern of responses varied over time. Responses to CO₂ decreased in the +175 μl l⁻¹ and increased in the +350 μl l⁻¹ after the first year, whereas responses to N decreased after the first year and became non-significant by year six. Foliar N concentrations were lower and tree C:N ratios were higher with elevated CO₂ in the early years, but this was offset by the increases in biomass, resulting in substantial increases in N uptake with elevated CO₂. Nitrogen budget estimates showed that the major source of the N for unfertilized trees, with or without elevated CO₂, was likely the soil organic N pool. There were no effects of elevated CO₂ on soil C, but a significant decrease in soil N and an increase in soil C:N ratio in year six. Nitrogen fertilization had no significant effect on tree C:N ratios, foliar N concentrations, soil C content, soil N content, or soil C:N ratios. There were no significant interactions between CO₂ and N treatments, indicating that N fertilization had no effect on responses to CO₂ and that CO₂ treatments had no effect on responses to N fertilization. These results illustrate the importance of long-term studies involving more than one level of treatment to assess the effects of elevated CO₂.     

Enhancement of [m-methoxy3H]MDL100907 binding to 5HT2A receptors in cerebral cortex and brain stem of streptozotocin induced diabetic rats

5-Hydroxytryptamine_2A (5-HT_2A) receptor kinetics was studied in cerebral cortex and brain stem of streptozotocin (STZ) induced diabetic rats. Scatchard analysis with [³H] (±) 2,3dimethoxyphenyl-1-[2-(4-piperidine)-methanol] ([³H]MDL100907) in cerebral cortex showed no significant change in maximal binding (B_max) in diabetic rats compared to controls. Dissociation constant (K_d) of diabetic rats showed a significant decrease (p < 0.05) in cerebral cortex, which was reversed to normal by insulin treatment. Competition studies of [³H]MDL100907 binding in cerebral cortex with ketanserin showed the appearance of an additional low affinity site for 5-HT_2A receptors in diabetic state, which was reversed to control pattern by insulin treatment. In brain stem, scatchard analysis showed a significant increase (p < 0.05) in B_max accompanied by a significant increase (p < 0.05) in K_d. Competition analysis in brain stem also showed a shift in affinity towards a low affinity State for 5-HT_2A receptors. All these parameters were reversed to control level by insulin treatment. These results show that in cerebral cortex there is an increase in affinity of 5-HT_2A receptors without any change in its number and in the case of brain stem there is an increase in number of 5HT_2A receptors accompanied by a decrease in its affinity during diabetes. Thus, from the results we suggest that the increase in affinity of 5-HT_2A receptors in cerebral cortex and upregulation of 5-HT_2A receptors in brain stem may lead to altered neuronal function in diabetes.     

Role of secretory granules in inositol 1,4,5-trisphosphate-dependent Ca(2+) signaling: from phytoplankton to mammals

The majority of secretory cell calcium is stored in secretory granules that serve as the major IP₃-dependent intracellular Ca²⁺ store. Even in unicellular phytoplankton secretory granules are responsible for the IP₃-induced Ca²⁺ release that triggers exocytosis. The number of secretory granules in the cell is directly related not only to the magnitude of IP₃-induced Ca²⁺ release, which accounts for the majority of the IP₃-induced cytoplasmic Ca²⁺ release in neuroendocrine cells, but also to the IP₃ sensitivity of the cytoplasmic IP₃ receptor (IP₃R)/Ca²⁺ channels. Moreover, secretory granules contain the highest IP₃R concentrations and the largest amounts of IP₃Rs in any subcellular organelles in neuroendocrine cells. Secretory granules from phytoplankton to mammals contain large amounts of polyanionic molecules, chromogranins being the major molecules in mammals, in addition to acidic intragranular pH and high Ca²⁺ concentrations. The polyanionic molecules undergo pH- and Ca²⁺-dependent conformational changes that serve as a molecular basis for condensation-decondensation phase transitions of the intragranular matrix. Likewise, chromogranins undergo pH- and Ca²⁺-dependent conformational changes with increased exposure of the structure and increased interactions with Ca²⁺ and other granule components at acidic pH. The unique physico-chemical properties of polyanionic molecules appear to be at the center of biogenesis, and physiological functions of secretory granules in living organisms from primitive to advanced species.     

2020-2050年CO2施肥效应促进全球陆地生态系统碳吸收

CO₂施肥效应是全球变绿的主要原因,随着大气中CO₂浓度的持续增加,预估未来气候变化条件下,CO₂施肥效应对陆地生态系统的影响对减缓全球气候变化具有重大意义。基于未来气候情景数据和Farquhar模型,并结合生态过程模型BEPS(Boreal Ecosystem Productivity Simulator),定量化研究2020—2050年CO₂施肥效应对全球叶面积指数(LAI)和总初级生产力(GPP)的影响。研究结果显示2020—2050年,在RCP2.6、RCP4.5和RCP8.5气候情景下,CO₂施肥效应导致的LAI年际变化趋势分别为0.002、0.003和0.005 m^-2m^-2a^-1;三个气候情景下CO₂施肥效应对LAI的影响为CO₂每增加0.1%,LAI平均增加约8.1%—9.2%,由此导致GPP对应增加7.9%—14.6%;由CO₂施...     

Inactivation of 5-HT1A and [3H]5-HT Binding Sites by N-Ethoxycarbonyl-2-Ethoxy-1, 2-Dihydroquinoline (EEDQ) in Rat Brain

Inactivation of 5-HT_1A and [³H]5-HT binding sites by N-Ethoxycarbonyl-2-ethoxy-1, 2-dihydro-quinoline (EEDQ) was studied in regions of rat brain. After exposure to EEDQ (4 mg/kg body wt.) for 7 days, it is observed that the density of 5-HT₁ receptor sites was decreased by nearly 20% in both cortex and hippocampus. The decrease, however, in 5-HT_1A sites was more significant (70%) in both the regions. The affinity of [³H]5-HT to 5-HT₁ sites was decreased significantly in both cortex and hippocampus after exposure to EEDQ, without affecting the Kd of 5-HT_1A sites. Displacement studies suggested that EEDQ has high affinity to 5-HT₁ sites with a Ki of 42.9 ± 2.4 nM. After exposure neither basal nor 5-HT stimulated adenylyl cyclase activity was changed in cortex. The results of this study suggest that EEDQ decreases the density of 5-HT₁ and 5-HT_1A receptor sites but does not cause functional downregulation of these sites in rat brain.     

Cortical 5-HT1A receptor downregulation by antidepressants in rat brain

Total 5-HT binding sites and 5-HT_1A receptor density was measured in brain regions of rats treated with imipramine (5 mg/kg body wt), desipramine (10 mg/kg body wt) and clomipramine (10 mg/kg body wt), for 40 days, using [³H]5-HT and [³H]8-OH-DPAT, respectively. It was observed that chronic exposure to tricyclic antidepressants (TCAs) results in significant downregulation of total [³H]5-HT binding sites in cortex (42–76%) and hippocampus (35–67%). The 5-HT_1A receptor density was, however, decreased significantly (32–60%) only in cortex with all the three drugs. Interestingly, in hippocampus imipramine treatment increased the 5-HT_1A receptor density (14%). The affinity of [³H]8-OH-DPAT was increased only with imipramine treatment both in cortex and hippocampus. The affinity of [³H]5-HT to 5-HT binding sites in cortex was increased with imipramine treatment and decreased with desipramine and clomipramine treatment. 5-HT sensitive adenylyl cyclase (AC) activity was significantly increased in cortex with imipramine (72%) and clomipramine (17%) treatment, whereas in hippocampus only imipramine treatment significantly increased AC activity (50%). In conclusion, chronic treatment with TCAs results in downregulation of cortical 5-HT_1A receptors along with concomitant increase in 5-HT stimulated AC activity suggesting the involvement of cortical 5-HT_1A receptors in the mechanism of action of TCAs.     

Movements of Echinochrome Granules during the Early Development of Sea Urchin Eggs

ECHINOCHROME pigment granules in unfertilized eggs of the sea urchin Arbacia punctulata undergo randomly-directed saltatory movements^1,2. After fertilization, nearly all these granules migrate to the egg cortex and become embedded. Subsequent pigment granule movements may represent mass cortical changes rather than independent granule movements^2,3. At the fourth cleavage, a quartet of micromeres containing little or no pigment forms at the vegetal pole. By the two or four-cell stage, pigment granules have begun to move out of this region, leaving a “clear area” on each blastomere (Fig. 1 and refs. 4, 5). To investigate possible mechanisms for these movements and their relation to cortical events     

Combined effects of CO2 concentration and nutrient status on the biomass production and nutrient uptake of birch seedlings (Betula pendula)

Birch seedlings (Betula pendula) were grown for four months in a greenhouse at three nutrient levels (fertilization of 0, 100 and 500 kg ha^-1 monthy) and at four CO₂ concentrations (350, 700, 1050 and 1400 ppm). The effect of CO₂ concentration on the biomass production depended on the nutrient status. When mineralization of the soil material was the only source of nutrients (0 kg ha^-1), CO₂ enhancement reduced the biomass production slightly, whereas the highest production increase occurred at a fertilization of 100 kg ha^-1, being over 100% between 350 and 700 ppm CO₂. At 500 kg ha^-1 the production increase was smaller, and the production decreased beyond a CO₂ concentration of 700 ppm. The CO₂ concentration had a slight effect on the biomass distribution, the leaves accounting for the highest proportion at the lowest CO₂ concentration (350 ppm). An increase in nutrient status led to a longer growth period and increased the nutrient concentrations in the plants, but the CO₂ concentration had no effect on the growth rhythm and higher CO₂ reduced the nutrient concentrations.     

Cyclic GMP-phosphodiesterase inhibition does not alter cerebral oxygen consumption

The effect of zaprinast, a cyclic guanosine monophosphate inhibitor, on the level of cyclic GMP and cerebral O₂ consumption was determined. Anesthetized male Long-Evans rats were divided into a control group (n=15) and a zaprinast treated group (n=15). Vehicle was applied topically to the left cortex and 3·10⁻³ M zaprinast was applied to the right cortex. A saline treated control group was also studied. Regional cerebral blood flow was determined by [¹⁴C]-iodoantipyrine and regional O₂ extraction was determined by microspectrophotometry. The level of cyclic GMP was measured by radioimmunoassay. There were no hemodynamic or blood gas differences between groups. The level of cyclic GMP was not significantly different between the right and left cerebral cortex of the control group (17.0±4.3 and 17.7±4.6 pmol/g). In the zaprinast treated group, there was a significant (46%) increase in the level of cyclic GMP in the zaprinast treated cortex (20.5±8.1) in comparison to the vehicle treated cortex (14.0±5.7). Zaprinast did not significantly alter cerebral blood flow. There were no significant differences in regional O₂ extraction. The O₂ consumption of the zaprinast treated cortex (8.0±3.3 ml O₂·min⁻¹·100 g⁻¹) was not different from that of the vehicle treated cortex (7.0±2.9) or those of the control group. Thus, our data indicated that the increased level of cyclic GMP had no significant effect on cerebral oxygen consumption.     

Modifications of the carbon and nitrogen allocations in the plant (Triticum aestivum L.) soil system in response to increased atmospheric CO2 concentration

The aim of this work was to examine the response of wheat plants to a doubling of the atmospheric CO₂ concentration on: (1) carbon and nitrogen partitioning in the plant; (2) carbon release by the roots; and (3) the subsequent N uptake by the plants. The experiment was performed in controlled laboratory conditions by exposing fast-growing spring wheat plants, during 28 days, to a ¹⁴CO₂ concentration of 350 or 700 L L^–1 at two levels of soil nitrogen fertilization. Doubling CO₂ availability increased total plant production by 34% for both N treatment. In the N-fertilized soil, the CO₂ enrichment resulted in an increase in dry mass production of 41% in the shoots and 23% in the roots; without N fertilization this figure was 33% and 37%, respectively. In the N-fertilized soil, the CO₂ increase enhanced the total N uptake by 14% and lowered the N concentration in the shoots by 23%. The N concentration in the roots was unchanged. In the N-fertilized soil, doubling CO₂ availability increased N uptake by 32% but did not change the N concentrations, in either shoots or roots. The CO₂ enrichment increased total root-derived carbon by 12% with N fertilization, and by 24% without N fertilization. Between 85 and 90% of the total root derived-¹⁴C came from respiration, leaving only 10 to 15% in the soil as organic ¹⁴C. However, when total root-derived ¹⁴C was expressed as a function of root dry weight, these differences were only slightly significant. Thus, it appears that the enhanced carbon release from the living roots in response to increased atmospheric CO₂, is not due to a modification of the activity of the roots, but is a result of the increased size of the root system. The increase of root dry mass also resulted in a stimulation of the soil N mineralization related to the doubling atmospheric CO₂ concentration. The discussion is focused on the interactions between the carbon and nitrogen allocation, especially to the root system, and the implications for the acquisition of nutrients by plants in response to CO₂ increase.Abbreviations N soil fertilization without nitrogen - N soil fertilization with nitrogen     

The effect of elevated carbon dioxide and fertilization on primary and secondary metabolites in birch,Betula pendula (Roth)

Seedlings of European white birch (Betula pendula Roth) were grown in growth chambers for one growth season under four carbon dioxide regimes (350, 700, 1050 and 1400 ppm) and at three fertilization levels (0, 100 and 500 kg ha^–1 monthly). The soluble carbohydrates and secondary phenolics in the leaves and stems were analysed. It was found that fertilizer addition reduced the amounts of glucose and fructose while sucrose remained almost unaffected. The sugar content of leaves increased at 700 ppm and 1050 ppm of CO₂ and decreased at the highest CO₂ concentration (1400 ppm). The amounts of proanthocyanidins and flavonoids in leaves decreased with fertilization addition and increased with CO₂ enrichment. The production of simple phenolic glucosides varied according to the fertilization and CO₂ treatments. The triterpenoid content of stems seemed to increase with fertilization and CO₂-addition. Our results indicate that the production of phytochemicals in the birch seedlings is very sensitive to both fertilization and CO₂ addition, which is in agreement with earlier studies, and thus provide some support for the hypothesis of carbon allocation to plant defence when there is an excess of carbon and nutrient. The considerable variation in the production of secondary components may indicate that the synthesis of these defensive metabolites can be regulated by a plant to certain extent, depending on the ability of the plant to acclimate to changes in the physical environment.     

The15N methodology in estimating N2 fixation by vetch and pea grown in pure stand or in mixtures with oat

Cereal-legume mixtures are frequently the best management decision for forage production instead of growing crops in pure stands. Nitrogen fertilization of cereal-legume mixtures is questionable since combined nitrogen could depress N₂ fixation by legumes. The objectives of this study were (1) to examine the effect of N fertilization on N₂ fixation by vetch and field peas in pure and in mixed stands with oats, and (2) to examine if there is any transfer of N from legumes to associated cereals. The field experiment was conducted for two growing seasons. The treatments were pure stands of vetch, pea and oats, and the mixtures of the two legumes with oats at the seeding ratios 90:10 and 75:25, fertilized with labelled¹⁵N at the rates of 15 and 90 kg N ha⁻¹. Nitrogen fertilization of 90 kg N ha⁻¹ suppressed N₂ fixation in both legumes grown in pure and in mixed stands. Crops grown in mixtures in many instances had lower atom %¹⁵N excess. Whether this was due to high N₂ fixation in the case of legume and transfer in the case of oat or the differences were due to practical problems of the¹⁵N technique is not clearly shown by the results, so based on the literature the aspect is discussed as well as the precautions which should be considered in using the¹⁵N technique in such studies.     

Imipramine-Induced Changes in 5-HT2 Receptor Sites and Inositoltrisphosphate Levels in Rat Brain

The effect of chronic administration of Imipramine on [³H]Spiperone binding to 5-HT₂ sites and inositoltrisphosphate (IP₃) levels in rat cerebral cortex was studied. Our data shows that treatment with imipramine (5 mg/kg body weight, intraperitoneally) for 30 days significantly down regulates 5-HT₂ receptors sites (262 ± 29 fmol/mg protein) in cerebral cortex (38%), compared to control rats (425 ± 60 fmol/mg protein., P < 0.001). However there was no significant change in the affinity of [³H]-Spiperone binding (kd) to 5-HT₂ sites in cerebral cortex after exposure to imipramine (Kd = 0.84 ± 0.11 nM). It is also observed that imipramine treatment significantly reduces 5-HT stimulated [³H]IP₃ formation in cerebral cortex (6,411 ± 708 dpm/mg protein), compared to the saline treated rats (12,238 ± 1,544 dpm/mg protein; P < 0.001), with concomitant decrease in Pdtlns-4–5-P₂. This study suggests that the therapeutic action of imipramine in brain might be by reducing hypersensitivity of 5-HT₂ receptors by down regulation, which leads to reduced levels of inositolphospholipids. This inturn reduces the levels of IP₃. In conclusion, imipramine acts at presynaptic site by blocking the reuptake of serotonin and at post synaptic site it downregulates 5-HT₂ sites with decreased IP₃ levels after chronic exposure.     

Subchronic Treatment with Methamphetamine and Phencyclidine Differentially Alters the Adenosine A1 and A2A Receptors in the Prefrontal Cortex,Hippocampus, and Striatum of the Rat

Subchronic treatment with MAP (4.6 mg/kg, i.p., once daily for 11 days) significantly decreased the K_d, but not B_max, values of [³H]1,3-dipropyl-8-cyclopentylxanthine ([³H]DPCPX) binding to adenosine A₁ receptors in the prefrontal cortex and hippocampus, but not striatum, of rat brain. However, subchronic treatment with PCP (10 mg/kg, i.p., once daily for 11 days) did not alter the K_d and B_max values of [³H]DPCPX binding to adenosine A₁ receptors in these three regions. Subchronic treatment with MAP or PCP did not alter the B_max and K_d values of [³H]2-p-(2-carboxyehyl)phenethylamino-5-N-ethylcarboxyamidoadenosine ([³H]CGS21680) binding to adenosine A_2A receptors in the striatum. Furthermore, subchronic treatment with MAP or PCP significantly decreased the specific binding of [³H]CGS21680 to adenosine A_2A receptors in the hippocampus, but not in the prefrontal cortex. Thus, these results suggest that MAP and PCP may produce differential effects on the adenosine A_2A receptors, but not adenosine A₁ receptors in rat brain.     

A rhizolysimeter facility for studying the dynamics of crop and soil processes: description and evaluation

Increased atmospheric carbon dioxide (CO₂) concentration will likely cause changes in plant productivity and composition that might affect soil decomposition processes. The objective of this study was to test to what extent elevated CO₂ and N fertility-induced changes in residue quality controlled decomposition rates. Cotton (Gossypium hirsutum L.) was grown in 8-l pots and exposed to two concentrations of CO₂ (390 or 722 μmol mol^-1) and two levels of N fertilization (1.0 or 0.25 g l^-1 soil) within greenhouse chambers for 8 wks. Plants were then chemically defoliated and air-dried. Leaf, stem and root residues were assayed for total non-structural carbohydrates (TNC), lignin (LTGA), proanthocyanidins (PA), C and N. Respiration rates of an unsterilized sandy soil (Lakeland Sand) mixed with residues from the various treatments were determined using a soda lime trap to measure CO₂ release. At harvest, TNC and PA concentrations were 17 to 45% higher in residues previously treated with elevated CO₂ compared with controls. Leaf and stem residue LTGA concentrations were not significantly affected by either the elevated CO₂ or N fertilization treatments, although root residue LTGA concentration was 30% greater in plants treated with elevated CO₂. The concentration of TNC in leaf residues from the low N fertilization treatment was 2.3 times greater than that in the high N fertilization treatment, although TNC concentration in root and stem residues was suppressed 13 to 23% by the low soil N treatment. PA and LTGA concentrations in leaf, root and stem residues were affected by less than 10% by the low N fertilization treatment. N concentration was 14 to 44% lower in residues obtained from the elevated CO₂ and low N fertilization treatments. In the soil microbial respiration assay, cumulative CO₂ release was 10 to 14% lower in soils amended with residues from the elevated CO₂ and low N fertility treatments, although treatment differences diminished as the experiment progressed. Treatment effects on residue N concentration and C:N ratios appeared to be the most important factors affecting soil microbial respiration. The results of our study strongly suggest that, although elevated CO₂ and N fertility may have significant impact on post-harvest plant residue quality of cotton, neither factor is likely to substantially affect decomposition. Thus, C cycling might not be affected in this way, but via simple increases in plant biomass production. This revised version was published online in June 2006 with corrections to the Cover Date.     

Nitric oxide (NO) increase at fertilization in sea urchin eggs upregulates fertilization envelope hardening

Previous studies indicate that the nitric oxide (NO) increase at fertilization in sea urchin eggs is Ca²⁺-dependent and attributed to the late Ca²⁺ rise. However, its role in fertilization still remains unclear. Simultaneous measurements of the activation current, by a single electrode voltage clamp, and NO, using the NO indicator DAF-FM, showed that the NO increase occurred at the time of peak current (t_p) which corresponds to peak [Ca²⁺]_i, suggesting that NO is not related to any other ionic changes besides [Ca²⁺]_i. We measured O₂ consumption by a polarographic method to examine whether NO regulated a respiratory burst for protection as reported in other biological systems. Our results suggested NO increased O₂ consumption. The fluorescence of reduced pyridine nucleotides, NAD(P)H was measured in controls and when the NO increase was eliminated by PTIO, a NO scavenger. Surprisingly, PTIO decreased the rate of the fluorescence change and the late phase of increase in NAD(P)H was eliminated. PTIO also suppressed the production of H₂O₂ and caused weak and high fertilization envelope (FE). Our results suggest that NO increase upregulates NAD(P)H and H₂O₂ production and consolidates FE hardening by H₂O₂.     

Fluxes of greenhouse gases from Andosols under coffee in monoculture or shaded by <Emphasis Type="Italic">Inga densiflora</Emphasis> in Costa Rica

The objective of this study was to evaluate the effect of N fertilization and the presence of N₂ fixing leguminous trees on soil fluxes of greenhouse gases. For a one year period, we measured soil fluxes of nitrous oxide (N₂O), carbon dioxide (CO₂) and methane (CH₄), related soil parameters (temperature, water-filled pore space, mineral nitrogen content, N mineralization potential) and litterfall in two highly fertilized (250 kg N ha⁻¹ year⁻¹) coffee cultivation: a monoculture (CM) and a culture shaded by the N₂ fixing legume species Inga densiflora (CIn). Nitrogen fertilizer addition significantly influenced N₂O emissions with 84% of the annual N₂O emitted during the post fertilization periods, and temporarily increased soil respiration and decreased CH₄ uptakes. The higher annual N₂O emissions from the shaded plantation (5.8 ± 0.3 kg N ha⁻¹ year⁻¹) when compared to that from the monoculture (4.3 ± 0.1 kg N ha⁻¹ year⁻¹) was related to the higher N input through litterfall (246 ± 16 kg N ha⁻¹ year⁻¹) and higher potential soil N mineralization rate (3.7 ± 0.2 mg N kg⁻¹ d.w. d⁻¹) in the shaded cultivation when compared to the monoculture (153 ± 6.8 kg N ha⁻¹ year⁻¹ and 2.2 ± 0.2 mg N kg⁻¹ d.w. d⁻¹). This confirms that the presence of N₂ fixing shade trees can increase N₂O emissions. Annual CO₂ and CH₄ fluxes of both systems were similar (8.4 ± 2.6 and 7.5 ± 2.3 t C-CO₂ ha⁻¹ year⁻¹, −1.1 ± 1.5 and 3.3 ± 1.1 kg C-CH₄ ha⁻¹ year⁻¹, respectively in the CIn and CM plantations) but, unexpectedly increased during the dry season.