Strains Of Fusarium Moniliforme Research Articles

Efficiency of a biological growth regulator in the cultivation of branched seedlings

The article presents the results of studies on the effectiveness of biological preparations Gibbersib - obtained on the basis of the Fusarium moniliforme strain - a polygibberellin preparation containing a set of gibberellic acids, as well as the preparation 6-Benzyladenine - a synthetic cytokinin intended to activate the vital activity of a plant associated with the growth and development of lateral shoots that improve the crowning of seedlings. According to the results of preparations testing with separate and combined use, the effectiveness of using each of the regulator separately, in comparison with the control, was confirmed. The greatest efficiency was obtained with the use of treatment with combined regulators, which makes it possible to enhance the branching of seedlings.

Evaluation of antagonistic activity and mechanisms of endophytic yeasts against pathogenic fungi causing economic crop diseases.

This work aimed to evaluate in vitro antagonistic activities and mechanisms of endophytic yeasts against phytopathogenic fungi. A total of 407 yeast strains isolated from tissue of rice, corn, and sugarcane leaves were evaluated for their antagonistic activities against ten phytopathogenic fungi. Only strains of Wickerhamomyces anomalus and Kodamaea ohmeri were found to inhibit the growth of phytopathogenic fungi. Wickerhamomyces anomalus (seven strains) showed antagonistic activity against Curvularia lunata (cause of dirty panicle disease of rice), three Fusarium moniliforme strains (cause of bakanae disease of rice, stalk rot disease of corn, and red rot disease of sugarcane), and Rhizoctonia solani (cause of sheath blight disease of rice). Whereas four Kodamae ohmeri strains inhibited growth of F. moniliforme (cause of bakanae disease of rice). Antagonistic mechanisms of W. anomalus were based on the production of volatile organic compounds (VOCs) (mainly 3-methyl-1-butyl acetate and 3-methyl-1-butanol), fungal cell wall-degrading enzymes (β-1,3-glucanase and chitinase), and siderophores as well as phosphate and zinc oxide solubilization. As for K. ohmeri, the production of VOCs (mainly 3-methyl-1-butanol), β-1,3-glucanase and chitinase were hypothesized as possible mechanisms. The antagonistic activity of W. anomalus against these phytopathogenic fungi and of K. ohmeri against F. moniliforme, and the analysis of the VOCs produced by K. ohmeri are reported for the first time. Two W. anomalus strains, DMKU-RE13 and DMKU-CE52, were evaluated for controlling rice sheath blight disease caused by R. solani in rice plants in the greenhouse and found to suppress the disease 55.2-65.1%, whereas 3% validamycin suppressed this disease by 88.5%.

Developmental and behavioural effects of the endophytic Fusarium moniliforme Fe14 towards Meloidogyne graminicola in rice

AbstractThe root‐knot nematode, Meloidogyne graminicola, is an important pest of rice in many rice production areas worldwide. The endophyte Fusarium moniliforme strain Fe14, isolated from a disinfected root of rice, has previously shown potential antagonistic activity against M. graminicola. This study shows the effects of Fe14 on M. graminicola behaviour, infection, development and reproduction. The endophyte Fe14 colonisation significantly reduced M. graminicola penetration into rice roots by 55% and increased the male to female ratio nine times. The endophyte also delayed juvenile development into female inside the rice root. These results suggest a suboptimal performance of the giant cell and a cumulative effect of the endophyte on the long‐term root‐knot nematode population development. In split‐root assays, the application of Fe14 at the inducer side significantly reduced nematode invasion at the responder side by 38% and 60% in two independent trials. This result suggests a systemic effect of the endophyte on rice plants. The root exudates from Fe14‐treated plants were either less attractive or had repellent effect on nematode movement. The results, when compared to what was described for other endophytic Fusarium against other nematode species, may indicate a basal response mechanism initiated in the plant by endophytic Fusarium spp. The present study may give leads for unravelling the molecular mechanisms responsible for the induced systemic defence responses in plants.

Development and validation of an inoculation method to assess the efficacy of biological treatments against Verticillium wilt in olive trees

Verticillium wilt in olive trees, caused by the soil-borne fungus Verticillium dahliae, is one of the most serious diseases of this crop due to high tree mortality and the difficulty of control. One of the major constraints to developing control measures against this disease is the lack of inoculation methods to evaluate the effectiveness of treatments. Here, we compared five inoculation methods for screening biological control agents (BCAs). The soil infested with cornmeal sand medium at 20 % w/w performed the best, and its effectiveness was further tested in olive plants treated with six BCAs, four strains of the fungal species Fusarium moniliforme, F. oxysporum, Gliocladium roseum and Phoma sp., and two fermented mixtures of several yeasts and bacteria. Strains of F. oxysporum, Phoma sp. and two mixtures of microorganisms significantly decreased the severity of the disease in potted plants of the susceptible olive cv. Picual.

Penicillium piceum: a potential source for antimicrobial agents and β-lactamase inhibitors

AbstractThere is an urgent need to discover new anti-microbial agents that can overcome the increasing antibiotic resistance posed by multidrug resistant, β-lactamase producing bacteria. β-Lactam antibiotics are structurally related through the presence of a core β-lactam ring. Resistant pathogens are able to produce β-lactamase enzymes that hydrolyze the β-lactam ring and inactivate the antibiotic. One approach to counteract this resistance is to develop β-lactamase inhibitors, which, on administration with β-lactams, restores the antibiotic activity for its continued clinical use. In the present study, a fungal Penicillium strain was isolated, identified and characterized for its antimicrobial and β-lactamase inhibition activities. The strain was identified as Penicillium piceum and was grown in suitable media under submerged cultivation. The antibacterial activity was observed against Bacillus subtilis, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. Higher antimicrobial activity was observed against gram-positive bacteria as compared to gram-negative bacteria. A certain amount of antifungal activity was also observed against the fungal strains of Penicilium chrysogenum, Rhizopus stolonifer, Fusarium moniliforme, Aspergillus niger, Giberella fujikuroi and Saccharomyces cerevisiae. Iodometric and bioassay analyses confirmed that P. piceum produced β-lactamase inhibitors having activity against penicillinase (Bacillus cereus). Solvent-solvent extraction with butanol yielded a three-fold increase in β-lactamase inhibition activity. High-performance liquid chromatography analysis of the concentrated product proved it to be a single compound. The infrared (IR) spectrum of the compound was compared with the reference IR spectra of sulbactam sodium, tazobactam and potassium clavulanate. It was found to be different thereby inferring the need for extensive structural elucidation for future applications.

STUDY THE EFFECT OF CARBON AND NITROGEN SOURCES ON GIBBERELLIC ACID PRODUCTION BY LOCAL STRAINS OF

Two local strains of Fusarium moniliforme, were isolated from leaves and roots of maize plant. The two strains were compared for gibberellic acid production. The production of gibberellic acid by the local strain (1) was higher than strain (2). The strain (1) of F. moniliforme was chosen for the later experiments. The effects of culture components and conditions were studied at aiming to improve gibberellic acid production by the local strain (1). The optimum incubation period for the highest production of gibberellic acid was 9 days. The glucose, as a carbon source, gave the highest production of gibberellic acid reached 91.13 mg/liter especially at the concentration 2%. Ammonium nitrate, as a nitrogen source, gave the higher production of gibberellic acid reached 94.55 mg/liter and at the concentration 3 gm/liter. The technology of the spectrophotometer was used for the quantitative evaluation for gibberellic acid in the samples and at the wase longitude of 254 nanometer.

EFFECT OF SOME GROWTH FACTORS ON GIBBERELLIC ACID PRODUCTION FROM WILD STRAIN OF Fusarium moniliforme

This study showed that the addition of certain compounds separately such as KH2PO4 , K2SO4 , corn steep liquor or yeast extract had affected gibberellic acid production from wild strain of Fusarium moniliforme. Corn steep liquor at the concentration 0.25 gm/L gave 92.31 mg/L of gibberellic acid, while potassium sulphate at the concentration 0.20 gm/L gave 91.95 mg/L of gibberellic acid, potassium dihydrogen phosphate gave the highest production where reached 93.48 mg/L especially at the concentration 0.5 gm/L, at the concentration 1.5 gm/L yeast extract gave 93.15 mg/L of gibberellic acid. The initial pH for highest production of gibberellic acid was 5.5 . The optimal incubation temperature to produce highest yield of acid was 30° C.

Synthesis, characterization and biocidal studies of new ferrocenyl thiadiazolo-triazinone complexes

One-pot synthesis of ferrocenyl ligand, 1,1′-bis[1,5-methyl-4H-(1,3,4)-thiadiazolo(2,3-c)(1,2,4)triazin-4-one]ferrocene was prepared from the reaction of 1,1′-diacetylferrocene with 4-amino-2,3-dihydro-6-methyl-3-thioxo[1,2,4]triazin-5(4H)one. The ligand, L, forms 1:1 complexes with Mn(II), Fe(III), Co(II), Ni(II), Cu(II) and Zn(II) in good yield. Characterization of the ligand and its complexes was carried out using IR, 1H NMR, magnetic susceptibility as well as elemental analysis. Biocidal activity of the ligand and its complexes were determined against the standard fungal strains of Aspergillus niger, Cladosporium herbarum and Fusarium moniliforme using the paper disc diffusion method; and against bacterial strains of Escherichia coli and Staphylococcus aureus using viable cell counting technique. The results indicated that the complexes are biologically more active than the free ligand. The biocidal activity depends on the metal ion, concentration as well as the tested fungi and bacteria. Copyright © 2006 John Wiley & Sons, Ltd.

Determination of fumonisin biosynthesis gene and toxicity of Fusarium moniliforme strains isolated in China

To study the relationship between fumonisin biosynthesis gene and toxicity of Fusarium moniliforme isolated in China. The toxigenic gene of 29 Fusarium moniliforme isolated from different provinces and varied food samples were determined. Eighteen fum5-positive strains were selected for biosyhesizing fumonisin and determined by high performance liquid chromatography (HPLC). Twenty-six isolates were identified as fum5 gene positive strains. And all of these strains produced FB(1), FB(2) and FB(3). The amount of FB(1), FB(2) and FB(3) was ranging from 0.41-140.20 mg/kg, 0.06-14.30 mg/kg to 0.30-58.00 mg/kg, except one strain produced a lower level of FB(1) only. It wight be the first report showing a high level fumonisin-producing strain isolated from the sesame sample and identified in the world. The amount of FB(1), FB(2) and FB(3) produced by the isolate was 128.84 mg/kg, 11.80 mg/kg and 14.88 mg/kg. It should have a close relationship between fumonisins biosynthesis gene and toxicity of Fusarium moniliforme isolated in China. The study demonstrated that strain of Fusarium moniliforme might contaminate the sesame sample and produce a high level of fumonisins.

Purification and biochemical characterization of polygalacturonase II produced in semi-solid medium by a strain of Fusarium moniliforme

A strain of Fusarium moniliforme isolated from a tropical mangrove ecosystem near Mumbai, India and deposited in the National Collection of Industrial Microorganisms (NCIM) as F. moniliforme NCIM 1276. The organism produced a single extracellular polygalacturonase (PG I) [EC 3.2.1.15] at pH 5 and a single pectate lyase (PL) [EC 4.2.2.2] at pH 8 in liquid medium containing 1% citrus pectin. Growth on semi-solid medium containing wheat bran and orange pulp resulted in a three-fold increase in PG production and a two-fold increase in PL production in comparison with that in liquid medium. The increased production of PG on semi-solid media, as compared to production in liquid media was investigated. The increased production of PG was partly due to the expression of a second polygalacturonase (PG II) isoenzyme by the fungus which was biochemically different from the one produced in liquid medium. The second PG II was a 30.6 kDa enzyme, had an alkaline p I of 8.6, the K m was 0.166 mg ml −1, V max 13.33 μmol min −1 mg −1 and the k cat was 403 min −1. It had a specific activity of 18.66 U mg −1. The differences between the PGs (PG I and PG II) suggest that the two enzymes are the products of different genes. The fungus also produced the same two PGs when it infected Lycopersicon esculentum (tomato). Only one PL was produced irrespective of growth conditions.

Production of fumonisins by Fusarium moniliforme strains isolated from corn grain

Fusarium moniliforme is the predominant fusarium species in the grain mycoflora of corn grown in Northern Caucasus, accounting for 95% of fusarium isolates. Eighty-five Fusarium moniliforme strains were grown on grain substrate and checked for the presence of fumonisins (B1 + B2 + B3) by indirect solid-phase enzyme immunoassay (EIA). All strains were capable of producing fumonisins (0.95 to 32,000 mg/kg). Strains sampled in the Krasnodar krai produced the highest fumonisin levels (averaging 5490 mg/kg).

Production of fumonisins by strains of Fusarium moniliforme according to temperature, moisture and growth period

Production of fumonisins B1 (FB1) and B2 (FB2) by two Brasilian strains (LAMIC 2999/96 and 113F) and one American strain (NRRL 13616) of Fusarium moniliforme were evaluated in laboratory cultures subjected to different temperatures (20, 25, and 30ºC), and moisture contents (25, 34, and 42%) on corn substrate. The cultures were grown during 10, 20, 30, 45, and 60 days, totalizing 135 treatments with two repetitions for each one. The fumonisins were extracted with acetonitrile/water. The clean-up with end-capped C18 silica (C18ec) cartridges and fumonisin derivatization with o-phtaldialdeyde were carried out through an automated sample processor system (ASPEC), followed by quantification of the toxins through HPLC. Fumonisin production varied widely, reaching average yields from 0.25 to 5515.45 µg/g of FB1 and from 0.15 to 3032.10 µg/g of FB2. In the present work, the factors strain, temperature, moisture and days of fungal culture were evaluated, and all of them had a bearing on the amounts of fumonisins produced. The highest FB1 average yields were obtained by the strain 113F, under the following conditions: 34% moisture content, 60 culture days, and temperature of 25ºC. The highest FB2 average yield was obtained by the same strain with cultures over 45 days, 42% moisture content, at the temperature of 25ºC. Via regression analysis, the ideal temperature for fumonisins production was, calculated as 24.5 and 24.3ºC (± 2ºC) for FB1 and FB2, respectively.

Cytotoxic and genotoxic effects of fumonisin B1 on rabbit kidney RK13 cell line.

Fumonisins, mycotoxins produced by certain strains of Fusarium moniliforme, could induce various diseases in animals and are suspected human carcinogens. Fumonisin B1 (FB1), the most commonly found fumonisin, has been characterised as a tumour initiator and a tumour promoter, a mitogen and an anti-proliferative agent. In this study we examined the cytotoxicity and genotoxicity of FB1 in rabbit kidney RK13 cells. To evaluate the effects of FB1 on survival of this cell line we analysed cell viability, membrane integrity, DNA fragmentation and overall morphology of the cells. The genotoxic potential of FB1 was estimated by monitoring the ability of this mycotoxin to induce micronuclei in RK13 cells. Exposure to FB1 caused a significant increase in micronucleus frequency in a concentration- and in a time-dependent manner. Nanomolar concentrations of FB1 decreased cell viability after 24 h and even more so after 48 h of exposure. The morphological changes observed suggested that an increased number of RK13 cells were dying by the process of apoptosis. However, FB1 also induced impairments of cell and mitochondrial membrane integrity, as assessed by lactate dehydrogenase and glutamate dehydrogenase leakage. These results could imply that nanomolar concentrations of FB1 induced apoptosis, which subsequently may proceed to secondary necrosis. In summary, our observations suggest that FB1 is genotoxic and cytotoxic to RK13 cells.

Characterization of gibberellin producing strains of Fusarium moniliforme based on DNA polymorphism.

The gibberellins are one of the major groups of growth promoting hormones and are secondary metabolites of the fungus Fusarium moniliforme (Perfect stage: Gibberella fujikuroi). Sixteen strains of Fusarium from different geographical regions and different hosts were analysed for their ability to produce gibberellins (GA) and for genetic relatedness by random amplified polymorphic DNA (RAPD). Range of gibberellin production varied between 28.9 to 600.0 mg g(-1) dry weight of mycelium in different strains of Fusarium. RAPD analysis showed completely different pattern between high, moderate and low producing strains. High producers formed nearly identical RAPD patterns, whereas the low and moderate producers gave heterologous amplification patterns. Since Fusarium pallidoroseum was in another group, it was possible to distinguish between different species of the genus Fusarium by RAPD. These investigations may find an application in the diagnosis of unknown Fusarium species and in distinguishing isolates of Gibberella fujikuroi within the section of Liseola.

Occurrence of Fusarium moniliforme and fumonisins in kernels of maize hybrids in Venezuela

Fusarium moniliforme (FM) incidence and fumonisins (FUM) content were determined in white kernel samples of maize hybrids, harvested in farms of Guarico and Portuguesa states, Venezuela. FM incidence was estimated in subsamples placed on a malt-salt-agar medium. Values in the range of 0-15 % were considered low, from 15-30% intermediate, and more than 30% high. FUM content was quantified through specific immunoassay (Fumonitest, VICAM Sci. Tech., USA) and values above 1 ppm were considered high. The hybrids were compared using a completely randomized design and results were subjected to analysis of variance and mean comparisons. The correlation between FUM content and FM incidence was also estimated. FM and FUM were detected in all of the 89 samples. Mean incidence of FM among hybrids ranged from 26.0 to 72.0% in Portuguesa state (PS) and from 28.7 to 55.7 in Guarico state (GS). Significant differences among hybrids were determined in farms of El Playon and Turen, PS, and El Sombrero and El Socorro, GS. The incidence of FM was high in all hybrids. FUM contents ranged from 0.80 to 10.00 ppm in PS, and from 1.50 to 13.00 ppm in GS. Significant differences among hybrids were found in El Playon, PS, and in both locations of GS. Only in Ospino, PS, the content of FUM was correlated with FM incidence. This indicates the natural occurence of FM strains with different toxigenic capabilities. The results show that Guarico and Portuguesa states are hazardous regions regarding FM and FUM in maize kernels.

Conversion of Various Aromatic Compounds by Resting Cells of Fusarium moniliforme Strain MS31.

Resting cells of Fusarium moniliforme strain MS31 convert propylbenzene to 1-phenylpropanol with high regio- and stereospecificity. To elucidate the scope of substrate acceptability by the fungus, we used various aromatic compounds for the bioconversion. The fungus hydroxylated various alkylbenzenes at the benzylic position to produce optically active alcohols. Butylbenzene was converted to nonbenzylic alcohols. In all cases, the R absolute configuration of products was more abundant. Aromatic compounds with linear side chains and (1-methylethyl)benzene were converted to their corresponding alcohols with an enantiomeric excess of 94% to 100%. Further oxidation of the alcohols was detected, but it was weak.

Optimal Conditions for Production of (R)-1-Phenylpropanol by Fusarium moniliforme Strain MS31.

Resting cells of Fusarium moniliforme strain MS31 produced (R)-1-phenylpropanol from propylbenzene. The components of the medium and the reaction conditions were adjusted to increase the specific activity of the hydroxylating enzyme involved. Glucose and sodium nitrate were selected as carbon and nitrogen sources, respectively. The substrate, propylbenzene, inhibited fungal growth and the activity of the enzyme. Acetoin added to the medium increased both growth and activity of the enzyme, and hydroxylation of propylbenzene increased by 1.4-fold. Maximum bioconversion of propylbenzene by resting cells of the fungus was at 25-30 degrees C and pH 7.0 with cells at concentration of 40 mg (dry) per milliliter of reaction mixture. Conversion was accelerated as soon as propylbenzene was added; slowing 2 h later. In the end, F. moniliforme strain MS31 produced (R)-1-phenylpropanol with an enantiomeric excess of 98% at the concentration of 16 mM (2.2 mg.ml(-1)).

Involvement of cytochrome P450 in hydroxylation of propylbenzene by Fusarium moniliforme strain MS31

Fusarium moniliforme strain MS31 can oxidize propylbenzene to ( R)-1-phenylpropanol with what may be a cytochrome P450. Hydroxylation of propylbenzene needed molecular oxygen, and NADPH as a coenzyme gave a higher yield than NADH. The hydroxylation proceeded further when FAD and FMN were added than in their absence, suggesting that the enzyme was a flavo-protein. Carbon monoxide inhibited the hydroxylation, as did other cytochrome P450 inhibitors such as SKF 525A and miconazole. These characteristics matched those of a microsomal cytochrome P450 monooxygenase system that contained NADPH-cytochrome P450 reductase.

Involvement of Cytochrome P450 in Hydroxylation of Propylbenzene by Fusarium moniliforme Strain MS31

Fusarium moniliforme strain MS31 can oxidize propylbenzene to (R)-1-phenylpropanol with what may be a cytochrome P450. Hydroxylation of propylbenzene needed molecular oxygen, and NADPH as a coenzyme gave a higher yield than NADH. The hydroxylation proceeded further when FAD and FMN were added than in their absence, suggesting that the enzyme was a flavo-protein. Carbon monoxide inhibited the hydroxylation, as did other cytochrome P450 inhibitors such as SKF 525A and miconazole. These characteristics matched those of a microsomal cytochrome P450 monooxygenase system that contained NADPH-cytochrome P450 reductase.

Optimal conditions for production of ( R)-1-phenylpropanol by Fusarium moniliforme strain MS31

Resting cells of Fusarium moniliforme strain MS31 produced ( R)-1-phenylpropanol from propylbenzene. The components of the medium and the reaction conditions were adjusted to increase the specific activity of the hydroxylating enzyme involved. Glucose and sodium nitrate were selected as carbon and nitrogen sources, respectively. The substrate, propylbenzene, inhibited fungal growth and the activity of the enzyme. Acetoin added to the medium increased both growth and activity of the enzyme, and hydroxylation of propylbenzene increased by 1.4-fold. Maximum bioconversion of propylbenzene by resting cells of the fungus was at 25–30°C and pH 7.0 with cells at concentration of 40 mg (dry) per milliliter of reaction mixture. Conversion was accelerated as soon as propylbenzene was added; slowing 2 h later. In the end, F. moniliforme strain MS31 produced ( R)-1-phenylpropanol with an enantiomeric excess of 98% at the concentration of 16 mM (2.2 mg·ml −1).