Production of Trichothecene Mycotoxins by Fusarium Species in Shake Culture

Twelve T-2 toxin-producing isolates and four fusarenon-X-producing isolates of Fusarium species were examined for their ability to produce trichothecene mycotoxins in shake culture and jar fermentation. T-2 toxin producers such as Fusarium solani, F. sporotrichiodes, and F. tricinctum produced T-2 toxin and neosolaniol in semisynthetic medium. F. solani M-1-1 produced the largest amount of the mycotoxins in a nutrient medium consisting of 5% glucose (or sucrose), 0.1% peptone, and 0.1% yeast extract in either shake culture or jar fermentation at 24 to 27 C for 5 days. None of the isolates produced significant amount of fusarenon-X in shake cultures.     

Mycotoxin Production by Fusarium Species Isolated from Bananas

The ability of Fusarium species isolated from bananas to produce mycotoxins was studied with 66 isolates of the following species: F. semitectum var. majus (8 isolates), F. camptoceras (3 isolates), a Fusarium sp. (3 isolates), F. moniliforme (16 isolates), F. proliferatum (9 isolates), F. subglutinans (3 isolates), F. solani (3 isolates), F. oxysporum (5 isolates), F. graminearum (7 isolates), F. dimerum (3 isolates), F. acuminatum (3 isolates), and F. equiseti (3 isolates). All isolates were cultured on autoclaved corn grains. Their toxicity to Artemia salina L. larvae was examined. Some of the toxic effects observed arose from the production of known mycotoxins that were determined by thin-layer chromatography, gas chromatography, or high-performance liquid chromatography. All F. camptoceras and Fusarium sp. isolates proved toxic to A. salina larvae; however, no specific toxic metabolites could be identified. This was also the case with eight isolates of F. moniliforme and three of F. proliferatum. The following mycotoxins were encountered in the corn culture extracts: fumonisin B(inf1) (40 to 2,900 (mu)g/g), fumonisin B(inf2) (150 to 320 (mu)g/g), moniliformin (10 to 1,670 (mu)g/g), zearalenone (5 to 470 (mu)g/g), (alpha)-zearalenol (5 to 10 (mu)g/g), deoxynivalenol (8 to 35 (mu)g/g), 3-acetyldeoxynivalenol (5 to 10 (mu)g/g), neosolaniol (50 to 180 (mu)g/g), and T-2 tetraol (5 to 15 (mu)g/g). Based on the results, additional compounds produced by the fungal isolates may play prominent roles in the toxic effects on larvae observed. This is the first reported study on the mycotoxin-producing abilities of Fusarium species that contaminate bananas.     

Zearalenone Production by Fusarium Species

One-hundred-and-thirteen isolates of Fusarium were tested for their ability to produce zearalenone on autoclaved corn. They belonged to the following species (number of producers per number tested): F. epispheria, (0/1); F. moniliforme, (0/8); Gibberella fujikuroi, (0/3); F. nivale, (0/7); F. oxysporum, (0/15); F. roseum, (31/51); F. solani, (0/9); F. tricinctum (3/19). The isolates of individual species produced the following amounts of zearalenone per gram of corn: 3 isolates of F. roseum (0.6 to 119 mug), 3 of F. roseum "Culmorum" (1 to 210 mug), 3 of F. roseum "Equiseti" (0.6 to 2.0 mug), F. roseum "Gibbosum" (115 to 175 mug), 21 of F. roseum "Graminearum" (0.2 to 230 mug), and 3 of F. tricinctum (0.2 to 6.0 mug). All isolates of F. roseum "Graminearum" which formed the perithecial stage of G. zeae (G. roseum) produced zearalenone. Production occurred by the wild but not the appressed cultural type. Zearalenone production by F. tricinctum was confirmed by a mouse bioassay.     

Beauvericin Production by Fusarium Species

Beauvericin is a cyclohexadepsipeptide mycotoxin which has insecticidal properties and which can induce apoptosis in mammalian cells. Beauvericin is produced by some entomo- and phytopathogenic Fusarium species (Fusarium proliferatum, F. semitectum, and F. subglutinans) and occurs naturally on corn and corn-based foods and feeds infected by Fusarium spp. We tested 94 Fusarium isolates belonging to 25 taxa, 21 in 6 of the 12 sections of the Fusarium genus and 4 that have been described recently, for the ability to produce beauvericin. Beauvericin was produced by the following species (with the number of toxigenic strains compared with the number of tested strains given in parentheses): Fusarium acuminatum var. acuminatum (1 of 4), Fusarium acuminatum var. armeniacum (1 of 3), F. anthophilum (1 of 2), F. avenaceum (1 of 6), F. beomiforme (1 of 1), F. dlamini (2 of 2), F. equiseti (2 of 3), F. longipes (1 of 2), F. nygamai (2 of 2), F. oxysporum (4 of 7), F. poae (4 of 4), F. sambucinum (12 of 14), and F. subglutinans (3 of 3). These results indicate that beauvericin is produced by many species in the genus Fusarium and that it may be a contaminant of cereals other than maize.     

Biological and Chemical Detection of Trichothecene Mycotoxins of Fusarium Species

The procedure for biological and chemical detection of trichothecene-type mycotoxins and its application to the screening of Fusarium for toxic strains were described.     

Microbial Production of Ursodeoxycholic Acid from Lithocholic Acid by Fusarium equiseti M41

A fungus identified as Fusarium equiseti was isolated from soil and found to carry out 7β-hydroxylation of lithocholic acid to ursodeoxycholic acid (35% yield; 350 mg/liter) in 112 h.     

Fusarium Species Colonizing Spears and Forming Mycotoxins in Field Samples of Asparagus from Germany and Poland

The occurrence of Fusarium spp. and associated mycotoxins in asparagus spears was evaluated in Poland in 2002 and 2003 and in Germany in 2002. Spears of two cultivars, Eposs and Gijnlim, were collected from two locations in Poland, Swidwowiec and Poznan, on sandy and sandy loam soil, respectively. Fusarium oxysporum and F. proliferatum were detected at an average incidence of 38.3% and 15.8% in the spear sections sampled, respectively. In stands of 11 (tested) cultivars of asparagus sampled in Germany on sandy soil, the same species dominated, however, they were less frequent than in Poland (26.6% and 5.6% of the spears infected with F. oxysporum and F. proliferatum, respectively). Chemical analyses revealed that fumonisin B₁ (FB₁) and moniliformin (MON) were present in some of the spears sampled in Poland. FB₁ was not found and MON was not assessed in spears sampled in Germany in 2002, but F. proliferatum was able to form the toxin in vitro in the range from 101.4 up to 205.8 μg/kg maize kernel substrate. Asparagus samples in Poland contained FB₁ at up to 5.6 μg/kg spear fresh weight. The highest MON concentration (1350 μg/kg) was detected in cultivar Eposs in Marcelin, Poland, in 2002. MON and FB₁ were found in spears infected by both F. oxysporum and F. proliferatum, however, only the latter fungus was able to synthesize both toxins.     

赤霉素和脱落酸对黑稻黄化幼苗中胚轴伸长生长的作用研究

研究了不同浓度的GA和ABA对三种黑稻黄化幼苗中胚轴伸长生长的影响。结果表明 :1.2 5 μmol/L的GA和ABA对三种黑稻黄化幼苗中胚轴的伸长生长有显著的促进作用 ,GA效应高于ABA ;三个供试稻种中 ,黑帅对GA和ABA的反应最为显著 ,且GA和ABA对其有叠加效应     

Microbiological Production of Gibberellic Acid in Glucose Media

Gibberellic acid production from various substrates was studied in 43 strains of Fusarium, among which F. moniliforme strain IOC-3326 was selected as the best producer. Experiments were carried out in shaker flasks and pilot plant fermentors. The results indicate that the best substrate for gibberellic acid production with this strain is composed of the following: glucose, 20 g; corn steep liquor, 25 g; ammonium nitrate, 2.6 g; monopotassium phosphate, 0.5 g; potassium sulfate, 0.2 g; and water, 1000 ml. Glucose, ammonium nitrate, and corn steep liquor were found to be critical. With this medium, maximal yields of 1196 mg per liter in shaker flasks and 997 mg per liter in fermentors were produced.     

Mycotoxins produced by Discolor section Fusarium species

An improved TLC method ofFusaria metabolites detection and quantitation has been elaborated. A total 92 isolates of Discolor sectionFusaria from cereals and potato have been examined from the point of view of cultures morphology and ability to produce characteristic mycotoxins. Low nutrient media (CLA, SNA) were found as suitable for production of uniform and typical macroconidia in studied cultures. All 26 isolates ofF. sambucinum Fuckel (=F.sulphureum, Schlecht) formed diacetoxyscirpenol in amount 20-1000 mg/kg and all 17.F. crookwellense Burgess N. & T. produced zearalenone (16-602 mg/kg).F. graminearum Schwabe produced: zearalenone 14/14 isolates, deoxynivalenol 11/14 isolates, both up to 77 mg/kg. Out of 26F. culmorum cultures originating from Poland 22 produced zearalenone up to 675 mg/kg, 17/26 3 acetyldeoxynivalenol up to 280 mg/kg and 16/26 deoxynivalenol up to 220 mg/kg. The difference in metabolism agrees with the difference in morphology of those species.     

Cellular Fatty Acid Composition of Selected Pseudomonas Species

The cellular fatty acid composition of 10 reference strains representing eight species of Pseudomonas was determined by gas-liquid chromatography. A variety of acids were detected in these organisms, including branched and straight-chained acids, cyclopropane, and hydroxy acids. Comparison of the presence and relative amounts of these acids among strains was useful for distinguishing various Pseudomonas species.     

Biological Assays for Two Mycotoxins Produced by Fusarium tricinctum

A survey was made to detect microorganisms useful for assaying butenolide [4-acetamido-4-hydroxy-2-butenoic acid gamma-lactone] and T-2 toxin [4beta, 15-diacetoxy-8alpha-(3-methylbutyryloxy)-12,13-epoxytricothec -9-en-3alpha-ol]. These mycotoxins produced by strains of Fusarium tricinctum have been implicated in mycotoxicosis of livestock. Although butenolide proved to be a very weak antibiotic, assay discs containing 100 mug of this toxin inhibited Sprillum serpens NRRL B-2052, Vibrio tyrogenus NRRL B-1033, and Xanthomonas campestris NRRL B-1459. T-2 toxin had no effect on 54 bacterial strains but inhibited 6 of 11 fungi. Growth of Rhodotorula rubra NRRL Y-7222 and Penicillium digitatum NRRL 1202 was retarded by assay discs containing 4 mug of T-2 toxin. Solutions with less than 1 mug of T-2 per ml toxin were readily detected by a pea seed germination test. Germination was reduced more than 50% when seeds imbibed solutions of 0.5 mug of T-2 toxin per ml. Butenolide had no effect on pea seed germination at concentrations as high as 200 mug/ml.     

Downregulation of the Basic Peroxidase Isoenzyme from Zinnia elegans by Gibberellic Acid

Hypocotyl formation during the epigeal germination of seedlings is under strict hormonal regulation. In a 3 d old Zinnia elegans seedling system, gibberellic acid (GA_3) exerts an opposite effect to that exerted by light on hypocotyl photomorphogenesis because GA_3 promotes an etiolated-like growth with an inhibition of radial (secondary) growth. For this reason, the effect of GA_3 on the basic peroxidase isoenzyme from Z. Elegans (ZePrx), an enzyme involved in hypocotyl lignin biosynthesis, was studied. The results showed that GA_3 reduces ZePrx activity, similarly to the way in which it reduces seedling secondary growth. This hormonal response is supported by the analysis of the ZePrx promoter, which contains four types of GA_3-responsive cis-elements: the W Box/O2S; the Pyr Box; the GARE; and the Amy Box. Taken together, these results suggest that ZePrx is directly regulated by GA_3, with this effect matching the inhibitory effect of GA on the hypocotyl secondary growth.