动物的共生菌固氮

介绍了共生菌固氮涉及的动物和微生物类群、动物共生菌固氮的性质和机理。应用乙炔还原法和固氮酶基因检测等研究表明,所涉及的动物有7门13纲23目50科99属174种。动物肠道具有丰富的微生境,供不同生理需求的固氮菌生长发育,所蕴含的共生固氮菌类群也十分丰富,涵盖植物共生固氮菌、植物内生固氮菌、植物根际固氮菌、自生固氮菌等生态类型。一般认为动物共生固氮菌来源于环境,其性质属于联合共生固氮。动物共生固氮菌一般与其他共生生物形成复合体,以满足固氮过程中对电子和质子供体、能量供给、固氮酶活性保护以及氨阻遏解除等方面的需求。动物共生菌固氮产物氨的同化也需要多种共生物的协同作用,可能通过谷氨酰胺合成酶/谷氨酸合成酶等途径。总体上,食物氮、非蛋白氮和共生菌固氮相互协调,形成营养和解毒的代谢网络,共同维持动物体内氮素营养的动态平衡,并对未来研究提出展望。     

蓝藻Anabaena 7120固氮的光调节

蓝藻Anabaena 7120固氮是一个依赖于光的过程,暗中乙炔还原活性检测不出来。预先以强光、弱光或黑暗对蓝藻进行短期处理,也会引起其固氮活性呈现相应的高低差异。在蓝藻固氮反应中加入NH_4Cl、KNO_3或尿素等结合态氮时,乙炔还原活性即受到抑制,弱光加剧这种抑制,受尿素抑制的蓝藻固氮活性恢复时,强光下比弱光下快。一系列光合抑制剂对蓝藻固氮活性都有抑制作用,弱光下此种抑制加剧。尿素和此类抑制剂对蓝藻固氮活性呈现协同性抑制,其效应和弱光下尿素对固氮抑制加剧是一致的。受尿素抑制的蓝藻固氮活性恢复过程中,加入光合抑制剂的恢复慢,而且只能维持在一个很低的水平上。     

不同时期艾比湖湿地盐角草群落土壤固氮微生物的多样性分析

【目的】研究新疆艾比湖湿地不同季节盐角草根际和非根际土壤固氮微生物的多样性和丰富度与环境因子的相关性,以期探究在荒漠化和盐渍化不断严重的艾比湖湿地中随着季节变化的固氮微生物群落对恢复生态功能起到的潜在作用,为后续的湿地保护和退化恢复工作提供理论支持和数据基础。【方法】应用Illumina HiSeq PE250测序技术,分析6个土壤样本固氮微生物的多样性,结合相关的理化因子并利用RDA分析法探究土壤理化性质和固氮微生物菌落结构及丰富度的相关性。【结果】艾比湖湿地盐角草植物根际土壤的固氮微生物多样性高于非根际土壤,7月的土壤固氮微生物多样性高于10月和4月的土壤。土杆菌属(Geobacter)、假单胞菌属(Pseudomonas)、固氮菌属(Azotobacter)和慢生根瘤菌属(Bradyrhizobium)等为盐角草根际和非根际土壤中的共同优势菌属。这些固氮微生物优势菌属隶属于变形菌门(Proteobacteria)和蓝藻门(Cyanobacteria),且相对丰富度占比为85%和10%,其余各菌门共占比较少,仅为5%。土壤中固氮微生物的优势菌群与碱解氮(AN)、全氮(TN)、速效钾(AK)和有效磷(TP)呈显著相关。【结论】随着时间的推移土壤样本中固氮微生物的多样性和群落结构也发了改变,同一时期植物根际与非根际土壤中固氮微生物的群落结构并不相同。土壤的环境因子与固氮细菌的群落结构和丰富度的相关性研究可以为艾比湖湿地的退化恢复提供数据基础和理论支持。     

铜尾矿废弃地生物土壤结皮固氮微生物多样性

生物土壤结皮能够有效提高矿业废弃地有机质和氮的积累,促进植被的恢复.本研究基于固氮微生物的nifH基因多样性,以铜陵铜尾矿废弃地3种类型的生物土壤结皮(藻结皮、藓结皮和藻-藓混合结皮)为对象,利用变性梯度凝胶电泳(DGGE)技术研究生物土壤结皮中固氮微生物的多样性以及废弃地植物群落发育对其产生的影响.结果表明： 尾矿库裸地表面的藻结皮的固氮微生物多样性最高,其次为维管植物群落下的藻-藓混合结皮,苔藓结皮的固氮蓝藻多样性最低;随着维管植物群落高度和盖度的增加,固氮微生物多样性降低,铜尾矿废弃地的pH、水分、有机质、养分含量(氮和磷)以及有效态和总重金属浓度对固氮微生物多样性的影响均不显著.测序和系统发育分析表明,废弃地结皮中固氮微生物以蓝藻为主,主要为不具有异形胞的丝状蓝藻.
     

宏基因组测序分析东寨港红树林淤泥和水体微生物的多样性

为深入了解海南东寨港红树林生态系统微生物多样性及其在氮、磷、硫等代谢循环中的功能特点,本研究采用宏基因组测序,从物种注释与丰度、群落功能及多样性指数等角度,分析红树林淤泥和水体生境中微生物群落结构及生态功能的特异性。结果显示,在淤泥中检测到53个门、909个属的微生物类群,有3个占比超过1%的优势门类,其中变形杆菌门为83.78%,处于绝对优势,其下的12个优势属全部来自变形杆菌门;不动杆菌属是聚磷微生物的主要类群,其在淤泥中含量是水体的107.7倍,硫氧化单胞菌属、脱硫杆菌属是硫化物代谢的主要菌属,主要存在于淤泥生境当中。在水体中检测到64个门、1 522个属,包括13个优势门类、7个优势属;Nitrospinae和硝化螺旋菌门是亚硝酸盐氧化代谢的关键类群,两者在水体中占比分别是淤泥中的28.1倍和6.8倍。多样性评估得知,水体样品中的Shannon Wiener指数和Simpson指数均高于淤泥样品,两样品在属分类学单元上的Simpson指数趋近于1,表明红树林生态系统具有非常高的微生物多样性,水体生境的微生物多样性高于淤泥;亚硝酸盐的微生物代谢循环主要发生在水体生境中,微生物对磷的富集作用和硫化合物的氧化还原代谢主要发生在淤泥生境中。本研究有助于认识东寨港红树林湿地生境中的微生物资源状况,为保护红树林生态系统和开发利用其中的微生物资源提供依据。     

三峡库区水体中固氮微生物多样性及其影响因素

【目的】研究分析不同时空条件下三峡库区水体固氮微生物多样性,并探讨其与地球化学参数的相关性。【方法】采集三峡库区不同时间(三月份和六月份)和空间(干流与支流)的水体样品,对其进行地球化学参数分析,并通过构建克隆文库分析样品中固氮功能基因(nifH)的多样性进而探讨其与水体地化参数的相关关系。【结果】统计分析显示三峡库区水体固氮微生物α-多样性和群落组成具有时空差异。支流水体样品的固氮微生物α-多样性高于干流水体样品;六月水体样品的固氮微生物α-多样性高于三月水体样品。三峡库区三月水体样品中的固氮微生物群落以Proteobacteria (50.3%)和Firmicutes (40.0%)为主;六月水体样品的固氮微生物群落以Proteobacteria(48.4%)、Firmicutes(25.4%)和Cyanobacteria(19.0%)为主。Mantel检验结果显示:固氮微生物群落结构的差异与温度、pH和DIC等地球化学参数具有显著(P0.05)相关性,其中温度和pH的相关性系数最大。【结论】三峡库区固氮微生物的种群结构和多样性具有时空差异,影响三峡水库水体中固氮微生物群落结构与多样性的主要环境因素为温度和pH,同时浊度、DIC、氨氮也对库区水体固氮微生物群落结构和多样性有一定的影响。     

亚热带分层水库固氮微生物时空分布格局

生物固氮作用是水生态系统氮元素的重要来源途径之一,通常通过固氮微生物实现。但是,目前人们对亚热带分层水库固氮微生物多样性、分布和丰度认识还非常有限。以厦门市汀溪水库为例,基于固氮基因(nifH)综合应用克隆文库、定量PCR、定量RT-PCR研究固氮微生物在不同季节和不同水层的时空分布格局与演替规律。结果表明,汀溪水库具有丰富多样的固氮微生物,包括蓝藻、α-变形菌、β-变形菌、γ-变形菌、厚壁菌,以及少量未知的固氮细菌和序列;固氮微生物的群落组成、丰度、多样性和活性均呈现显著的时空差异。春、夏和秋3季表层和底层蓝藻nifH基因序列所占比例均超过50%,其中表层高于底层;冬季表层和底层蓝藻OTU数目比例超过50%。聚类分析表明,冬季表层和底层群落汇聚为一类;春、夏和秋三个季节表层首先聚为一类,然后与底层分别汇为一支。汀溪水库热分层时期的固氮微生物群落组成的空间差异大于季节差异,而且表层水体蓝藻在所有固氮微生物中占据绝对优势地位。相关分析表明,固氮微生物RNA丰度和RNA/DNA分别与氨氮、水温显著负相关;固氮微生物DNA丰度与溶解氧、pH、叶绿素a显著负相关,与硝氮显著正相关。综上所述,亚热带水库热分层对固氮微生物的群落结构具有显著的影响,在水库环境保护和生态管理中,特别是蓝藻水华防控时,要充分考虑水体热分层的生态效应。     

三种荒漠灌木根际可培养固氮细菌类群及其固氮和产铁载体能力

【目的】揭示西鄂尔多斯荒漠孑遗灌木四合木(Tetraena mongolica)、沙冬青(Ammopiptanthus mongolicus)、白刺(Nitraria tangutorum)根际可培养固氮细菌类群,分析固氮酶活性和产铁载体能力,以期为认识和利用荒漠植物根际促生细菌提供依据。【方法】以Ashby无氮培养基、采用涂布划线法分离纯化根际固氮细菌;基于16S r RNA基因分析类群组成;以乙炔还原方法测定固氮酶活性;以铬天青S蓝色平板定性筛选产铁载体菌株,以分光光度计法定量产铁载体能力。【结果】共分离出固氮细菌22株,分别属于3个门与9个属,其中变形菌门(Proteobacteria,82%)为绝对优势门,假单胞菌属(Pseudomonas,27.27%)为优势属;Rhizobium和Bacillus分别是沙冬青和四合木的独有属,而白刺独有属有Enterobacter、Stenotrophomonas和Paenibacillus;10株在无氮培养基上生长迅速,它们的固氮酶活性在871.71-3 383.09 nmol C2H4/(H·Culture)之间,并且其中有7株具有产铁载体能力,其产铁载体的As/Ar值范围为0.35-0.79。【结论】鄂尔多斯荒漠珍稀孑遗灌木植物根际固氮细菌类群多样,植物间差异明显,包含多种高固氮酶活性和产铁载体能力的固氮细菌,可作为植物生长促进根际细菌的重要来源。     

宝天曼落叶阔叶林土壤细菌多样性

土壤微生物在森林生态系统中起着重要作用。高通量测序方法的出现为进一步认识土壤微生物提供了契机。本文利用Illumina Miseq高通量测序技术对宝天曼森林土壤的细菌多样性进行了初步研究。结果显示: 在31个采样点内, 随着采样点增加, 检测出不同分类水平的土壤细菌类群也在增多, 当采样点达到31个时, 检测出的土壤细菌类群达到45门163纲319目495科785属和42,632个OTU; 31个土壤样品中所检测出的细菌类群平均有34.2门114.7纲215.2目323.7科446.6属5,924.7个OTU, 其中门、纲、目分类水平上的优势类群(所占比例)分别为变形菌门(Proteobacteria)(38.30%)、α-变形菌纲(α-Proteobacteria)(18.08%)、根瘤菌目(Rhizobiales)(10.62%)。这些初步研究结果表明在一定程度上宝天曼森林土壤有较高的细菌多样性水平, 为进一步认识森林土壤细菌多样性与植物多样性关系等奠定了基础。     

豆科作物间作燕麦对土壤固氮微生物丰度和群落结构的影响

利用荧光定量PCR和高通量测序的研究方法,比较了3种种植模式(燕麦单作,O;大豆/燕麦,OSO;绿豆/燕麦,OMO)对燕麦土壤固氮微生物数量和群落组成的影响.结果表明: 与大豆和绿豆间作显著改变了燕麦土壤的理化性质.燕麦土壤固氮微生物nifH基因拷贝数为每克干土1.75×10¹⁰～7.37×10¹⁰,拔节期和成熟期OSO和OMO中nifH基因拷贝数分别是O中的2.18、2.64和1.92、2.57倍,且各处理成熟期nifH基因拷贝数显著低于拔节期.样品稀释性曲线和覆盖度结果表明,各样品nifH基因测序结果可靠.与绿豆间作显著提高了燕麦土壤nifH基因的多样性.各样品固氮微生物属水平上优势类群均为Azohydromonas、固氮菌属、慢生根瘤菌属、Skermanella和在属水平上无法归类的固氮微生物,但各优势类群相对丰度存在差异.样品OTU的venn分布和主成分分析显示,拔节期和成熟期nifH基因群落结构存在差异,两个生育时期OSO和OMO具有更相似的nifH基因群落结构.表明与大豆和绿豆间作可显著提高燕麦土壤固氮微生物的数量,并影响固氮微生物的群落组成.     

A numerical model of nitrogen flxation and its application to Lake Valencia,Venezuela*

SUMMARY. 1. A numerical model for calculation of daily and annual nitrogen fixation in lakes is presented. The model is based on empirically-derived equations for the rates of nitrogen fixation by heterocysts (nitrogen-fixing cells) in relation to light and on functions for the vertical and tetnporal distributions of heterocysts and light in a lake. 2. Applications of the model to Lake Valencia, Venezuela, between December 1980 and December 1981 indicated that nitrogen fixation is largely a surface phenomenon in this lake: 80% of diurnal fixation occurred within 1m of the water surface. 3. Nitrogen fixation is largely restricted to periods of lake stratification, when the phytoplankton have sufficient light for growth, but dissolved inorganic nitrogen is scarce. Nitrogen fixation was maximal late in the stratification period of 1981: 85 % of fixation occurred within the last 3 months of the 9-month period. 4. The annual nitrogen fixation in Lake Valencia is 26 kg ha^?1, which is comparable to the nitrogen fixation in temperate eutrophic lakes with seasonal blue-green algal blooms. However, nitrogen fixation accounted for only 23% of the total nitrogen supply to Lake Valencia in 1981.     

ROLE OF THE NITROGEN-FIXING BACTERIAL MICROFLORA IN THE EPIPHYTISM OF TILLANDSIA (BROMELIACEAE)

Nitrogen-fixing activity in the phyllosphere of 12 species of Tillandsia from different Mexican habitats was evaluated by the acetylene reduction assay, and nitrogen-fixing microorganisms were isolated and characterized. The leaves from eight of the 12 Tillandsia species examined exhibited nitrogenase activity in enrichment cultures. Among the microorganisms implicated—Agrobacterium, Bacillus, Erwinia, Pseudomonas, Rahnella, Vibrio, and Xanthomonas—only Bacillus megatherium reduced acetylene in pure culture. Our findings suggest that nitrogen fixation in the phyllosphere of the sampled epiphytes occurs under suitable conditions and that most of the bacteria involved are primarily soil and water inhabitants. The results also suggest a relationship between the composition of the nitrogen-fixing microbial communities grown on the leaf and the different development of the leaf area in Tillandsia due to the aerial components (wings) of the trichomes.     

固氮蓝细菌束毛藻生物固氮策略研究进展

固氮蓝细菌束毛藻(Tricodesmium)是海洋中丰度最高的固氮微生物,贡献了约42%的海洋生物固氮,为海洋生态系统提供了新的氮源,驱动海洋初级生产力和食物网,在海洋生物地球化学循环中发挥重要作用。作为海洋中“新氮”主要贡献者,束毛藻是一种不产生异形胞的丝状固氮蓝细菌。因为生物固氮的关键酶固氮酶对氧气十分敏感,一般固氮蓝细菌通常产生异形胞或采用夜间固氮的方式进行生物固氮,避免氧气对固氮酶的抑制作用。近年来研究发现,束毛藻具有一套独特的生物固氮体系,能够使同一藻丝在白天同时完成光合作用和生物固氮,并具有复杂的调控机制。本文综述了近年来束毛藻生物固氮策略的最新研究进展,介绍了其生物固氮和光合作用之间的精密调控机制,对拓展固氮微生物尤其是海洋蓝细菌固氮机制的认识具有借鉴意义。     

Developing a two-step method for retrieving cyanobacteria abundance from inland eutrophic lakes using MERIS data

Cyanobacteria are the main dominant species in inland eutrophic lakes during algae blooms, and measures of cyanobacteria abundance can be used for monitoring and early detection of algal blooms by remote sensing. During May 2013 and August 2016, a total 137 water samples were collected from Lake Taihu and Lake Chaohu. Remote-sensing reflectance was measured, surface water was collected in the field, and chlorophyll-a concentration, phycocyanin concentration, suspended-matter concentration and phytoplankton pigment absorption parameters were measured in the laboratory. The composition and density of planktonic algae were also detected by microscope examination. The remote-sensing reflectance at 15 MERIS bands was simulated based on our measured spectral data, and a two-step method for detecting cyanobacteria abundance using the partial least squares model based on 5 MERIS bands was developed. The results showed that the estimation algorithm can predict cyanobacteria abundance in inland eutrophic lakes with satisfactory accuracy, with RMSE of 7.56 and MAPE of 13.44 %. This algorithm was successfully applied to the MERIS image acquired on August 12, 2010, and showed a reasonable spatial distribution of cyanobacteria abundance in Lake Taihu. It demonstrated that the developed estimation method was an effective way to monitor cyanobacteria abundance in water with a potential to be successfully applied to Sentinel-3 images.     

Do top-down and bottom-up controls interact to exclude nitrogen-fixing cyanobacteria from the plankton of estuaries? An exploration with a simulation model

Explaining the nearly ubiquitous absence of nitrogen fixation by planktonic organisms in strongly nitrogen-limited estuaries presents a major challenge to aquatic ecologists. In freshwater lakes of moderate productivity, nitrogen limitation is seldom maintained for long since heterocystic, nitrogen-fixing cyanobacteria bloom, fix nitrogen, and alleviate the nitrogen limitation. In marked contrast to lakes, this behavior occurs in only a few estuaries worldwide. Primary production is limited by nitrogen in most temperate estuaries, yet no measurable planktonic nitrogen fixation occurs. In this paper, we present the hypothesis that the absence of planktonic nitrogen fixers from most estuaries is due to an interaction of bottom-up and top-down controls. The availability of Mo, a trace metal required for nitrogen fixation, is lower in estuaries than in freshwater lakes. This is not an absolute physiological constraint against the occurrence of nitrogen-fixing organisms, but the lower Mo availability may slow the growth rate of these organisms. The slower growth rate makes nitrogen-fixing cyanobacteria in estuaries more sensitive to mortality from grazing by zooplankton and benthic organisms.We use a simple, mechanistically based simulation model to explore this hypothesis. The model correctly predicts the timing of the formation of heterocystic, cyanobacterial blooms in freshwater lakes and the magnitude of the rate of nitrogen fixation. The model also correctly predicts that high zooplankton biomasses in freshwaters can partially suppress blooms of nitrogen-fixing cyanobacteria, even in strongly nitrogen-limited lakes. Further, the model indicates that a relatively small and environmentally realistic decrease in Mo availability, such as that which may occur in seawater compared to freshwaters due to sulfate inhibition of Mo assimilation, can suppress blooms of heterocystic cyanobacteria and prevent planktonic nitrogen fixation. For example, the model predicts that at a zooplankton biomass of 0.2 mg l^–1, cyanobacteria will bloom and fix nitrogen in lakes but not in estuaries of full-strength seawater salinity because of the lower Mo availability. Thus, the model provides strong support for our hypothesis that bottom-up and top-down controls may interact to cause the absence of planktonic nitrogen fixation in most estuaries. The model also provides a basis for further exploration of this hypothesis in individual estuarine systems and correctly predicts that planktonic nitrogen fixation can occur in low salinity estuaries, such as the Baltic Sea, where Mo availability is greater than in higher salinity estuaries.     

The Potential for Nitrogen Fixation in a Lake Receiving Sewage Effluent (Grasmere, English Lake District)

The potential for nitrogen fixation in the water column and sediment of Grasmere (English Lake District) was investigated using the acetylene reduction technique, with reference to seasonal changes and concentrations of dissolved oxygen and inorganic nitrogen in the epilimnion and hypolimnion. Potential rates of nitrogen fixation and MPN counts of nitrogen-fixing bacteria correlated with each other and with decreases in dissolved oxygen concentration. The results suggested that selective changes in the nitrogen-fixing microflora took place during the period of thermal stratification. Neither light-dependent fixation, nor cyanobacteria could be detected in water column samples, and acetylene reduction was detected, for most of the season, only in samples of water and sediment to which a source of carbon and phosphorus had been added. The possible role of the sewage effluent in suppressing nitrogen fixation is discussed.     

Targeted deep sequencing reveals high diversity and variable dominance of bloom-forming cyanobacteria in eutrophic lakes

Cyanobacterial blooms in eutrophic lakes are severe environmental problems worldwide. To characterize the spatiotemporal heterogeneity of cyanobacterial blooms, a high-throughput method is necessary for the specific detection of cyanobacteria. In this study, the cyanobacterial composition of three eutrophic waters in China (Taihu Lake, Dongqian Lake, and Dongzhen Reservoir) was determined by pyrosequencing the cpcBA intergenic spacer (cpcBA-IGS) of cyanobacteria. A total of 2585 OTUs were obtained from the normalized cpcBA-IGS sequence dataset at a distance of 0.05. The 238 most abundant OTUs contained 92% of the total sequences and were classified into six cyanobacterial groups. The water samples of Taihu Lake were dominated by Microcystis, mixed Nostocales species, Synechococcus, and unclassified cyanobacteria. Besides, all the samples from Taihu Lake were clustered together in the dendrogram based on shared abundant OTUs. The cyanobacterial diversity in Dongqian Lake was dramatically decreased after sediment dredging and Synechococcus became exclusively dominant in this lake. The genus Synechococcus was also dominant in the surface water of Dongzhen Reservoir, while phylogenetically diverse cyanobacteria coexisted at a depth of 10 m in this reservoir. In summary, targeted deep sequencing based on cpcBA-IGS revealed a large diversity of bloom-forming cyanobacteria in eutrophic lakes and spatiotemporal changes in the composition of cyanobacterial communities. The genus Microcystis was the most abundant bloom-forming cyanobacteria in eutrophic lakes, while Synechococcus could be exclusively dominant under appropriate environmental conditions.     

Field measurements of nitrogen-fixing activity of intact saplings ofAlnus maximowiczii in the subalpine zone of Mt Fuji

Nitrogen-fixing (acetylene-reducing) activity of intact saplings ofAlnus maximowiczii was measured under natural conditions in the subalpine zone of Mt Fuji. The nitrogen-fixing activity was detected from the middle of June when expansion of leaves had just begun to the end of October when the shedding of leaves was almost completed. Diurnal changes in the activity were almost parallel with those of ground temperature. The measured nitrogen-fixing activity was related to ground temperature and total leaf area. Using this relation, annual nitrogen fixation was estimated from the data of ground temperature and leaf area measured in the field. The amount of annual nitrogen fixation was almost the same as that of nitrogen used for annual growth. It was concluded that nitrogen fixation by nodules made a considerable contribution to the nitrogen economy in the saplings ofA. maximowiczii.