Quantification of Aspergillus carbonarius in grapes using a real time PCR assay

Paschalitsa Tryfinopoulou; Dimosthenis Kizis; George-John E Nychas; Efstathios Z Panagou

doi:10.1016/j.fm.2015.05.009

Quantification of Aspergillus carbonarius in grapes using a real time PCR assay

Food Microbiol. 2015 Oct:51:139-43. doi: 10.1016/j.fm.2015.05.009. Epub 2015 May 27.

Authors

Paschalitsa Tryfinopoulou¹, Dimosthenis Kizis², George-John E Nychas³, Efstathios Z Panagou³

Affiliations

¹ Laboratory of Microbiology and Biotechnology of Foods, Department of Food Science and Human Nutrition, Agricultural University of Athens, Iera Odos 75, Athens 11855, Greece. Electronic address: ptryf@aua.gr.
² Laboratory of Microbiology and Biotechnology of Foods, Department of Food Science and Human Nutrition, Agricultural University of Athens, Iera Odos 75, Athens 11855, Greece; Benaki Phytopathological Institute, Department of Phytopathology, Laboratory of Mycology, St. Delta 8, 14561 Athens, Greece.
³ Laboratory of Microbiology and Biotechnology of Foods, Department of Food Science and Human Nutrition, Agricultural University of Athens, Iera Odos 75, Athens 11855, Greece.

PMID: 26187838
DOI: 10.1016/j.fm.2015.05.009

Abstract

A study on the representation of Aspergillus carbonarius in the vineyards of the Mesogeia geographical region of Attica, Greece, was conducted. One hundred and twenty five samples of the indigenous drought and disease resistant Savatiano wine grape variety, the most widely planted in Greece, were collected. The sample's total DNA extracts were initially tested for fungal DNA presence by targeting the Internal Transcribed Spacer (ITS) region in end-point Polymerase Chain Reactions. Samples which were proved positive were further subjected to PCR analysis using specific primers targeting an A. carbonarius polycetide synthase (pks) gene. Among ITS positive samples (70%), A. carbonarius was represented in 42% of them. Furthermore, a SYBR Green I Real Time PCR method was used to quantify the amount of this species in the grape samples. The values of the positive samples were estimated in the range of 13 to 50 × 10(3) fungal haploid genomes/g grapes. The significance of this study lies in the applicability of a rapid and culture-independent method to detect and quantify A. carbonarius on grapes.

Keywords: Aspergillus carbonarius; Real time PCR; Wine grapes.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Aspergillus / genetics*
Aspergillus / isolation & purification*
DNA Primers
DNA, Fungal / genetics
Food Contamination*
Food Microbiology / methods
Genome, Fungal
Greece
Haploidy
Real-Time Polymerase Chain Reaction / methods*
Vitis / microbiology*

Substances

DNA Primers
DNA, Fungal