Quantification of Aspergillus carbonarius in grapes using a real time PCR assay. (October 2015)

• Record Type: Journal Article
• Authors: Tryfinopoulou, Paschalitsa; Kizis, Dimosthenis; Nychas, George-John E.; Panagou, Efstathios Z.
• Is Part Of: Food microbiology. Volume 51(2015:Oct.)
• Page Start: 139
• Page End: 143
• Publication Date: 2015-10
• Subjects: Aspergillus carbonarius -- Real time PCR -- Wine grapes; Food Microbiology -- Periodicals; Aliments -- Microbiologie -- Périodiques; Food -- Microbiology; Periodicals; Food -- Microbiology -- Periodicals; Food contamination -- Periodicals; 664.001579
• DOI: 10.1016/j.fm.2015.05.009
• Languages: English
• ISSNs: 0740-0020

Abstract

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. Abstract : RT PCR used in spiked and native grapes for A. carbonarius quantification. Standard curve (E = 104.2% and R 2 > 0.99) used for A. carbonarius DNA and RT PCR Ct. qPCR of A. carbonarius DNA (native grapes) resulted from 13 to 50 × 10 3 haploids/g.