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Acta Alimentaria
Authors: E. Sedaghati, M. Nikkhah, R. Zare, K. Fotuhifar, S. Kocsubé, Cs. Vágvölgyi, and J. Varga

Ochratoxin A is a mycotoxin produced by Aspergillus and Penicillium species. This mycotoxin is a common contaminant of various foods including cereal products, spices, dried fruits, coffee, beer and wine. Besides species assigned to Aspergillus section Circumdati, black Aspergilli including A. niger, A. carbonarius and A. sclerotioniger are also able to produce this mycotoxin. Black Aspergilli have been found to be the predominant fungi contaminating pistachio nuts worldwide. We examined the species distribution of black Aspergilli on Iranian pistachio nuts. Sequence-based identifications have been carried out using partial calmodulin sequence data. Our data indicate that instead of the potential ochratoxin and fumonisin producing A. niger species, A. tubingensis dominates on Iranian pistachio nuts. This species is unable to produce either of these mycotoxins, consequently do not contribute to mycotoxin contamination of pistachio nuts in Iran. Further studies are in progress to clarify the role of other Aspergilli in ochratoxin contamination of pistachio in Iran.

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Acta Biologica Hungarica
Authors: Z. Farkas, J. Márki-Zay, Judit Kucsera, Cs. Vágvölgyi, W. Golubev, and Ilona Pfeiffer

Wickerhamomyces anomalus VKM Y-159 strain produces two types of toxin designated as WAKT a and WAKT b, encoded by chromosomal genes. The WAKT a toxin is heat-labile, pronase sensitive acting in pH range 3–4 affecting on several yeasts including pathogenic Candida species while the WAKT b toxin is protease- and thermo-resistant, acting in pH range 3–7 on two species, Candida alai and Candida norvegica. The rapid decrease of the number of viable cells after toxin treatment demonstrates that both toxins have cytocidic effect.

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Acta Biologica Hungarica
Authors: L. Kredics, Kata Terecskei, Zsuzsanna Antal, A. Szekeres, L. Hatvani, L. Manczinger, and Cs. Vágvölgyi

Eleven cold-tolerant Trichoderma isolates were screened for the production of proteolytic activities at 10 °C. Based on the activity profiles determined with paranitroanilide substrates at 5 °C, strain T221 identified as Trichoderma atroviride was selected for further investigations. The culture broth of the strain grown at 10 °C in casein-containing culture medium was concentrated by lyophilization and subjected to gel filtration, which was followed by chromatofocusing of the fraction showing the highest activity on N -benzoyl-Phe-Val-Arg-paranitroanilide. The purified enzyme had a molecular weight of 24 kDa, an isoelectric point of 7.3 and a pH optimum of 6.2. The temperature optimum of 25 °C and the low thermal stability suggested that it is a true cold-adapted enzyme. Substrate specificity data indicate that the enzyme is a proteinase with a preference for Arg or Lys at the P1 position. The effect of proteinase inhibitors suggests that the enzyme has a binding pocket similar to the one present in trypsin.

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Acta Biologica Hungarica
Authors: E. Horváth, G. Papp, Z. Gazdag, J. Belágyi, Á. Blaskó, J. Deli, Cs. Vágvölgyi, and M. Pesti

A carotenoid-less Phaffia rhodozyma mutant (MCP 325) exhibited significantly higher resistance to oxidative stressors such as menadione, H2O2 and K2Cr2O7 than its astaxanthin-producing parental strain (MCP 324). The absence of carotenoids in the mutant did not explain this phenomenon. The cause of the decreased superoxide, hydroxyl radical and glutathione contents, the increased peroxide concentration and the elevated specific activity of catalase under uninduced conditions may be a second mutation. Peroxide treatment induced specific catalase activity in the mutant but not in the parental strain. Regulation of these processes led to the result that, in spite of the mutations, the two strains exhibited the same multiplication rate and generation time.

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Acta Microbiologica et Immunologica Hungarica
Authors: L. Kredics, Zsuzsanna Antal, A. Szekeres, L. Hatvani, L. Manczinger, Cs. Vágvölgyi, and Erzsébet Nagy

Cellulolytic, xylanolytic, chitinolytic and b-1,3-glucanolytic enzyme systems of species belonging to the filamentous fungal genus Trichoderma have been investigated in details and are well characterised. The ability of Trichoderma strains to produce extracellular proteases has also been known for a long time, however, the proteolytic enzyme system is relatively unknown in this genus. Fortunately, in the recent years more and more attention is focused on the research in this field. The role of Trichoderma proteases in the biological control of plant pathogenic fungi and nematodes has been demonstrated, and it is also suspected that they may be important for the competitive saprophytic ability of green mould isolates and may represent potential virulence factors of Trichoderma strains as emerging fungal pathogens of clinical importance. The aim of this review is to summarize the information available about the extracellular proteases of Trichoderma. Numerous studies are available about the extracellular proteolytic enzyme profiles of Trichoderma strains and about the effect of abiotic environmental factors on protease activities. A number of protease enzymes have been purified to homogeneity and some protease encoding genes have been cloned and characterized. These results will be reviewed and the role of Trichoderma proteases in biological control as well as their advantages and disadvantages in biotechnology will be discussed.

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Extracellular β-glucosidase activity of 94 strains, representing 24 species of the genera Gilbertella, Mucor, Rhizomucor , and Rhizopus was evaluated in submerged culture and under solid state fermentation on wheat bran. Gilbertella persicaria G1 isolate showed the highest activity (70.9 U ml −1 ) followed by other Gilbertella (58.6–59.0 U ml −1 ) and Rhizomucor miehei isolates (29.2–42.0 U ml −1 ). Optimum temperature for enzyme production was 25 °C for Gilbertella and Mucor , and 30 °C for Rhizomucor and Rhizopus strains. Enzymes of R. miehei strains proved to be thermotolerant preserving up to 92.8% residual activity after heating to 75 °C in the presence of cellobiose substrate. Enzymes of Mucor racemosus f. chibinensis, R. miehei and Rhizopus microsporus var. oligosporus strains were activated at acidic condition (pH 4). Glucose was a strong inhibitor for each fungal β-glucosidase tested but some of them showed ethanol tolerance up to 20% (v/v). Ethanol also activated the enzyme in these strains suggesting glycosyl transferase activity.

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Acta Microbiologica et Immunologica Hungarica
Authors: A. Szekeres, B. Leitgeb, L. Kredics, Zsuzsanna Antal, L. Hatvani, L. Manczinger, and Cs. Vágvölgyi

Peptaibols and the related peptaibiotics are linear, amphipathic polypeptides. More than 300 of these secondary metabolites have been described to date. These compounds are composed of 5-20 amino acids and are generally produced in microheterogeneous mixtures. Peptaibols and peptaibiotics with unusual amino acid content are the result of non-ribosomal biosynthesis. Large multifunctional enzymes known as peptide synthetases assemble these molecules by the multiple carrier thiotemplate mechanism from a remarkable range of precursors, which can be N-methylated, acylated or reduced. Peptaibols and peptaibiotics show interesting physico-chemical and biological properties including the formation of pores in bilayer lipid membranes, as well as antibacterial, antifungal, occasionally antiviral activities, and may elicit plant resistance. The three-dimensional structure of peptaibols and pep­taibiotics is characterized predominantly by one type of the helical motifs a-helix, 310-helix and b-bend ribbon spiral. The aim of this review is to summarize the data available about the biosynthesis, biological activity and conformational properties of peptaibols and peptaibiotics described from Trichoderma species.

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Acta Biologica Hungarica
Authors: Á. Horváth, P. Sántha, V. Horváth, Nóra Török, I. Nagy, G. Jancsó, Cs. Vágvölgyi, and F. Somogyvári

A new, rapid method is described which permits the genotyping of genetically modified animals from a microlitre volume of whole blood samples via one step polymerase chain reaction amplification. The major advantage of the presented method is the exclusion of a DNA preparation step, which significantly reduces the time expenditure and work load of the genetic testing. Pilot studies indicate, that this method is efficient and applicable also on tissue biopsies and larger amount of blood providing a rapid and reliable new technique over conventional genotyping approaches.

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Acta Alimentaria
Authors: J. Varga, S. Kocsubé, Gy. Szigeti, V. Man, B. Tóth, Cs. Vágvölgyi, and T. Bartók

Black mould rot caused by black Aspergilli is an important post-harvest disease of onion worldwide. Usually Aspergillus niger is cited as the causative agent based on morphological criteria. In this study, the mycobiota and fumonisin contamination of mouldy onion bulbs purchased in Hungary were examined. All except one of the examined mouldy samples were found to be contaminated with black Aspergilli, which could be isolated both from the outer dry and the inner fleshy scales of onion bulbs. Species assignment of the isolates was carried out using sequence analysis of part of the calmodulin gene. Sequence data revealed that all 35 black Aspergilli isolated from onions belong to the Aspergillus awamori species. The range of fumonisin isomers present in the onion samples was also examined using reversed-phase high-performance liquid chromatography/electrospray ionization-ion trap mass spectrometry. Two of the examined onion samples were found to be contaminated with fumonisins at a rate of about 0.3 mg kg−1. This is the first report on fumonisin contamination of onion bulbs. The fumonisin isomers observed include fumonisins B2–4, 3-epi-FB4, iso-FB1 (FB6) and an iso-FB2,3 form. The range of fumonisin isomers detected in the onion bulbs indicates that probably A. awamori is responsible both for mould rot and fumonisin contamination of onions in Hungary.

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The paper contains an overview of the results of the studies made on the truffle Terfezia terfezioides, particularly the investigations related to the associations of this fungus with plants. Twelve plant species originated from a natural habitat of the fungus were supposed to be connected with T. terfezioides based on the anatomy of the endogenous fungal structures in their roots. Aseptic experiments were carried out on modified MMN substrates with different phosphate concentrations to study the interaction of T. terfezioides with Robinia pseudoacacia and Helianthemum ovatum. The colonization of the roots of black locust was always weaker than that of Helianthemum. The main characteristics were the intracellular coiled, branched, frequently septated hyphae in dead root cells. The intercellular hyphae formed Hartig-net with finger like structures only in Helianthemum. the interactions could not be considered unambiguously as mycorrhizae. There was no difference between the RFLP profiles of the nr DNA ITS of nineteen fruit bodies collected at the same time from the habitat and the ITS of three randomly chosen specimens were identical on sequence level, too. These invariability makes to design species specific PCR primers possible to check unambiguously the host plants.

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