Anabaena Oryzae Research Articles

Use of Cyanobacteria for Controlling Flax Seedling Blight

Cyanobacteria are one of important groups of prokaryotes due to their effects on growth and development of plants and its role in biological control of phytopathogenic fungi such as : Fusarium oxysporum and Rhizoctonia solani on flax by producing various biologically active substances . In the present study , fourteen cyanobacterial strains were isolated from rice rhizosphere in kafr El–sheikh (k) ( North Delta Region ) and El- Dakahliya (D) ( East Delta Region ) . Only five cyanobacterial strains showed antagonistic effects against the both pathogenic fungi and were indentified as Nostoc muscorum k , Anabaena oryzae D , Anabaena oryzae k , Nostoc pruniforme D and Oscillatoria brevis D. The strains were analysed for phosphate solubilization , production of IAA , ammonia , HCN , production of some enzymes , and effect of their filterates on seed germination of flax seeds . Flax seeds treated with cyanobacterial filtrates germinated faster and produced higher seedlings compared with the nontreated ones .

Algal Diversity in the Rice Field of Parsa District (Nepal)

Twenty two species of algae were collected from rice field of Parsa district, Nepal and identified and recorded. Three Chlorophyceae, ten heterocystous cyanophceae and nine non-heterocystous cyanophyceae. Oscillatoria was most common in all the three selected rice field. Other species like Nostoc linkia, Anabaena oryzae and Merismopedia elegans have been also found as dominant taxa. Six cyanophyta and two chlorophyta are newly reported for Nepal.

Isolation and Identification of Dominant N2 Fixing Cyanobacterial Strains from Different Locations

Isolation of cyanobacteria dominated in the soil samples collected from different sites in Kafr El- Sheikh, and El-Dkahlia governorates isolates were successfully obtained as bacterial free cyanobacteria. They were identified cyanobacteria was carried out using the following: thallus color, thallus morphology and size of heterocyst, vegetative and reproductive cells. Heterocyst-forming cyanobacteria were also cultured as Anabaena sp., Nostoc sp., Oscillatoria sp., and Chroococcus sp. Nostoc sp . were isolated at high frequency; Nostoc sp. and Anabaena sp. isolates representing in kafr El-Sheikh and El-Dakahlia Governorates. Less frequency of cyanobacteria belonging to different genera namely Oscillatoria sp. and Chroococcus sp. Nostoc spp.were recorded highest amount of fixed nitrogen and mass production with in increasing incubation period of interacellular and /or extracellular nitrogenfixation. While,the most active strains for nitrogenase activity were Nostoc mucorum K and Anabaena oryzae D.

TLC Bioautographic Detection and Characterization of Antibacterial Compound from the Cyanobacterium Anabaena oryzae

TLC Bioautographic Detection and Characterization of Antibacterial Compound from the Cyanobacterium Anabaena oryzae

Bioremediation of heavy metals from municipal sewage by cyanobacteria and its effects on growth and some metabolites of Beta vulgaris

ABSTRACTThe present research was done to study the ability of cyanobacterial species for removing heavy metals from sewage. As well, to estimate the growth and some metabolites of Beta vulgaris irrigated with sewage treated by cyanobacterial species. The best removal results were obtained by Anabaena oryzae compared to the other studied cyanobacteria. Whereas A. oryzae showed high removal efficiency for cadmium (Cd2+) followed by lead (Pb2+), zinc (Zn2+), iron (Fe2+), copper (Cu2+) and manganese (Mn2+) (88.5, 83.1, 68.8, 62.0, 55.2 and 42.4%, respectively). Irrigation of plants by untreated or treated sewage generally caused stimulation in the total proteins, proline, carbohydrates and ascorbic acid. B. vulgaris grown in soil irrigated with untreated sewage showed maximum catalase, peroxidase activity, hydrogen peroxide (H2O2) and lipid peroxidation compared to the other treatments. The heavy metals availability was relatively low in the plant irrigated with treated sewage by cyanobacterial species, so the antioxidants requirement was low and hence the induction of antioxidants was lower compared to the plant irrigated with untreated sewage.

Approach to improve the productivity of bioactive compounds of the cyanobacterium Anabaena oryzae using factorial design

Cyanobacteria are one of the richest sources of biomedical relevant compounds with extensive therapeutic pharmaceutical applications and are also known as producer of intracellular and extracellular metabolites with diverse biological activities. The genus Anabaena sp. is known to produce antimicrobial compounds, like phycocyanin and others. The goal of this study was to optimize the production of these bioactive compounds. The Plackett–Burman experimental design was used to screen and evaluate the important medium components that influence the production of bioactive compounds. In this present study, eight independent factors including NaNO3, K2HPO4, MgSO4·7H2O, CaCl2, citric acid, ammonium ferric citrate, ethylene diamine tetraacetic acid disodium magnesium salt (EDTA-Na2Mg) and Na2CO3 were surveyed and the effective variables for algal components production of Anabaena oryzae were determined using two-levels Plackett–Burman design. Results analysis showed that the best medium components were NaNO3 (2.25gl−1); K2HPO4 (0.02gl−1); MgSO4 (0.0375gl−1); CaCl2 (0.018gl−1); citric acid (0.009gl−1); ammonium ferric citrate (0.009gl−1) and EDTA-Na2 (0.0015gl−1) respectively. The total chlorophyll-a, carotenoids, phenol, tannic acid and flavonoid contents in crude extract of Anabaena oryzae were determined. They were 47.7, 4.11, 0.256, 1.046 and 1.83μg/ml, respectively. The antioxidant capacity was 62.81%.

RESPONSE OF SOYBEAN PLANTS TO INOCULATION WITH RHIZOBIA AND CYANOBACTERIA

Field experiments were carried out on a clayey alluvial soil to evaluate the effect of (Bradyrhizobium japonicum (B.j), ,Nostoc muscorum, Anabaena oryzae, B.j. +Nostc, B.j.+Anabaena, and B.j.+Nostc+Anabaena , on soybean (Glycine max L.) vegetative growth, root nodulation status , yield and its components. Also, microbial activity in soybean rhizosphere soil and technology characters of seeds (protein, oil, and carbohydrate content) were determined. Results indicated that, inoculation with Brady rhizobium or cyanobacteria improved plant growth parameters, i.e., plant height, number of branches, number and dry weight of pods above the control (60kgN) . However, co-inoculation of Brady rhizobium with the two cyanobacteria significantly enhanced those parameters, where they scored significant increases in number and dry weight of root nodules, as well as nitrogenase activity of the root system, as compared with the single inoculation with Brady rhizobium alone or the control. The mixed inoculation improved dehydrogenase activity in the rhizosphere soil, as well as the seed content of oil, protein and carbohydrate were achieved. Also, the highest yield and its components were also recorded with the mixed inoculation.

Screening of norharmane from seven cyanobacteria by high-performance liquid chromatography

Background:Cyanobacteria, including pharmaceutically and medicinally valuable compounds attract the great attention lately. Norharmane (9H-pyrido (3,4-b) indole found in some cyanobacteria revealed a great number of biological effects.Objective:Seven cyanobacteria were isolated and identified from Yesilirmak River and Gaziosmanpasa University Campus to determine the norharmane content.Materials and Methods:Cyanobacteria collected from Tokat, Turkey were isolated and identified by morphologically. Norharmane (9H-pyrido [3,4-b] indole) quantities were presented for seven cyanobacteria, Chroococcus minutus (Kütz.) Nägeli, Geitlerinema carotinosum (Geitler) Anagnostidis, Nostoc linckia Bornet ex Bornet and Flahault, Anabaena oryzae F. E. Fritsch, Oscillatoria limnetica Lemmermann, Phormidium sp. Kützing ex Gomont, and Cylindrospermum sp. Kutzing ex E. Bornet and C. Flahault by high-performance liquid chromatography.Results:The norharmane amount indicated for cyanobacterial culture media altered in a species-dependent kind in the range of 0.81–10.87 μg/g. C. minutus produced the most norharmane among the investigated cyanobacteria as 10.87 μg/g.Conclusion:Cyanobacteria could be an important source of norharmane as well as pharmaceutically valuable compounds.SUMMARY Seven cyanobacteria were isolated and identified from Yesilirmak RiverQuantitative analysis of norharmane was executed on isolated cyanobacteriaFour cyanobecteria species included the norharmaneChroococcus minutus contained the most norharmane (10.87 μg/g). Abbreviations used: HPLC: High performance liquid chromatograph.

Biodegradation of crude oil by Anabaena oryzae, Chlorella kessleri and its consortium under mixotrophic conditions

Oil spill occurs frequently all around the world which certainly impacts adversely the environment. Thus a low cost and environmentally friendly bioremediation could be an alternative way to solve the oil pollution problem. The present study presents a bioremediation attempt using the microalgae/cyanobacterium consortium of Anabaena oryzae and Chlorella kessleri. Each of A. oryzae and C. kessleri as well as its consortium could grow at different concentrations of crude oil (0.5, 1, and 1.5%) under mixotrophic conditions. However, 1% of crude oil was the optimum concentration for their maximum growth. Chlorophyll-a content of C. kessleri, A. oryzae and its consortium increased from the first day of incubation with 0.5 and 1% crude oil concentration up to 14 days. While, chlorophyll-b content of the consortium decreased at different concentration of crude oil after 14 days. The carotenoids content of A. oryzae and C. kessleri increased with increasing the oil concentration from 0.5 to 1% with increasing the incubation time until day seven. GC/MS analysis showed that some aliphatic compounds such as, 3-methyl-decane, heptadecane, octadecane, nonadecane, docosane and tetracosane completely disappeared when A. oryzae or C. kessleri was incubated mixotrophically with 1% crude oil. Aromatic compounds such as, naphthalene decahydro-2-methyl, benzene-decyl, cyclohexanebutyl- completely disappeared when the consortium was incubated mixotrophically with 1% crude oil for 30 days. Overall, our results indicate that C. kessleri, A. oryzae and its consortium can grow mixotrophically, and enhance the crude oil biodegradation.

Siderophore mediated attenuation of cadmium toxicity by paddy field cyanobacterium Anabaena oryzae

The efficacy of dihydroxamate siderophore in attenuation/mitigation of cadmium toxicity, an optimum level of cadmium under which cyanobacterium can grow and produce siderophore, complexation behaviour of cadmium and siderophore and kinetics of complexation along with in silico analysis have been investigated in paddy field cyanobacterium Anabaena oryzae under iron replete and deplete conditions. Results showed that cadmium concentration as 1.0μM is optimum for growth as well as siderophore production in iron replete and deplete culture conditions. The complexation between cadmium–siderophore was evident in spectral scan. Kinetic studies as well as in silico docking analysis indicate that like iron, cadmium also forms a thermodynamically stable complex with siderophore. Thus our results suggest that the siderophore produced by Anabaena oryzae can act as a biological sequestering agent for the mitigation of cadmium metal ions from the paddy fields to improve crop productivity.

Fungicidal activity of extracellular products of cyanobacteria against Alternaria porri

ABSTRACTAlthough cyanobacteria are recognized as renewable sources of biomass for bioactive compounds, they have received little attention as potential biocontrol agents of foliar plant diseases. The purpose of this study was to evaluate the fungicidal efficacy of cyanobacterial extracellular products against Alternaria porri, which causes onion purple blotch disease, in vitro and under greenhouse conditions. Among the tested cyanobacteria (Anabaena oryzae, Arthrospira sp., Nostoc minutum, N. muscorum and Oscillatoria sp.), extracellular metabolites of N. muscorum and Oscillatoria sp. reduced the linear growth of A. porri by 20.37 and 36.34%, respectively. Culture filtrates of both Oscillatoria sp. and N. muscorum contained high concentrations of phenolic compounds (104.33 and 145.0 mg l−1, respectively), and alkaloids (473.31 and 378.12 mg l−1, respectively). GC-Mass analysis revealed that the most prevalent component in culture filtrates of N. muscorum and Oscillatoria sp. was beta ionone (7.21 and 7.17% of total analytes, respectively). Norharmane and α-iso-methyl ionone constituted 7.08 and 6.83% in N. muscorum and Oscillatoria sp., respectively. Components identified in the two culture filtrates included piperazine derivatives, isocyclocitral, α-trans-seequicyclocitral, phytol, oleic acid, fatty acids esters (methyl palmitate and linoleic acid mythyl ester), myristic alcohol and palmityl chloride. Application of culture filtrates of N. muscorum and Oscillatoria sp. under greenhouse conditions reduced disease severity by 55.1–66.5%. In conclusion, extracellular metabolites of N. muscorum and Oscillatoria sp. have potential as effective fungicides to control purple blotch disease of onion.

Influence of Various Levels of Iron and Other Abiotic Factors on Siderophorogenesis in Paddy Field Cyanobacterium Anabaena oryzae.

Siderophore production in Anabaena oryzae was investigated under the influence of various levels of iron and other abiotic factors such as pH, temperature, light and different nitrogen sources. Optimization of culture conditions under controlled mechanisms of these abiotic factors lead to the siderophore production in significant amount. Under iron-starved condition, A. oryzae extracellularly releases 89.17% hydroxymate-type siderophore. Slightly alkaline pH and 30 °C temperature was found stimulatory for the cyanobacterial growth and siderophorogenesis (88.52% SU and 83.87% SU, respectively). Excess iron loading had a negative impact on siderophore production along with the alterations in the morphology and growth. Further, scanning electron microphotographs signified that higher concentrations of iron lead to complete damage of the cells and alterations in membrane proteins possibly transporters responsible for exchange of siderophore complex from environment to the cell. SDS-PAGE analysis of whole cell proteins showed overexpression of low molecular weight proteins ranges between 20.1 to 29.0 kDa up to 100-μM iron concentrations. These polypeptides/proteins might be involved in maintaining iron homeostasis by regulating siderophore production. Results suggest that lower concentrations of iron ≤ 50 μM along with other abiotic factors are stimulatory, whereas higher concentrations (>50 μM) are toxic. Data further suggested that cyanobacterium A. oryzae can serve as a potential biofertilizer especially in iron-rich soil through sequestration by the power of natural Fe(III)-siderophore complex formation.

Sex hormonal disruption by cyanobacterial bioactive compounds

This study investigated the effect of some cyanobacterial exudates on the level of sex hormones in a mammalian model and to identify the molecules that could act as endocrine disruptor. In this study, the cytotoxicity of purified exudates of some axenic cyanobacterial cultures (Nostoc ellipsosporum, Nostoc muscorum, Anabaena oryzae & Anabaena. sp.) was evaluated against normal melanocyte cell line (HFB4). The intraperitoneal injection of the cyanobacterial exudates demonstrated a marked disturbance in the serum level of testosterone, progesterone, follicular stimulating hormone (FSH) and luteinizing hormone (LH) of male mice. GC-MS and LC-MS/MS analysis showed the presence of some sterol-like compounds in the cyanobacterial filtrates. This work demonstrated that the nontoxic cyanobacterial species have the ability to produce some bioactive compounds into their surroundings that can disrupt the mammalian reproductive hormones. The cyanobacterial extracellular bioactive molecules can affect on the production of the sex hormones via positive and negative feedback and may be a risk to human beings.

MICROCYSTINS IN RAW WATER, DRINKING WATER, AND SOME ISOLATES OF CYANOPROKARYOTES

Raw water samples of two drinking water treatment plants (named New Mansoura and Bilqas stations) were collected for a full annual cycle. Analyses of total soluble inorganic N, dissolved reactive phosphorus (DRP) and Total dissolved phosphorus (TDP) were carried out. Qualitative and quantitative analysis of Cyanoprokaryotes and photosynthetic pigments of phytoplankton were investigated. Total microcystins of raw water supply and potable water were seasonally analyzed using ELISA. Seven Cyanobprokaryotes species were isolated, cultured under lab conditions and biomass contents of microcystin–LR and -YR were analyzed using HPLC. Physico-chemical characteristics and photosynthetic pigments displayed pronounced periodic variations. Species composition and abundance of Cyanoprokaryotes exhibited marked seasonal variation during the period of study. Cyanoprokaryotes biovolume peaked in summer at New Mansoura (214.3mm3m-3) and Bilqas (161.3 mm3m-3) water treatment stations. The minimum mean biovolume of 25.2 and 30.8 mm3m-3 were recorded in winter at New Mansoura and in autumn at Bilqas station, respectively. Total microcystins in raw water maintained seasonal variations. Microcystins values were 2.2 and 2.0 µg l-1 during summer at Bilqas and New Mansoura respectively. Theses values of 1µg l-1 exceeding the WHO guidelines. Total microcystins of potable water were below the WHO guideline except in summer at New Mansoura (1.2µg l-1). Microcystins in biomass of different species of Cyanoprokaryotes varied between 3.0 mg g-1 (8% LR and 92%YR) and 0.3 mg g-1 (100% LR) recorded for Anabaena oryzae and Anabaena khannae, respectively.

Phylogenetic comparison among the heterocystous cyanobacteria based on a polyphasic approach

Phylogenetic comparison has been done among the selected heterocystous cyanobacteria belonging to the sections IV and V. The hierarchical cluster analysis based on antibiotics sensitivity showed a distant relationship between the members of Nostocales and Stigonematales. Thus, multiple antibiotic resistance pattern used as marker provide easy, fast, and reliable method for strain discrimination and genetic variability. However, morphological, physiological (both based on principal component analysis) and biochemical analysis grouped true branching cyanobacteria along with the members of section IV. Molecular analysis based on 16S rRNA gene sequences revealed that Hapalosiphon welwitschii and Westiellopsis sp. were grouped in cluster I whereas Scytonema bohnerii, a false branching genera showed a close proximity with Calothrix brevissima in cluster II. Cluster III of clade 2 included Nostoc calcicola and Anabaena oryzae which proved the heterogeneity at the generic level. Cluster IV the largest group of clade 2 based on 16S rRNA gene sequences includes six strains of the genera Nostoc, Anabaena, and Cylindrospermum showing ambiguous evolutionary relationship. In cluster IV, Anabaena sp. and Anabaena doliolum were phylogenetically linked by sharing 99% sequence similarity. Probably, they were of the same genetic makeup but appear differently under the diverse physiological conditions. Section IV showed polyphyletic origin whereas section V showed monophyletic origin. Results suggested that either morphological or physiological or biochemical or molecular attribute is not sufficient to provide true diversity and phylogeny of the cyanobacteria at the generic level and thus, a polyphasic approach would be more appropriate and reliable.

ENHANCEMENT OF SUGAR BEET SEED GERMINATION, PLANT GROWTH, PERFORMANCE AND BIOCHEMICAL COMPONENTS AS CONTRIBUTED BY ALGAL EXTRACELLULAR PRODUCTS

Effect of culture filtrates of nine algal strains belonging to Nostocales and Chlorellales on seeds germination, growth and certain biochemical characteristics of sugar beet was tested for their contents of Indole acetic, gibberellic and abscisic acids. Plant seeds soaked in algal culture filtrates for 24 hrs showed maximum germination percentage when soaked in filtrates of Spirulina platensis (72.9%), Anabaena oryzae (70.0%), Oscillatoria sp (69.8%) and Nostoc muscorum (68.7%) over the counterparts which were soaked in water. Greenhouse pot experiments were conducted during the winter growing seasons of 2005/2006 and 2006/2007 to investigate the effect of algal culture filtrates application methods as phytohormones source on growth stimulation and yield quality of sugar beet. Results revealed that different application methods had no significant effect on leaves and roots dry weight as well as total soluble solids, sucrose and purity percentages in root juice during the two seasons, while the positive significancy was conducted by the treatments of Nostoc muscorum, Anabaena oryzae, Spirulina platensis, Nostoc humifusum, Anabaena flous aquae and Phormedium fragile culture filtrates. The combination of seed-presoaking and foliar application method significantly increased leaves chlorophyll contents, as well as, nitrogen, phosphorus and potassium percentages in roots over the control in both seasons.

Treatment of mixed domestic–industrial wastewater using cyanobacteria

Alexandria Sanitary Drainage Company (ASDCO), Alexandria, Egypt has two primary treatment plants, the eastern and the western wastewater treatment plants (EWTP and WWTP) that receive mixed domestic-industrial influents and discharge into L. Mariut. The lake is subjected therefore to severe levels of pollution and dominated by members of cyanobacteria that can cope with the high pollution load in the lake water. Isolation and utilization of the locally generated cyanobacterial biomass for remediation processes of highly toxic pollutants offers a very efficient and cheap tool for governmental or private industrial activities in Alexandria and will generate a source of revenue in Egyptian localities. The main objective of the present study was to investigate the biodegradation and biosorption capacity of some potential cyanobacterial species dominating the lake ecosystem toward organic and inorganic contaminants polluting the primary-treated effluents of the EWTP and WWTP. The primary effluents were subjected to biological treatment using three axenic cyanobacterial strains (Anabaena oryzae, Anabaena variabilis and Tolypothrix ceytonica) as batch system for 7 days. Removal efficiencies (RE) of the different contaminants were evaluated and compared. Results confirmed the high efficiencies of the investigated species for the removal of the target contaminants which were species and contaminant-dependent. BOD5 and COD recorded 89.29 and 73.68% as maximum RE(s) achieved by Anabaena variabilis and Anabaena oryzae, respectively. The highest RE of the TSS recorded 64.37% achieved by Tolypothrix ceytonica, while 38.84% was recorded as the highest TSD RE achieved by Anabaena variabilis. Tolypothrix ceytonica also exhibited the highest RE for FOG recorded 93.75%. Concerning the contaminant metals, Tolypothrix ceytonica showed the highest biosorption capacity where 86.12 and 94.63% RE were achieved for Zn and Cu, respectively. In conclusion, results of the present study confirmed the advantageous potential of using the tested cyanobacterial species for the treatment of contaminated wastewater. Results also clearly showed the quality improvement of the discharged wastewater which in turn will eliminate or at least minimize the expected deterioration of the receiving environment.

English

Phosphorous (P) starved cells of the cyanobacterium Anabaena oryzae showed higher phosphate uptake rates than P-sufficient cells. The P-uptake obeyed saturation kinetics. The Km value for P-deficient cells was lower (54.34 mM) than P-sufficient cells (82.64mM) while Vmax was higher in P-deficient and lower in P-sufficient cells. Salinity (NaCl) stimulated phosphate uptake significantly in the cyanobacterium which is followed by greater amount of P-accumulation in the form of polyphosphate bodies. Inhibition of P-uptake in P-deficient cells was 45% in dark grown compared to light grown cells. P-uptake was inhibited 52 and 85% in culture treated with 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU; 10 mM) and carbonyl cyanide m-chloro phenylhydrazone (CCCP; 100 mM), respectively, suggesting that energy for uptake could be derived from oxidative phosphorylation and photophosphorelation.   Key words: Anabaena oryzae, metabolic inhibitors, phosphorus-uptake, salinity.

A rapid and cost-effective method of genomic DNA isolation from cyanobacterial culture, mat and soil suitable for genomic fingerprinting and community analysis

This study presents a phenol and lysozyme free protocol for genomic DNA isolation of cyanobacteria from culture, mats and soil. For an efficient and pure DNA isolation from cyanobacteria having tough cell wall, extra steps of glass beading and Sepharose 4B purification were added. The modified method gave a higher yield of DNA than the phenol: chloroform extraction method. Four parameters selected for purity testing of the isolated DNA were: (i) restriction digestion with Hind III, (ii) randomly amplified polymorphic DNA-PCR of axenic culture of cyanobacteria to assess phylogenetic relatedness, (iii) denaturing gradient gel electrophoretic (DGGE) analysis of cyanobacterial mat and soil to ascertain the applicability of the isolated DNA for community analysis, and (iv) sequencing of partial 16S rDNA of Hapalosiphon intricatus BHULCR1, Anabaena doliolum LCR1, Anabaena oryzae LCR2, Aulosira fertilissima LCR4, and Tolypothrix tenuis LCR7 and BLAST analysis to confirm their cyanobacterial identity. Data generated from above analyses lead us to conclude that the modified method in question is rapid, cost effective, health and time conscious and promising for genetic fingerprinting and community analysis of cyanobacteria from diverse habitats.

Factors modulating alkaline phosphatase activity in the diazotrophic rice-field cyanobacterium, Anabaena oryzae

Alkaline phosphatases (APase), both phosphomonoesterase (PMEase) and phosphodiesterase (PDEase) were studied in the cyanobacterium Anabaena oryzae for their specific requirements of temperature, pH, micro- and macronutrients and their activities in the presence of salinity and heavy metal stress. The alkaline phosphatases (PMEase and PDEase) are quite stable enzymes and require a narrow range of pH (pH 10–10.2) and temperature (35–40 °C) for their optimal activity.