Abstract

The wine sector requires quick and reliable methods for Vitis vinifera L. varietal identification. The number of V. vinifera varieties is estimated in about 5,000 worldwide. Single Nucleotide Polymorphisms (SNPs) represent the most basic and abundant form of genetic sequence variation, being adequate for varietal discrimination. The aim of this work was to develop DNA-based assays suitable to detect SNP variation in V. vinifera, allowing varietal discrimination. Genotyping by sequencing allowed the detection of eleven SNPs on two genes of the anthocyanin pathway, the flavanone 3-hydroxylase (F3H, EC: 1.14.11.9), and the leucoanthocyanidin dioxygenase (LDOX, EC 1.14.11.19; synonym anthocyanidin synthase, ANS) in twenty V. vinifera varieties. Three High Resolution Melting (HRM) assays were designed based on the sequencing information, discriminating five of the 20 varieties: Alicante Bouschet, Donzelinho Tinto, Merlot, Moscatel Galego and Tinta Roriz. Sanger sequencing of the HRM assay products confirmed the HRM profiles. Three probes, with different lengths and sequences, were used as bio-recognition elements in an optical biosensor platform based on a long period grating (LPG) fiber optic sensor. The label free platform detected a difference of a single SNP using genomic DNA samples. The two different platforms were successfully applied for grapevine varietal identification.

Details

Title
Alternative SNP detection platforms, HRM and biosensors, for varietal identification in Vitis vinifera L. using F3H and LDOX genes
Author
Gomes, Sónia 1 ; Castro, Cláudia 2 ; Barrias, Sara 2 ; Pereira, Leonor 1 ; Jorge, Pedro 3 ; Fernandes, José R 4   VIAFID ORCID Logo  ; Martins-Lopes, Paula 1   VIAFID ORCID Logo 

1  University of Trás-os-Montes and Alto Douro, P.O. Box 1013, Vila Real, Portugal; University of Lisboa, Faculty of Sciences, BioISI – Biosystems & Integrative Sciences Institute, Campo Grande, Lisboa, Portugal 
2  University of Trás-os-Montes and Alto Douro, P.O. Box 1013, Vila Real, Portugal 
3  INESC TEC, Rua do Campo Alegre n. 687, Porto, Portugal 
4  University of Trás-os-Montes and Alto Douro, P.O. Box 1013, Vila Real, Portugal; INESC TEC, Rua do Campo Alegre n. 687, Porto, Portugal; CQVR and Department of Physics, University of Trás-os-Montes & Alto Douro, Vila Real, Portugal 
Pages
1-12
Publication year
2018
Publication date
Apr 2018
Publisher
Nature Publishing Group
e-ISSN
20452322
Source type
Scholarly Journal
Language of publication
English
DOI
https://doi.org/10.1038/s41598-018-24158-9
ProQuest document ID
2023993703
Copyright
© 2018. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.