蓝细菌16S rRNA双甲基化酶基因ksgA的进化分析、蛋白结构预测及功能验证

为研究核糖体小亚基rRNA双甲基化酶基因ksgA的进化关系、蛋白结构和功能,利用MEGA 5.2软件构建了37种蓝细菌ksgA基因和16S rDNA的系统进化树,通过SWISS-MODEL分析和预测KsgA的蛋白质结构模型,并通过克隆集胞蓝细菌Synechocystis sp.strain PCC 6803、鱼腥蓝细菌Anabaena sp.strain PCC7120和聚球蓝细菌Synechococcus elongates sp.strain PCC 7942的ksgA基因对大肠杆菌ksgA突变体进行了异源互补.结果表明:ksgA基因在蓝细菌中与16S rDNA进化分支高度相似,且体现出蓝细菌进化的聚类特征;蓝细菌KsgA甲基化酶与该蛋白家族的其他蛋白质拥有共同的保守结构、催化位点和配体结合位点;克隆的3种蓝细菌ksgA基因均能在一定程度上互补大肠杆菌ksgA基因的功能.     

Nitrogen and Phosphorus in Coastal Sediments Covered by Cyanobacteria Mats (9 pp)

Background   Biocommunities of phototrophic microorganisms (principally cyanobacteria) developing on the surface sediments of shallow, warm and semi-isolated coastal environments, bind or trap small particles on their polysaccharide sheaths creating organosedimentary structures. Those structures are called algal mats and they are commonly flat and laminar in shape. They are also called recent stromatolites due to their remarkable similarity to the fossil ones. Objectives   Not many geochemical studies exist concerning cyanobacterial mats in Mediterranean coasts, in comparison with the biological ones. Considering the importance of nitrogen and phosphorus in the cyanobacteria mat development, the present study aims to contribute to the knowledge of their chemical behaviour in such environments. Methods   Sediment cores of about 10 cm length along with water samples were collected during the course of a year from two Greek coastal areas. Sediments were fractionated in three distinct layers according to their structure. A wet oxidation method for the simultaneous determination of total nitrogen and total phosphorus was applied to all sediment samples. The total organic carbon (TOC), the total inorganic phosphorus and the natural pigment content in the sediment samples were also determined. Standard spectrophotometric methods were used for the determination of the concentration of dissolved nutrients. In situ measurements of pH, temperature and salinity were also carried out. Results and Discussion   The sampling areas were found to be in the mesotrophic level. The group of aerobic filamentous cyanobacteria dominated the sediment biocommunities. The pH of the sediment showed a gradual reduction downwards. The highest content of TOC, total nitrogen and total phosphorus were detected in the surface layer of the sediment samples. The contribution of organic phosphorus to the total phosphorous ranged between 0% and 50% with an average value of 26.6%. A depletion of organic phosphorus in the anoxic middle layer of the sediment was detected. A good positive correlation between total nitrogen and chlorophyll-a, as well as total nitrogen and TOC in the surface sediment layer, was observed. A good negative correlation between total phosphorous in sediments and dissolved phosphates in seawater was also observed. Conclusions   The high content of total nitrogen recorded in the surface sediments indicate that cyanobacteria accumulate nitrogen acquiring it from the sediment, the marine environment and the air (in the form of N2). A proportional accumulation in phosphorus also happens, although to a lesser degree. The temporal fluctuation of nitrogen content in the sediment's surface layer agrees with the temporal fluctuation of primary production. The cyanobacterial mat acts as a natural pump that transfers phosphorus from seawater to the surface of the sediment. Recommendations and Outlook   A further, more extensive, multidisciplinary study of cyanobacterial mats is needed for the determination of the biogeochemical processes that take place there and for the effective environmental management of the areas where these forms develop.     

土壤生物结皮在干旱半干旱地区退化生态系统恢复中的作用(英文)

 土壤生物结皮是由细菌、真菌、藻类、地衣和苔藓等形成的一种混合体,作为干旱半干旱地区生态系统的组成部分,它对生态系统镶嵌格局和生态过程有不可忽视的影响;同时,土壤生物结皮通过影响局部水分条件,可以起到稳定土壤表层、减少土壤侵蚀、增加土壤氮养分的作用:因此,土壤生物结皮能够为土壤表层提供一种天然保护的作用,对干旱半干旱地区退化生态系统的恢复具有非常重要的作用。     

Haematological indices are modulated in juvenile carp, Cyprinus carpio L., exposed to microcystins produced by cyanobacterial water bloom

This study evaluated the influence of toxic cyanobacterial water blooms on the blood indices of the common carp, Cyprinus carpio L. Experimental fish were exposed to a natural population of cyanobacterial water blooms (mainly Microcystis aeruginosa and M. ichthyoblabe), which contained microcystins [total concentration 133–284 μg g^?1 (DW), concentration in water 2.8–7.4 μg L^?1]. Haematological indices showed marked changes in fish exposed to the cyanobacterial population in comparison with the control group. Statistical evaluation of the influence of cyanobacterial water blooms on biochemical indices of the juvenile carp showed a distinct decrease in albumin, alanine aminotransferase, total bilirubin, calcium, cholesterol, glucose, phosphorus and iron when compared to controls. Values of red blood counts [haemoglobin, haematocrit (PCV), mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration] and lactate were significantly increased compared to controls. After exposure to cyanobacterial water bloom, the carp were kept in clean water to monitor the persistence of biochemical indices. The influence of cyanobacterial populations on calcium, cholesterol, glucose, lactate, phosphorus and PCV persisted up to 28 days after conclusion of the experiment. Duration of exposure, toxicity and density of cyanobacterial water blooms had an important impact on individual haematological indices.     

Cyanotoxins: bioaccumulation and effects on aquatic animals

Cyanobacteria are photosynthetic prokaryotes with wide geographic distribution that can produce secondary metabolites named cyanotoxins. These toxins can be classified into three main types according to their mechanism of action in vertebrates: hepatotoxins, dermatotoxins and neurotoxins. Many studies on the effects of cyanobacteria and their toxins over a wide range of aquatic organisms, including invertebrates and vertebrates, have reported acute effects (e.g., reduction in survivorship, feeding inhibition, paralysis), chronic effects (e.g., reduction in growth and fecundity), biochemical alterations (e.g., activity of phosphatases, GST, AChE, proteases), and behavioral alterations. Research has also focused on the potential for bioaccumulation and transferring of these toxins through the food chain. Although the herbivorous zooplankton is hypothesized as the main target of cyanotoxins, there is not unquestionable evidence of the deleterious effects of cyanobacteria and their toxins on these organisms. Also, the low toxin burden in secondary consumers points towards biodilution of microcystins in the food web as the predominant process. In this broad review we discuss important issues on bioaccumulation and the effects of cyanotoxins, with emphasis on microcystins, as well as drawbacks and future needs in this field of research.     

Bioactivity of benthic and picoplanktonic estuarine cyanobacteria on growth of photoautotrophs: inhibition versus stimulation

Understanding potential biochemical interactions and effects among cyanobacteria and other organisms is one of the main keys to a better knowledge of microbial population structuring and dynamics. In this study, the effects of cyanobacteria from benthos and plankton of estuaries on other cyanobacteria and green algae growth were evaluated. To understand how the estuarine cyanobacteria might influence the dynamics of phytoplankton, experiments were carried out with the freshwater species Microcystis aeruginosa and Chlorella sp., and the marine Synechocystis salina and Nannochloropsis sp. exposed to aqueous and organic (70% methanol) crude extracts of cyanobacteria for 96 h. The most pronounced effect observed was the growth stimulation. Growth inhibition was also observed for S. salina and M. aeruginosa target-species at the highest and lowest concentrations of cyanobacterial extracts. The methanolic crude extract of Phormidium cf. chalybeum LEGE06078 was effective against S. salina growth in a concentration-dependent manner after 96 h-exposure. All of the cyanobacterial isolates showed some bioactivity on the target-species growth, i.e., inhibitory or stimulating effects. These results indicate that the analyzed cyanobacterial isolates can potentially contribute to blooms' proliferation of other cyanobacteria and to the abnormal growth of green algae disturbing the dynamic of estuarine phytoplankton communities. Since estuaries are transitional ecosystems, the benthic and picoplanktonic estuarine cyanobacteria can change both freshwater and marine phytoplankton succession, competition and bloom formation. Furthermore, a potential biotechnological application of these isolates as a tool to control cyanobacteria and microalgae proliferation can be feasible. This work is the first on the subject of growth responses of photoautotrophs to cyanobacteria from Atlantic estuarine environments.     

Preparation of calibration standards of N1-H paralytic shellfish toxin analogues by large-scale culture of cyanobacterium Anabaena circinalis (TA04)

Mouse bioassay is the official testing method to quantify paralytic shellfish toxins (PSTs) in bivalves. A number of alternative analytical methods have been reported. Some methods have been evaluated by a single laboratory validation. Among the different types of methods, chemical analyses are capable of identifying and quantifying the toxins, however a shortage of the necessary calibration standards hampers implementation of the chemical analyses in routine monitoring of PSTs in bivalves. In our present study, we studied preparation of major PST analogues as calibrants by large-scale cultivation of toxic freshwater cyanobacteria Anabaena circinalis TA04. The cells were steadily grown in 10 L bottle for 28 days. The primary N1-H toxins, C1/C2, were produced at a concentration of 1.3 ± 0.1 μmol/L. The intracellular and extracellular toxins occupied 80% and 20%, respectively. Over 220 μmol of the toxins was obtained from approximately 200 L of the culture over six months, demonstrating that it is sufficient to prepare saxitoxin analogues. The toxins were chemically converted to six N1-H analogues. Preparation of the analogues was carried out at relatively high yields (50-90%). The results indicate that our preparation method is useful to produce N1-H toxins. In our present study, detailed conditions for preparation of one of the rare N1-H analogues, gonyautoxin-5, were investigated.     

Evaluating the efficacy of cyanobacterial formulations and biofilmed inoculants for leguminous crops

Our investigation was aimed towards evaluating the agronomic potential of biofilmed preparations (developed using Anabaena/Trichoderma as matrices with different agriculturally useful bacteria/fungi as partners) and selected cyanobacterial strains (Anabaena laxa (T7) RP8/Calothrix sp.). The formulations were prepared using paddy straw compost:vermiculite (1:1) as carrier and tested as inoculants in mungbean and soybean. The effects of the formulations were evaluated in terms of microbiological, nutrient availability, and plant biometric parameters. The Trichoderma viride–Bradyrhizobium biofilm exhibited 20–45% enhancement in fresh/dry weight of plants over other microbial treatments, while the T. viride–Azotobacter biofilm exhibited highest dehydrogenase activity in the soil and nitrogen fixation. T7 RP8 recorded statistically at par yield values with the T. viride–Bradyrhizobium (T5) biofilm treatment in mungbean. In soybean, among all the treatments, the T5 biofilm recorded the highest fresh weight of plants and available N in soil at harvest. The Anabaena–T. viride biofilmed formulations proved to be the most promising for soybean, recording 12–25% enhanced yield and microbial activity (measured as dehydrogenase activity). This study highlights the promise of cyanobacterial inoculants and biofilmed biofertilizers as promising inputs for integrated nutrient management strategies in agriculture.     

Fluorescence induction characteristics of Anacystis nidulans during recovery from iron deficiency

Cultures of the cyanobacterium, Anacystis nidulans, were grown in iron deficient medium and examined for fluorescence emission at room temperature. Iron deficiency induced several alterations in fluorescence emission kinetics which were reversed when iron was restored. When excited by 450 nm light (absorbed by chlorophyll), iron stressed cells showed an enhanced maximum fluorescence yield (Fm), due in large part to an increase in original fluorescence (Fo) and a depressed variable fluorescence. Both Fm and Fo declined during the early stages of recovery from iron deficiency. Inhibitors of chlorophyll biosynthesis (such as levulinic acid and gabaculine) had little influence on these early stages of recovery suggesting that newly synthesized chlorophyll is not responsible for the decline in Fm. In contrast, when excited by 550 or 600 nm light (absorbed by phycocyanin), iron deficient cells showed no increase in Fm. A fast rise fluorescence transient was observed which was absent in both normal or fully recovered cells. This transient was attenuated by preillumination (600 nm light), and recovered in darkness with a half time of 2–4 minutes. These results suggest that the reoxidation of acceptors on the oxidizing side of PS II is considerably slower in iron deficient cells.