海洋固氮生物多样性及其对海洋生产力的氮、碳贡献

固氮生物及其共生体在自然生态系统的氮素供给和生产力的持续发展中有重要作用。是贫营养盐海域和海洋生态系统中新生产力的主要贡献者。就海洋固氮生物研究的历史和现状,以及在世界不同海域和海洋生态环境条件下,海洋固氮生物的物种多样性和在海洋生态环境中的生存方式、共生互作关系多样化,作较全面的概述。综合分析了海洋固氮生物的固氮能力（活性）、在光合固碳中的作用和对初级生产力的贡献。介绍了固氮生物在海洋生物链和海洋不同生态系统和生物群落中的作用及其对环境的影响的最新研究概况。     

N2-fixation by methanotrophs sustains carbon and nitrogen accumulation in pristine peatlands

Symbiotic relationships between N₂-fixing prokaryotes and their autotrophic hosts are essential in nitrogen (N)-limited ecosystems, yet the importance of this association in pristine boreal peatlands, which store 25 % of the world’s soil (C), has been overlooked. External inputs of N to bogs are predominantly atmospheric, and given that regions of boreal Canada anchor some of the lowest rates found globally (~1 kg N ha^?1 year^?1), biomass production is thought to be limited primarily by N. Despite historically low N deposition, we show that boreal bogs have accumulated approximately 12–25 times more N than can be explained by atmospheric inputs. Here we demonstrate high rates of biological N₂-fixation in prokaryotes associated with Sphagnum mosses that can fully account for the missing input of N needed to sustain high rates of C sequestration. Additionally, N amendment experiments in the field did not increase Sphagnum production, indicating that mosses are not limited by N. Lastly, by examining the composition and abundance of N₂-fixing prokaryotes by quantifying gene expression of 16S rRNA and nitrogenase-encoding nifH, we show that rates of N₂-fixation are driven by the substantial contribution from methanotrophs, and not from cyanobacteria. We conclude biological N₂-fixation drives high sequestration of C in pristine peatlands, and may play an important role in moderating fluxes of methane, one of the most important greenhouse gases produced in peatlands. Understanding the mechanistic controls on biological N₂-fixation is crucial for assessing the fate of peatland carbon stocks under scenarios of climate change and enhanced anthropogenic N deposition.     

Ecological consequences of the expansion of N2-fixing plants in cold biomes

Research in warm-climate biomes has shown that invasion by symbiotic dinitrogen (N₂)-fixing plants can transform ecosystems in ways analogous to the transformations observed as a consequence of anthropogenic, atmospheric nitrogen (N) deposition: declines in biodiversity, soil acidification, and alterations to carbon and nutrient cycling, including increased N losses through nitrate leaching and emissions of the powerful greenhouse gas nitrous oxide (N₂O). Here, we used literature review and case study approaches to assess the evidence for similar transformations in cold-climate ecosystems of the boreal, subarctic and upper montane-temperate life zones. Our assessment focuses on the plant genera Lupinus and Alnus, which have become invasive largely as a consequence of deliberate introductions and/or reduced land management. These cold biomes are commonly located in remote areas with low anthropogenic N inputs, and the environmental impacts of N₂-fixer invasion appear to be as severe as those from anthropogenic N deposition in highly N polluted areas. Hence, inputs of N from N₂ fixation can affect ecosystems as dramatically or even more strongly than N inputs from atmospheric deposition, and biomes in cold climates represent no exception with regard to the risk of being invaded by N₂-fixing species. In particular, the cold biomes studied here show both a strong potential to be transformed by N₂-fixing plants and a rapid subsequent saturation in the ecosystem’s capacity to retain N. Therefore, analogous to increases in N deposition, N₂-fixing plant invasions must be deemed significant threats to biodiversity and to environmental quality.     

Performance of an age series of alnus-cardamom plantations in the Sikkim Himalaya: productivity,energetics and efficiencies

Biomass, net primary productivity, energetics and energy efficiencies were estimated in an age series of Alnus-cardamom plantations in the eastern Himalaya. The impact of stand age (5, 10, 15, 20, 30 and 40 years) on the performance of mixtures of N2-fixing (Alnus nepalensis) and non-N2-fixing (large cardamom) plants was studied. Large cardamom (Amomum subulatum) is the most important perennial cash crop in the region and is cultivated predominantly under Alnus trees. Net primary productivity was lowest (7 t ha(-1) per year) in the 40-year-old stand and was more than three times higher (22 t ha(-1) per year) in the 15-year-old stand. Agronomic yield of large cardamom peaked between 15 and 20 years of age. Cardamom productivity doubled from the 5- to the 15-year-old stand, and then decreased with plantation age to reach a minimum in the 40-year-old stand. Performance of cardamom in association of N2-fixing Alnus remained beneficial until 20 years of age. Annual net energy fixation was highest (444 x 10(6) kJ ha(-1) per year) in the 15-year-old stand, being 1.4 times that of the 5-year-old stand and 2.9-times that of the 40-year-old stand. Inverse relationships of production efficiency, energy conversion efficiency and energy utilized in N2-fixation against stand age, and a positive relationship between production efficiency and energy conversion efficiency suggest that the younger plantations are more productive. The Alnus-cardamom plantation system will be sustainable by adopting a rotational cycle of 15 to 20 years.     

Natural15N abundance as a method of estimating the contribution of biologically fixed nitrogen to N2-fixing systems: Potential for non-legumes

The¹⁵N abundance of plants usually closely reflects the¹⁵N abundance of their major immediate N source(s); plant-available soil N in the case of non-N₂-fixing plants and atmospheric N₂ in the case of N₂ fixing plants. The¹⁵N abundance values of these sources are usually sufficiently different from each other that a significant and systematic difference in the¹⁵N abundance between the two kinds of plants can be detected. This difference provides the basis for the natural¹⁵N abundance method of estimating the relative contribution of atmospheric N₂ to N₂-fixing plants growing in natural and agricultural settings. The natural¹⁵N abundance method has certain advantages over more conventional methods, particularly in natural ecosystems, since disturbance of the system is not required and the measurements may be made on samples dried in the field. This method has been tested mainly with legumes in agricultural settings. The tests have demonstrated the validity of this method of arriving at semi-quantitative estimates of biological N₂-fixation in these settings. More limited tests and applications have been made for legumes in natural ecosystems. An understanding of the limits and utility of this method in these systems is beginning to emerge. Examples of systematic measurements of differences in¹⁵N abundance between non-legume N₂-fixing systems and neighbouring non-fixing systems are more unusual. In principle, application of the method to estimate N₂-fixation by nodulated non-legumes, using the natural¹⁵N abundance method, is as feasible as estimating N₂-fixation by legumes. Most of the studies involving N₂-fixing non-legumes are with this type of system (e.g., Ceanothus, Chamabatia, Eleagnus, Alnus, Myrica, and so forth). Resuls of these studies are described. Applicability for associative N₂-fixation is an empirical question, the answer to which probably depends upon the degree to which fixed N goes predominantly to the plant rather than to the soil N pool. The natural¹⁵N abundance method is probably not well suited to assessing the contribution of N₂-fixation by free-living microorganisms in their natural habitat, particularly soil microorganisms.This work was supported in part by subcontracts under grants from the US National Science Foundation (DEB79-21971 and BSR821618)     

入侵种加拿大一枝黄花对土壤特性的影响

生物入侵已给人类社会带来了巨大的环境和经济损失,但关于植物入侵对生态系统的影响后果还知之甚少。土壤是生态系统的重要组成部分,植物入侵对土壤特性的影响已引起了生态学家的普遍关注。我们将原产北美的入侵杂草加拿大一枝黄花(Solidagocanadensis)和同属土著植物一枝黄花(S.decurrens)种植在具有相同利用历史的土壤中,通过检测生长后期不同植物下土壤特性的差异来研究加拿大一枝黄花的入侵后果。测定的土壤特性包括pH值、总碳、总氮、土壤有机质、硝态氮、铵态氮、净铵化速率、净硝化速率和净矿化速率等。野外选取了加拿大一枝黄花的典型入侵地上海江湾绿地进行土壤特性调查作为辅证。移栽实验和野外调查得到了基本一致的结果:加拿大一枝黄花调节了土壤pH值,增加了总碳、氮库和有机质库,降低了铵氮库和硝氮库。加拿大一枝黄花还促进了微生物的矿化速率和铵化速率,这表明该物种提高了土壤无机氮和铵氮的供给。控制无机氮浓度和无机氮形式的独立实验表明,高氮供给和富铵氮条件有利于加拿大一枝黄花的生长。本研究不仅为外来植物入侵后果的评估提供实验证据,而且为植物成功入侵的机制探索提供思路。     

Characterization of a Mannitol-Utilizing, Nitrogen-Fixing Bradyrhizobium japonicum USDA 110 Derivative

We have isolated a colonial derivative of Bradyrhizobium japonicum USDA 110 (designated MN-110) that is both mannitol utilizing and N(2) fixing. Derivative MN-110 showed growth on mannitol and glucose similar to that of non-N(2)-fixing, mannitol-utilizing L2-110. Derivative MN-110 showed high constitutive and induced d-mannitol dehydrogenase activity (similar to L2-110) relative to N(2)-fixing, non-mannitol-utilizing I-110. Hybridization to EcoRI and HindIII total DNA digests with cloned USDA 110 nif DK and nif H genes revealed similar patterns for non-N(2)-fixing mannitol-utilizing derivative L1-110 and derivative MN-110. Symbiotic tests with soybean cultivars Ransom and Lee indicate MN-110 to be a superior N(2)-fixing derivative compared with derivative I-110 and the parent strain USDA 110. However, these differences were not revealed when comparing 28-day-old soybean-B. japonicum associations but were apparent in 49-day-old associations. It was apparent from this work that mannitol utilization was not necessarily correlated to symbiotic effectiveness in B. japonicum and that gene rearrangements were not responsible for differences in N(2) fixation between L1-110 or L2-110 and MN-110.     

入侵与本地植物对气候变暖和氮沉降交互效应的动态响应

入侵与本地植物对气候变暖和氮沉降交互效应的动态响应 在全球气候变暖背景下,对入侵物种扩张的预测往往并未考虑到同时出现的氮沉降变化。因此,气候变暖和氮沉降的复杂交互将如何改变入侵物种和本地物种的生长动态尚需进一步探索。在此,本研究假设氮沉降和温度的同时增加可能对入侵植物的生长促进效应大于本地植物。本研究在模拟气候变暖、氮沉降及其交互处理下,对入侵植物加拿大一枝黄花(Solidago canadensis L.)及其本地共存物种艾草(Artemisia argyi Levl. et Van)的生长响应进行温室对照试验。结果表明：由于氮沉降对物种生长的显著促进效应,温度升高和氮沉降的交互作用导致入侵物种和本地物种的生长适应性显著提高,即温氮交互可能使区域微生境更加有利于植物生长。然而,在生物量、高度和直径等生态适应特征方面,入侵物种加拿大一枝黄花的相对增加幅度显著低于本地物种艾草,这表明入侵物种加拿大一枝黄花相对于本地物种艾草的生长优势会在未来气候变暖与氮沉降持续增强的背景下逐渐减弱。因此,纳入氮沉降因素可能会缓解入侵物种加拿大一枝黄花在气候变暖条件下的入侵扩张。     

Potential to improve transfer of N in intercropped systems by optimising host-endophyte combinations

Possibilities for improving N transfer from N2-fixing plants to non-N2-fixing plants by mycorrhiza have been investigated. Initially, the genetic variability with respect to N uptake was assessed by screening five varieties of chicory (Cichorium intybus L.), four of peas (Pisum sativum L.) and three of red clover (Trifolium pratense L.) in combination with eight isolates of arbuscular mycorrhizal fungi. The most promising plant - fungi combinations identified through the cultivar screening were used to optimise conditions for N transfer between intercropped N2-fixing plants (peas and clover) and non-N2-fixing chicory. In the first experiment, the recovery of fixed legume N was investigated using three cultivars, of chicory intercropped with pea variety, and inoculated with one of four mycorrhizal isolates. Roots of the N2-fixing pea and the non-N2-fixing chicory were separated by a root-free soil layer in a three-compartment container. A section of the legume roots was forced to grow into a separate compartment which received four split applications of ¹⁵N. The percentage of N in the chicory derived from transfer ranged between 3% and 50%. In a second experiment one chicory variety was intercropped with one red clover variety and inoculated with four mycorrhizal isolates respecetively. A harvest regime was chosen in which the shoots were harvested from intercropped plants at 3,4.5 and 6 months of age. At three months the percentage of N in the chicory derived from transfer ranged between 15% and 18% and at a plant age of 4.5 months from 46 to 77%. At six months the percentage of N in the chicory roots derived from transfer of legume N ranged from 20 to 34% and varied with fungal isolate. Our results show that there is potential for improving N transfer in intercropped plant systems through the methodological selection of suitable plant and mycorrhizal partners.     

Responses of soil microorganisms to resource availability in urban, desert soils

Terrestrial desert ecosystems are strongly structured by the distribution of plants, which concentrate resources and create islands of fertility relative to interplant spaces. Atmospheric nitrogen (N) deposition resulting from urbanization has the potential to change those spatial patterns via resource inputs, resulting in more homogeneous soil resource availability. We sampled soils at 12 desert remnant sites around Phoenix, Arizona along a model-predicted gradient in N deposition to determine the degree to which deposition has altered spatial patterns in soil resource availability and microbial activity. Soil microbial biomass and abundance were not influenced by atmospheric N deposition. Instead, plant islands remained strong organizers of soil microbial processes. These islands of fertility exhibited elevated pools of resources, microbial abundance, and activity relative to interspaces. In both plant islands and interspaces, soil moisture and soil N concentrations predicted microbial biomass and abundance. Following experimental wetting, carbon dioxide (CO₂) flux from soil of interspaces was positively correlated with N deposition, whereas in plant islands, soil CO₂ flux was positively correlated with soil moisture content and soil organic matter. Soil CO₂ flux in both patch types showed rapid and short-lived responses to precipitation, demonstrating the brief time scales during which soil biota may process deposited materials. Although we observed patterns consistent with N limitation of microbes in interspaces, we conclude that atmospheric N deposition likely accumulates in soils because microbes are primarily limited by water and secondarily by carbon or nitrogen. Soil microbial uptake of atmospherically deposited N likely occurs only during sparse and infrequent rainfall.     

The invasive Sorghum halepense harbors endophytic N2-fixing bacteria and alters soil biogeochemistry

Exotic plants invading new habitats frequently initiate broad changes in ecosystem functioning. Sorghum halepense is an invasive grass capable of growing in nitrogen (N)-poor tallgrass prairie soils that creates near monocultures in once phylogenetically diverse-communities. The biogeochemistry of soils invaded by S. halepense was compared to that of un-invaded native prairie soils. Invaded soils contained two to four times greater concentrations of alkaline metals, micronutrients, and essential plant nutrients than native prairie soils. The notable exception was Ca⁺², which was always significantly lower in invaded soils. The N-content of S. halepense above-ground biomass was 6.4 mg g^?1 (320 mg N plant^?1) and suggested a supplemental N source supporting plant growth. Altered soil biogeochemistry in invaded areas coupled with high above-ground biomass in N-poor soils suggested N₂-fixing activity associated with S. halepense. Nitrogenase activity of plant tissues indicated that N₂-fixation was occurring in, and largely restricted to, S. halepense rhizomes and roots. A culture approach was used to isolate these N₂-fixing bacteria from plant tissues, and 16S rRNA gene sequencing was used to identify these bacterial isolates. Nitrogenase activity of bacterial isolates indicated several were capable of N₂-fixation. In addition to N₂-fixation, other roles involved in promoting plant growth, namely mobilizing phosphorus and iron chelation, are known for closest matching relatives of the bacterial isolates identified in this work. Our results indicate that these plant growth-promoting bacteria may enhance the ability of S. halepense to invade and persist by altering fundamental ecosystem properties via significant changes in soil biogeochemistry.     

INTERACTION OF NITROGEN FIXATION AND PHOSPHORUS LIMITATION IN APHANIZOMENON FLOS-AQUAE (CYANOPHYCEAE)1

The effect of simultaneous nitrogen fixation and phosphorus limitation on the physiological adaptation and growth performance of Aphanizomenon flos-aquae (L.) Ralfs PCC 7905 was studied in continuous culture. In the absence of ammonia, N₂ fixation occurred and the maximum growth rate (as determined in diluted batch cultures) was lower. However, no distinction could be made between the steady-state N uptake rates (based on cellular N contents) of N₂-fixing cells and cells grown with ammonia. At the higher dilution rates, the residual P concentration increased with increasing dilution rate, more so under N₂-fixing conditions, compared to the cultures grown in the presence of ammonia. More generally, the yield of biomass per consumed P, as the biomass concentration itself, decreased with increasing dilution rate, and both were lower under N₂-fixing conditions. The restricted biomass production under N₂-fixing conditions suggests that reduction of N loading may benefit lake restoration projects. The influence of N₂-fixation on the severity of P limitation is discussed in terms of metabolic control analysis. From the increase of the residual P concentration on switching from ammonium to N₂-fixing conditions, it is deduced that under N₂-fixing and P-limited conditions, control of growth is shared by N and P metabolism.     

Nitrogen and phosphorus retranslocation and N:P ratios of litterfall in three tropical plantations: luxurious N and efficient P use by Acacia mangium

Some tropical N₂-fixing trees exhibit specific characteristics for phosphorus (P) acquisition and utilisation that contrast with the large nitrogen (N) fluxes in their litterfall. To investigate differences in N and P cycling in N₂-fixing plantations, litterfall and fresh leaf quality of a N₂-fixing Acacia mangium plantation were compared with that of a non-N₂-fixing Swietenia macrophylla plantation and a coniferous Araucaria cunninghamii plantation. The N concentration in the A. mangium litterfall was higher than that in the litterfall of the two other species, whereas the P concentration in the A. mangium leaf litterfall was 0.16 mg g^–1, which was only 12–22% of that of the other species. The P concentration in the reproductive parts of A. mangium was markedly higher (16.1 mg g^–1) than those in the other fractions. The N:P ratio was higher in the leaf fall (81) compared to the fresh leaves (29) of A. mangium, in contrast to the N:P ratios in the leaf samples of the other two species. An analysis of a global litterfall dataset of tropical plantations indicated that N:P ratios in litterfall were significantly higher in N₂-fixers than in non-N₂-fixers, and those of A. mangium were high among species in the N₂-fixer group. These results indicated that A. mangium efficiently retranslocated P in contrast to very large N cycling, under field conditions. These differences may be related to other physiological characteristics of A. mangium.     

Quantitative shotgun proteomics of enriched heterocysts from Nostoc sp. PCC 7120 using 8-plex isobaric peptide tags

The filamentous cyanobacterium Nostoc sp. strain PCC 7120 is capable of fixing atmospheric nitrogen. The labile nature of the core process requires the terminal differentiation of vegetative cells to form heterocysts, specialized cells with altered cellular and metabolic infrastructure to mediate the N2-fixing process. We present an investigation targeting the cellular proteomic expression of the heterocysts compared to vegetative cells of a population cultured under N2-fixing conditions. New 8-plex iTRAQ reagents were used on enriched replicate heterocyst and vegetative cells, and replicate N2-fixing and non-N2-fixing filaments to achieve accurate measurements. With this approach, we successfully identified 506 proteins, where 402 had confident quantifications. Observations provided by purified heterocyst analysis enabled the elucidation of the dominant metabolic processes between the respective cell types, while emphasis on the filaments enabled an overall comparison. The level of analysis provided by this investigation presents various tools and knowledge that are important for future development of cyanobacterial biohydrogen production.     

δ<Superscript>15</Superscript>N constraints on long-term nitrogen balances in temperate forests

Biogeochemical theory emphasizes nitrogen (N) limitation and the many factors that can restrict N accumulation in temperate forests, yet lacks a working model of conditions that can promote naturally high N accumulation. We used a dynamic simulation model of ecosystem N and δ¹⁵N to evaluate which combination of N input and loss pathways could produce a range of high ecosystem N contents characteristic of forests in the Oregon Coast Range. Total ecosystem N at nine study sites ranged from 8,788 to 22,667 kg ha⁻¹ and carbon (C) ranged from 188 to 460 Mg ha⁻¹, with highest values near the coast. Ecosystem δ¹⁵N displayed a curvilinear relationship with ecosystem N content, and largely reflected mineral soil, which accounted for 96–98% of total ecosystem N. Model simulations of ecosystem N balances parameterized with field rates of N leaching required long-term average N inputs that exceed atmospheric deposition and asymbiotic and epiphytic N₂-fixation, and that were consistent with cycles of post-fire N₂-fixation by early-successional red alder. Soil water δ¹⁵NO₃ ⁻ patterns suggested a shift in relative N losses from denitrification to nitrate leaching as N accumulated, and simulations identified nitrate leaching as the primary N loss pathway that constrains maximum N accumulation. Whereas current theory emphasizes constraints on biological N₂-fixation and disturbance-mediated N losses as factors that limit N accumulation in temperate forests, our results suggest that wildfire can foster substantial long-term N accumulation in ecosystems that are colonized by symbiotic N₂-fixing vegetation.     

Two-year interactions between invasive Solidago canadensis and soil decrease its subsequent growth and competitive ability

Aims Plant–soil interaction (PSI) has been implicated as a causative mechanism promoting plant invasions, and some mechanisms underlying PSI effects remain unclear. Here, we attempted to address how altered soil microbes and nutrients influence PSI effects.Methods Soil was cultured by an invasive forb Solidago canadensis for two years. We conducted an experiment, in which S. canadensis and Chinese natives were grown either alone or together in control and cultured soils, and determined the growth of S. canadensis and five natives and the competitive ability of S. canadensis. We analyzed the microbial community composition and nutrients of two types of soils.Important findings Compared to the control soil, the soil cultured by S. canadensis decreased the subsequent growth of S. canadensis and five Chinese natives, as well as the competitive ability of S. canadensis against Chinese natives. Soil microbial community composition was significantly altered due to soil culturing. Total fatty acids, bacteria, Gram-negative bacteria and Gram-positive bacteria had no responses to soil culturing; fungi, aerobic bacteria and fungi/bacteria ratio significantly decreased with soil culturing; anaerobes and Gram-negative/positive bacteria ratio greatly increased with soil culturing. Soil nitrogen (N) dramatically decreased with soil culturing, whereas soil phosphorus (P) was unchanged. These results suggest that negative PSI effects may be linked to decreases in soil fungi, aerobic bacteria and soil N and increases in soil anaerobic bacteria and the ratio of Gram-negative/positive bacteria. Our findings provide an initial indication that S. canadensis– soil interaction alone could exhibit limited contributions to its success in the early stage of invasion.     

The impact of simulated chronic nitrogen deposition on the biomass and N2-fixation activity of two boreal feather moss–cyanobacteria associations

Bryophytes achieve substantial biomass and play several key functional roles in boreal forests that can influence how carbon (C) and nitrogen (N) cycling respond to atmospheric deposition of reactive nitrogen (N_r). They associate with cyanobacteria that fix atmospheric N₂, and downregulation of this process may offset anthropogenic N_r inputs to boreal systems. Bryophytes also promote soil C accumulation by thermally insulating soils, and changes in their biomass influence soil C dynamics. Using a unique large-scale (0.1 ha forested plots), long-term experiment (16 years) in northern Sweden where we simulated anthropogenic N_r deposition, we measured the biomass and N₂-fixation response of two bryophyte species, the feather mosses Hylocomium splendens and Pleurozium schreberi. Our data show that the biomass declined for both species; however, N₂-fixation rates per unit mass and per unit area declined only for H. splendens. The low and high treatments resulted in a 29% and 54% reduction in total feather moss biomass, and a 58% and 97% reduction in total N₂-fixation rate per unit area, respectively. These results help to quantify the sensitivity of feather moss biomass and N₂ fixation to chronic N_r deposition, which is relevant for modelling ecosystem C and N balances in boreal ecosystems.     

Role of the Azotobacter vinelandii nitrogenase-protective shethna protein in preventing oxygen-mediated cell death

Azotobacter vinelandii strains lacking the nitrogenase-protective Shethna protein lost viability upon carbon-substrate deprivation in the presence of oxygen. This viability loss was dependent upon the N(2)-fixing status of cultures (N(2)-fixing cells lost viability, while non-N(2)-fixing cells did not) and on the ambient O(2) level. Supra-atmosheric O(2) tensions (40% partial pressure) decreased the viable cell number of the mutant further, and the mutant had a slightly higher spontaneous mutation frequency than the wild type in the high-O(2) conditions. Iron starvation conditions, which resulted in fourfold-reduced superoxide dismutase levels, were also highly detrimental to the viability of the protective protein mutants, but these conditions did not affect the viability of the wild-type strain. Nitrogenase or other powerful reductants associated with N(2) fixation may be sources of damaging partially reduced oxygen species, and the production of such species are perhaps minimized by the Shethna protein.