Penicillium Expansum Strains Research Articles

First report of post-harvest tuber rot of Apios americana Medikus caused by Penicillium expansum in South Korea.

Apios americana Medikus, a perennial vine legume native to North America, is known as 'Indian potato' or 'Apios' for their underground tubers that are used for human consumption in Korea (Choi et al., 2017). These tubers are known to be rich in isoflavones as well as other secondary products to have known several medicinal properties (Chu et al., 2019). The harvested tubers in 2020 were observed to rot during storage of tubers for 4 months at 4°C in the genetic resource storage room at the Bioenergy Crop Research Institute, National Institute of Crop Science, Muan, Jeollanam-do, South Korea. The incidence of rot symptoms with blue mold was less than 1% per 20 kg box. Ten infected tubers were collected from several boxes and the lesions were cut into small pieces, and then surface sterilized in 1.5% NaClO for 2 minutes, followed by rinsing 3 times with sterilized water. To investigate their morphological characteristics, ten isolates were cultured in Malt Extract Agar (MEA) medium at 25° C for 5 days (Pitt and Hocking, 1988). The surface morphology of the mycelium had white or light green fluffy, and completely blue spores were formed after about 5 days. The conidia were one-stage branched with an elliptical shape, about 3.5 to 4.3 × 2.9 to 3.6 (mean 3.8 ± 0.3 × 3.2 ± 0.2) ㎛ in diameter (n=30). Genomic DNAs of the isolates were extracted using Solgent DNA Extraction Kit (Solgent, Daejeon, Korea), and then PCR products of the internal transcribed spacer (ITS1/ITS4) region and the beta-tubulin gene (Bt2a/Bt2b) were sequenced and analyzed (Glass and Donaldson, 1995). The BLASTn showed that the representative isolate had 99% homology with reference Penicillium expansum strain ICMP 2708 (ITS region and TUB2 gene) in NCBI GenBank. The sequences of the isolate were deposited in GenBank as accession numbers MZ636667 and MZ702813 for ITS and TUB2 genes, respectively. Based on the morphological characteristics and molecular analysis, the isolate was identified as P. expansum. Pathogenicity assays of the isolate were also performed using three tubers in three replicates inoculated with spore suspension (concentration, 1×106 conidia/mL) and compared with a control group inoculated with sterilized water. The inoculated Apios tubers were placed in a plastic box maintained in conditions of high humidity at 25°C. Five days after inoculation, the typical symptoms were observed on inoculated tubers, and no symptoms were observed in the control one. P. expansum was again isolated from artificially inoculated tubers to complete Koch's assumption. This is the first report of P. expansum causing tuber rot in A. americana in South Korea. As the cultivated area of Apios is increasing in Korea, it will be necessary to develop effective storage methods and management strategies for the control of storage diseases such as blue mold.

Nature-Inspired Chemistry of Complex Alkaloids: Combining Targeted Molecular Networking Approach and Semisynthetic Strategy to Access Rare Communesins in a Marine-Derived Penicillium expansum.

Communesins are rare alkaloids isolated from fungi of the genus Penicillium. In this work, the extract of a marine-derived Penicillium expansum strain was studied using targeted molecular networking approach allowing to detect 65 communesins including 55 new ones. A fragmentation pattern for dimethylvinyl communesins was established and a script was implemented allowing to predict the structure and map all communesins in a global molecular network. A semisynthetic strategy was carried out to obtain some minor congeners from the two isolated communesins A and B. Nine communesins were then synthetised: two of them were already described as produced by the studied strain; four are new natural products which occurrence in the extracts was confirmed; three are new semi-synthetic analogues never described so far. These communesins were evaluated for their cytotoxicity on two human cancer cell lines KB and MCF-7 leading to a preliminary study of their structure-activity relationships.

In vivo evaluation of Aggressiveness, pathogenicity and patulin accumulation by three Penicillium expansum strains isolated from Algerian apples

Among the postharvest pathogens of apple fruit, Penicillium expansum (P.expansum) is considered to be one of the most common fungal pathogens worldwide. It is the causal agent of blue mold decay; the main serious economically disease of apple fruits, and the main producer of patulin (PAT). The principal objective of this work was to evaluate the aggressiveness and the PAT production of three isolated strains from apples produced in the East of Algeria. These strains were isolated then screened for their inhibitory activity against Escherichia coli and Bacillus subtilis (18 mm and 18 mm for S7; 10 mm and 11mm for S3), as well as their ability to cause blue rot (S4). The three strains S7, S3, and S4 (MN904448, MT023795 and MN904449, respectively) were identified as P. expansum by ITS1, ITS4, and β-tubulin gene sequences analysis. The in vitro ability to produce PAT was confirmed by Gas chromatography-mass spectrometry of strains’ crude extracts cul-tured on YES (Yeast-Extract-Sucrose) medium. Indeed, these strains exhibited a great capacity to cause blue mold decay for all tested apples (26 to 52 mm lesion diameter), after nine days of incubation at 25°C. Apples rotten area analysis that was performed by UPLC analysis revealed that these three strains were able to produce PAT at different concentrations ranging from 0.57 to 469 μg/mL. Thus, these strains constitute a risk to consumer health because these amounts exceed the permissive level of PAT.

An eco-friendly approach to textile and tannery wastewater treatment using maghemite nanoparticles (γ-Fe2O3-NPs) fabricated by Penicillium expansum strain (K-w)

Wastewater treatment is one of the vital solutions to the global water shortage and many related environmental problems. The present study aimed at biosynthesizing maghemite nanoparticles (γ-Fe2O3-NPs) using Penicillium expansum (K-w) for the purpose of wastewater treatment. The γ-Fe2O3-NPs resultant was characterized using fourier transform infrared (FT-IR) spectroscopy, X-ray diffraction (XRD), transmission electron microscope (TEM), dynamic light scattering (DLS), and X-ray photoelectron spectroscopy (XPS). The maghemite nanoparticles capacity to mediate decolorization and degradation of textile and tanning wastewater effluents was investigated. Results verified that the successful synthesis of spherical shaped Fe2O3-NPs of 15.0–66.0 nm size, with XRD peaks that match the cubic structure of the γ-Fe2O3. XPS analysis confirmed the presence of iron as Fe2O3. Decolorization and degradation analyses indicated that γ-Fe2O3-NPs are effective biocatalysts for dye degradation under dose- and time-dependent manner. The highest decolorization gain (89 %–90 %) occurred after 6.0–8.0 h of incubation at 1.0 mg mL-1 of γ-Fe2O3-NPs. Quality of the treated textile wastewater was improved, as indicated by the reduction in chemical oxygen demand (COD), total dissolved solids (TDS), and total suspended solids (TSS) by 86.8, 71.8, and 89.3 %, respectively. GC–MS results showed the complete disappearance or reduction of peaks, or peak areas in treated textile wastewater in comparison with the untreated samples. In addition, treatment of tanning wastewater with γ-Fe2O3-NPs has also resulted in decolorization, as well as reduction in COD, TDS, and TSS levels by 93.1, 40.5, and 95.9 %, respectively. The treated tanning wastewater showed low heavy metal levels, with removal percentages ranging from 66 %–98 % for Co, Cr, Pb, Ni, and Cd, indicating the high efficiency of γ-Fe2O3-NPs in improving treated wastewater quality.

Penicillium expansum strain isolated from indoor building material was able to grow on gypsum board and emitted guttation droplets containing chaetoglobosins and communesins A, B and D.

AimsEmission of toxic metabolites in guttation droplets of common indoor fungi is not well documented. The aims of this study were (i) to compare mycotoxins in biomass and guttation droplets from indoor fungi from a building following health complaints among occupants, (ii) to identify the most toxic strain and to test if mycotoxins in guttation liquids migrated trough air and (iii) to test if toxigenic Penicillium expansum strains grew on gypsum board.Methods and ResultsBiomass suspensions and guttation droplets from individual fungal colonies representing Aspergillus, Chaetomium, Penicillium, Stachybotrys and Paecilomyces were screened toxic to mammalian cells. The most toxic strain, RcP61 (CBS 145620), was identified as Pen. expansum Link by sequence analysis of the ITS region and a calmodulin gene fragment, and confirmed by the Westerdijk Institute based on ITS and beta‐tubulin sequences. The strain was isolated from a cork liner, was able to grow on gypsum board and to produce toxic substances in biomass extracts and guttation droplets inhibiting proliferation of somatic cells (PK‐15, MNA, FL) in up to 20 000‐fold dilutions. Toxic compounds in biomass extracts and/or guttation droplets were determined by HPLC and LC‐MS. Strain RcP61 produced communesins A, B and D, and chaetoglobosins in guttation droplets (the liquid emitted from them) and biomass extracts. The toxins of the guttation droplets migrated c. 1 cm through air and condensed on a cool surface.ConclusionsThe mycotoxin‐containing guttation liquids emitted by Pen. expansum grown on laboratory medium exhibited airborne migration and were >100 times more toxic in bioassays than guttation droplets produced by indoor isolates of the genera Aspergillus, Chaetomium, Stachybotrys and Paecilomyces. Significance and Impact of the StudyToxic exudates produced by Pen. expansum containing communesins A, B and D, and chaetoglobosins were transferable by air. This may represent a novel mechanism of mycotoxin dispersal in indoor environment.

Detoxification and anti-mycotoxigenic effect of Ochratoxin A produced by Penicillium expansum species utilizing probiotic Pichia kudriavzevii and biogenic-selenium nano-formulations

Ochratoxins are overall spread fungal secondary metabolites combined chiefly by some toxigenic types of Aspergillus and Penicillium. A few physical, chemical and biological techniques were found among literary reports to wipe out, inactivate or decrease the bioavailability of mycotoxins counting ochratoxin A (OTA). Concerning the creation of OTA, Penicillium expansum strains inspected in this examination using TLC were recorded surprisingly for the first time. They were subjected to the beforehand announced probiotic yeast Pichia kudriavzevii, biosynthesized Selenium nanoparticles (SeNPs) and consolidated plans of both to look at their capability to detoxificate or lessening of this mycotoxin. Every single tried experiment was found to be capable for diminishing OTA as confirmed by the quantitative assessment using HPLC. Where, the blend of Pichia kudriavzevii and SeNPs-plans demonstrated that most of OTA evacuation rate created by P.expansum5 (50.18 %). While the least OTA expulsion rate was watched delivered by P.expansum3 (76.08%) treated with SeNPs as it were. The ultrastructural investigations of P.expansum5 cells completed with TEM demonstrated the continuous harm of the fungal cells constituents as it appeared from the investigations. The most elevated impacts were seen among the cytoplasmic constituents of P.expansum5 cells treated with the previous blend of probiotic and SeNPs that were totally contracted and pulled back far from the cell membrane leaving a gigantic separation region of no organelles. Remarkable cytoplasmic ultrastructure's deformations of P.expansum5 cells with appearance of some vacuolation as a survival adaptation mode of the fungal cells were additionally recorded. Conclusively, this is the primary statement of OTA biodegradation using a combination strategy of probiotic yeast Pichia kudriavzevii and biogenic-SeNPs which endorsing the impact of utilizing these novel natural blends for the detoxification, anti-mycotoxigenic and/or diminution of OTA production by mycotoxegenic fungal species. This natural mixture could be recommended to be utilized as a food/feed additive, during storage and as prophylactic agent in pre-and post-harvest crops.

Evaluation of antifungal activity of three essential oils against selected isolates of Penicillium, Purpureocillium and Acremonium

Storage is a critical process of food production because incorrect conditions during this process lead to the growth of economic costs caused by material loss. It is important to maintain adequate microbiological purity of storage areas. The development of microbes present in warehouses may lead to secondary contamination of stored materials and reduce their quality. The aim of the study was to evaluate the effect of essential oils on the growth of Acremonium strictum , Penicillium citrinum, Penicillium expansum and Purpureocillium lilacinum . Fungal strains were isolated from the food warehouse. Studies on the influence of selected mix of phenolic substances were performed using three commercially available essential oils, namely rosewood, rosemary and thyme. Evaluation of antifungal activity of these essential oils was done by plate dilution method and expressed as inhibition rate of mycelial growth and index of mycelial growth, and by minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) assays. Thyme oil showed the highest antifungal activity and inhibited growth of all fungal strains at the lowest tested concentration (1 μL mL-1). Penicillium expansum strain was found to be the most resistant to the tested essential oils.

Biocontrol Agents Increase the Specific Rate of Patulin Production by Penicillium expansum but Decrease the Disease and Total Patulin Contamination of Apples

Synthetic fungicides are commonly employed for the control of postharvest diseases of fruits. However, due to health concerns about the use of these chemicals, alternative control methods including biocontrol based on antagonistic yeasts are gaining in popularity. In this study, we investigated the effects of two biocontrol yeasts, Rhodotorula mucilaginosa strain 3617 and Rhodotorula kratochvilovae strain LS11, on blue mold and patulin (PAT) contamination caused by Penicillium expansum strains PY and FS7 in artificially inoculated Fuji apples stored at 20°C for 9 days. To correlate the development of the P. expansum strains in yeast-treated and untreated apples with PAT production, we quantified their biomass in the infected fruits using a recently published quantitative real-time polymerase chain reaction method based on specific primers for patF, a gene from P. expansum that is involved in PAT biosynthesis. Both yeasts significantly reduced the disease incidence caused by the two strains of P. expansum up to 5–7 days of incubation, and lowered their biomass and the progression of symptoms up to 9 days. Interestingly, both yeasts strains increased the rate of PAT production (expressed as ng patulin/μg fungal DNA) by the two pathogenic strains. Nevertheless, both biocontrol agents reduced the total PAT contamination, especially in the case of P. expansum strain FS7, the higher PAT producer of the two tested P. expansum strains. Comparing between the yeast strains, R. kratochvilovae LS11 was more effective than R. mucilaginosa 3617 for the control of P. expansum.

A gene encoding a new cold-active lipase from an Antarctic isolate of Penicillium expansum

Cold-active lipases are of significant interest as biocatalysts in industrial processes. We have identified a lipase that displayed activity towards long carbon-chain-p-nitrophenyl substrates (C12-C18) at 25 °C from the culture supernatant of an Antarctic Penicillium expansum strain assigned P. expansum SM3. Zymography revealed a protein band of around 30 kDa with activity towards olive oil. DNA fragments of a lipase gene designated as lipPE were isolated from the genomic DNA of P. expansum SM3 by genomic walking PCR. Subsequently, the complete genomic lipPE gene was amplified using gene-specific primers designed from the 5'- and 3'-regions. Reverse transcription PCR was used to amplify the lipPE cDNA. The deduced amino acid sequence consisted of 285 residues that included a predicted signal peptide. Three peptides identified by LC/MS/MS analysis of the proteins in the culture supernatant of P. expansum were also present in the deduced amino acid sequence of the lipPE gene suggesting that this gene encoded the lipase identified by initial zymogram activity analysis. Full analysis of the nucleotide and the deduced amino acid sequences indicated that the lipPE gene encodes a novel P. expansum lipase. The lipPE gene was expressed in E. coli for further characterization of the enzyme with a view of assessing its suitability for industrial applications.

Genetic structure and natural variation associated with host of origin in Penicillium expansum strains causing blue mould

Blue mould, caused by Penicillium expansum, is one of the most economically damaging postharvest diseases of pome fruits, although it may affect a wider host range, including sweet cherries and table grapes. Several reports on the role of mycotoxins in plant pathogenesis have been published, but few focussed on the influence of mycotoxins on the variation in host preference amongst producing fungi. In the present study the influence of the host on P. expansum pathogenicity/virulence was investigated, focussing mainly on the relationship with patulin production. Three P. expansum strain groups, originating from apples, sweet cherries, and table grapes (7 strains per host) were grown on their hosts of isolation and on artificial media derived from them. Strains within each P. expansum group proved to be more aggressive and produced more patulin than the other two groups under evaluation when grown on the host from which they originated. Table grape strains were the most aggressive (81% disease incidence) and strongest patulin producers (up to 554μg/g). The difference in aggressiveness amongst strains was appreciable only in the presence of a living host, suggesting that the complex pathogen–host interaction significantly influenced the ability of P. expansum to cause the disease. Incidence/severity of the disease and patulin production proved to be positively correlated, supporting the role of patulin as virulence/pathogenicity factor. The existence of genetic variation amongst isolates was confirmed by the High Resolution Melting method that was set up herein, which permitted discrimination of P. expansum from other species (P. chrysogenum and P. crustosum) and, within the same species, amongst the host of origin. Host effect on toxin production appeared to be exerted at a transcriptional level.

A new improved strategy for the selection of cold-adapted antagonist yeasts to control postharvest pear diseases

Postharvest diseases cause considerable losses of harvested fruits during transportation and storage. Many yeast species have been reported as good antagonists against postharvest pear pathogens. In this work, we used a novel selection strategy that involves the isolation of yeasts from washing fluids, showing biocontrol activity against a regional Penicillium expansum strain (primary screening), originally obtained from fruit wounds after long time storage at −1/0°C. About 26 isolates representative of the 11 yeast species identified in the 27 selected washing waters were chosen to be evaluated in a secondary screening against a regional Botrytis cinerea strain on pear wounds. Among yeasts tested, 38% showed complete control of P. expansum, but only 15% reduced the decay incidence of B. cinerea to 60–80% at −1/0°C. These results reveal that some of the yeasts found can be biological alternatives to fungicides in the control of P. expansum and B. cinerea infections. Based on the data obtained, our strategy seems to be much more effective than the previously reported methods in obtaining successful biocontrol agents.

Modelling the inhibitory effect of copper sulfate on the growth of Penicillium expansum and Botrytis cinerea

This study aimed to investigate the effect of copper sulfate (from 0 to 8 mmol kg(-1)) on radial growth rate and lag time of two moulds responsible for vine grapes spoilage: Penicillium expansum strain 25·03 and Botrytis cinerea, strains BC1 and BC2. A new model was developed to describe tailing and shoulders in the inhibition curves. Because of tailing, the minimum inhibitory concentration (MIC), was not defined as the concentration at which no growth was observed, but as the concentration at which the lag time was infinite. The concentrations at which μ = μ(opt)/2, (Cu50), were in the range of 2·2-2·6 mmol kg(-1). Radial growth rate of P. expansum and the reciprocal of the lag time were linearly correlated (r = 0·84). In contrast, in the range 0-4 mmol kg(-1), an inhibition of growth of B. cinerea was observed whereas germination remained unaffected (i.e. the lag time was constant). In the range 4-8 mmol kg(-1) , the radial growth rate of B. cinerea was almost constant (c. 1 mm day(-1)), but germination was inhibited (i.e. the lag time was increased). The MIC values were 4·7 mmol kg(-1) for P. expansum, 8·2 and 7·3 mmol kg(-1) for B. cinerea strain BC1 and BC2, respectively, demonstrating that some isolates of these moulds are resistant to copper. Copper concentrations at 4 mmol kg(-1) would be sufficient to control the development of these isolates, but the toxicity of copper should be extended to other isolates and evaluated in vineyards.

Potential of patulin production by Penicillium expansum strains on various fruits

In this study, we investigated the pathogenicity and patulin production by ten strains of Penicillium expansum on various fruits (apples, apricots, kiwis, plums and peaches) at two (4°C and 25°C) different temperature regimes. All strains caused the infectious rots on all fruits at 4 and 25°C except one strain (PEX 09) at 4°C. Two strains (PEX 20 and PEX 12) out of ten produced the highest amounts of patulin on all fruits tested. The patulin production by P. expansum is high at 25°C compared to 4°C. All strains of P. expansum accumulated patulin ranging from 100-13,200μg/kg and nine strains ranging from 100-12,100μg/kg in all fruits at 25°C and 4°C, respectively. Among ten strains of P. expansum, strain PEX 20 produced the greatest amount of patulin on apricots (13,200μg/kg of rotten fruit) and on apples (12,500μg/kg) at 25°C after 9days of incubation. At 4°C, this strain produced 12,100, 12,000, 2,100 and 1,200μg/kg of patulin on apricots, apples, plums and peaches, respectively, after 45days of incubation. Strain PEX 12 produced the highest amount of patulin on kiwis (10,700μg/kg) at 25°C and 10,300μg/kg at 4°C. Patulin production by P. expansum on peaches and plums at both temperatures were lower than other fruits. The results of this study showed that careful removal of rotten fruits is essential to produce patulin-free fruit juice, since high patulin levels in apricots, apples and kiwis could result in a level greater than 50μg/kg of this mycotoxin in finished fruit juices, when one contaminated fruit occurs in 264, 250 and 214 fruits, respectively. So, the fruit processors should take care in not using rotten fruits for juice production to avoid the patulin problem worldwide, since this study proved that most important fruits being used for juice production and direct human consumption are susceptible to P. expansum and subsequent patulin production even at low temperatures. This is the first comprehensive report regarding patulin production by different strains of P. expansum on various fruits from Italy at different temperature regimes.

Crystal structure of a triacylglycerol lipase from Penicillium expansum at 1.3 Å determined by sulfur SAD

Triacylglycerol lipases (EC 3.1.1.3) are present in many different organisms including animals, plants, and microbes. Lipases catalyze the hydrolysis of long-chain triglycerides into fatty acids and glycerol at the interface between the water insoluble substrate and the aqueous phase. Lipases can also catalyze the reverse esterification reaction to form glycerides under certain conditions. Lipases of microbial origin are of considerable commercial interest for wide variety of biotechnological applications in industries, including detergent, food, cosmetic, pharmaceutical, fine chemicals, and biodiesel. Nowadays, microbial lipases have become one of the most important industrial enzymes. PEL (Penicillium expansum lipase) is a fungal lipase from Penicillium expansum strain PF898 isolated from Chinese soil that has been subjected to several generations of mutagenesis to increase its enzymatic activity. PEL belongs to the triacylglycerol lipases family, and its catalytic characteristics have been studied. The enzyme has been used in Chinese laundry detergent industry for several years (http://www.leveking.com). However, the poor thermal stability of the enzyme limits its application. To further study and improve this enzyme, PEL was cloned and sequenced. Furthermore, it was overexpressed in Pichia pastoris. PEL contains GHSLG sequence, which is the lipase consensus sequence Gly-X1-Ser-X2-Gly, but has a low amino acid sequence identities tomore » other lipases. The most similar lipases are Rhizomucor miehei (PML) and Rhizopus niveus (PNL) with a 21% and 20% sequence identities to PEL, respectively. Interestingly, the similarity of PEL with the known esterases is somewhat higher with 24% sequence identity to feruloyl esterase A. Here, we report the 1.3 {angstrom} resolution crystal structure of PEL determined by sulfur SAD phasing. This structure not only presents a new lipase structure at high resolution, but also provides a structural platform to analyze the published mutagenesis results. The structure may also open up new avenues for future protein engineering study on PEL.« less

Purification of Candida guilliermondii and Pichia ohmeri killer toxin as an active agent against Penicillium expansum

An antifungal assay with cell-free culture supernatant of Pichia ohmeri 158 and Candida guilliermondii P3 was tested against Penicillium expansum strain #2 at 25°C by measuring hyphal length and percentage conidia germination. C. guilliermondii was more effective against P. expansum conidia germination (58.15% inhibition), while P. ohmeri showed higher inhibition of mycelial growth (66.17%), indicating a probable mechanism associated with killer activity. This killer toxin (molecular mass <3 kDa) was partially purified by normal phase HPLC, using TSKgel Amide-80 analytical and preparative columns. Compared with crude extract, the killer toxin eluted from the post analytical column significantly inhibited P. expansum:% inhibition rose from 42.16 to 90.93% (C. guilliermondii) and 39.32 to 91.12% (P. ohmeri) (p < 0.05). The one-step purification process was adequate in isolating killer toxin from culture supernatant and also increased anti-Penicillium activity.

Modeling the effect of temperature on the growth rate and lag phase of Penicillium expansum in apples

The objective of the present study was to develop validated models that describe the effect of storage temperature on the growth rate and lag phase of six Penicillium expansum strains. The growth of the selected strains was therefore studied on Apple Puree Agar Medium (APAM) at 30, 25, 16, 10, 4 and 2 °C. Growth rates and lag phases were estimated using linear regression. Several secondary models were evaluated and for the growth rate, a modification of the extended Ratkowsky model was selected. Regarding the lag phase, the Arrhenius–Davey model provided the best adjustment to the observed data. Model validation was performed in two steps. Firstly, the developed models were validated on APAM. The obtained bias factors ( B f) ranged from 0.91 to 1.14 and the accuracy factors ( A f) were < 1.2 for the validation performed on APAM, indicating that the models were good predictors of the true mean colony growth rate and lag phase. Afterwards, an external validation was carried out in apples. For the growth rate, B f ranged from 0.64 to 0.81 and A f < 1.39, indicating conservative predictions. On the contrary for the lag phase, a clear deviation was observed between predictions and observed values on apples (0.35 < B f < 0.7 and A f > 1.6). These results highlight that the use of simulation or synthetic media for the development of predictive models for the lag phase of moulds can lead to inadequate predictions and that a validation on the real food matrix is necessary. Application of the developed models is possible in the framework of Quantitative Risk Assessment to develop control strategies against blue mould rot in apple and enables the inclusion of strain variability. However, possible underestimation of the lag phase should be taken into account.

Some fungicides and growth inhibitor/biocontrol-enhancer 2-deoxy- d-glucose increase patulin from Penicillium expansum strains in vitro

This study assessed the effect of disease control agents, fungicides and 2-deoxy- d-glucose (DOG), on a mycotoxin, patulin, production by strains of Penicillium expansum. Fungicides and DOG were added individually to agar on which the fungus was then grown. In an initial screen of six fungicides, three indicated that patulin was stimulated. These were tested further at a range of concentrations and colony diameters (cd) and patulin production were measured. Although patulin production appeared similar at each concentration in most cases, cd were reduced by fungicide in two out of three cases. So patulin was stimulated in relation to the diameter of the fungal colonies. This is interesting from an academic standpoint and also has applied relevance to fruit producers. In addition, the growth inhibitor and biocontrol-enhancer DOG appeared to stimulate patulin production although results were not statistically significant. The effect of fungal control agents on mycotoxin production requires consideration.

The biosynthesis of low-molecular-weight nitrogen-containing secondary metabolite-alkaloids by the resident strains of Penicillium chrysogenum and Penicillium expansum isolated on the board of the Mir space station

The analysis of the absorption spectra of the low-molecular-weight nitrogen-containing secondary metabolites--alkaloids--of 4 Penicillium chrysogenum strains and 6 Penicillium expansum strains isolated on board the Mir space station showed that all these strains synthesize metabolites of alkaloid origin (roquefortine, 3,12-dihydroroquefortine, meleagrin, viridicatin, viridicatol, isorugulosuvin, rugulosuvin B, N-acetyl-tryptamine, and a "yellow metabolite" containing the benzoquinone chromophore).

Structural investigation of two cell-wall polysaccharides of Penicillium expansum strains

The structure of two cell-wall polysaccharides isolated from three different strains of Penicillium expansum, the type species of the genus, have been established by 1D and 2D NMR spectroscopy, and also by methylation analyses. The water-soluble polysaccharide F1S-B consisted of a linear tetrasaccharide repeating unit with the following structure: [ → 6)-β- d-Gal f-(1 → 5)-β- d-Gal f-(1 → 5)-β- d-Gal f-(1 → 5)-β- d-Gal f-(1 → ] n The alkali-soluble polysaccharide F1I is a (1 → 3)-α- d-glucan.

Citrinin-producing capacity of Penicillium expansum strains from apple packinghouses of Lerida (Spain)

A study of the citrinin producing capacity in 122 Penicillium expansum strains isolated from apples and various locations of apple packaging premises Lleida (Spain) was conducted. Among the strains examined, 46% produced citrinin in a laboratory substrate and 73.2% of these strains were isolated from decayed apples. All citrinin producing strains produced metabolites which inhibited Bacillus cereus and Bacillus subtilis. The majority of the strains producing citrinin in solid substrate also produced citrinin in apples and apple juices.