Citrinin as an accessory establishment factor of P. expansum for the colonization of apples.

Penicillium expansum is the causal agent of blue mold decay of apples. This fungal species can produce the two important mycotoxins patulin and citrinin. It was previously shown that patulin represents a colonization factor for the infection of apples. No definitive information about the importance of citrinin for the colonization of apples is currently available. The pksCT gene of the citrinin cluster codes for the citrinin polyketide synthase. Mutants of P. expansum in which the pksCT was inactivated showed a drastic decrease in the citrinin production. In addition, the pksCT mutants were also reduced in the ability to colonize apples. Externally added citrinin restored the capacity of the mutants to colonize apples roughly to that of the wild type. A kinetic analysis of the expression of the two respective pks genes of patulin (patK) and citrinin (pksCT) revealed that both genes are highly expressed in the first phase during the colonization process. The production of patulin in the apple matrix coincides with the expression of the patK gene. Almost no citrinin could be identified analytically during the first phase but only at a later stage of the colonization. It could be demonstrated that citrinin is degraded in apples and can tightly be bound to pectin. Overall the results suggest that citrinin may have an accessory function for the establishment of the colonization guided by other factors.

Arginine acts as an inhibitor of the biosynthesis of several mycotoxins.

It is well known that the type and the availability of nitrogen have a great influence on the biosynthesis of certain mycotoxins. Here it is shown that some amino acids have no influence, some others strongly support and a third group inhibits the biosynthesis of ochratoxin (OTA) by Penicillium nordicum even in a complex medium, such as PDA. Arginine (Arg) is one of the strong OTA inhibiting amino acids. It was shown that Arg not only inhibits OTA in Penicillium but also citrinin (CIT) biosynthesis in Penicillium verrucosum, Penicillium expansum and Penicillium citrinum and alternariol (AOH), alternariol monomethylether (AME) and tenuazonic acid (TeA) biosynthesis in Alternaria alternata. The minimal inhibitory concentration of Arg differs depending on the mycotoxin and the species analysed. However, the OTA biosynthesis by P. verrucosum and P. nordicum was most sensitive. Growth, on the other hand, was much less affected by Arg. Urea, a metabolite of Arg catabolism, shows a similar inhibitory activity. In wheat medium containing 50mM Arg almost no OTA was produced by Penicillium, in contrast to plain wheat medium.