Novel Ergot Alkaloids Production from Penicillium citrinum Employing Response Surface Methodology Technique.

Ergot alkaloids are novel pharmaceutical and therapeutic agents synthesized in this study using fungal species Penicillium citrinum. To get the maximum yield of ergot alkaloids a statistical process of response surface methodology was employed using surface culture fermentation technique. Initially, the strain of Penicillium was improved using physical (ultraviolet (UV) and chemical (ethyl methane sulfonate (EMS) treatments to get the maximum yield of ergot alkaloids through surface culture fermentation technique. After improving the strain, survival rate of colonies of Penicillium citrinum treated with UV and EMS was observed. Only 2.04% living colonies were observed after 150 min of exposure of Penicillium citrinum in UV light and 3.2% living colonies were observed after 20 min of the exposure in EMS. The mutated strains of Penicillium citrinum were screened for their production of ergot alkaloids and after fermentation experiments, maximum yield was obtained from PCUV-4 and PCEMS-1 strains. After strain improvement, Plackett-Burman design (PBD) and Box-Behnken design (BBD) of RSM were employed and 10-fold yield enhancement (35.60 mg/100 mL) of ergot alkaloids was achieved. This enhancement in yield of ergot alkaloids proved the positive impacts of RSM and UV on the yield of ergot alkaloids. The study provides a cost effective, economical and sustainable process to produce medically important ergot alkaloids which can be used in various pharmaceutical formulations to treat human diseases.

Effects of UV Irradiation on Penicillium Strains Isolated from a Bread Plant and the Application to Bakery Products.

We tested treatement with UV irradiation for controlling the growth of bread mold. First, we analyzed the sterilizing effect of a dose of approximately 25 mJ/cm2 radiation on nine Penicillium and two Talaromyces strains that were isolated from a bread-manufacturing plant. The P. chermesinum and P. paneum strains were sterilized completely at that dose, while it was only partially effective against P. corylophilum. P. chrysogenum and P. decumbens were sterilized at a dose of approximately 120 mJ/cm2, while T. amestolkiae was sterilized at approximately 150 mJ/cm2. Sterilization of T. cecidicola and P. hispanicum required more than 200 mJ/cm2 of radiation. These results suggest that UV resistance varies depending on the species and the strains. We also carried out UV irradiation of bread at 70 mJ/cm2: a dose at which the taste of bread is not affected; we observed that mold growth was delayed visibly compared to the non-irradiated bread. These results suggest that UV irradiation at 70 mJ/cm2 is effective at delaying mold growth, though it does not cause complete sterilization. This method should prove useful for extending the shelf-life of bread.

Metagenomics-Guided Survey, Isolation, and Characterization of Uranium Resistant Microbiota from the Savannah River Site, USA.

Despite the recent advancements in culturomics, isolation of the majority of environmental microbiota performing critical ecosystem services, such as bioremediation of contaminants, remains elusive. Towards this end, we conducted a metagenomics-guided comparative assessment of soil microbial diversity and functions present in uraniferous soils relative to those that grew in diffusion chambers (DC) or microbial traps (MT), followed by isolation of uranium (U) resistant microbiota. Shotgun metagenomic analysis performed on the soils used to establish the DC/MT chambers revealed Proteobacterial phyla and Burkholderia genus to be the most abundant among bacteria. The chamber-associated growth conditions further increased their abundances relative to the soils. Ascomycota was the most abundant fungal phylum in the chambers relative to the soils, with Penicillium as the most dominant genus. Metagenomics-based taxonomic findings completely mirrored the taxonomic composition of the retrieved isolates such that the U-resistant bacteria and fungi mainly belonged to Burkholderia and Penicillium species, thus confirming that the chambers facilitated proliferation and subsequent isolation of specific microbiota with environmentally relevant functions. Furthermore, shotgun metagenomic analysis also revealed that the gene classes for carbohydrate metabolism, virulence, and respiration predominated with functions related to stress response, membrane transport, and metabolism of aromatic compounds were also identified, albeit at lower levels. Of major note was the successful isolation of a potentially novel Penicillium species using the MT approach, as evidenced by whole genome sequence analysis and comparative genomic analysis, thus enhancing our overall understanding on the uranium cycling microbiota within the tested uraniferous soils.

Inactivation of conidia from three Penicillium spp. isolated from fruit juices by conventional and alternative mild preservation technologies and disinfection treatments.

Fungi are able to grow on diverse food products and contribute to food spoilage worldwide causing food loss. Consumers prefer freshly squeezed fruit juices, however, the shelf life of these juices is limited due to outgrowth of yeast and fungi. The shelf life of pulsed electric field (PEF) treated juice can be extended from 8 days up to a few weeks before spoilage by moulds becomes apparent. Conidia produced by three Penicillium ssp. (Penicillium expansum, Penicillium buchwaldii and Penicillium bialowiezense), previously isolated from spoiled PEF treated fruit juice and smoothie, were characterized for resistance towards selected mild physical processing techniques in orange juice and toward sanitizers on surfaces. The results show that Penicillium spp. conidia are susceptible to mild heat, high pressure pasteurization (HPP), PEF, cold atmospheric plasma (CAP), UV, and chemical sanitizers chlorine dioxide and hypochlorite albeit with different susceptibility. Treatment with mild heat, HPP, PEF, or chlorine dioxide reduced conidia by more than 5 log. For hypochlorite, UV, and CAP the reduction was between 1 and 3 log. Together, this study provides data for the development of intervention strategies to eliminate spoilage mould conidia in fruit juices.

Assessment of lead tolerance in gamma exposed Aspergillus niger van Tieghem & Penicillium cyclopium Westling.

Purpose: Present study deals with the role of gamma irradiation in modulating lead (Pb) tolerance of Aspergillus niger van Tieghem. and Penicillium cyclopium Westling. Materials and methods: After being exposed to gamma absorbed doses those fungal strains were subjected to heavy metal uptake efficacies and anti-oxidative study. Fourier Transform Infrared (FTIR) spectra and Scanning Electron Microscopic (SEM) studies were also evaluated. Result: Gamma exposed A. niger & P. cyclopium showed enhanced growth in terms of colony forming unit (CFU) and more Pb uptake efficacies compared to their un-irradiated counterparts. FTIR spectra illustrated the involvement of functional groups in Pb biosorption. SEM photographs revealed the structural deformities in both the fungal strains after being exposed to Pb and gamma. Upregulated anti-oxidative defense system (super oxide dismutase, catalase, total glutathione) in gamma exposed fungal groups are accountable for enhanced Pb tolerance and removal than that of their un-irradiated counterparts. Conclusion: The outcomes of this study exhibit a light towards a new step of heavy metal bioremediation.

Fungicidal Effects of Ultraviolet Light (254 nm) Irradiation on Contaminated Museum Packing and Storing Materials.

In storage of modern museums, collections are packed and stored with acid-free paper-based materials for keeping safe and stable conditions. Direct contact of fungal contaminated packing and storing materials with the collections is concerned about expanding of infection in storage facilities. In this study, fungicidal effects of UV light irradiation on the materials such as archival board and Japanese tissue paper contaminated with Penicilliun commune and Chaetomium globosum were tested. The analyzed materials were divided into two groups; Group 1 was examined with 20 µl of spore suspensions of fungi (106 cfu/ml) ; and Group 2 was tested on Czapek- Dox agar medium modified without sugar and inoculated with 100 µl of the spore suspensions of fungi (106 cfu/ml) . Six doses of UV irradiation were examined on Group 1 and five doses on Group 2 in addition to control. The assessment was done by using 1) adenosine triphosphate (ATP) bioluminescence assay and double staining to determine the cell viability; 2) observation under light microscope to evaluate morphophysiological change of tested fungi (spores and hyphae) . Because of the thinness and high transparency of tissue paper, UV irradiations were highly efficient to fungicide its fungal contamination compared with archival board. In spite of the high resistance of C. globosum spores, the rate of growth was slow, and with a little amount of perithecia or fruiting bodies and a high amount of ycelium (which damaged rapidly through UV irradiation) . This may be due to a low relative humidity of the incubation environment. Minimum dosage of UV irradiation with fungicidal effectiveness against all fungal contamination was estimated as 118 J/cm2.

Feasibility of 3D UV-C treatment to reduce fungal growth and mycotoxin loads on maize and wheat kernels.

Fungal disease of grain crops is a concern for the agricultural industry, resulting in economic losses. Aside from severe yield losses, mycotoxigenic fungi such as Penicillium and Fusarium can produce harmful mycotoxins, including deoxynivalenol (DON), zearalenone (ZEN), and ochratoxin A (OTA). This proof-of-concept study explored the feasibility and effects of ultraviolet (UV) C light at 253.7 nm to reduce fungal and mycotoxin loads on model surfaces as well as on maize and wheat kernels using benchtop 2D and 3D illumination strategies. Reduction of Penicillium verrucosum (98.6%) and Fusarium graminearum (88.8%) on agar was achieved using a UV-C dose of 100 mJ cm-2. Naturally occurring fungal growth resembling P. verrucosum on maize was reduced by 79% after exposure to 5000 mJ cm-2. Similarly, fungal growth resembling F. graminearum on maize was reduced by 60% with 1000 mJ cm-2. On wheat, significant reduction of fungal growth was not observed. Maximal reduction of DON (97.3%), ZEN (75.4%), and OTA (91.2%) on filter paper was obtained using 15,000 mJ cm-2. The overall reduction of DON (30%; 14%), ZEN (52%; 42%), and OTA (17%; 6%) on maize and wheat, respectively, was lower than on filter paper. Moisture and crude protein content as well as percent germination of maize kernels were not affected by UV-C treatment up to 5000 mJ cm-2. This study has shown that 3D UV-C treatment is a feasible option for reducing Fusarium and Penicillium growth on maize kernels and, at higher doses, decreasing ZEN by ~ 50%.

Light-emitting Diode Blue Light Alters the Ability of Penicillium digitatum to Infect Citrus Fruits.

Penicillium digitatum (Pers.:Fr.) Sacc. is the main fungus causing postharvest losses in citrus fruits. Previous work showed the potential of LED blue light (LBL) in controlling P. digitatum growth. Here, we have investigated whether LBL alters the ability of this fungus to infect citrus fruits. Before fruit infection, Petri plates inoculated with the same conidia concentration were held under darkness (control) or LBL (100 µmol m-2 s-1 ) for 8 d (continuous light), or were treated with the same LBL for 3 d and then shifted to darkness for 5 d (non-continuous light). Spores from cultures exposed to continuous light showed very low capacity to germinate (1.8% respect to control) but a high viability and a similar morphology and ability to infect the fruits than spores from control cultures. The number of spores produced in plates exposed to non-continuous light was slightly lower than in control plates, but they showed much lower viability and lower capacity to infect the fruits. This effect was more likely related to aberrant morphology of spores, which formed aggregates, than to its metabolic activity or its ability to produce ethylene that might contribute to destroy natural defense barriers from the fruit.

Inhibitory effect of gamma irradiation and its application for control of postharvest green mold decay of Satsuma mandarins.

Gamma irradiation has been shown to be effective for the control of postharvest fungi in vitro, but little is known regarding antifungal action, responses to gamma irradiation, and its application to fresh produce. Gamma irradiation was evaluated for its in vitro and in vivo antifungal activity against Penicillium digitatum on Satsuma mandarin fruits. Green mold was inhibited in a dose-dependent manner. Gamma irradiation showed a complete inhibition of spore germination, germ tube elongation, and mycelial growth of P. digitatum, particularly at 1.0kGy. To further investigate the mechanisms by which gamma irradiation inhibits fungal growth, the membrane integrity and cellular leakage of conidia were tested, indicating that gamma irradiation results in the loss of plasma membrane integrity, causing the release of intracellular contents such as soluble proteins. In vivo assays demonstrated that established doses can completely inhibit the growth of fungal pathogens, but such high doses cause severe fruit damage. Thus, to eliminate the negative impact on fruit quality, gamma irradiation at lower doses was evaluated for inhibition of P. digitatum, in combination with a chlorine donor, sodium dichloro-s-triazinetrione (NaDCC). Interestingly, only a combined treatment with 0.4kGy of gamma irradiation and 10ppm of NaDCC exhibited significant synergistic antifungal activity against green mold decay. The mechanisms by which the combined treatment decreased the green mold decay of mandarin fruits can be directly associated with the disruption of cell membrane of the fungal pathogen, which resulted in a loss of cytoplasmic material from the hyphae. These findings suggest that a synergistic effect of combining treatment with gamma irradiation with NaDCC has potential as an antifungal approach to reduce the severity of green mold in mandarin fruits.

Comparison of Free and Immobilized L-asparaginase Synthesized by Gamma-Irradiated Penicillium cyclopium.

Gamma irradiation is used on Penicillium cyclopium in order to obtain mutant cells of high L-asparaginase productivity. Using gamma irradiation dose of 4 KGy, P. cyclopium cells yielded L-asparaginase with extracellular enzyme activity of 210.8 ± 3 U/ml, and specific activity of 752.5 ± 1.5 U/mg protein, which are 1.75 and 1.53 times, respectively, the activity of the wild strain. The enzyme was partially purified by 40-60% acetone precipitation. L-asparaginase was immobilized onto Amberlite IR-120 by ionic binding. Both free and immobilized enzymes exhibited maximum activity at pH 8 and 40 degrees C. The immobilization process improved the enzyme thermal stability significantly. The immobilized enzyme remained 100% active at temperatures up to 60 degrees C, while the free asparaginase was less tolerant to high temperatures. The immobilized enzyme was more stable at pH 9.0 for 50 min, retaining 70% of its relative activity. The maximum reaction rate (V(max)) and Michaelis-Menten constant (K(m)) of the free form were significantly changed after immobilization. The K(m) value for immobilized L-asparaginase was about 1.3 times higher than that of free enzyme. The ions K+, Ba2+ and Na+ showed stimulatory effect on enzyme activity with percentages of 110%, 109% and 106% respectively.

Short-term UV-B exposure induces metabolic and anatomical changes in peel of harvested lemons contributing in fruit protection against green mold.

UV-B radiation (UVBR) is a small fraction of the solar spectrum from 280 to 315nm. UVBR produces photomorphogenic acclimation responses in plants, modulating their cellular structure and physiology. Here, changes in the peel of harvested lemons after short time exposure to UVBR were analyzed and its potential effects against fungal infection were studied. In the flavedo, UVBR treatment induced variations in the respiratory profiles and increased the phenolic compound contents. Final products of the flavonoid pathway (flavones, flavonols and anthocyanins) increased more markedly than their precursors (flavanones and dihydroflavonols). The increased accumulation of soluble phenolics in the flavedo of treated lemons is associated with the high antioxidant activity found in the flavedo of these samples. Supporting the biochemical determinations, anatomical observations showed abundant intravacuolar deposits of phenolic compounds and an increase in the cell wall thickness in UVBR-treated samples. Metabolic and anatomical modifications associated to UVBR improved natural defenses against Penicillium digitatum, the causal agent of green mold disease. Our results suggest that mature postharvest lemons exposed to the artificial radiation showed phenotypic plasticity, allowing an acclimation response to UVBR which confers fruit resistance to pathogens. Thus, combination of UVBR with other treatments could represent an important improvement to control postharvest diseases on citrus.

Disruption of ku70 involved in non-homologous end-joining facilitates homologous recombination but increases temperature sensitivity in the phytopathogenic fungus Penicillium digitatum.

The dominant mechanism to repair double-stranded DNA breaks in filamentous fungi is the non-homologous end joining (NHEJ) pathway, and not the homologous recombination (HR) pathway that operates in the mutation of genes by replacement of target DNA for selection cassettes. The key to improve HR frequency is the inactivation of the NHEJ pathway by eliminating components of its Ku70/80 heterodimeric complex. We have obtained ku70 mutants of Penicillium digitatum, the main citrus postharvest pathogen. The increased efficiency of HR in Δku70 strains was demonstrated by the generation of mutants in two different chitin synthase genes (PdchsII and PdchsV). P. digitatum Δku70 strains showed no differences from the parental strain in vegetative growth, asexual development or virulence to citrus fruit, when experiments were conducted at the optimal temperature of 24°C. However, growth of Δku70 strains at temperatures higher than 24°C demonstrated a detrimental effect in axenic growth and conidia production. These observations are in agreement with previous studies describing differences between ku70 mutants and their parental strains in some fungal species, and must be taken into account for future applications of the Δku approach to increase HR efficiency in fungi.

UV Tolerance of Spoilage Microorganisms and Acid-Shocked and Acid-Adapted Escherichia coli in Apple Juice Treated with a Commercial UV Juice-Processing Unit.

The enhanced thermal tolerance and survival responses of Escherichia coli O157:H7 in acid and acidified foods is a major safety concern for the production of low-pH products, including beverages. Little is known about this phenomenon when using UV light treatments. This study was conducted to evaluate the effects of strain (E. coli O157:H7 strains C7927, ATCC 35150, ATCC 43895, and ATCC 43889 and E. coli ATCC 25922) and physiological state (control-unadapted, acid adapted, and acid shocked) on the UV tolerance of E. coli in apple juice treated under conditions stipulated in current U.S. Food and Drug Administration regulations. A greater than 5-log reduction of E. coli was obtained under all tested conditions. A significant effect of strain (P < 0.05) was observed, but the physiological state did not influence pathogen inactivation (P ≥ 0.05). The UV sensitivity of three spoilage microorganisms (Aspergillus niger, Penicillium commune, and Alicyclobacillus acidoterrestris) was also determined at UV doses of 0 to 98 mJ/cm(2). Alicyclobacillus was the most UV sensitive, followed by Penicillium and Aspergillus. Because of the nonsignificant differences in UV sensitivity of E. coli in different physiological states, the use of an unadapted inoculum would be adequate to conduct challenge studies with the commercial UV unit used in this study at a UV dose of 14 mJ/cm(2). The high UV tolerance of spoilage microorganisms supports the need to use a hurdle approach (e.g., coupling of refrigeration, preservatives, and/or other technologies) to extend the shelf life of UV-treated beverages.

Effect of LED Blue Light on Penicillium digitatum and Penicillium italicum Strains.

Studies on the antimicrobial properties of light have considerably increased due in part to the development of resistance to actual control methods. This study investigates the potential of light-emitting diodes (LED) blue light for controlling Penicillium digitatum and Penicillium italicum. These fungi are the most devastating postharvest pathogens of citrus fruit and cause important losses due to contaminations and the development of resistant strains against fungicides. The effect of different periods and quantum fluxes, delaying light application on the growth and morphology of P. digitatum strains resistant and sensitive to fungicides, and P. italicum cultured at 20°C was examined. Results showed that blue light controls the growth of all strains and that its efficacy increases with the quantum flux. Spore germination was always avoided by exposing the cultures to high quantum flux (700 µmol m(-2) s(-1) ) for 18 h. Continuous light had an important impact on the fungus morphology and a fungicidal effect when applied at a lower quantum flux (120 µmol m(-2) s(-1) ) to a growing fungus. Sensitivity to light increased with mycelium age. Results show that blue light may be a tool for P. digitatum and P. italicum infection prevention during handling of citrus fruits.

Antifungal effect of gamma irradiation and sodium dichloroisocyanurate against Penicillium expansum on pears.

UNLABELLED: Gamma irradiation (GI) was evaluated for its in vitro and in vivo antifungal activity against Penicillium expansum on pear fruits. GI showed a complete inhibition of spore germination, germ tube elongation and mycelial of P. expansum, especially 1·8 kGy. GI affected the membrane integrity and cellular leakage of conidia in a dose-dependent manner. Furthermore, the leakage of protein and sugar from mycelia increased along with the dose. GI was evaluated at lower doses in combination with a chlorine donor, sodium dichloro-s-triazinetrione (NaDCC), to examine the inhibition of P. expansum. Interestingly, only a combined treatment with 0·2 kGy of GI and 70 ppm of NaDCC exhibited significant synergistic antifungal activity. The mechanisms by which the combined treatment decreased the blue mould decay of pear fruits could directly associated with the disruption of the cell membrane of the fungal pathogen, resulting in a loss of cytoplasmic materials from the hyphae. SIGNIFICANCE AND IMPACT OF THE STUDY: Gamma irradiation (GI) is used as an effective nonchemical approach to inactive pathogens. This study investigated the antifungal effect of gamma irradiation and its combined treatment with a chlorine donor on this fungal pathogen, both in vitro and in vivo. This study emphasized that the integration of low-dose GI and a chlorine donor, NaDCC, exhibited a significant antifungal effect, and that its mechanisms are directly associated with membrane integrity of fungal spores, promising that GI has the potential to be an antifungal approach.

Immobilization technique for enhanced production of the immunosuppressant mycophenolic acid by ultraviolet and gamma-irradiated Penicillium roqueforti.

AIMS: Different entrapment matrices were screened to immobilize two strains of Penicillium roqueforti (AG101 and LG109) for more effective production of mycophenolic acid (MPA). Further improvement in the MPA productivity from immobilization of spores and mycelia was adopted by UV and gamma irradiation. METHODS AND RESULTS: Penicillium roqueforti strains were immobilized in different entrapping carriers and used for MPA production in shake flask cultures. Maximum MPA production was achieved on using an alginate concentration of 3·0% (w/v) and a mycelial fresh weight of 10% (w/v). MPA produced by alginate-immobilized spores and mycelia was almost double in comparison to the free system. The MPA-producing ability of immobilized AG101 and LG109 strain was significantly enhanced by mutagenesis through irradiation by UV (254 nm) for 120 and 90 min, respectively and gamma rays at 0·75 KGy. The feasibility of MPA production in a semi-continuous form by immobilized cells as affected by irradiation was adopted. CONCLUSIONS: MPA production by immobilized spores and mycelia was more intensified by UV and gamma irradiation. Moreover, the immobilized cell culture was superior to free-cell culture. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings indicate the future possibility to reduce the cost of producing fermentation-based drugs.

Characterization of sakA gene from pathogenic dimorphic fungus Penicillium marneffei.

Eukaryotes utilize stress activated protein kinase (SAPK) pathways to adapt to environmental stress, including heat, osmotic, oxidative or nutrient stresses. Penicillium marneffei (Talaromyces marneffei), the dimorphic pathogenic fungus that can cause disseminated mycosis in HIV-infected patients, has to encounter various types of stresses both outside and inside host cells. However, the strategies used by this fungus in response to these stresses are still unclear. In this report, the stress-activated kinase (sakA) gene of P. marneffei was characterized and the roles of this gene on various stress conditions were studied. The sakA gene deletion mutant was constructed using the split marker method. The phenotypes and sensitivities to varieties of stresses, including osmotic, oxidative, heat and cell wall stresses of the deletion mutant were compared with the wild type and the sakA complemented strains. Results demonstrated that the P. marneffei sakA gene encoded a putative protein containing TXY phosphorylation lip found in the stress high osmolarity glycerol 1 (Hog1)/Spc1/p38 MAPK family, and that this gene was involved not only in tolerance against oxidative and heat stresses, but also played a role in asexual development, chitin deposition, yeast cell generation in vitro and survival inside mouse and human macrophages.

Optimization of submerged fermentation conditions for immunosuppressant mycophenolic acid production by Penicillium roqueforti isolated from blue-molded cheeses: enhanced production by ultraviolet and gamma irradiation.

Mycophenolic acid (MPA) is a promising drug owing to its immunosuppressive and biological activities. In this study, two strains of Penicillium roqueforti designated as AG101 and LG109 were selected among several strains isolated from Roquefort cheese samples on the basis of their activity for MPA-producing ability. The appropriate fermentation conditions necessary for MPA biosynthesis by the two respective fungal strains were investigated. These conditions included selection of the cultivation medium, agitation rate, incubation temperature, fermentation time, pH value, inoculum size, and fermentation medium volume. Maximum MPA productivities were maintained when the fermentation process was carried out using a medium composed of (g l(-1)): Sucrose, 30; peptone, 5.0; KH2PO4, 1.0; MgSO4·7H2O, 0.5 and KCl, 0.5; pH 6.0, inoculated with an inoculum size of 6.0 % (v/v), and incubated at 25 °C for 10 days at 120 rpm. The potentiality of both P. roqueforti strains for further improvement of MPA production was applied by mutagenesis through exposure to irradiation by ultraviolet rays (UV, 254 nm) for different periods of time and gamma rays at various doses (KGy). The dry cell weight of both irradiated fungal strains showed a greater reduction when irradiated either with UV or gamma rays. However, the MPA yield of both strains was increased by 1.27-1.39 fold when irradiated with UV rays and by 2.11-2.33 fold when irradiated with gamma rays, as compared with the respective controls (non-irradiated cultures). These findings indicate the future possibility to reduce the cost of producing fermentation-based drugs.

Inactivation effect of electron beam irradiation on fungal load of naturally contaminated maize seeds.

BACKGROUND: This work focuses on the effect of accelerated electrons (0.1-6.2 kGy) on naturally attached fungi on maize seeds. The fungal viability and corresponding inactivation kinetics were determined. The inactivation and radiosensitivity of the most abundant species in the contaminant fungi detected on maize seeds (Aspergillus spp., Penicillium spp. and Fusarium spp.) are discussed. RESULTS: Fungal contamination of maize seeds decreased significantly with increasing irradiation dose. The survival curve of total fungi determined by the blotter test showed a sigmoidal pattern that can be attributed to the mixture of fungal subpopulations with different radiation sensitivities. This behaviour could be modelled well (R² = 0.995) with a modified Gompertz equation. The predicted values for shoulder length and inactivation rate were 0.63 ± 0.10 kGy and 0.44 ± 0.04 kGy⁻¹ respectively. The sensitivity of the most common fungi to electron beam treatment followed the order Penicillium spp. > Fusarium spp. > Aspergillus spp., with total inactivation at irradiation doses of 1.7, 2.5 and 4.8 kGy respectively. CONCLUSION: The effect of electron beam treatment against fungi on naturally contaminated maize seeds depended on irradiation dose, allowing the control of maize fungal load.

Comparative proteome analysis of Penicillium verrucosum grown under light of short wavelength shows an induction of stress-related proteins associated with modified mycotoxin biosynthesis.

In this study the differentially expressed protein population of Penicillium verrucosum grown either in the dark or under light with a wavelength of 450nm has been analyzed. Light of short wavelength led to oxidative stress in the fungal cell; under this condition the mycotoxin biosynthesis revealed a mutual shift from ochratoxin A to citrinin. Using a proteomic approach combining an optimized protein extraction method with 2-dimensional SDS-PAGE followed by HPLC-ESI-TOF-MS/MS mass spectrometric analysis, initially 56 significantly differential proteins (light vs. dark) were detected comprising proteins of a broad range of isoelectric points and molecular masses. In total, 46 proteins could be identified further by database query, most of these proteins are assumed to be involved in response to stress (e.g. antioxidative proteins, heat shock proteins) and general metabolic processes (e.g. glycolysis, ATP supply). Proteome analyses are necessary to unravel the regulation of secondary metabolite biosynthesis at a translational level. This may enable identification of proteins which are involved in mycotoxin biosynthesis, adaption processes or even stress compensation mechanisms. This study depicts the first proteome analysis of P. verrucosum.