水分对发状念珠藻生理活性的影响

研究了水分对发状念珠藻(Nostor flagelliforme Born.et Flah.)生理活性的影响作用。结果表明：干藻体在湿润的过程中,呼吸、光合和固氮活性依次恢复;且随水分含量的增加,光合活性和固氮活性逐渐增强,呼吸作用缓慢减弱并在一定水平上保持相对稳定,自由水是束缚水8倍左右时发菜生理活性全面恢复。吸水饱和的藻体在干燥过程中,光合、呼吸、固氮作用依次停止;呼吸作用随水分的丧失逐渐下降;固氮活性、光合活性在水分丧失20％～40％时有一定程度的增强,出现活性高峰;此后,生理活性下降,水分完全丧失时,光合作用终止,呼吸和固氮作用极其微弱。试验证明,水分是发状念珠藻生理活性的重要限制因子,适宜的水分有助于发菜维持正常的生理代谢和生长。     

话生物肥源——固氮蓝藻的识别和利用

蓝藻能光合自养,属古老的单细胞原核生物。目前已知的有二千余种,其中固氮蓝藻一百多种,常见的有念珠藻属（Ｎｏｓｔｏｃ）、单歧藻属（Ｔｏｌｙｐｏｔ－ｒｉｘ）、鱼腥藻属（Ａｎａｂａｅｎａ）等俗称“青苔”。蓝藻含有藻蓝素,故多为蓝绿色,手感微带粘滑;显微镜下见不到叶绿体和细胞核;用碘液染色,没有蓝色淀粉出现,可视为蓝藻共有的特性;是识别蓝藻并和其它藻类区别开来的方法。鉴别蓝藻有无固氮能力,可看它能否在缺氮的营养液中生长;根据这个原理,就可判断：凡能在潮湿,贫瘠缺氮的表土、岩石、墙基、屋顶、树皮、荒漠、火…     

不同温度对念珠藻生理生化特性的影响

文章尝试比较不同温度对念珠藻（Nostocsp.）细胞生长和生理特性的影响。主要内容是分别在10℃、25℃、35℃和40℃环境中对念珠藻进行9 d的室内培养，然后测定不同温度下念珠藻的生物量、光合活性、总多糖、总蛋白质和总脂质含量。研究结果表明，低温（10℃）对念珠藻细胞生长的促进与光合活性的提高基本无影响；而高温（35℃、40℃）会促进藻细胞的生长和光合活性的提升。也就是说，低温时念珠藻细胞的多糖含量几乎不变，蛋白质含量与脂质含量先升高后降低；而高温时多糖含量和蛋白质含量增加，脂质含量降低。以上结果说明温度胁迫会使念珠藻对生物大分子物质进行调控用于抵抗环境变化。     

黄土高原水蚀风蚀交错区固氮微生物群落多样性在生物结皮中的演变规律

生物土壤结皮在黄土高原的水土保持、养分补给等方面发挥着重要生态功能。固氮微生物是固氮过程的关键功能群,影响着土壤氮素平衡,然而黄土高原水蚀风蚀交错区严重的水土流失导致土壤养分大量损失,尤其是氮素。该区域生物结皮广泛发育,它们在干旱半干旱区土壤氮素积累过程中起着重要作用。但是该区域生物结皮不同发育阶段中的固氮潜力、固氮微生物群落多样性及其关键环境控制因子尚不清楚。以黄土高原水蚀风蚀交错区为研究区,三种生物结皮类型和裸地对照为研究对象,采用Illumina HiSeq平台对nifH基因扩增产物进行生物信息学分析,揭示了固氮潜力及微生物群落多样性的分布规律。结果表明,生物结皮演替显著影响固氮酶活性、nifH基因丰度和固氮微生物α-多样性,各值均在苔藓结皮最高,显著高于其它演替阶段,且生物结皮层高于结皮下层土壤。在固氮微生物群落组成上,裸地以变形菌门的红螺菌目、酸硫杆菌目和根瘤菌目为主;而在藻结皮、地衣结皮和藓结皮阶段,均表现为蓝藻门的念珠藻目占绝对优势,是固氮微生物的关键类群,其中Trichormus、念珠藻属是优势固氮物种;与藻结皮、地衣结皮相比,藓结皮中念珠藻目的相对丰度显著降低,而变形菌门的红螺菌目、酸硫杆菌目和根瘤菌目的丰度明显上升。除此之外,蓝杆藻属、鱼腥藻属、斯克尔曼氏菌属、瘤胃梭菌属在藻结皮、地衣结皮、藓结皮也占明显优势。念珠藻目以及Trichormus的丰度与固氮酶活性、nifH基因丰度呈显著正相关关系,而与红螺菌目、酸硫杆菌目、梭菌目、根瘤菌目呈显著负相关;Trichormus相关的物种在黄土高原生物结皮氮输入过程中可能发挥着关键作用。生物结皮演替通过改变土壤pH值、有机质、全氮、含水量等理化特征而影响了微生境,通过环境筛选和物种重排作用对固氮微生物群落进行综合调控,其中pH与土壤全氮(TN)发挥着关键作用。     

柄杆菌对固氮蓝藻生物量及色素的影响

柄杆菌对固氮蓝藻生物量和色素的影响研究结果表明:多态柄杆菌(Caulobacter polymorphus)017-41或新月柄杆菌(Caulobacter creseentus)CB_2的活菌、死菌及破碎细胞悬液分别与鱼腥藻(Anabaena)、念珠藻(Nostoc)不同藻珠混合培养时,试验组生长量均优于对照组;对衰老黄化的藻培养物的生长刺激作用尤为显著;试验组藻培养物的藻蓝或藻红素含量亦明显高于对照组。其作用机理尚待进一步阐明。     

普通念珠藻营养物质及其利用

本文测定了普通念珠藻的粗蛋白、粗脂肪、糖及矿物质营养元素的含量.并研究普通念珠藻的提取物作为植物生长促进剂. 普通念珠藻的提取物用于小白菜、萝卜、生菜、绿豆及水稻种子能提高发芽率.例如,提取物粉末1g溶于1升水中,这种溶液稀释到1.25—5ppm,并浸种,在发芽率方面可观测到提高.     

异形胞分化相关基因在点形念珠藻厚壁孢子中的转录表达

点形念珠藻(Nostoc punctiforme)ATCC29133是一种丝状固氮蓝藻,能分化产生异形胞、厚壁孢子、藻殖段等细胞类型。异形胞是一种提供微氧条件以进行固氮作用的特化细胞,厚壁孢子是在逆境下产生的能耐受干旱、冰冻等恶劣环境的细胞1。与异形胞相似,厚壁孢子的外被也具有多糖和糖脂成分2,3。厚壁孢子在某些蓝藻藻丝       

3种水稻土中7株固氮蓝细菌的分离与特征

【背景】蓝细菌是水生和陆地生态系统中生物固氮的主要贡献者。【目的】增加对稻田土壤固氮蓝细菌的了解,获得用于进一步研究的可培养固氮蓝细菌菌株。【方法】选择3种具有不同固氮能力的水稻土,采用BG11-N培养基分离培养固氮蓝细菌菌株,对新分离菌株进行形态特征观察,通过基因组DNA的nifH基因扩增明确其固氮潜力,进一步采用乙炔还原法和~(15)N_2示踪法定量测定其固氮能力,通过基因组DNA的16SrRNA基因序列比对进行鉴定。【结果】在光照培养条件下,采用BG11-N培养基共分离纯化得到自养菌株7株,细胞呈圆形或椭圆形、单列、无分枝、丝状和念珠状,在固体培养基上形成团垫状菌落。新分离菌株在BG11-N培养基中生长状况良好,以基因组DNA为模板可扩增出nifH基因,乙炔还原法和~(15)N_2示踪法测定结果显示具有较高固氮能力,同时具有铁载体生成能力。结合16S rRNA基因序列比对和形态特征,7株菌被初步鉴定隶属于念珠藻科(Nostocaceae)。【结论】从水稻土中分离到在稻田生物固氮中发挥重要作用的蓝细菌(念珠藻科)菌株,可培养固氮蓝细菌菌株固氮能力较高,兼具铁载体生成能力,可作为进一步深入研究的微生物资源,具有潜在的研究应用价值。     

发状念珠藻藻殖段的分化及其光合特性的研究

发状念珠藻 (NostocflagelliformeBorn .etFlah .)存在着两个重要而明显的个体发育阶段 ,即营养藻丝体和藻殖段。采用弱光 (铺垫砂粒遮光 ) ,红光或在白光下向培养基中加入DCMU (3,4_dichlorophenyl_1,1_dimethylurea)等方法 ,可促进营养藻丝体转变成藻殖段。用可见光吸收光谱、低温荧光发射光谱和光合放氧活性表示发状念珠藻藻丝体与藻殖段的光合特性 ,表明营养藻丝体和藻殖段的可见光吸收光谱和色素含量差别不大。而两者在不同光强范围 (110～ 12 0 0 μmol·m-2 ·s-1)和不同温度 (15～ 45℃ )下的光合放氧活性 ,表明发状念珠藻的藻殖段比营养藻丝体可能更适合在低光强下和较高的温度下生长。从荧光发射光谱可以看出 ,在光合能量传递中营养藻丝体比藻殖段在两个光系统之间的光能分配上更加均衡 ;但是藻殖段中藻胆体吸收光能向两个光系统的传递比营养藻丝体的更加有效。可以认为藻殖段的形成对光合作用的结构与功能产生影响。     

念珠藻属植物nifH基因的适应性进化分析

念珠藻(Nostoc)固氮过程关键在于固氮酶的催化,而固氮酶复合物中的铁蛋白(NifH)是由高度保守的nifH基因编码的,该基因是进化史上现存最古老的功能基因之一。该研究选取念珠藻属及近缘类群的nifH基因序列共40条,采用最大似然法构建系统发育树;运行PAML4.9软件,对nifH基因编码蛋白进行生物信息学分析,并使用分支模型、位点模型和分支-位点模型检测该基因的选择位点,探讨nifH基因的适应性进化特征。结果表明:(1)最大似然树显示内类群中该研究物种共分为6个分支(A、B、C、D、E和F),其中D和E是2个大的分支,每个大分支中又各包含2个特殊的小分支A、F和B、C,其中F分支包含新疆古尔班通古特沙漠采集到的9株念珠藻,A分支包含F分支及该研究测定序列的4株葛仙米,B分支包含本研究测定序列的4株地皮菜和3株未定种的念珠藻,C分支包含NCBI数据库中下载的5株念珠藻、鱼腥藻序列和本研究测定序列的1株念珠藻。(2)在所分析的3种进化模型中,仅通过分支-位点模型检测出14个统计学上显著的正选择位点,即1F、2S、3S、4T、5A、6F、7F、8I、9S、10C、17I、27Y、29D和31R位点,表明念珠藻属植物的nifH基因发生了适应性变化,分支-位点模型是研究藻类基因适应性进化较好的模型。     

Physiological and biochemical studies on nitrogen fixing blue-green algae

Summary Experiments were made to investigate the influence exerted by calcium, strontium, cobalt and molybdenum on the growth and nitrogen fixation of a local strain of Nostoc commune.Calcium, cobalt and molybdenum at least in micro amounts were found to be required for the achievement of best growth and highest nitrogen fixation.Calcium seems to be irreplaceable by strontium, the latter element exerted a strong inhibitory action on both growth and nitrogen fixation processes.     

Physiological and biochemical studies on nitrogen fixation by blue-green algae

Summary The influence of some physiological and environmental conditions on the growth and nitrogen fixation by a local strain of Nostoc commune revealed that the variation of hydrogen ion concentration of the nutritive medium, light intensity and temperature greatly affected the growth rate, cellular nitrogen and extracellular nitrogen of the experimental organism. It was possible however, to formulate the optimum conditions for growth and nitrogen fixation by this organism.     

光强和氮源对念珠藻胞外多糖分泌的影响

胞外多糖（EPS）是结皮蓝藻形成生物结皮的胶结剂,为了理解常球状存在的丝状蓝藻Nostoc胶结沙粒的机理,探讨了光强40、80 E/（m2s）和氮源（气态氮,硝态氮）对结皮优势种Nostoc sp.分泌EPS（包括荚膜多糖CPS和释放多糖RPS）的影响规律及其内在机理。结果发现:Nostoc sp.在气态氮和硝态氮下都有相似的快速生长,但其分泌的RPS、CPS及EPS量,在硝态氮下均随光强的增加而增加,在气态氮下却与光强没有关系。相关代谢研究发现,在硝态氮下细胞内有更高含量的可溶性糖和蔗糖。进一步的相关分析发现,在两种氮源下,蔗糖量与RPS量或CPS量间的显著正相关都只发生在80 E/（m2s）下,在气态氮中,两光强下的胞内总糖量都与CPS量显著负相关。以上结果说明,Nostoc sp.在氮源利用和光强适应方面都有明显优势,它即使在快速生长的对数期,也可同时分泌相当量的EPS,这使其在球状藻殖段形成之前胶结沙粒成为可能。由此可推知,Nostoc sp.在贫瘠沙土表面的最初生长过程中,其胞外的EPS均来自胞内的固碳产物,在高光强下,蔗糖很可能是其EPS合成的原料。       

Cyanobacterial Microbiotic Crusts in Eroded Soils of a Tropical Dry Forest in the Baja California Peninsula,Mexico

This study deals with the cyanobacterial composition, and the nitrogen fixation of four members, of cryptobiotic crusts collected from eroded soils in a transitional area between arid and tropical climatic environments. Identification was based on microscopic analyses. Morphotypes were identified directly from reactivated natural crusts and from cultured strains. The identified morphotypes were Scytonema cf. ocellatum, Scytonema sp., Microcoleus cf. paludosus, M. cf. sociatus, Calothrix cf. elenkinii, C. cf. marchica, Nostoc cf. microscopicum, and Phormidium sp. Results show that the cyanobacterial composition of the microbiotic crusts studied is different from those in warmer and cooler deserts, particularly the absence of Microcoleus cf. vaginatus and Nostoc cf. commune in our samples. Such differences could be caused by the transitional character of the area. The results of the acetylene reduction assay show that the capacity of nitrogen fixation of some morphotypes is limited to the heterocyst-forming morphotypes.     

Electron transfer protein complexes in the thylakoid membranes of heterocysts from the cyanobacterium Nostoc punctiforme

Filamentous, heterocystous cyanobacteria are capable of nitrogen fixation and photoautotrophic growth. Nitrogen fixation takes place in heterocysts that differentiate as a result of nitrogen starvation. Heterocysts uphold a microoxic environment to avoid inactivation of nitrogenase, e.g. by downregulation of oxygenic photosynthesis. The ATP and reductant requirement for the nitrogenase reaction is considered to depend on Photosystem I, but little is known about the organization of energy converting membrane proteins in heterocysts. We have investigated the membrane proteome of heterocysts from nitrogen fixing filaments of Nostoc punctiforme sp. PCC 73102, by 2D gel electrophoresis and mass spectrometry. The membrane proteome was found to be dominated by the Photosystem I and ATP-synthase complexes. We could identify a significant amount of assembled Photosystem II complexes containing the D1, D2, CP43, CP47 and PsbO proteins from these complexes. We could also measure light-driven in vitro electron transfer from Photosystem II in heterocyst thylakoid membranes. We did not find any partially disassembled Photosystem II complexes lacking the CP43 protein. Several subunits of the NDH-1 complex were also identified. The relative amount of NDH-1M complexes was found to be higher than NDH-1L complexes, which might suggest a role for this complex in cyclic electron transfer in the heterocysts of Nostoc punctiforme.     

Physiological evidence indicates microcystin-LR to be a part of quantitative chemical defense system

The functional role of microcystins (MC) is poorly understood. Here, we investigated the effect of pure MC-LR recovered from the freshwater planktonic cyanobacterium Nostoc sp. BHU001 on five closely related cyanobacteria (Nostoc muscorum, Nostoc commune, Anabaena fertilissima, Anabaena doliolum, and Cylinderospermum majus) isolated from different habitats as well as on the producer itself (Nostoc sp. BHU001). MC-LR was found to be a general growth inhibitor active at nanomolar range (25–100 μg L^?1). It inhibited the growth of all cyanobacterial strains in a concentration-dependent manner, except the producer. A. fertilissima was the most sensitive species. MC-LR affected vital metabolic processes such as photosynthesis, respiration, and nitrogen fixation. Nitrogenase activity showed maximum sensitivity, followed by respiration, photosynthesis, and general growth. The photosynthetic electron transport activity was maximally inhibited at PSI, followed by whole chain and PSII activities. Thus, MC-LR is active at multiple sites causing energy constraint to the vital metabolic processes of the target organisms. However, its requirement at high concentration, which is environmentally irrelevant, and lack of quantitative information on the extracellular release of MC-LR suggest that MC-LR has no allelopathic function and could be a part of a quantitative chemical defense system.     

A simple method for culturing neonatal Biomphalaria glabrata snails

A simplified method for the culture of neonatal Biomphalaria glabrata snails on a diet of the cyanobacteria Nostoc sp. has been developed. Previous studies reported using Nostoc sp. grown on a mud-based medium to feed neonatal snails; this medium is difficult to prepare and gives inconsistent growth between laboratories due to variations in the soil used in its preparation. Here, we report the use of Bg-11 medium supplemented with sodium nitrate for the culture of Nostoc sp. as a food source for neonatal B. glabrata snails. Bacteria grown in the medium may be maintained in continuous culture and used exclusively as the nutrient source for neonatal snails. Neonatal snails fed with these bacteria exhibit a growth rate of 1-2 mm per week and may be transferred to standard culture conditions after 2-3 wk. Survival of neonatal snails after 2 wk on the Nostoc sp. diet was 60-80%, while survival after switching to standard culture conditions approached 100%. Analysis of the growth rate and survival of neonatal snails maintained on Nostoc sp., as well as the growth rate, survival, and sexual maturation of the resulting juvenile snails, showed that snails maintained using this method are phenotypically comparable to those maintained under other standard laboratory conditions.     

Isolation and purification of an axenic diazotrophic drought-tolerant cyanobacterium, Nostoc commune, from natural cyanobacterial crusts and its utilization for field research on soils polluted with radioisotopes

Nitrogen fixation and drought tolerance confer the ability to grow on dry land, and some terrestrial cyanobacteria exhibit these properties. These cyanobacteria were isolated in an axenic form from Nostoc commune clusters and other sources by modifying the method used to isolate the nitrogen-fixing and drought-tolerant cyanobacterium Nostoc sp. HK-01. Of these cyanobacteria, N. commune, which is difficult to isolate and purify, uses polysaccharides to maintain water, nitrogen fertilizers for nitrogen fixation, and can live in extreme environments because of desiccation tolerance. In this study, we examined the use of N. commune as biosoil for space agriculture and possible absorption of radioisotopes ((134)Cs, (137)Cs). This article is part of a Special Issue entitled: Photosynthesis Research for Sustainability: from Natural to Artificial.     

Rewetting of drought-resistant blue-green algae: Time course of water uptake and reappearance of respiration,photosynthesis, and nitrogen fixation

Summary The response of the terrestrial blue-green algae Nostoc flagelliforme, Nostoc commune, and Nostoc spec. to water uptake has been investigated after a drought period of approximately 2 years. Rapid half-times of rewetting (0.6, 3.3, and 15.5 min, respectively) are found. The surfaceto-mass ratio of the three species is inversely correlated to the speed of water uptake and loss. The ecological relevance of these different time courses is discussed.Respiration starts immediately after a 30-min rewetting period, whereas photosynthetic oxygen evolution reaches its maximum activity after 6 and 8 h with N. commune and N. flagelliforme, respectively. In the dark, recovery of oxygen uptake by N. commune is somewhat impaired, while slightly stimulated with N. flagelliforme. With both species, recovery of photosynthesis is inhibited by darkness.Using colonies kept dry for two years, nitrogenase activity of N. commune attains its maximum 120 to 150 h after rewetting, while only 50 h were needed with algal mats kept dry for two days.Thus, after a 2-year drought period, the physiological sequence of reactivation is respiration—photosynthesis—nitrogen fixation. Respiration and photosynthesis precede growth and are exhibited by existing vegetative cells, whereas recovery of nitrogen fixation is dependent on newly differentiated heterocysts.     

An iTRAQ-based quantitative analysis to elaborate the proteomic response of Nostoc sp. PCC 7120 under N2 fixing conditions

Nostoc sp. PCC 7120 is an oxygen-evolving photoautotrophic N2 fixing filamentous cyanobacterium. Upon nitrogen starvation, a range of processes are initiated, such as differentiation of the heterocysts, specific cells where N2 fixation takes place. We have characterized and quantified the proteome of the Nostoc sp. PCC 7120 wild-type strain grown under N2 fixing and non-N2 fixing conditions. To assess global proteome changes in response to environmental changes, measurements were made using the quantitative proteomics tool, iTRAQ, on a whole cell digest. From this approach, a total of 486 different proteins was accurately identified across 2 biological replicate experiments, where 226 identifications contained 2 or more distinct peptides. Results of metabolic regulation will be discussed to demonstrate that proteomics represents an important tool for the development of heterocystous cyanobacteria for future biological H2 production.