Activity of trans-2-hexenal against Penicillium expansum in 'Conference' pears

AIMS: To investigate the effects of trans-2-hexenal on blue mould disease, patulin content and fruit quality in 'Conference' pears. METHODS AND RESULTS: Fruits, wounded and inoculated with Penicillium expansum or non-inoculated, were exposed to trans-2-hexenal vapour treatment (12.5 microl l(-1)) at 20 degrees C. A greater reduction of decay was obtained by treatment application 24 or 48 h after inoculation, in contrast trans-2-hexenal application 2 h after inoculation was ineffective. Fruit storage temperature (-1 degrees C) after treatment did not affect the antifungal activity. Although 2-h exposure to trans-2-hexenal was effective in reducing blue mould, an exposure of at least 8 h was required to reduce fruit patulin content. Treatments did not affect fruit physical-chemical characteristics. After 6 days at 20 degrees C following exposure, trans-2-hexenal residue in treated fruits was less than the natural content of the compound in unripe fruits. CONCLUSIONS: trans-2-Hexenal treatment is effective in the reduction of blue mould infections and patulin content in Conference pears when applied 24-48 h after pathogen inoculation. SIGNIFICANCE AND IMPACT OF THE STUDY: trans-2-Hexenal could be a natural alternative to fungicides in the control of P. expansum infections. Further work is needed to study the methods and conditions avoiding the persistence of off-odours and off-flavours in pears after their exposure to trans-2-hexenal vapours.     

LaeA regulation of secondary metabolism modulates virulence in Penicillium expansum and is mediated by sucrose

Penicillium expansum, the causal agent of blue mould rot, is a critical health concern because of the production of the mycotoxin patulin in colonized apple fruit tissue. Although patulin is produced by many Penicillium species, the factor(s) activating its biosynthesis are not clear. Sucrose, a key sugar component of apple fruit, was found to modulate patulin accumulation in a dose‐responsive pattern. An increase in sucrose culture amendment from 15 to 175 mm decreased both patulin accumulation and expression of the global regulator laeA by 175‐ and five‐fold, respectively, whilst increasing expression of the carbon catabolite repressor creA. LaeA was found to regulate several secondary metabolite genes, including the patulin gene cluster and concomitant patulin synthesis in vitro. Virulence studies of ΔlaeA mutants of two geographically distant P. expansum isolates (Pe‐21 from Israel and Pe‐T01 from China) showed differential reduction in disease severity in freshly harvested fruit, ranging from no reduction for Ch‐Pe‐T01 strains to 15%–25% reduction for both strains in mature fruit, with the ΔlaeA strains of Is‐Pe‐21 always showing a greater loss in virulence. The results suggest the importance of abiotic factors in LaeA regulation of patulin and other secondary metabolites that contribute to pathogenicity.     

Patulin accumulation in apples by Penicillium expansum during postharvest stages

AIMS: The aim of this study was to assess the opportunities of Penicillium expansum to develop and produce patulin in apples during cold storage and in the steps prior to processing of apple products. METHODS AND RESULTS: Two lots of apples var. Golden with different ripeness degree were used. Half of each lot was fungicide treated. Apples were inoculated with P. expansum and stored at 1 degrees C for 6 weeks. The extent of lesions and patulin accumulation both at the end of cold storage and after 3 days at 20 degrees C were assessed. Short storage at 20 degrees C aimed to simulate the transport and storage steps at room temperature before processing. Lesion size significantly increased during the storage at 20 degrees C. An interaction between fungicide treatment and ripeness degree was found; efficiency of fungicide treatment was higher for ripe apples. Although lesions were evident after cold storage, no patulin was detected. Patulin was detected only when fruits were further stored at 20 degrees C. Neither ripeness degree nor fungicide treatment affected patulin accumulation. CONCLUSIONS: Cold storage periods of 6 weeks do not lead to patulin accumulation. SIGNIFICANCE AND IMPACT OF THE STUDY: Shortening preprocessing times at warm temperatures would result into a reduction in patulin content at initial steps of fruits entering the processing plants.     

Patulin in grape must and new, still fermenting wine (Federweißer)

Extreme differences in temperature were anticipating a high mycotoxin contamination of the 2006 vintage. 96 samples of different grapes and must qualities of the producing region Mosel-Saar-Ruwer as well as Federweißer from retail sales were analyzed for patulin according to EN 14177, whereat the fermentation of the must was stopped by addition of sodium azide.     

Determination of patulin by online-SPE-LC

The mycotoxin patulin mainly occurs in fruits and fruit-derived products. For its determination a newly developed method employing a simplified liquid-liquid partitioning step followed by an online-SPE-LC analysis with UV detection is presented. The sample is diluted with phosphate buffer and extracted with ethyl acetate. The extract is evaporated and re-dissolved. The online-SPE-LC analysis employs a styrene-divinylbenzene copolymere phase for the SPE step, on which the carbonate washing step is carried out as well. The final LC analysis with UV detection uses a Polaris C18A column. This method reaches a limit of quantification of 15 μg/kg (clear apple juice), with a standard deviation of 10.7% (matrix calibration, n=5; c (patulin)=50 μg/kg). Presented at the 28^th Mykotoxin-Workshop, Bydgoszcz, Poland, May 29–31, 2006     

Blistering1 Modulates Penicillium expansum Virulence Via Vesicle-mediated Protein Secretion

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Highlights

• •The Blistering1 locus in Penicillium expansum encodes a novel protein with a DnaJ domain.
• •A single T-DNA insertion was generated in the blistering1 gene which manifested in reduced decay in apple and nearly undetectable levels of the mycotoxin patulin in the T625 mutant.
• •T625 failed to secrete a set of enzymes that degrade plant cell walls and the three final biosynthetic enzymes of patulin synthesis.
• •Quantitative mass spectrometry of mycelial proteins revealed that the mutant had altered cellular networks controlling protein processing in the endoplasmic reticulum, protein export, vesicle-mediated transport, and endocytosis.
• •Transmission electron microscopy of hyphal cross sections confirmed that the mutant formed abnormally enlarged endosomes or vacuoles.
• •These data reveal that Blistering1 affects internal and external protein processing involving vesicle-mediated transport in a family of fungi with medical, commercial, and agricultural importance.

     

Effect of patulin on the interdigitating dendritic cells (IDCs) of rat thymus

Patulin is a common fungal contaminant of ripe apples used for the production of apple juice concentrates and it is also present in other fruits, vegetables and food products. Patulin is a secondary metabolite produced by species of the genera Penicillium, Aspergillus and Byssochlamys. Patulin has been reported to be mutagenic, carcinogenic and teratogenic. Antigen-presenting cells (APCs) are of prime importance in the innate immune response; they capture antigen in tissues and then migrate to the lymphoid organs to present the antigen to T lymphocytes. Thus, they are crucial for the initiation of immunity. Interdigitating dendritic cells (IDCs) are a subset of APCs that are present at the lymphatic organs. In the thymus, they act in positive and negative selection during T cell development. In the present study, patulin was administered orally to growing male rats aged 5-6 weeks. A dose of 0.1 mg kg(-1) bw day(-1) was given to rats for a period of 60 or 90 days daily. The effect of patulin on the IDCs of thymus was investigated by transmission electron microscopy (TEM), and the results were evaluated in terms of cell destruction. In the rats of the control group, it was observed that the IDCs had an indented nucleus, a clear cytoplasm and numerous membrane extensions. In the cytoplasm, a well-developed golgi complex, mitochondria, granular endoplasmic reticulum and a small number of lysosomal structures were observed. At day 60 of patulin-treated rat groups (P-60), loss of cristae in mitochondria and chromatin margination and lysis in the nucleus were found. It was observed that the IDCs had a perinuclear area of cytoplasm surrounded by a peripheral electron-lucent zone. In the cytoplasm of the 90-day patulin-treated rat group (P-90), a peripheral electron-lucent zone was also found, similar to the P-60 group. Additionally increase in vesicular and lysosomal structures, increase in apoptotic bodies and condensation of chromatin in the nucleus were noted. It was observed that patulin leads to apoptotic body formation and cell apoptosis in the IDCs of rat thymus especially in the P-90-treated groups.     

Removal of patulin from apple juice using inactivated lactic acid bacteria

Aims: Apples and apple products are the most notably commodities contaminated with patulin (PAT), which cause detrimental effects on human health and economic problems. The primary objective of this study was to investigate the removal of PAT contamination from apple juice using 10 different inactivated lactic acid bacteria (LAB) strains. Methods and Results: Significant quantities of PAT ranging from 47 to 80% were bound to all tested bacterial strains, whereas Lactobacillus rhamnosus 6224 and Enterococcus faecium 21605 caused a decrease of PAT by 80·4 and 64·5%, respectively. The results showed that the binding of PAT depends on the initial concentration of toxin and the adsorption temperature, also the differences in biomass existed among the 10 bacterial strains. IR analysis was performed to identify potential functional groups and the possible binding sites related to PAT adsorption. Conclusions: The removal of PAT was observed to be strain specific. The results indicated that the biosorption process did not affect the quality of juice. FTIR analysis showed that the cell wall plays a key role in PAT adsorption. Significance and Impact of the Study: Our results proof that inactivated LAB have the potential as a novel and promising adsorbent to bind PAT effectively.