Sorgoleone,a sorghum root exudate: Algicidal activity and acute toxicity to the ricefish Oryzias latipes

The discovery of natural and natural-based compounds has resulted in its application as an alternative to synthetic algicides to control harmful algae in aquatic systems. Of the many natural-product-based algicides, sorgoleone, a natural plant product from Sorghum bicolor root exudates has been investigated for its controlling effect on different algal species and its acute fish toxicity. Growth of the blue green algal species Microcystis aeruginosa Kützing was completely inhibited by the crude methanol extract of sorghum root at 20 μg mL⁻¹. The most noticeable inhibition was observed in the bioassay of n-hexane soluble extract, where 98% growth inhibition occurred in M. aeruginosa at the concentration of 1.25 μg mL⁻¹. Sorgoleone very effectively controlled blue green algae inhibiting 97% of M. aeruginosa at 0.5 μg mL⁻¹ and 99% of Anabaena affinis Lemmermann at 4 μg mL⁻¹. In contrast, inhibition of the green algae species Chlorella vulgaris Beijerinck and Scenedensmus spp. at 16 μg mL⁻¹ sorgoleone was 87 and 68%, respectively. There were no mortalities or adverse effects observed in any of the fish exposed to water control, solvent control, and a nominal concentration of 1 μg mL⁻¹ during the test period. The no observed effect concentration (NOEC) value was 1.5 μg mL⁻¹ for the tested fish (O. latipes). Sorgoleone can be considered as an effective and an ecologically and environmentally sustainable approach to controlling harmful algae.     

蓝细菌断裂DNA聚合酶DnaE的体内/外反式剪接分析

蓝细菌Anabaena PCC7120的DNA聚合酶DnaE由2个断裂基因dnaENI和dnaECI编码.通过纯化它们的部分基因在大肠杆菌中的表达产物DnaENI′和DnaECI′来免疫家兔获得抗体,用于对断裂的DnaE在体内和体外的反式剪接活性进行鉴定.在体外实验中,将2个断裂的DNA聚合酶DnaENI′和DnaECI′混合,进行反应,并利用它们的抗体对反应产物进行鉴定;在体内实验中,利用以上抗体对Anabaena PCC7120细胞总蛋白进行免疫杂交分析.实验结果表明,蓝细菌AnabaenaPCC7120中断裂的2个DnaE多肽在体内和体外均能进行反式剪接.体外实验中,纯化的DnaENI′和DnaECI′的混合反应产生新的蛋白产物,既有成熟的反式剪接产物DnaEN′-,又有剪切了内含肽后的副产物DnaEN′和DnaE,且DTT的存在对剪接反应具有促进作用;此外体内实验中,在Anabaena PCC7120细胞中既能检测到完整的DnaE,也能检测到未作用的DnaENI,但未能检测到DnaECI.     

Calmodulin in heterocystous cyanobacteria: biochemical and immunological evidence

Abstract The occurrence activity and localization of calmodulin in three heterocystous cyanobacteria of the genus Anabaena were studied. Boiled crude extracts caused a Ca²⁺-dependent stimulation of NAD kinase. Such a stimulation was blocked by EGTA and chlorpromazine. SDS-PAGE and Western blot analysis using antiserum against eukaryotic spinach calmodulin, revealed a polypeptide of about 17 kDa. Immunogold localization of calmodulin gave a dense gold label both vegetative cells and heterocysts. The label was mainly confined to the centroplasm in vegetative cells, while it was evenly distributed in the cytoplasm of mature heterocysts.     

Physiological and Biochemical Alterations in a Diazotrophic Cyanobacterium <Emphasis Type="Italic">Anabaena cylindrica</Emphasis> Under NaCl Stress

Growth, morphological variation, and liquid chromatography–photodiode array detection–mass spectrometric analysis of pigments have been studied in a diazotrophic cyanobacterium Anabaena cylindrica in response to NaCl stress. The chlorophyll and cellular protein contents increased initially in response to 50 mM NaCl. Further increment in NaCl concentration, however, resulted in a significant decrease in both chlorophyll and cellular protein. A. cylindrica cells subjected to NaCl stress also showed morphological variations by having alteration in their size and volume. A. cylindrica cells subjected to NaCl stress also exhibited altered plastoquinone and chlorophyll-a (chl a) levels in comparison to its NaCl-untreated counterpart. Furthermore, a relative increase in plastoquinone level and a subsequent decrease in chl a level were recorded in NaCl adapted cells of A. cylindrica in response to NaCl stress. These results suggest that owing to adaptation various morphological, physiological, and biochemical changes occur in the cyanobacterium A. cylindrica in response to NaCl stress.     

Impact of different abiotic stresses on growth, photosynthetic electron transport chain, nutrient uptake and enzyme activities of Cu-acclimated Anabaena doliolum

This study provides a comparative account of the effects of cadmium, temperature, ultraviolet-B and sodium chloride on the growth, photosynthesis, nutrient uptake and enzyme activities of untreated control and copper-acclimated Anabaena doliolum. Reduction in all the studied parameters, except carotenoids, was maximum for sodium chloride followed by ultraviolet-B, temperature and cadmium treatments, the reduction being greater in control than acclimated A. doliolum. Among the various parameters, photosystem II was most sensitive for all the stresses in both control and acclimated A. doliolum. Likewise, O2 evolution was more susceptible to various stressors than 14C uptake. Ammonium uptake and glutamine synthetase (GS, EC 6.3.1.2) were the least affected parameters. As compared to control, acclimated Anabaena exhibited higher ATP content under normal conditions. These results attest our hypotheses that acclimated Anabaena was physiologically more robust than control and that salinity was more injurious to the test organism than other abiotic stresses investigated.     

PROTEIN TURNOVER AND HETEROCYST DIFFERENTIATION IN THE CYANOBACTERIUM ANABAENA VARIABILIS1

Under conditions of starvation for fixed nitrogen, cells of the filamentous cyanobacterium Anabaena variabilis Kütz, degrade much of their protein prior to heterocyst differentiation. Cells starved for a source of fixed nitrogen initially degraded about 2% of their protein per hour; by 24 h after nitrogen stepdown about 40% of the protein was degraded. Most of the acid-soluble radiolabeled material was excreted into the medium. Proteolysis was completely inhibited by chloramphenicol, by cyanide, or in the dark, hut was only partially inhibited in the presence of dichlorophenyl dimethylurea. Methionine sulfoximine (MSX) (an inhibitor of glutamine synthetase) in the presence of ammonia caused heterocysts to form. MSX treated cells degraded protein; however, the amount of protein degraded was much less than in cells starved for ammonia. Glutamine, which can serve as a nitrogen source for this strain, did not prevent starvation-induced proteolysis and did not prevent the differentiation of heterocysts.     

Comparison of changes in toxicity during growth of Anabaena circinalis (cyanobacteria) determined by mouse neuroblastoma bioassay and HPLC

Changes in toxicity were studied during batch culture of a strain ofAnabaena circinalis (AWQC ANA-311F) producing paralyticshellfish toxins (PSTs). PSTs were extracted from cells and culture mediumduring the culture period of 29 days. Samples were analysed by HPLC withpost-column oxidation coupled to fluorescence detection, and tested with themouse neuroblastoma bioassay (MNB). Seven PSTs were detected, but only fivewerequantified. Based on chemical analysis of PSTs by HPLC, atheoretical saxitoxin concentration related to mouse toxicity wascalculated. Neuroblastoma cell survival was measured during the MNB andcorrelated to total toxicity of the A. circinalis samples.Comparison was made between the changes in total toxicity shown by MNB and thetheoretical saxitoxin concentration shown by HPLC. There was asignificant positive correlation between the two in the culture medium, but notthe cells.     

Inhibition of cell division blocks the synthesis of the second nitrogenase (Nif2) in the cyanobacterium Anabaena variabilis

Anabaena variabilis ATCC 29413 belongs to the cyanobacteria that use a specific cell type, heterocysts, for fixation of atmospheric nitrogen under aerobic conditions. Nitrogen fixation under anaerobic conditions is catalyzed by a Mo-dependent nitrogenase (Nif2) that is expressed in the vegetative cells. We demonstrate here using immunolocalization/light microscopy (LM) that the synthesis of NifH2 is mainly initiated in dividing vegetative cells along the trichomes. Blocking cell division by cephalexin abolished nitrogenase synthesis under anaerobic conditions.     

IRON-MEDIATED CHANGES IN THE GROWTH OF LAKE ERIE PHYTOPLANKTON AND AXENIC ALGAL CULTURES1

The effect of iron enrichment on algal growth and photosynthesis was investigated using natural assemblages of Lake Erie phytoplankton and axenic cultures of Anabaena, Scenedesmus and Selenastrum. Cell yield and photosynthesis were frequently inhibited in the presence of unchelated iron over the range of 3.6 to 53.7 μM iron as FeCl₃. In lake water and in a defined medium with low nutrient concentrations, the degree of inhibition by iron could be reduced by chelating the iron with EDTA or by enriching the cultures with phosphorus. Chemical analyses revealed that the EDTA efectively reduced the ability of the ferric iron to remove soluble phosphorus from the media. EDTA was also observed to reduce rather than enhance iron uptake by axenic cultures of A. flos-aquae. These data support the hypothesis that additions of EDTA to low-nutrient media may serve to stimulate algal growth in the presence of iron by preventing the iron from altering extracellular concentrations of soluble ions essential for algal metabolism. In medium with high nutrient concentrations, the soluble phosphorus concentration was not appreciably altered by either EDTA-chelated or unchelated iron enrichment (0.9 to 53.7 μM). Instead, the observed enhancement of cell yield by EDTA-chelated iron in nutrient-rich media appeared to be due to the direct effect of iron on intracellular metabolic processes.