Localized Tetrazolium Reduction in Relation to N2 Fixation, CO2 Fixation, and H2 Uptake in Aquatic Filamentous Cyanobacteria

The aquatic filamentous cyanobacteria Anabaena oscillarioides and Trichodesmium sp. reveal specific cellular regions of tetrazolium salt reduction. The effects of localized reduction of five tetrazolium salts on N₂ fixation (acetylene reduction), ¹⁴CO₂ fixation, and ³H₂ utilization were examined. During short-term (within 30 min) exposures in A. oscillarioides, salt reduction in heterocysts occurred simultaneously with inhibition of acetylene reduction. Conversely, when salts failed to either penetrate or be reduced in heterocysts, no inhibition of acetylene reduction occurred. When salts were rapidly reduced in vegetative cells, ¹⁴CO₂ fixation and ³H₂ utilization rates decreased, whereas salts exclusively reduced in heterocysts were not linked to blockage of these processes. In the nonheterocystous genus Trichodesmium, the deposition of reduced 2,3,5-triphenyl-2-tetrazolium chloride (TTC) in the internal cores of trichomes occurs simultaneously with a lowering of acetylene reduction rates. Since TTC deposition in heterocysts of A. oscillarioides occurs contemporaneously with inhibition of acetylene reduction, we conclude that the cellular reduction of this salt is of use in locating potential N₂-fixing sites in cyanobacteria. The possible applications and problems associated with interpreting localized reduction of tetrazolium salts in cyanobacteria are presented.     

Mesopelagic N2 Fixation Related to Organic Matter Composition in the Solomon and Bismarck Seas (Southwest Pacific)

Dinitrogen (N₂) fixation was investigated together with organic matter composition in the mesopelagic zone of the Bismarck (Transect 1) and Solomon (Transect 2) Seas (Southwest Pacific). Transparent exopolymer particles (TEP) and the presence of compounds sharing molecular formulae with saturated fatty acids and sugars, as well as dissolved organic matter (DOM) compounds containing nitrogen (N) and phosphorus (P) were higher on Transect 1 than on Transect 2, while oxygen concentrations showed an opposite pattern. N₂ fixation rates (up to ~1 nmol N L^-1 d^-1) were higher in Transect 1 than in Transect 2, and correlated positively with TEP, suggesting a dependence of diazotroph activity on organic matter. The scores of the multivariate ordination of DOM molecular formulae and their relative abundance correlated negatively with bacterial abundances and positively with N₂ fixation rates, suggesting an active bacterial exploitation of DOM and its use to sustain diazotrophic activity. Sequences of the nifH gene clustered with Alpha-, Beta-, Gamma- and Deltaproteobacteria, and included representatives from Clusters I, III and IV. A third of the clone library included sequences close to the potentially anaerobic Cluster III, suggesting that N₂ fixation was partially supported by presumably particle-attached diazotrophs. Quantitative polymerase chain reaction (qPCR) primer-probe sets were designed for three phylotypes and showed low abundances, with a phylotype within Cluster III at up to 10³ nifH gene copies L^-1. These results provide new insights into the ecology of non-cyanobacterial diazotrophs and suggest that organic matter sustains their activity in the mesopelagic ocean.     

极端干旱对贵州省喀斯特地区植物的影响

基于2010年贵州省旱灾中喀斯特地区受损植被的调查资料,分析受损植被的物种组成、植物区系、生活型、叶质及其立地环境条件.结果表明:调查的6个样地中,因旱受损植物达31种,杉木、南蛇藤、密花树、齿叶铁仔、青冈和云南樟的受损数量最多,受损物种以热带中型革质单叶的常绿小乔木为主.受损植物主要分布在坡度较陡的顺倾坡中上部和水平产状碳酸盐岩发育的薄层石灰土上,土厚＜30 cm和石土面小生境植物受损最多,分别占87.4%和40.0%.建议喀斯特地区适当增加温带性质小型叶耐旱落叶乔木及灌木的种植面积,植树造林应选择石沟和土面等土层较厚、立地条件较好的小生境.     

Defoliation in White Clover: Regrowth, Photosynthesis and N2 Fixation

Single plants of white clover, grown in a controlled environmentand dependent for nitrogen on fixation in their root nodules,were defoliated once by removing approximately half their shoottissue. Their regrowth was compared with the growth of comparableundefoliated plants. Two similar experiments were carried out:in the first, plants were defoliated at 2.5 g, and in the secondat 1.2 g total plant d. wt. Defoliation reduced rate of N₂ fixation by > 70 per cent,rate of photosynthesis by 83–96 per cent, and rate ofplant respiration by 30–40 per cent. Nodule weights initiallydeclined following defoliation as a result of loss of carbohydratesand other unidentified components. No immediate shedding ofnodules was observed but nodules on the most severely defoliatedplants exhibited accelerated senescence. The original rates of N₂ fixation were re-attained after 5–6or 9 d regrowth, with increase in plant size at defoliation.In general, the rate of recovery of N₂ fixation was relatedto the re-establishment and increase of the plant's photosyntheticcapacity. Throughout the growth of both defoliated and undefoliatedplants nodule respiration (metabolism) accounted for at least23 ± 2 per cent of gross photosynthesis. The unit ‘cost’of fixing N₂ in root nodules, in terms of photosynthate, appearedto be unaffected by defoliation, except perhaps for plants veryrecently defoliated. Similarly, the percentage nitrogen contentsof shoot, root and nodules of defoliated plants became adaptedwithin a few days to those characteristic of undefoliated plants. Trifolium repens, white clover, N₂ fixation, defoliation, photosynthesis, respiration     

N2O, CH4 and CO2 emissions from seasonal tropical rainforests and a rubber plantation in Southwest China

The main focus of this study was to evaluate the effects of soil moisture and temperature on temporal variation of N₂O, CO₂ and CH₄ soil-atmosphere exchange at a primary seasonal tropical rainforest (PF) site in Southwest China and to compare these fluxes with fluxes from a secondary forest (SF) and a rubber plantation (RP) site. Agroforestry systems, such as rubber plantations, are increasingly replacing primary and secondary forest systems in tropical Southwest China and thus effect the N₂O emission in these regions on a landscape level. The mean N₂O emission at site PF was 6.0 ± 0.1 SE μg N m⁻² h⁻¹. Fluxes of N₂O increased from <5 μg N m⁻² h⁻¹ during dry season conditions to up to 24.5 μg N m⁻² h⁻¹ with re-wetting of the soil by the onset of first rainfall events. Comparable fluxes of N₂O were measured in the SF and RP sites, where mean N₂O emissions were 7.3 ± 0.7 SE μg N m⁻² h⁻¹ and 4.1 ± 0.5 SE μg N m⁻² h⁻¹, respectively. The dependency of N₂O fluxes on soil moisture levels was demonstrated in a watering experiment, however, artificial rainfall only influenced the timing of N₂O emission peaks, not the total amount of N₂O emitted. For all sites, significant positive correlations existed between N₂O emissions and both soil moisture and soil temperature. Mean CH₄ uptake rates were highest at the PF site (−29.5 ± 0.3 SE μg C m⁻² h⁻¹), slightly lower at the SF site (−25.6 ± 1.3 SE μg C m⁻² h⁻¹) and lowest for the RP site (−5.7 ± 0.5 SE μg C m⁻² h⁻¹). At all sites, CH₄ uptake rates were negatively correlated with soil moisture, which was also reflected in the lower uptake rates measured in the watering experiment. In contrast to N₂O emissions, CH₄ uptake did not significantly correlate with soil temperature at the SF and RP sites, and only weakly correlated at the PF site. Over the 2 month measurement period, CO₂ emissions at the PF site increased significantly from 50 mg C m⁻² h⁻¹ up to 100 mg C m⁻² h⁻¹ (mean value 68.8 ± 0.8 SE mg C m⁻² h⁻¹), whereas CO₂ emissions at the SF and RP site where quite stable and varied only slightly around mean values of 38.0 ± 1.8 SE mg C m⁻² h⁻¹ (SF) and 34.9 ± 1.1 SE mg C m⁻² h⁻¹ (RP). A dependency of soil CO₂ emissions on changes in soil water content could be demonstrated for all sites, thus, the watering experiment revealed significantly higher CO₂ emissions as compared to control chambers. Correlation of CO₂ emissions with soil temperature was significant at the PF site, but weak at the SF and not evident at the RP site. Even though we demonstrated that N and C trace gas fluxes significantly varied on subdaily and daily scales, weekly measurements would be sufficient if only the sink/ source strength of non-managed tropical forest sites needs to be identified.     

西南岩溶地区黄荆叶片碳酸酐酶的稳定性

研究表明西南岩溶地区几种植物叶片中含有明显活性的碳酸酐酶(carbonic anhydrase,CA),从黄荆(Vitex negundo)叶片提取碳酸酐酶,该酶具有良好的热稳定性.阴离子及岩溶环境主要金属离子对碳酸酐酶活性有一定的影响,Ca2 、Mg2 低于10 mmol·L-1,Mn2 、Zn2 低于0.01 mmol·L-1,Co2 、Cu2 低于0.001 mmol·L-1,NO3-、C1-、Br-、I-、NO2-低于0.01 mmol·L-1时,酶活性较稳定.0.001～0.01 mmol·L-1的Ca2 、Mg2 对该酶有激活作用.研究结果表明,该酶比较适应岩溶土壤水体的离子环境从而保持酶活性的相对稳定.     

天津近海自由生活线虫多样性分析

2007年4月在面临高度开发的天津近岸海域,对沉积物中自由生活的线虫样品进行了采集,并分析了其多样性,为生物多样性研究和环境监测提供科学数据.结果表明,该海域自由生活线虫有80余种,生物丰度以拉氏矛咽线虫Dorylaimopsis rabalais的优势最突出.调查海域的沉积物虽然受到了一定的污染,但其污染状况并不恶劣.垂直的层次分析表明一些多样性指数受采样深度影响,分析时应关注采样深度.     

Nitrogen Fixation by Free-living Bacteria in the Soil and in the Canopy of Douglas Fir

Nitrogen fixing bacteria have been isolated from the soil andthe leaves of Douglas fir. Measurements, taken at monthly intervalsusing ¹⁵N, have shown that small quantities of nitrogen arefixed on the leaves and in the various soil layers and thatthe highest rates of fixation occur in the spring. Further studieswith ¹⁵N have shown that the products of nitrogen fixation inDouglas fir soils are available for growth of seedlings andfor denitrification.     

Environmental Factors Controlling N2 Fixation in Mediterranean Rice Fields

Abstract The objectives of this study were to analyze the environmental controls on N₂ fixation in Spanish rice fields. Nitrogenase activity, measured as the acetylene-reducing activity (ARA), was estimated in situ during different intervals of the cropping period. At the same time, physical and chemical variables and cyanobacterial occurrence were determined in water and soil. Nitrogen fixation was measurable at all sampling sites, being higher in July and lower in June after a short dry period. The ARA values ranged from 0.23 to 75.5 kg N Na⁻¹ year⁻¹. Because blooms or other conspicuous cyanobacterial forms were not included in the measurements, maximum rates of nitrogen fixation may have been higher. Environmental variables that correlated with ARA varied on a seasonal basis. Water properties such as calcium, hardness, or conductivity, and soil properties such as conductivity and sodium correlated positively with N₂ fixation; however, nutrient parameters such as dissolved inorganic nitrogen or soluble reactive phosphorus were negatively correlated. Cyanobacterial abundance, in general, did not correlate with ARA. The overall conclusion is that nitrogen fixation may be an important N input in the N cycle of rice fields, and could lessen pollution problems by lowering the demand for chemical fertilizers. Received 24 January 1996; Accepted: 11 June 1996     

Localized Tetrazolium Reduction in Relation to N(2) Fixation, CO(2) Fixation, and H(2) Uptake in Aquatic Filamentous Cyanobacteria

The aquatic filamentous cyanobacteria Anabaena oscillarioides and Trichodesmium sp. reveal specific cellular regions of tetrazolium salt reduction. The effects of localized reduction of five tetrazolium salts on N(2) fixation (acetylene reduction), CO(2) fixation, and H(2) utilization were examined. During short-term (within 30 min) exposures in A. oscillarioides, salt reduction in heterocysts occurred simultaneously with inhibition of acetylene reduction. Conversely, when salts failed to either penetrate or be reduced in heterocysts, no inhibition of acetylene reduction occurred. When salts were rapidly reduced in vegetative cells, CO(2) fixation and H(2) utilization rates decreased, whereas salts exclusively reduced in heterocysts were not linked to blockage of these processes. In the nonheterocystous genus Trichodesmium, the deposition of reduced 2,3,5-triphenyl-2-tetrazolium chloride (TTC) in the internal cores of trichomes occurs simultaneously with a lowering of acetylene reduction rates. Since TTC deposition in heterocysts of A. oscillarioides occurs contemporaneously with inhibition of acetylene reduction, we conclude that the cellular reduction of this salt is of use in locating potential N(2)-fixing sites in cyanobacteria. The possible applications and problems associated with interpreting localized reduction of tetrazolium salts in cyanobacteria are presented.     

Malonate Catabolism Does Not Drive N2 Fixation in Legume Nodules

Malonyl-coenzyme A (CoA) decarboxylase, malonyl-CoA synthetase, and malonate transporter mutants of Rhizobium leguminosarum bv. viciae and trifolii fixed N₂ at wild-type rates on pea and clover, respectively. Thus, malonate does not drive N₂ fixation in legume nodules.     

1, N2-Ethenoguanosine: Three Methods of Synthesis

Abstract

Three novel approaches to synthesis of 1, N²-ethe-noguanosine from guanosine: i) reaction with aq. chloroacet-aldehyde at pH 9–10 ii) application of anhydrous chloro- and bromoacetaldehydes iii) 1-alkylation with bromoacetaldehyde diethyl acetal, subsequent hydrolysis to aldehyde on the nucleoside level and cyclization-afforded the title compound in satisfactory yields.     

N2 Fixation (C2H2 reduction) by epiphylls on coffee,Coffea arabica

Nitrogen fixation (C₂H₂ reduction) by epiphylls on coffee,Coffea arabica, grown in sites with different degrees of shade, was determined. Coffee leaves with nitrogen-fixing epiphylls were found in all sites in approximately equal numbers. Rates of C₂H₂ reduction were similar for all sites and throughout the year, averaging 3.21 nmoles C₂H₂ reduced leaf with epiphylls^–1 day^–1. Apparently, neither rates of activity nor abundance of leaves with nitrogen-fixing epiphylls is related to the degree of shade in a site. No correlation was found between percent epiphyll cover and the presence or magnitude of nitrogen-fixing activity. Calculated annual fixation by epiphylls on coffee was low, ranging from 0.7 g N₂ ha^–1 year^–1 for the shadeless site to 1.4 g N₂ ha^–1 year^–1 for the site withIngajinicuil shade trees. These results suggest that epiphyll fixation is not an important source of nitrogen for the coffee ecosystem studied.     

N2 Fixation,Carbon Metabolism,and Oxidative Damage in Nodules of Dark-Stressed Common Bean Plants

Common beans (Phaseolus vulgaris L.) were exposed to continuous darkness to induce nodule senescence, and several nodule parameters were investigated to identify factors that may be involved in the initial loss of N2 fixation. After only 1 d of darkness, total root respiration decreased by 76% and in vivo nitrogenase (N2ase) activity decreased by 95%. This decline coincided with the almost complete depletion (97%) of sucrose and fructose in nodules. At this stage, the O2 concentration in the infected zone increased to 1%, which may be sufficient to inactivate N2ase; however, key enzymes of carbon and nitrogen metabolism were still active. After 2 d of dark stress there was a significant decrease in the level of N2ase proteins and in the activities of enzymes involved in carbon and nitrogen assimilation. However, the general collapse of nodule metabolism occurred only after 4 d of stress, with a large decline in leghemoglobin and antioxidants. At this final senescent stage, there was an accumulation of oxidatively modified proteins. This oxidative stress may have originated from the decrease in antioxidant defenses and from the Fe-catalyzed generation of activated oxygen due to the increased availability of catalytic Fe and O2 in the infected region.     

N2 Fixation in Feather Mosses is a Sensitive Indicator of N Deposition in Boreal Forests

Nitrogen (N) fixation in the feather moss–cyanobacteria association represents a major N source in boreal forests which experience low levels of N deposition; however, little is known about the effects of anthropogenic N inputs on the rate of fixation of atmospheric N₂ in mosses and the succeeding effects on soil nutrient concentrations and microbial community composition. We collected soil samples and moss shoots of Pleurozium schreberi at six distances along busy and remote roads in northern Sweden to assess the influence of road-derived N inputs on N₂ fixation in moss, soil nutrient concentrations and microbial communities. Soil nutrients were similar between busy and remote roads; N₂ fixation was higher in mosses along the remote roads than along the busy roads and increased with increasing distance from busy roads up to rates of N₂ fixation similar to remote roads. Throughfall N was higher in sites adjacent to the busy roads but showed no distance effect. Soil microbial phospholipid fatty acid (PLFA) composition exhibited a weak pattern regarding road type. Concentrations of bacterial and total PLFAs decreased with increasing distance from busy roads, whereas fungal PLFAs showed no distance effect. Our results show that N₂ fixation in feather mosses is highly affected by N deposition, here derived from roads in northern Sweden. Moreover, as other measured factors showed only weak differences between the road types, atmospheric N₂ fixation in feather mosses represents a highly sensitive indicator for increased N loads to natural systems.     

Conservation in Soil of H2 Liberated from N2 Fixation by Hup- Nodules

Pigeon peas (Cajanus cajan) were grown in large soil columns (90-cm length by 30-cm diameter) and inoculated with four different strains of cowpea rhizobia, which varied with respect to hydrogen uptake activity (Hup). Despite the profuse liberation of H₂ from Hup^- nodules in vitro, H₂ gas was not detected in any of the soil columns. When H₂ was injected into the columns, the rates of consumption were highest in the treatments (including control) containing Hup^- nodules (218 and 177 nmol · h⁻¹ · cm⁻²) and lowest in the Hup⁺ treatments (158, 92, and 64 nmoles · h⁻¹ · cm⁻²). In situ H₂ uptake rates in small soil cores at fixed distances from the nodules decreased exponentially with distance from the nodule (R² = 0.99). This decrease in H₂ consumption was associated with a similar decrease in numbers of H₂-oxidizing chemolithotrophic bacteria as determined by the most-probable-number method. On the basis of two equations derived separately upon diffusive theory (Fix's Law) and kinetic theory (Michaelis-Menten), the empirically derived rate constants and coefficients indicated that all of the H₂ emitted from Hup^- nodules would be consumed by H₂-oxidizing bacteria within a 3- to 4.5-cm radius of the nodule surface. It is concluded that H₂ is not lost from the soil-plant ecosystem during N₂ fixation in C. cajan but is conserved by H₂-oxidizing bacteria.     

Azolla-Anabaena Relationship : XIII. Fixation of [N]N(2)

The major radioactive products of the fixation of [¹³N]N₂ by Azolla caroliniana Willd.-Anabaena azollae Stras. were ammonium, glutamine, and glutamate, plus a small amount of alanine. Ammonium accounted for 70 and 32% of the total radioactivity recovered after fixation for 1 and 10 minutes, respectively. The presence of a substantial pool of [¹³N]N₂-derived ¹³NH₄⁺ after longer incubation periods was attributed to the spatial separation between the site of N₂-fixation (Anabaena) and a second, major site of assimilation (Azolla). Initially, glutamine was the most highly radioactive organic product formed from [¹³N]N₂, but after 10 minutes of fixation glutamate had 1.5 times more radiolabel than glutamine. These kinetics of radiolabeling, along with the effects of inhibitors of glutamine synthetase and glutamate synthase on assimilation of exogenous and [¹³N]N₂-derived ¹³NH₄⁺, indicate that ammonium assimilation occurred by the glutamate synthase cycle and that glutamate dehydrogenase played little or no role in the synthesis of glutamate by Azolla-Anabaena.     

Sucrose Catabolism in Clostridium pasteurianum and Its Relation to N2 Fixation

The growth constant and Y (sucrose) (grams of cells per mole of sucrose) for NH(3)-grown cultures of Clostridium pasteurianum were 1.7 times those of N(2)-grown cultures, whereas the rate of sucrose utilized per gram of cells per hour was similar for both conditions. The Y (sucrose) of chemostat cultures grown on limiting NH(3) under argon at generation times equal to those of N(2)-fixing cultures was less than that of cultures grown on excess NH(3), but cells of NH(3)-limited cultures contained the N(2)-fixing system in high concentration. The concentration of the N(2)-fixing system in whole cells, when measured with adenosine triphosphate (ATP) nonlimiting, was more than twofold greater than the amount needed for the N(2) actually fixed. Thus, energy production from sucrose, and not the concentration of the N(2)-fixing system nor the maximal rate at which N(2) could be fixed, was the limiting factor for growth of N(2)-fixing cells. Either NH(3) or some product of NH(3) metabolism partially regulated the rate of sucrose metabolism since, when cultures fixing N(2), growing on NH(3), or growing on limiting NH(3) in the absence of N(2) were deprived of their nitrogen source, the rate of sucrose catabplism decreased. Calculations showed that the rate of ATP production was the growth rate-limiting factor in cells grown on N(2), and that the increased sucrose requirement of N(2)-fixing cultures in part reflected the energy demand of N(2) fixation. Calculations indicated that whole cells require about 20 moles of ATP for the fixation of 1 mole of N(2) to 2 moles of NH(3).     

Effects of Nitrogen Addition on Nitrogen Resorption in Temperate Shrublands in Northern China

Nutrient resorption from senescing leaves is a key mechanism of nutrient conservation for plants. The nutrient resorption efficiency is highly dependent on leaf nutrient status, species identity and soil nutrient availability. Nitrogen is a limiting nutrient in most ecosystems, it is widely reported that nitrogen resorption efficiency (NRE) was highly dependent on the soil nitrogen availability and vary with N deposition. The effects of nitrogen deposition on NRE and nitrogen concentration in green and senescing leaves have been well established for forests and grasslands; in contrast, little is known on how plants in shrublands respond to nitrogen deposition across the world. In this study, we conducted a two-year nitrogen addition manipulation experiment to explore the responses of nitrogen concentration in green and senescing leaves, and NRE of seven dominant species, namely, Vitex negundo, Wikstroemia chamaedaphne, Carex rigescens and Cleistogenes chinensis from the Vitex negundo community, and Spirea trilobata, Armeniaca sibirica, V. negundo, C. rigescens and Spodiopogon sibiricus from the Spirea trilobata community, to nitrogen deposition in two typical shrub communities of Mt. Dongling in northern China. Results showed that NRE varied remarkably among different life forms, which was lowest in shrubs, highest in grasses, and intermediate in forbs, implying that shrubs may be most capable of obtaining nitrogen from soil, grasses may conserve more nitrogen by absorption from senescing leaves, whereas forbs may adopt both mechanisms to compete for limited nitrogen supply from the habitats. As the N addition rate increases, N concentration in senescing leaves ([N]_s) increased consistent from all species from both communities, that in green leaves ([N]_g) increased for all species from the Vitex negundo community, while no significant responses were found for all species from the Spirea trilobata community; NRE decreased for all species except A. sibirica from the Vitex community and W. chamaedaphn from the Spirea community. Given the substantial interspecific variations in nutrient concentration, resorption and the potentially changing community composition, and the increased soil nutrient availability due to fertilization may indirectly impact nutrient cycling in this region.