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. 2023 Aug 17;12(16):3091.
doi: 10.3390/foods12163091.

A Molecular Toolbox to Identify and Quantify Grape Varieties: On the Trace of "Glera"

Ilaria Carrara  1 , Valeria Terzi  1 , Roberta Ghizzoni  1 , Stefano Delbono  1 , Giorgio Tumino  2 , Manna Crespan  3 , Massimo Gardiman  3 , Enrico Francia  4 , Caterina Morcia  1
Affiliations

A Molecular Toolbox to Identify and Quantify Grape Varieties: On the Trace of "Glera"

Ilaria Carrara et al. Foods. .

Abstract

A pillar of wine authenticity is the variety/ies used. Ampelographic descriptors and SSR markers, included in several national and international databases, are extensively used for varietal identification purposes. Recently, SNP markers have been proposed as useful for grape varietal identification and traceability. Our study has been directed toward the development of a molecular toolbox able to track grape varieties from the nursery to the must. Two complementary approaches were developed, exploiting SNP markers with two different technologies, i.e., a high-throughput platform for varietal identification and a digital PCR system for varietal quantification. As proof-of-concept, the toolbox was successfully applied to the identification and quantification of the "Glera" variety along the Prosecco wine production chain. The assays developed found their limits in commercial, aged wines.

Keywords: chip digital PCR; grape chain authenticity; high-throughput SNP genotyping; varietal traceability.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Grape genomic regions carrying polymorphisms (highlighted in yellow) on which the two digital PCR-based assays ID AN33KY7 and ID AN49FJ4 aimed at “Glera” traceability were designed.
Figure 2
Figure 2
“Glera”-rooted dendrogram obtained after neighbor-joining clustering analysis.
Figure 3
Figure 3
Chip digital PCR analysis of three “Glera” accessions (A1–A3; B1–B3) and one non-“Glera” variety (A4,B4) with the two assays ID AN33KY7 (samples A1–4) and ID AN49FJ4 (samples B1–4). Samples A1–3, in which 100% “Glera” DNA was added as a template, show a VIC signal, visualized as red dots (the “Glera” allele was marked with VIC), whereas the A4 sample (i.e., non-“Glera” variety DNA) gave a FAM signal (blue dots). Samples B1–3, in which 100% “Glera” DNA was added as a template, show a FAM signal, visualized as blue dots (the “Glera” allele was marked with FAM), whereas the B4 sample (i.e., non-“Glera” variety DNA) gave a VIC signal (red dots). All of the patterns have a yellow cloud due to DNA-empty wells.
Figure 4
Figure 4
Two-dimensional scatter graphs generated by chip digital PCR analysis of “Glera” and non-“Glera” mixed DNA (85% “Glera” and 15% non-“Glera”) with the two assays ID AN33KY7 (left panel) and ID AN49FJ4 (right panel). In the left panel, red dots stand for “Glera” and blue dots for non-“Glera” varieties. On the other hand, in the right panel, blue dots stand for “Glera” and red dots for non-“Glera” varieties. All of the patterns have a yellow cloud due to DNA-empty wells.
Figure 5
Figure 5
Two-dimensional scatter graphs generated by chip digital PCR analysis of DNA extracted from leaves (A1,B1), berries (A2,B2), and musts (A3,B3) of “Glera” with the two assays ID AN33KY7 (samples A1–3) and ID AN49FJ4 (samples B1–3).
Figure 6
Figure 6
Output of a dPCR analysis carried out with the assay ID AN49FJ4 on extracts from Prosecco commercial wine sample. Yellow dots stand for no amplification.
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