Distribution of Plasmids in Heterocystous Cyanobacteria

The distribution of plasmids was studied in 15 different heterocystous cyanobacteria using agarose gel electrophoresis. Out of these, six were found to possess only one plasmid DNA of size ranging from 30–33 mD. A simplified and less time consuming protocol for studying the plasmids has been suggested.     

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.     

Epiphytic Cyanobacteria on Chara vulgaris Are the Main Contributors to N2 Fixation in Rice Fields

The distribution of nitrogenase activity in the rice-soil system and the possible contribution of epiphytic cyanobacteria on rice plants and other macrophytes to this activity were studied in two locations in the rice fields of Valencia, Spain, in two consecutive crop seasons. The largest proportion of photodependent N₂ fixation was associated with the macrophyte Chara vulgaris in both years and at both locations. The nitrogen fixation rate associated with Chara always represented more than 45% of the global nitrogenase activity measured in the rice field. The estimated average N₂ fixation rate associated with Chara was 27.53 kg of N ha⁻¹ crop⁻¹. The mean estimated N₂ fixation rates for the other parts of the system for all sampling periods were as follows: soil, 4.07 kg of N ha⁻¹ crop⁻¹; submerged parts of rice plants, 3.93 kg of N ha⁻¹ crop⁻¹; and roots, 0.28 kg of N ha⁻¹ crop⁻¹. Micrographic studies revealed the presence of epiphytic cyanobacteria on the surface of Chara. Three-dimensional reconstructions by confocal scanning laser microscopy revealed no cyanobacterial cells inside the Chara structures. Quantification of epiphytic cyanobacteria by image analysis revealed that cyanobacteria were more abundant in nodes than in internodes (on average, cyanobacteria covered 8.4% ± 4.4% and 6.2% ± 5.0% of the surface area in the nodes and internodes, respectively). Epiphytic cyanobacteria were also quantified by using a fluorometer. This made it possible to discriminate which algal groups were the source of chlorophyll a. Chlorophyll a measurements confirmed that cyanobacteria were more abundant in nodes than in internodes (on average, the chlorophyll a concentrations were 17.2 ± 28.0 and 4.0 ± 3.8 μg mg [dry weight] of Chara⁻¹ in the nodes and internodes, respectively). These results indicate that this macrophyte, which is usually considered a weed in the context of rice cultivation, may help maintain soil N fertility in the rice field ecosystem.     

Iron-Stimulated N2 Fixation and Growth in Natural and Cultured Populations of the Planktonic Marine Cyanobacteria Trichodesmium spp

In light of recent proposals that iron (Fe) availability may play an important role in controlling oceanic primary production and nutrient flux, its regulatory impact on N₂ fixation and production dynamics was investigated in the widespread and biogeochemically important diazotrophic, planktonic cyanobacteria Trichodesmium spp. Fe additions, as FeCl₃ and EDTA-chelated FeCl₃, enhanced N₂ fixation (nitrogenase activity), photosynthesis (CO₂ fixation), and growth (chlorophyll a production) in both naturally occurring and cultured (on unenriched oligotrophic seawater) Trichodesmium populations. Maximum enhancement of these processes occurred under FeEDTA-amended conditions. On occasions, EDTA alone led to enhancement. No evidence for previously proposed molybdenum or phosphorus limitation was found. Our findings geographically extend support for Fe limitation of N₂ fixation and primary production to tropical and subtropical oligotrophic ocean waters often characterized by Trichodesmium blooms.     

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.     

Nitrogen Fixation and Hydrogen Metabolism in Cyanobacteria

Summary: This review summarizes recent aspects of (di)nitrogen fixation and (di)hydrogen metabolism, with emphasis on cyanobacteria. These organisms possess several types of the enzyme complexes catalyzing N₂ fixation and/or H₂ formation or oxidation, namely, two Mo nitrogenases, a V nitrogenase, and two hydrogenases. The two cyanobacterial Ni hydrogenases are differentiated as either uptake or bidirectional hydrogenases. The different forms of both the nitrogenases and hydrogenases are encoded by different sets of genes, and their organization on the chromosome can vary from one cyanobacterium to another. Factors regulating the expression of these genes are emerging from recent studies. New ideas on the potential physiological and ecological roles of nitrogenases and hydrogenases are presented. There is a renewed interest in exploiting cyanobacteria in solar energy conversion programs to generate H₂ as a source of combustible energy. To enhance the rates of H₂ production, the emphasis perhaps needs not to be on more efficient hydrogenases and nitrogenases or on the transfer of foreign enzymes into cyanobacteria. A likely better strategy is to exploit the use of radiant solar energy by the photosynthetic electron transport system to enhance the rates of H₂ formation and so improve the chances of utilizing cyanobacteria as a source for the generation of clean energy.     

Response of N2-Fixing Cyanobacteria to Salt

The effect of salt on photosynthetic activity, acetylene reduction, and related activities was examined in two species of cyanobacteria, Nostoc muscorum and Calothrix scopulorum. Photosynthesis was more resistant to high salt concentration than was N₂ fixation. The salt resistance of both activities increased after a period of exposure of the cells to salinity. The transfer of electrons via ferredoxin and ferredoxin-nicotinamide adenine dinucleotide phosphate reductase was found to be extremely sensitive to salt. In comparison, the transfer of reducing power by glucose-6-phosphate dehydrogenase, isocitric dehydrogenase, and photosystem 1 was less affected by NaCl, whereas glutamine synthetase exhibited higher tolerance to salt.     

Iron-Stimulated N(2) Fixation and Growth in Natural and Cultured Populations of the Planktonic Marine Cyanobacteria Trichodesmium spp

In light of recent proposals that iron (Fe) availability may play an important role in controlling oceanic primary production and nutrient flux, its regulatory impact on N(2) fixation and production dynamics was investigated in the widespread and biogeochemically important diazotrophic, planktonic cyanobacteria Trichodesmium spp. Fe additions, as FeCl(3) and EDTA-chelated FeCl(3), enhanced N(2) fixation (nitrogenase activity), photosynthesis (CO(2) fixation), and growth (chlorophyll a production) in both naturally occurring and cultured (on unenriched oligotrophic seawater) Trichodesmium populations. Maximum enhancement of these processes occurred under FeEDTA-amended conditions. On occasions, EDTA alone led to enhancement. No evidence for previously proposed molybdenum or phosphorus limitation was found. Our findings geographically extend support for Fe limitation of N(2) fixation and primary production to tropical and subtropical oligotrophic ocean waters often characterized by Trichodesmium blooms.     

Phylogenomics Uncovers Evolutionary Trajectory of Nitrogen Fixation in Cyanobacteria

Biological nitrogen fixation (BNF) by cyanobacteria is of significant importance for the Earth’s biogeochemical nitrogen cycle but is restricted to a few genera that do not form monophyletic group. To explore the evolutionary trajectory of BNF and investigate the driving forces of its evolution, we analyze 650 cyanobacterial genomes and compile the database of diazotrophic cyanobacteria based on the presence of nitrogen fixation gene clusters (NFGCs). We report that 266 of 650 examined genomes are NFGC-carrying members, and these potentially diazotrophic cyanobacteria are unevenly distributed across the phylogeny of Cyanobacteria, that multiple independent losses shaped the scattered distribution. Among the diazotrophic cyanobacteria, two types of NFGC exist, with one being ancestral and abundant, which have descended from diazotrophic ancestors, and the other being anaerobe-like and sparse, possibly being acquired from anaerobic microbes through horizontal gene transfer. Interestingly, we illustrate that the origin of BNF in Cyanobacteria coincide with two major evolutionary events. One is the origin of multicellularity of cyanobacteria, and the other is concurrent genetic innovations with massive gene gains and expansions, implicating their key roles in triggering the evolutionary transition from nondiazotrophic to diazotrophic cyanobacteria. Additionally, we reveal that genes involved in accelerating respiratory electron transport (coxABC), anoxygenic photosynthetic electron transport (sqr), as well as anaerobic metabolisms (pfor, hemN, nrdG, adhE) are enriched in diazotrophic cyanobacteria, representing adaptive genetic signatures that underpin the diazotrophic lifestyle. Collectively, our study suggests that multicellularity, together with concurrent genetic adaptations contribute to the evolution of diazotrophic cyanobacteria.     

几种光合作用光响应典型模型的比较研究

光响应曲线是判定植物光合效率的重要方法,通过曲线可以获得植物光合特性的相关生理参数,但不同模型提取的光响应参数和指标存在差异。本文选择直角双曲线、非直角双曲线和两种指数曲线模型,分别对3个品系常绿杨光响应数据进行拟合,提取了各光响应曲线模型的主要特征参数,对比分析了各模型参数问的差异,并对光饱和点（LSP）的不同计算方法进行了讨论。最后用巨尾桉光响应数据对分析结果作了进一步验证。结果表明,直角和非直角双曲线模型拟合的最大净光合速率（P＇max）、表观量子效率（a）及暗呼吸速率（Rd）值高于指数模型拟合值,且直角双曲线拟合的各参数均比非直角双曲线拟合的各参数的值大,而两指数模型各参数拟合值基本一致;在LSP计算方法中,用光通量200μmol·m^-2·s^-1以下的点拟合的Pn-I直线与其它模型相结合得到光饱和点的方法不可靠,会使计算结果明显偏小,用接近最大总光合速率Pmax一定比例的方法估计LSP也存在较大偏差,而P＇max由于消除了Rd的影响,计算光饱和点时各模型的估计比例相对固定,是一个比较理想的LSP估计方法,初步得出直角、非直角及指数模型用P＇max来估计光饱和点时应选取的比例分别为（78±1）％、（82±1）％及（96±1）％。     

Light Harvesting and State Transitions in Cyanobacteria

Cyanobacteria are oxygenic phototrophic prokaryotes and are considered to be the ancestors of chloroplasts. Their photosynthetic machinery is functionally equivalent in terms of primary photochemistry and photosynthetic electron transport. Fluorescence measurements and other techniques indicate that cyanobacteria, like plants, are capable of redirecting pathways of excitation energy transfer from light harvesting antennae to both photosystems. Cyanobacterial cells can reach two energetically different states, which are defined as “State 1” (obtained after preferential excitation of photosystem I) and “State 2” (preferential excitation of photosystem II). These states can be distinguished by static and time resolved fluorescence techniques. One of the most important conclusions reached so far is that the presence of both photosystems, as well as certain antenna components, are necessary for state transitions to occur. Spectroscopic evidence suggests that changes in the coupling state of the light harvesting antenna complexes (the phycobilisomes) to both photosystems occur during state transitions. The finding that the phycobilisome complexes are highly mobile on the surface of the thylakoid membrane (the mode of interaction with the thylakoid membrane is essentially unknown), has led to the proposal that they are in dynamic equilibrium with both photosystems and regulation of energy transfer is mediated by changes in affinity for either photosystem.     

Effect of Indoleacetic Acid on Growth and Dinitrogen Fixation in Cyanobacteria

The effect of IAA on growth, dinitrogen fixation, and heterocystsfrequency of Anabaena PCC 7119 and Nodularia sp. have been investigated.Concentrations of IAA ranging from 10^–10 to 10^–4M did not change the growth of Anabaena PCC 7119. Concentrationshigher than 10^–4 M were inhibitory. Similar results werefound in Nodularia sp. although in this case the inhibitoryeffect appeared with 10^–5M of IAA. Neither the nitrogenaseactivity nor the heterocysts frequency were enhanced by IAAtreatment. (Received June 17, 1986; Accepted January 22, 1987)     

Microscale Vertical Profiles of N2 Fixation, Photosynthesis, O2, Chlorophyll a, and Light in a Cyanobacterial Assemblage

Profiles of ¹⁵N₂ fixation, O₂ production (gross photosynthesis), O₂ concentration, chlorophyll a concentration, and photon fluence rates were measured with 50-μm resolution in colonies of the heterocyst-forming cyanobacterium Nostoc parmelioides. Microelectrode measurements were made after 20 h of incubation under ¹⁵N₂ gas. Colonies were frozen, and 50-μm sections were prepared by using a freezing microtome and analyzed for ¹⁵N enrichment and chlorophyll a concentration. Colonies exhibited steep spatial gradients in rates of gross photosynthesis, O₂ concentration, and irradiance, with the highest values generally occurring at the surface. O₂ concentration, photosynthesis, and irradiance all showed positive correlations, but chlorophyll a concentrations varied independently of photosynthesis and irradiance. Forty-four percent of the variation in ¹⁵N incorporation was explained by gross photosynthesis (a positive correlation) when incorporation of ¹⁵N was expressed per unit of biomass (chlorophyll a).     

Green Light Drives CO2 Fixation Deep within Leaves

Maximal ^l4CO₂-fixation in spinach occurs in the middle of thepalisade mesophyll [Nishio et al. (1993) Plant Cell 5: 953],however, ninety percent of the blue and red light is attenuatedin the upper twenty percent of a spinach leaf [Cui et al. (1991)Plant Cell Environ. 14: 493]. In this report, we showed thatgreen light drives ¹⁴CO₂-fixation deep within spinach leavescompared to red and blue light. Blue light caused fixation mainlyin the palisade mesophyll of the leaf, whereas red light drovefixation slightly deeper into the leaf than did blue light.¹⁴CO₂-fixation measured under green light resulted in less fixationin the upper epidermal layer (guard cells) and upper most palisademesophyll compared to red and blue light, but led to more fixationdeeper in the leaf than that caused by either red or blue light.Saturating white, red, or green light resulted in similar maximal¹⁴CO₂-fixation rates, whereas under the highest irradiance ofblue light given, carbon fixation was not saturated, but itasymptotically approached the maximal ¹⁴CO₂-fixation rates attainedunder the other types of light. The importance of green lightin photosynthesis is discussed. ¹Supported in part by grants from Competitive Research GrantsOffice, U.S. Department of Agriculture (Nos. 91-37100-6672 and93-37100-8855).     

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     

大鼠吗啡依赖模型的建立

本实验采用剂量递增法,吗啡的给药剂量从10 mg/kg递增至80 mg/kg,用药5天停药,并经腹腔注射纳洛酮2 mg/kg催促后,实验观察大鼠主要行为学和植物神经症状表现.结果表明戒断症状明显,成功建立了大鼠吗啡依赖催促戒断模型.     

Neighborhood Dependence in Bayesian Spatial Models

The conditional autoregressive model and the intrinsic autoregressive model are widely used as prior distribution for random spatial effects in Bayesian models. Several authors have pointed out impractical or counterintuitive consequences on the prior covariance matrix or the posterior covariance matrix of the spatial random effects. This article clarifies many of these puzzling results. We show that the neighborhood graph structure, synthesized in eigenvalues and eigenvectors structure of a matrix associated with the adjacency matrix, determines most of the apparently anomalous behavior. We illustrate our conclusions with regular and irregular lattices including lines, grids, and lattices based on real maps.