Fumonisin production and other traits of Fusarium moniliforme strains from maize in northeast Mexico.

Strains of Fusarium moniliforme from maize seed collected in four fields in northeast Mexico were tested for fumonisin production in culture, for sexual compatibility, and for vegetative compatibility by using non-nitrate-utilizing mutants. The test results indicate that a diverse population of fumonisin-producing strains of F. moniliforme (Gibberella fujikuroi) mating population A predominates and that a potential exists for production of fumonisins in Mexican maize.     

Genetic analysis of fumonisin production and virulence of Gibberella fujikuroi mating population A (Fusarium moniliforme) on maize (Zea mays) seedlings.

The phytopathogenic fungus Gibberella fujikuroi mating population A (anamorph, Fusarium moniliforme) produces fumonisins, which are toxic to a wide range of plant and animal species. Previous studies of field strains have identified a genetic locus, designated fum1, that can determine whether fumonisins are produced. To test the relationship between fumonisin production and virulence on maize seedlings, a cross between a fum1+ field strain that had a high degree of virulence and a fum1- field strain that had a low degree of virulence was made, and ascospore progeny were scored for these traits. Although a range of virulence levels was recovered among the progeny, high levels of virulence were associated with production of fumonisins, and highly virulent, fumonisin-nonproducing progeny were not obtained. A survey of field strains did identify a rare fumonisin-nonproducing strain that was quite high in virulence. Also, the addition of purified fumonisin B1 to virulence assays did not replicate all of the seedling blight symptoms obtained with autoclaved culture material containing fumonisin. These results support the hypothesis that fumonisin plays a role in virulence but also indicate that fumonisin production is not necessary or sufficient for virulence on maize seedlings.     

Survey of fumonisin production by Fusarium species

Fumonisins B1 (FB1) and B2 (FB2), two structurally related mycotoxins with cancer-promoting activity, were recently isolated from corn cultures of Fusarium moniliforme MRC 826. These toxins have been reported to be produced also by isolates of F. proliferatum. Contamination of foods and feeds by F. moniliforme has been associated with human esophageal cancer risk, and FB1 has been shown to be the causative agent of the neurotoxic disease leukoencephalomalacia in horses. Because of the toxicological importance of the fumonisins, the potential to produce FB1 and FB2 was determined in a study of 40 toxic Fusarium isolates representing 27 taxa in 9 of the 12 sections of Fusarium, as well as two recently described species not yet classified into sections. With the exception of one isolate of F. nygamai, fumonisin production was restricted to isolates of F. moniliforme and F. proliferatum, in the section Liseola. The F. nygamai isolate produced 605 micrograms of FB1 g-1 and 530 micrograms of FB2 g-1, and the identity of the toxins was confirmed by capillary gas chromatography-mass spectrometry. This is the first report of the production of the fumonisins by F. nygamai.     

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).     

Study of the biosynthesis of fumonisins B1, B2 and B3 by different strains of Fusarium moniliforme

The relationship between fungal growth and the production of fumonisin on maize grain by 25 strains of Fusarium moniliforme of different origins has been investigated. Although sporulation was essentially the same for all the strains (about 10⁸ propagules g⁻¹ dry matter), ergosterol assays revealed marked variations in fungal biomass. All strains studied produced highly variable amounts of fumonisin B1, the highest levels being observed in strains of ergosterol content above 400 μg g⁻¹. However, no correlation could be established between the synthesized biomass and the quantity of fumonisins produced. We verified that ergosterol is an indicator of mycelial growth, and therefore of the potential toxicity of the analysed grain.     

Influence of kernel age on fumonisin B1 production in maize by Fusarium moniliforme.

Production of fumonisins by Fusarium moniliforme on naturally infected maize ears is an important food safety concern due to the toxic nature of this class of mycotoxins. Assessing the potential risk of fumonisin production in developing maize ears prior to harvest requires an understanding of the regulation of toxin biosynthesis during kernel maturation. We investigated the developmental-stage-dependent relationship between maize kernels and fumonisin B1 production by using kernels collected at the blister (R2), milk (R3), dough (R4), and dent (R5) stages following inoculation in culture at their respective field moisture contents with F. moniliforme. Highly significant differences (P 相似文献   

Production of fumonisins by Fusarium moniliforme strains from various substrates and geographic areas.

Strains of Fusarium moniliforme from different geographic areas and from corn and other substrates were tested for the ability to produce fumonisins in culture. The test results indicate that the potential exists for production of fumonisins by such strains in agricultural commodities and other substrates in widespread geographic areas.     

Production of fumonisins by Fusarium moniliforme strains from various substrates and geographic areas.

Strains of Fusarium moniliforme from different geographic areas and from corn and other substrates were tested for the ability to produce fumonisins in culture. The test results indicate that the potential exists for production of fumonisins by such strains in agricultural commodities and other substrates in widespread geographic areas.     

Dimorphic fungus characteristic of fumonisin-producing strains of Fusarium moniliforme from Zhejiang

Fusarium moniliforme and its fumonisins have been shown to be carcinogenic in lab animals and have been linked to high incidences of human esophageal cancer. In this study we report the dimorphic fungus characteristic of fumonisin-producing strains of F. moniliforme from foodstuffs in Zhejiang, China. All of the twenty strains of F. moniliforme shown produce fumonisin B₁ 475.9–6322.2 μg/g in corn medium. These strains of F. moniliforme form yeast-like colonies in Sabouraud's agar plates contained 9% NaCl at 37 °C incubator and shows mostly budding reproduction. In blood agar plates these strains of F. moniliforme appear grass-green haemolytic reactions. This is the first report that yeast-like growth, dimorphic pathogenic fungus feature is found in F. moniliforme. These results suggest that it is also important to program epidemiological surveys of F. moniliforme as a primary pathogenic fungus, while proceeding to produce mycotoxins of F. moniliforme in food hygiene. This revised version was published online in August 2006 with corrections to the Cover Date.     

Fumonisin contamination of food: progress in development of biomarkers to better assess human health risks.

Fumonisins, fungal toxins produced by Fusarium moniliforme, contaminate maize based foods and feeds throughout the world. They cause liver and kidney toxicity in animals in addition to leukoencephalomalacia in horses and pulmonary edema in pigs. Fumonisin B(1) is carcinogenic in rats and mice. Ecological studies have linked consumption of fumonisin contaminated maize with oesophageal cancer in human populations in South Africa and China. This review discusses the potential health risks for people exposed to the fumonisins, and describes how mechanistic studies of toxicity in animal models have allowed the development of putative biomarkers of fumonisin exposure at the individual level. The requirements for an applicable biomarker include sample availability as well as a high specificity and sensitivity for the exposure of interest. Most environmental toxic insults involve complex exposures both to other toxins and to infections; these confounding factors need to be considered in assessing both the validity of the biomarker and the exposure-disease associations. Fumonisins can be detected in the urine of animals in feeding studies but the sensitivity of the current methodology means only highly exposed people could be monitored. Mechanistic studies indicate that ceramide synthase, an enzyme involved in sphingolipid synthesis, is one cellular target for fumonisin toxicity and carcinogenicity, and this disruption to sphingolipid metabolism increases the ratio of two sphingoid precursors, sphinganine and sphingosine. The altered ratio has been observed in tissues, serum and urine for a number of animal models suggesting it as a good candidate marker of fumonisin exposure. Despite development of analytical methods to measure this biomarker there have been no studies to date correlating it to fumonisin intake in people. Given the toxic effects of fumonisins in animals and the widespread human exposure, which has been calculated to reach 440 micrograms kg(-1) body weight day(-1) in a population consuming high quantities (460 g day(-1)) of contaminated maize, then the development of biomarkers and their application in epidemiological studies should be a priority for research on these toxins.     

Occurrence of fumonisins (B<Subscript>1</Subscript>, B<Subscript>2</Subscript>, B<Subscript>3</Subscript>) in maize-based food and feed samples from Indonesia

A survey to evaluate the contamination level of total fumonisins in maize-based foodstuffs, maize and feed from Indonesia is described. The analyses were carried out by enzyme-linked immunosorbent assay (ELISA). Samples were collected from local retail stores around Yogyakarta, Indonesia between February and May 2001. The 101 samples were classified into six categories, i.e. industrially-produced food (n=24), products of small food manufacturers (n=17), maize flour (n=4), maize for food (n=9), maize for feed (n17), and formulated feed (n30). Control of the method showed that the detection limit was 8.7 μg/kg and repeatability is shown by relative standard deviation (RSD) of analyses of contaminated maize (n=5) of 10 %. Results of analyses indicate that 80 samples analysed were contaminated over a large range from 10.0-3307 pg/kg, and the concentration of fumonisins depended on the type of sample. Of four samples of maize flour, none were contaminated (below detection limit). Of 24 samples of industrially produced food, 14 were contaminated in the range 22.8 - 105 μg/kg and 18 of 19 food samples from small manufacturers were contaminated ranging from 12.9 to 234 μg/kg. The highest contamination was observed in maize samples: six of ten samples of maize for food were contaminated between 68.0 - 2471 μg/kg and 16 of 17 samples for feed contained fumonisins over a large range from 17.6 to 3306 μg/kg.     

HPLC/MS analysis of fusarium mycotoxins, fumonisins and deoxynivalenol

Fusarium fungi are widely found in agricultural products, worldwide and can produce a great variety of mycotoxins. Fumonisins, produced by F. moniliforme, and deoxynivalenol, produced by F. graminearum, are two such mycotoxins that have received considerable attention as food safety concerns by regulatory agencies. High Performance Liquid Chromatography/Mass Spectrometry (HPLC/MS) was found to be a convenient analytical method to detect and quantify the naturally occurring fumonisin homologs and deoxynivalenol in extracts from grains and food products. The fumonisins are detected primarily as protonated molecules in the positive ion electrospray ionization (ESI) mode as they elute from a C-18 reverse phase column during a methanol water gradient containing acetic acid to facilitate chromatography. Deoxynivalenol can be detected as positive or negative ions in the atmospheric pressure chemical ionization (APCI) mode or in the negative ion ESI mode. One nanogram amounts of fumonisins or deoxynivalenol injected into the HPLC system are easily detected with signal to noise allowing detection limits of 1 microg g(-1) or better to easily be achieved with minimal clean-up of grain extracts.     

Detection of fumonisin B1: comparison of flow-injection liposome immunoanalysis with high-performance liquid chromatography

Fumonisins are secondary metabolites of the fungus Fusarium moniliforme, a common mycotoxin in corn, which are known to cause cancer in a number of experimental animals and have been linked to human esophageal cancer in China and South Africa. A high-performance liquid chromatographic (HPLC) method is currently the most widely used method for the quantitative determination of fumonisins. This method utilizes precolumn derivatization with o-phthaldialdehyde, isocratic elution, and fluorescence detection. In this study, the HPLC method was chosen as the reference method to evaluate the reproducibility and accuracy of FILIA (flow-injection liposome immunoanalysis) for the detection of the fumonisin B1 (FmB1). Studies indicate that a recovery of 86-90% could be obtained when commercial yellow cornmeal spiked with FmB1 was extracted in 75% methanol, which correlated favorably (correlation coefficient, r(2)=0.945) with the result of 80-92% obtained using the flow-injection liposome immunoanalysis (FILIA) system. The data suggest that the FILIA method is comparable to HPLC for the detection of fumonisins in corn, animal feeds, and human foods. Important features of FILIA as compared to HPLC are, most importantly, lower detection limit (ca. 25 x lower), and also less complex and faster sample preparation and therefore increased analytical throughput. In addition, 24 human corn-based foods and 6 animal feeds were examined for the presence of FmB1 using HPLC and FILIA.     

Effect of commercial processing on fumonisin concentrations of maize-based foods

Fumonisin-contaminated maize was used to study the effect of three cooking and food processing methods and residual contamination of the food product. Frying, autoclaving and extrusion were examined with naturally-contaminated maize meal, maize flour and sweet maize kernels, all at two fumonisin concentrations. High Pressure Liquid Chromatography determination of fumonisins B1 and B2 and hydrolized fumonisin B1 (AP1) were performed in unprocessed materials and at the end of the experimental runs. Reductions of fumonisins concentration in processed products were obtained for fried polenta and from one of the two runs of extruded maize batter. These reductions were consistent with previous studies of the thermal degradation of fumonisins. Autoclaving sweet maize kernels apparently resulted in reductions of fumonisin concentrations that were highly temperature dependent, but this needs further study. The unexpectedly large reduction in fumonisin concentrations in the extrusion processing of batter with high initial concentrations also needs further investigation. There was no evidence that AP1 was formed under any of the conditions tested.     

Inhibition of sphingolipid biosynthesis by fumonisins. Implications for diseases associated with Fusarium moniliforme

Culture materials and grains contaminated with certain isolates of Fusarium moniliforme cause equine leucoencephalomalacia, porcine pulmonary edema syndrome, and liver cancer in rats. The causative agents are thought to be a family of compounds called fumonisins, which bear considerable structural similarity to the long-chain (sphingoid) base backbones of sphingolipids. Incubation of rat hepatocytes with fumonisins inhibited incorporation of [14C]serine into the sphingosine moiety of cellular sphingolipids with an IC50 of 0.1 microM for fumonisin B1. In contrast, fumonisin B1 increased the amount of the biosynthetic intermediate sphinganine, which suggests that fumonisins inhibit the conversion of [14C]sphinganine to N-acyl-[14C]sphinganines, a step that is thought to precede introduction of the 4,5-trans double bond of sphingosine (Merrill, A.H., Jr. and Wang, E. (1986) J. Biol. Chem. 261, 3764-3769). In agreement with this mechanism, fumonisin B1 inhibited the activity of sphingosine N-acyltransferase (ceramide synthase) in rat liver microsomes with 50% inhibition at approximately 0.1 microM and reduced the conversion of [3H]sphingosine to [3H]ceramide by intact hepatocytes. As far as we are aware, this is the first discovery of a naturally occurring inhibitor of this step of sphingolipid metabolism. These findings suggest that disruption of the de novo pathway of sphingolipid biosynthesis may be a critical event in the diseases that have been associated with consumption of fumonisins.     

Induction of mRNA accumulation corresponding to a gene encoding a cell wall hydroxyproline-rich glycoprotein by fungal elicitors

The Hrgp (hydroxyproline-rich glycoprotein) gene codes in maize for one of the most abundant proteins of the cell wall. HRGPs may contribute to the structural support of the wall and they have also been involved in plant defense mechanisms. This second aspect has been tested for the Hrgp gene in maize where, in contrast with the situation in dicot species, the gene is encoded by a single-copy sequence. Hrgp mRNA accumulation is induced in maize suspension-cultured cells by elicitors, isolated either from maize pathogenic or non-pathogenic fungi. The induction of Hrgp mRNA accumulation by elicitor extracted from Fusarium moniliforme has been studied in detail. The level of induction depends on elicitor concentration and remains high until at least 24 h. Ethylene and protein phosphorylation appear to be involved in the transduction pathway of Hrgp gene activation by the F. moniliforme elicitor but not by 5 µM methyl jasmonate or 1 mM salycilic acid. Different compounds known to participate in plant stress responses such as ascorbic acid or reduced glutathione have also a positive effect on Hrgp mRNA accumulation.     

Use of PCR for detection of maize seeds infected with the fungus Fusarium moniliforme Sheldon var.lactis

The procedure of the PCR for the detection of Fusarium moniliforme Sheldon var. lactis on the maize seeds has been elaborated. The based on a highly sensitive molecular genetics technology method of fungal pathogen detection allows to reveal its presence before the visual symptoms of seed damage.     

Heritability of fumonisin B1 production in Gibberella fujikuroi mating population A.

Fumonisins are mycotoxins produced by strains belonging to several different mating populations of Gibberella fujikuroi (anamorphs, Fusarium section Liseola), a major pathogen of maize and sorghum worldwide. We studied the heritability of fumonisin production in mating population A by crossing fumonisin-producing strains collected from maize and sorghum in the United States with fumonisin-nonproducing strains collected from maize in Nepal. Random ascospore and tetrad progeny from three of these crosses were analyzed by gas chromatography-mass spectrometry and high-performance liquid chromatography for their ability to produce fumonisins on autoclaved cracked maize. In all three crosses, the ability to produce fumonisins, predominately fumonisin B1, segregated as a single gene or group of closely linked genes. Intercrosses between appropriate progeny and parents were poorly fertile, so we could not determine if the apparent single genes that were segregating in each of these crosses were allelic with one another. Mating type and spore-killer traits were scored in some crosses, and each segregated, as expected, as a single gene that was unlinked to the ability to produce fumonisins. We conclude that G. fujikuroi mating population A provides a powerful genetic system for the study of this important fungal toxin.