红树林放线菌及其天然产物研究进展北大核心CSCD

红树林是热带亚热带潮间带的木本植物群落。为从海洋环境寻找新的天然产物,红树林已经成为放线菌资源收集、天然产物分离鉴定及其生物合成机制研究的热点。从巴哈马红树林分离的盐孢菌所产生的盐孢菌素(Salinosporamide A)已经进入临床研究。从红树林分离的放线菌类群已经达到8个亚目11科24属,并发现新属3个,新种31个。从红树林放线菌分离的新天然产物包括生物碱、喹啉等芳香类、阿扎霉素等大环类脂及吲哚衍生物等,大多数天然产物来源于红树林链霉菌。新型吲哚咔唑、吲哚倍半萜及厦霉素等以新颖的结构备受关注,相关生物合成途径已被揭示。     

新疆艾比湖沉积物中免培养放线菌的多样性北大核心CSCD

【目的】采用免培养的方法研究新疆艾比湖湖底沉积物放线菌组成及多样性。【方法】采集并混合5份艾比湖湖底沉积物样本,并提取其总DNA,采用放线菌通用引物对其16S r RNA基因序列进行Touchdown PCR,构建放线菌16S r RNA基因文库。蓝白斑筛选后随机挑选白色克隆分析,利用限制性内切酶HhaⅠ进行酶切分型,挑选具有独特限制性片段差异的阳性克隆进行测序分析。序列经Chimera Check检测,BLAST同源比对及构建16S r RNA基因序列系统发育树。【结果】随机挑选192个白色克隆,其中166个为阳性克隆,选取51个具有独特限制性片段差异的克隆进行序列分析。测序结果进行比对以及Chimera Check检测后,共获得36个可操作单元(Operational Taxonomic Units,OTUs),Gen Bank注册号为KR182090-KR182131。文库覆盖度结果表明克隆文库涵盖了本环境中90.4%的放线菌类群。聚类结果显示,艾比湖湖底沉积物中放线菌分为2个类群,第1个类群属于放线菌门(Actinobacteria),放线菌纲(Actinobacteria)中的放线菌目(Actinomycetales)、丙酸杆菌目(Propionibacteriales)、微球菌目(Micrococcales)和棒杆菌目(Corynebacteriales)4个目,该类群占克隆文库18.1%;另外1个类群属于Unclassified Actinobacteria的类群,分为3个不同的group,占整个克隆文库的81.9%。【结论】新疆艾比湖湖底沉积物中存在多种未知的放线菌类群。     

地嗜皮菌科放线菌的研究进展北大核心CSCD

地嗜皮菌科(Geodermatophilaceae)是放线菌中一个年轻的分类单元。早在1996年Normand曾提出过地嗜皮菌科,但一直未能得到公认;直到2006年,Normand综合了地嗜皮菌属(Geodermatophilus)、芽生球菌属(Blastococcus)和贫养杆菌属(Modestobacter)等3个属的共同特征,全面概括了地嗜皮菌科的典型特征,终于使地嗜皮菌科被正式编入放线菌的一个科。到目前为止,地嗜皮菌科涵盖了地嗜皮菌属、芽生球菌属和贫养杆菌属等3个属共25个有效描述种。地嗜皮菌科菌株被视为极端环境的先锋生物之一,在抗逆机制研究、沙漠治理、环境修复等方面初现优势。本文就地嗜皮菌科的建立、分类学特征、科内各属的研究现状、以及它们在生态学与应用研究方面的进展和前景进行了综述。     

一株分解鸡毛角蛋白的放线菌

自土壤中分离到放线苗SS-1菌株。该菌株能分解鸡毛角蛋白;生长最适温度为35℃,最适pH7.5;分解人发及羊毛的活性不显著。     

一株降解纤维素放线菌的分类研究

从高温稻草堆肥中分离到1株能降解纤维素的放线菌CN9,分别对其表型特征、化学特征和16SrRNA基因序列进行了分析,根据实验结果,CN9属于高温链霉菌。     

一株能分解纤维素放线菌的固态发酵研究

对一株从食草动物的粪便中分离出来的放线菌菌株(XW5),以秸杆粉和麸皮为底物,进行固态发酵,并提取纤维素酶粗酶。采用DNS法,进行酶活测定,研究发现该菌株的固态发酵酶活为4-7u/mg。并对酶活的影响因子、pH、温度以及金属离子等的影响进行了探讨,同时发现该酶为碱性纤维素酶。     

一株放线菌代谢产物除草活性的初步研究

从土壤中分离筛选出1株编号为Z40的链霉菌菌株,其代谢产物具有明显的除草活性。室内生物测定结果表明,Z40菌株发酵液干物质的除草活性主要表现为抑制生长作用。对反枝苋的主根和主茎生长抑制中浓度(EC50)分别为276.76 mg/L和1 609.37 mg/L,对狗尾草的主根和主茎生长抑制中浓度分别为501.81mg/L和629.01 mg/L。浓度为10 000 mg/L对油菜的萌发抑制率和主茎生长抑制率均为100.00%,而对小麦的生长发育没有明显影响。田间试验结果表明,Z40菌株代谢产物对稗、狗尾草、铁苋菜的防治效果均在80%以上,抑制率分别为80.30%,81.59%和88.71%。     

一株海洋放线菌的鉴定及其促生作用机理

【背景】海洋放线菌BM-2是本实验室从连云港海域分离得到的一株具有抗菌和促生作用的优良菌株,具有良好的开发应用前景。【目的】明确海洋放线菌BM-2的分类地位,揭示该菌株的促生作用机理,为菌株的开发应用提供理论依据。【方法】通过形态观察、生理生化特性和16S rRNA基因序列分析,对海洋放线菌BM-2菌株进行种属鉴定;采用透明圈法、平板划线法测定BM-2菌株解磷、解钾作用、固氮作用和产植酸酶、1-氨基环丙烷-1-羧基(1-aminocyclopropane-1-carboxylate,ACC)脱氨酶的能力;运用沙尔科夫斯基反应(Salkowski法)和铬天青(chromeazurol S,CAS)法分别测定菌株产吲哚乙酸(indole acetic acid,IAA)和产铁载体的能力。【结果】培养特征、菌落形态观察及生理生化试验结果表明,BM-2菌株符合链霉菌属(Streptomyces)的特征,16SrRNA基因序列与GenBank中栗褐链霉菌(Streptomycesbadius)的序列相似性为99.72%;BM-2菌株具有固氮和解有机磷活性,能够产生ACC脱氨酶、铁载体和IAA。【...     

一株放线菌次生代谢产物化学成分的研究

目的：研究放线菌这类重要的可再生资源的次生代谢产物的化学成分。方法：通过对一株采集于云南西部土壤放线菌的发酵培养,发酵液经TLC分离纯化、高效液相分析检测、核磁共振和质谱测定。结果：从其次生代谢产物中分离获取了8个化合物,其中4个化合物的结构已经初步确定。结论：4个化合物中两个具有抗肿瘤活性,两个具有抑菌活性。     

一株产纤溶酶放线菌的分离和鉴定

目的：从黄精根际土壤筛选产纤溶酶的放线菌,并对其进行鉴定。方法：采用稀释法分离放线菌,利用纤维蛋白平板法筛选能产生纤溶酶的菌株,根据16SrDNA序列、形态学观察和生理生化特征对纤溶活性高的菌株进行鉴定。结果：菌株XZNUM 00004代谢产物的纤溶酶活性为69U/ml,该酶加热到65℃仍能保留85%的活性。经形态学观察,培养特征、生理生化和分子分类学分析鉴定,与胶样链霉菌（Streptomyces gelaticus）相似性为99%。结论：初步推断XZNUM 00004属于链霉菌属（Streptomyces）。     

Bio-deposition of a calcium carbonate layer on degraded limestone by Bacillus species

To obtain a restoring and protective calcite layer on degraded limestone, five different strains of the Bacillus sphaericus group and one strain of Bacillus lentus were tested for their ureolytic driven calcium carbonate precipitation. Although all the Bacillus strains were capable of depositing calcium carbonate, differences occurred in the amount of precipitated calcium carbonate on agar plate colonies. Seven parameters involved in the process were examined: calcite deposition on limestone cubes, pH increase, urea degrading capacity, extracellular polymeric substances (EPS)-production, biofilm formation, ζ-potential and deposition of dense crystal layers. The strain selection for optimal deposition of a dense CaCO₃ layer on limestone, was based on decrease in water absorption rate by treated limestone. Not all of the bacterial strains were effective in the restoration of deteriorated Euville limestone. The best calcite precipitating strains were characterised by high ureolytic efficiency, homogeneous calcite deposition on limestone cubes and a very negative ζ-potential.     

Calcium carbonate precipitation by heterotrophic bacteria isolated from biofilms formed on deteriorated ignimbrite stones: influence of calcium on EPS production and biofilm formation by these isolates

Heterotrophic CaCO₃-precipitating bacteria were isolated from biofilms on deteriorated ignimbrites, siliceous acidic rocks, from Morelia Cathedral (Mexico) and identified as Enterobacter cancerogenus (22e), Bacillus sp. (32a) and Bacillus subtilis (52g). In solid medium, 22e and 32a precipitated calcite and vaterite while 52g produced calcite. Urease activity was detected in these isolates and CaCO₃ precipitation increased in the presence of urea in the liquid medium. In the presence of calcium, EPS production decreased in 22e and 32a and increased in 52g. Under laboratory conditions, ignimbrite colonization by these isolates only occurred in the presence of calcium and no CaCO₃ was precipitated. Calcium may therefore be important for biofilm formation on stones. The importance of the type of stone, here a siliceous stone, on biological colonization is emphasized. This calcium effect has not been reported on calcareous materials. The importance of the effect of calcium on EPS production and biofilm formation is discussed in relation to other applications of CaCO₃ precipitation by bacteria.     

Seawater requirement for the production of lipoxazolidinones by marine actinomycete strain NPS8920

A novel marine actinomycete strain NPS8920 produces a new class of 4-oxazolidinone antibiotics lipoxazolidinone A, B and C. Lipoxazolidinone A possesses good potency (1-2 microg/mL) against drug-resistant pathogens methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus faecium (VRE). Strain NPS8920 exhibits different morphologies in both agar and submerged cultures. The ability of strain NPS8920 to sporulate on saline-based agar media but not on deionized water-based agar medium supported that strain NPS8920 is a marine actinomycete. While strain NPS8920 does not require seawater for growth, the production of lipoxazolidinones by strain NPS8920 can only be detected in the seawater-based media. The optimal production of lipoxazolidinones was observed in the natural seawater-based medium. Strain NPS8920 produced 10-20% of lipoxazolidinones in the synthetic sea salt Instant Ocean-based medium and no production in the sodium chloride-based and deionized water-based media.     

Crosstalk between cellular morphology and calcium oscillation patterns Insights from a stochastic computer model

Agonist-induced oscillations in the concentration of intracellular free calcium ([Ca²⁺]₁) display a wide variety of temporal and spatial patterns. In non-excitable cells, typical oscillatory patterns are somewhat cell-type specific and range from frequency-encoded, repetitive Ca²⁺ spikes to oscillations that are more sinusoidal in shape. Although the response of a cell population, even to the same stimulus, is often extremely heterogeneous, the response of the same cell to successive exposures can be remarkably similar. We propose that such ‘Ca ²⁺ fingerprints’ can be a consequence of cell-specific morphological properties. The hypothesis is tested by means of a stochastic computer simulation of a two-dimensional model for oscillatory Ca ²⁺ waves which encompasses the basic elements of the two-pool oscillator introduced by Goldbeter et al. (Goldbeter A., Dupont G., Berridge M.J. Minimal model for signal-induced Ca²⁺-oscillations and for their frequency encoding through protein phosphorylation. Proc Natl Acad Sci USA 1990; 87: 1461–1465). In the framework of our extended spatiotemporal model, single cells can display various oscillation patterns which depend on the agonist dose, Ca²⁺ diffusibility, and several morphological parameters. These are, for example, size and shape of the cell and the cell nucleus, the amount and distribution of Ca²⁺ stores, and the subcellular location of the inositol(1,4,5)-trisphosphate-generating apparatus.     

Comparative proteomic analysis revealed the metabolic mechanism of excessive exopolysaccharide synthesis by Bacillus mucilaginosus under CaCO3 addition

Abstract

The metabolic mechanism of excessive exopolysaccharide (BMPS) synthesis by Bacillus mucilaginosus CGMCC5766 under CaCO₃ addition was investigated. Under CaCO₃ (5?g/L), the maximum BMPS concentration reached 28.4?g/L, which was 11.2 folds higher than that of the control. Proteomics was then used to analyze the proteins with substantial differences expressed by B. mucilaginosus with and without CaCO₃ addition. The proteomic results revealed that the enzymes related to the central metabolic pathway, amino acid biosynthesis, and nucleotide metabolism were depressed. By contrast, the UDP–glucose pyrophosphorylase involved in BMPS biosynthesis was overexpressed and converted metabolic flux from the biomass accumulation to the biosynthesis of BMPS. This research provides a new and widened perspective into understanding the mechanism of BMPS biosynthesis and applying theoretical and practical significance for the improvement of BMPS production from B. mucilaginosus.     

Effect of selected soil factors on chlorosis of peanut plants grown in calcareous soils in Cyprus

In 1986 and 1987 surveys were conducted of 34 (1986) and 35 (1987) peanut (Arachis hypogaea L.) fields in which the plants showed various degrees of chlorosis. In the areas concerned, plant appearance was classified according to a chlorotic index and corresponding soil samples were taken and analysed for CaCO₃, pH, NO₃–N and DTPA-extractable Fe in 1986 and for CaCO₃, NO₃–N and active lime in 1987.Regression analyses showed that CaCO₃, active lime and NO₃–N were positively correlated, while DTPA-extractable Fe was negatively correlated, with the chlorosis problem. The critical levels above which plants were chlorotic were 20 to 25% CaCO₃ and 10% active lime. Plants began to be chlorotic when DTPA-extractable Fe was below 2.5 mg·kg^–1. The soil factors examined explained about 60% of the variability in plant chlorosis.     

Effect of changes of pHi on intracellular calcium in a smooth muscle-like cell line

The effect of changes of pH_i on Ca_i were studied using fluorescent dyes in cells of the cultured smooth muscle-like line, BC3H-1. Resting Ca_i in these cells was 182 ± 12 nM (n = 74) at pH_o of 7.4. Upon exposure to NH₄Cl, which rapidly alkalinized cells, a transient increase of Cai to 349 ± 55 nM (n = 29) was observed. The peak of the transient occurred within 30 s of exposure to NH₄Cl and returned to baseline within 1 minute. Two other procedures which resulted in rapid cellular alkalinization also caused a transient rise in Ca_i: exposure to and then removal of CO₂ (Ca_i increased from 182 ± 22 to 248 ± 28 nM; n = 8); and exposure to and then removal of Na propionate (Ca_i increased from 242 ± 32 to 456 ± 71 nM; n = 9). The NH4Cl-induced Ca_i transient was eliminated by exposure to 0.2 mM TMB8 and to Ca-free solutions, but not by exposure to 0.5 mM LaCl₃. Sustained changes of pH_i can be induced by varying pH_o. When pH_o was lowered to 6.9, Ca_i fell by 49 ± 11 nM but increased by 203 ± 51 nM (n = 6) when pH_o was raised to 7.9. These data indicate that rapid alkalinization of BC3H-1 cells results in a rapid transient rise of Ca_i. This transient is most likely due to the release of Ca from intracellular stores but may also involve an increase of Ca influx. Steady state values of Cai are positively correlated with steady state pH_i. These data may have implications for the contractile state of smooth muscle during periods of acid/base disturbances and relate to the role of elevated pH_i in cells from hypertensive animals.     

Biomass, carbonate standing stock and production of the mediterranean coralCladocora caespitosa (L.)

Summary The Mediterranean coralCladocora caespitosa often occurs in large beds, i.e. populations of hemispherical clonies with stock densities varying between 1.9 and 4 coloneis ·m⁻². Laboratory measurements of volume, skeleton weight, surface and number of corallites per colony, coupled with mean annual growth rates evaluated through sclerochronology, allowed for the estimation of biomass, skeleton bulk density, calcimass (carbonate standing stock) and secondary production (both organic and inorganic) of twoC. caespitosa beds at 4 and 9 m depth. The mean colony biomass varied between 0.73 and 0.99 kg dw ·m⁻², corresponding to a calcimass between 2 and 5 kg CaCO₃·m⁻². Organic secondary production was 215.5–305.4 g dw of polyps ·m⁻²·y⁻¹, while the potential (mineral) production was 1.1–1.7 kg CaCO₃·m⁻²·y⁻¹, for the year 1996–1997. These values show thatC. caespitosa is one of the major carbonate producers within the Mediterranean and one of the major epibenthic species originating stable carbonate frameworks both in recent and past times.     

Conditioning film and initial biofilm formation on electrochemical CaCO3 deposition on a metallic net in the marine environment

Electrochemical deposition of minerals is a unique technology for artificial reef constructions, relying on calcium carbonate (CaCO₃) build-up over metallic structures through electrolysis of seawater. The present study traces the first 72 h following electric current termination on bacterial biofilm build-up on a metallic net covered with CaCO₃. 16S rRNA clone libraries indicated a dynamic succession. Proteobacteria and Bacteroidetes were evident at all sampling times while Cyanobacteria appeared only within the first 8 h. A significant increase in total organic carbon (TOC) and total protein was observed after 48 h with a significant correlation (R ² = 0.74), indicating TOC is a good tool for characterizing initial biofilm formation. 18S rRNA gene sequences obtained 72 h following current termination indicated a significant presence of Cnidarians (51%). Understanding the dynamics among primary bacterial settlers is important because they play a crucial role in driving the colonization of sessile invertebrate communities on artificial, as well as natural surfaces.     

Yield of submerged paddy and uptake of Zn,P and N as affected by liming and Zn fertilizers

The effects of CaCO₃, Zn sources and levels on the yield of submerged paddy and uptake of Zn, P and N to paddy were studied in green-house at Haryana Agricultural University, Hissar. Powdered CaCO₃ was mixed at 0,4 and 8 per cent and Zn was added at 0,5 and 10 ppm through ZnSO₄.7H₂O, ZnO and Zn EDTA separately. Dry weight at tillering and heading and grain and straw at maturity decreased significantly with 4 and 8 per cent CaCO₃ in comparison to the control. Increasing Zn application increased the dry weight and grain yield. Zn EDTA gave highest yield of paddy followed by ZnSO₄.7H₂O and ZnO.Increasing the application of CaCO₃ from 0–8 per cent decreased the concentration and uptake of Zn and increasing Zn application from 0–10 ppm increased concentration and uptake of Zn in paddy at tillering, heading and maturity. Zn EDTA gave the highest concentration and uptake of Zn followed by ZnSO₄.7H₂O and ZnO. There was interaction between Zn sources and CaCO₃.The concentration and uptake of N and P in paddy dry matter at tillering and heading and straw and grain at maturity decreased as compared to control with increasing CaCO₃ addition. The concentration and uptake of N increased and that of P decreased in paddy dry matter straw and grain with increasing Zn application. The highest concentration of N was observed with ZnO, followed by ZnSO₄.7H₂O and Zn EDTA. But highest uptake of N was observed with Zn EDTA followed by ZnSO₄.7H₂O and ZnO. As regards concentration and uptake of P, it was highest with ZnO followed by ZnSO₄.7H₂O and Zn EDTA.