Analysis of Pectic Enzyme Zymograms of Fusarium Species I. Fusarium lateritium and Related Species

Abstract Zymograms of the extracellular pectic enzymes (pectinesterase, polygalacturonase) of Fusarium lateritium and related species were prepared by electrophoresis from untreated culture filtrates. The zymogram technique gave evidence to conclude that F. lateritium, F. stilboides and F. xylarioides are distinct species. "pini" isolates are rather "subglutinans"-like fusaria than "lateritium"-like, while F. udum represents a genetical variant of F. oxysprorum .     

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.     

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.     

Distribution of Zearalenone-Producing Fusarium Species in Japan

One hundred sixty-six isolates of Fusarium spp. from domestic cereal grains, feed, and other sources were examined for their ability to produce zearalenone on autoclaved moist rice grains. They belonged to the following species (number of producers/number tested): F. roseum (9/28), F. roseum (Culmorum) (3/4), F. roseum (Gibbosum) (2/5), F. roseum (Avenaceum) (1/2), F. roseum (Scirpi) (0/1), F. tricinctum (1/4), F. tricinctum (Sporotrichiella) (0/7), F. lateritium (1/1), F. episphaeria (0/2), F. moniliforme (0/3), F. oxysporum (0/12), F. rigidiusculum (0/4), F. solani (0/4), F. splendens (0/1), F. nivale (0/2), and Fusarium spp. (15/86). Zearalenone was isolated from molded rice by ethanol extraction and purified by column chromatography. Selected isolates of F. roseum M-3-2 and F. roseum (Gibbosum) A-O-2 produced 50 to 100 mg of zearalenone per kg of rice. Increased yields (250 to 407 mg/kg of rice) were obtained by F. roseum M-3-2 when the substrate was supplemented with 1% peptone.     

Fusarium Species from the Fusarium fujikuroi Species Complex Involved in Mixed Infections of Maize in Northern Sinaloa,Mexico

Fusarium species belonging to the Fusarium fujikuroi species complex (FFSC) are associated with maize in northern Mexico and cause Fusarium ear and root rot. In order to assess the diversity of FFSC fungal species involved in this destructive disease in Sinaloa, Mexico, a collection of 108 fungal isolates was obtained from maize plants in 2007–2011. DNA sequence analysis of the calmodulin and elongation factor 1α genes identified four species: Fusarium verticillioides, F. nygamai, F. andiyazi and F. thapsinum (comprising 79, 23, 4 and 2 isolates, respectively). Differential distribution of Fusarium species in maize organs was observed, that is F. verticillioides was the most frequently isolated species from maize seeds, while F. nygamai predominated on maize roots. Mixed infections with F. verticillioides/F. thapsinum and F. verticillioides/F. nygamai were detected in maize seeds and roots, respectively. Pathogenicity assay demonstrated the ability of the four species to infect maize seedlings and induce different levels of disease severity, reflecting variation in aggressiveness, plant height and root biomass. Isolates of F. verticillioides and F. nygamai were the most aggressive. These species were able to colonize all root tissues, from the epidermis to the vascular vessels, while infection by F. andiyazi and F. thapsinum was restricted to the epidermis and adjacent cortical cells. This is the first report of F. nygamai, F. andiyazi and F. thapsinum infecting maize in Mexico and co‐infecting with F. verticillioides. Mixed infections should be taken into consideration due to the production and/or accumulation of diverse mycotoxins in maize grain.     

中国的膨孢组镰孢菌(英文)

膨孢组镰孢菌Fusarium在自然界中广泛分布。该组包括4个种:木贼镰孢菌F.equiseti,藨草镰孢菌F.scirpi,长脚镰孢菌F.longipes和紧致镰孢菌F.compactum。这4个种均产生腹背不平行弯曲的大孢子。分离获得并描述了其中的3个种。木贼镰孢菌F.equiseti是镰孢菌中最常见的种之一,它产生典型的腹背不平行弯曲的大孢子,大孢子的顶细胞和足跟状基细胞明显伸长,菌落因缺乏红色素而呈黄褐色。藨草镰孢菌F.scirpi是比较少见的种类,它的典型特征是在典型的十字形产孢细胞上产生大量的小型分生孢子。由于它在PDA培养基上容易发生小孢子缺乏型的变异,因此,常常被错误地鉴定为木贼镰孢菌F.equiseti。长脚镰孢菌F.longipes的大孢子最容易与其他种的大孢子区分,它的顶细胞和基细胞均极度延长。当长脚镰孢菌F.longipes菌落因为变异而失去产生红色素的能力时,也容易与木贼镰孢菌F.equiseti混淆。基于该组大孢子的典型特征,作者将锐顶镰孢菌F.acuminatum排除在该组之外。     

Identification of Toxigenic Fusarium Species using PCR Assays

Isolates of the toxigenic cereal pathogens Fusarium culmorum, Fusarium graminearum, Fusarium crookwellense and Fusarium avenaceum, from Poland (48 isolates) and 12 from England, New Zealand, Italy and Canada, were examined using random amplified polymorphic DNA (RAPD)-polymerase chain reaction (PCR), sequence-characterized amplified regions (SCARs), morphology and mycotoxin production under laboratory conditions. Their DNA products were compared by RAPD-PCR, which showed species-specific bands and the greatest diversity among isolates of F. avenaceum. PCR using three 20-mer-primer-pairs that are reported to be useful for identification of F. culmorum and F. graminearum group 2 confirmed their species-specificity. The same species-specific PCR product was observed in isolates of both nivalenol and deoxynivalenol chemotypes of F. culmorum or F. graminearum. A clear relationship was found between morphological and species-specific PCR identification of F. culmorum and F. graminearum isolates. However, F. avenaceum can be confused when using primers FA-ITS F/R (SCAR 2-14) with Fusarium tricinctum because the same band 272 bp appears in the gel, in both species probes.     

镰刀菌属一个中国新记录种

从山西省13个主要土壤类型的1 012个土壤样品中分离出镰刀菌333株,依据布斯的镰刀菌属分类系统鉴定其中1株为中国新记录种——柔毛镰刀菌(Fusarium flocciferum Corda),并对其进行描述和讨论。菌种保存于山西农业大学农学院植物病理实验室。     

小麦赤霉病镰孢菌毒素的生物合成及其分子调控

禾谷镰刀菌是小麦赤霉病的主要致病菌,其真菌次生代谢产生的单端孢霉烯类B型毒素,如雪腐镰刀菌烯醇(nivalenol,NIV)、脱氧雪腐镰刀菌烯醇(deoxynivalenol,DON)和其它乙酰化衍生物等污染小麦籽粒后对人畜健康构成威胁。综述了近年来国内外对小麦赤霉病镰孢菌单端孢霉烯类B型毒素生物合成的主要途径及分子调控研究进展,对毒素合成过程中的重要调控基因如TRI5、TRI7和TRI13在农业中的应用进行了阐述。     

Analysis of Pectic Enzyme Zymograms of Fusarium Species.

Zymograms of the extracellular polygalacturonase (PG), produced by isolates of F. culmorum and F. graminearum originating from different geographic locations and different sources, were compared. PG patterns were prepared by polyacrylamide gel electrophoresis (PAGE) and isoelectric focusing (IEF) of untreated fluid from liquid pectin salts cultures. There was no intraspecific variability between isolates of both Fusarium species. Electrophoretic and isofocusing PG patterns were species specific. On the basis of IEF patterns, F. culmorum (4 isozymes, estimated pI's 6.4, 6.6, 6.9, 7.1) and F. graminearum (5 isozymes, estimated pI's 6.4, 6.6, 6.9, 7.1, 7.5) could be separated from one another by the pH 7.5 PG isozyme.     

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.     

Occurrence and Pathogenicity of Fusarium Species in Banana Fruits

Banana fruits were studied over a six-month period in order to determine the incidence of species of the Fusarium genus and assess their potential pathogenicity. The 72 samples studied were commercially available in Italy and Spain, where they were brought from Panama, Ecuador and Canary Islands. Among the species detected in the fruits, Fusarium semitectum var. majus Wollenw. was predominant, followed by F. moniliforme Sheld., F. solani (Mart.) Appel & WoUenw., F. oxysporum Schlecht., F. proliferatum (Matsushima) Nirenberg, F. graminearum Schw., F. camptoceras WoUenw. &C Reinking, F. subglutinans (WoUenw. & Reinking) Nelson et al., F. dimerum Penzig in Sacc, F. acuminatum EU. & Ev., and F. equiseti (Corda) Sacc. Fusarium proliferatum had never been reported to occur as contaminating fungi in banana fruits to date. Fusarium subglutinans, F. acuminatum and F. graminearum were found to be the most markedly pathogenic of all. The lack of noticeable differences in relation to the incidence of the different species isolated from the samples indicates that the mycoflora found is typical of this fruit and does not depend on its origin.     

Effects of Temperature on Viability of Pathogenic Fusarium Species

The viability of three species of Fusarium pathogenic to winter cereals was differently affected by temperature. F. culmorum survived in vitro in soil for 4 months at 8, 20 and 30 °C, the number of colony forming units (CFU) recorded at these temperatures after incubation being 125, 185 and 624 % respectively when compared with the number present at the beginning of the experiment. F. avenaceum and F. heterosporum barely survived at 8 °C after incubation for 4 months but at 20 °C the numbers of CFU were 314 and 380 % respectively, while at 30 °C the corresponding number tor each of these species was 200 %.
At natural soil temperatures in winter, the number of CFU of F. culmorum after 4.5 months decreased to 60, 70 and 87 % of the number present at the beginning of this experiment when infested soil was buried at depths of 7–10, 15—20 and 30 cm respectively. In the upper soil layer where winter temperatures were lowest, survival of F. avenaceum and F. heterosporum did not occur. Survival rates for these two species at a depth of 15—20 cm were 20 and 5 % respectively. In the deepest layer of soil (30 cm) survival of these two species was 30 % for F. avenaceum and 10 % for F. heterosporum of the number present at the beginning of this experiment.
These results demonstrate that during the early spring stages of growth of winter wheat, inoculum of F. culmorum , unlike that of F. avenaceum and F. heterosporum , is also a source of infection in the top soil layer in our climatic conditions.     

Mating Type Sequences in Asexually Reproducing Fusarium Species

To assess the potential for mating in several Fusarium species with no known sexual stage, we developed degenerate and semidegenerate oligonucleotide primers to identify conserved mating type (MAT) sequences in these fungi. The putative α and high-mobility-group (HMG) box sequences from Fusarium avenaceum, F. culmorum, F. poae, and F. semitectum were compared to similar sequences that were described previously for other members of the genus. The DNA sequences of the regions flanking the amplified MAT regions were obtained by inverse PCR. These data were used to develop diagnostic primers suitable for the clear amplification of conserved mating type sequences from any member of the genus Fusarium. By using these diagnostic primers, we identified mating types of 122 strains belonging to 22 species of Fusarium. The α box and the HMG box from the mating type genes are transcribed in F. avenaceum, F. culmorum, F. poae, and F. semitectum. The novelty of the PCR-based mating type identification system that we developed is that this method can be used on a wide range of Fusarium species, which have proven or expected teleomorphs in different ascomycetous genera, including Calonectria, Gibberella, and Nectria.     

Phylogenetic Analysis of Clinically Relevant Fusarium Species in Iran

Mycopathologia - Fungi of the genus Fusarium are well known as major plant pathogens but also cause a broad spectrum of human infections. Sixty-three clinical isolates, collected during...     

Fusarium eumartii Growth in Resistant and Susceptible Oak Species

Fusarium eumartii is a fungus associated with declining Quercus robur , in which it is found in the vessels. The response of oak species to infection is known to vary: Q. robur is susceptible , but Quercus cerris and Quercus pubescens are resistant. An experiment was carried out in 1996 and repeated in 1997, to examine how F. eumartii colonization differed in oak species that were susceptible or resistant to the fungus by counting the number of vessels with mycelium at various distances from the inoculation site in infected seedlings and by determining the amount of viable fungus in infected tissue. Infected vessels with mycelium were counted on sections (10  μ m thick) cut at 0, 2, 4, 6, 8 and 10 cm from the inoculation site on 1-year-old inoculated seedlings as well as on sections cut every 2 cm to the seedling tip. The amount of viable fungus was determined by counting the colony forming units (CFUs) in stem segments from the same seedlings. Quercus robur seedlings had the greatest number of infected vessels and the greatest number of CFUs. Forty days after inoculation, the extent of vertical fungal spread was 28.12 cm in Q. robur , 3.15 cm in Q. cerris and 3.00 cm in Q. pubescens . The greatest number of CFUs was found in Q. robur at day 5 after inoculation. Analysis of variance confirmed the results.