Development of a novel quantitative PCR assay as a measurement for the presence of geosmin-producing fungi.

Aims: To provide an efficient technique for monitoring the off-flavoured fungal compound geosmin.

Methods And Results: Geosmin-associated gpe1 gene of Penicillium expansum displayed ≥99% similarity to cytochrome P450 gene of geosmin-producing P. restrictum, but ≤40% similarities to geosmin biosynthesis, non-cytochromic gene of Streptomyces avermitilis and cytochrome P450 genes of non-geosmin-producing Neotyphodium lolii, Phoma betae and P. paxilli. Serial 10-fold dilutions of P. expansum's DNA was subjected to a previously reported qPCR assay (Atoui et al. 2007), utilizing gpe1 specific primer pair 'SNgpe1F/SNgpe1R'. A linear relationship between DNA quantity and Cycle Threshold (Ct ), with strong correlative coefficient, was observed. Using the available physico-chemical method, geosmin was quantified in 188 grape samples. Penicillium spp's DNA was quantified in these samples, utilizing the developed qPCR assay. A strong positive correlation (R(2)  = 0·97) between Penicillium's DNA and geosmin concentration was observed. Furthermore, <50 ng μl(-1) Penicillium's DNA corresponds to geosmin level below the permitted intensity limit i.e. 4, for 'Flavour Profile Analysis'.

Conclusions: Penicillium spp., genomic DNA level can provide an efficient way to quantify geosmin.

Significance And Impact Of The Study: This particular qPCR technique can be utilized in numerous food industries, for the timely detection and monitoring of geosmin contamination.