High Resolution Melting DNA analysis for the traceability of plants and juices of blond and pigmented sweet orange

The economic relevance of sweet orange [Citrus sinensis (L.) Osbeck] raised the interest for the set-up of robust and scalable methods to assess the origin of plants and derived products. Molecular markers, such as single-nucleotide polymorphisms (SNPs), represent a robust tool for the univocal iden...

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Main Authors: Sebastiano Seminara, Stefania Bennici, Francesco Scollo, Mario Di Guardo, Pablo Aleza, Stefano La Malfa, Alessandra Gentile, Gaetano Distefano
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:Journal of Agriculture and Food Research
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Online Access:http://www.sciencedirect.com/science/article/pii/S2666154325001267
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author Sebastiano Seminara
Stefania Bennici
Francesco Scollo
Mario Di Guardo
Pablo Aleza
Stefano La Malfa
Alessandra Gentile
Gaetano Distefano
author_facet Sebastiano Seminara
Stefania Bennici
Francesco Scollo
Mario Di Guardo
Pablo Aleza
Stefano La Malfa
Alessandra Gentile
Gaetano Distefano
author_sort Sebastiano Seminara
collection DOAJ
description The economic relevance of sweet orange [Citrus sinensis (L.) Osbeck] raised the interest for the set-up of robust and scalable methods to assess the origin of plants and derived products. Molecular markers, such as single-nucleotide polymorphisms (SNPs), represent a robust tool for the univocal identification of the genetic origin of a specific cultivar (or group of accessions) with relevant application for traceability.In this work, the whole-genome sequencing of 29 accessions representing the four varietal groups of sweet orange (i.e.: Common, Navel, Acidless and Pigmented) allowed the in silico detection of unique SNPs to discriminate each group (group-specific SNPs) and ten pigmented orange accessions (genotype-specific SNPs).A subset of the SNPs detected in silico was then validated through a high-resolution melting assay (HRM) on additional 81 genotypes held in three ex-situ collections in Italy and Spain and on several matrices: leaves, hand‐squeezed and commercial fresh-squeezed juices, processed orange juices, and beverages. The HRM assay allowed the identification of 13 group-specific and 30 genotype-specific SNPs showing consistent results across the whole germplasm and the different food matrices tested. The HRM assay proved its efficacy also for the identification of juice blending with other sweet orange varieties (with a detection limit of 5 %). The identified unique SNPs represent a valuable tool to trace the varietal correspondence of plants and fruit-derived products. Their implementation with a scalable and robust HRM assay could be readily employed by growers and the food processing industry to ensure traceability along the chain.
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spelling doaj-art-b7c7500424364848b1b01c8b33bff2032025-08-20T03:25:08ZengElsevierJournal of Agriculture and Food Research2666-15432025-06-012110175510.1016/j.jafr.2025.101755High Resolution Melting DNA analysis for the traceability of plants and juices of blond and pigmented sweet orangeSebastiano Seminara0Stefania Bennici1Francesco Scollo2Mario Di Guardo3Pablo Aleza4Stefano La Malfa5Alessandra Gentile6Gaetano Distefano7Department of Agriculture, Food and Environment (Di3A), University of Catania, Via Santa Sofia 100, 95123, Catania, ItalyDepartment of Agriculture, Food and Environment (Di3A), University of Catania, Via Santa Sofia 100, 95123, Catania, ItalyDepartment of Agriculture, Food and Environment (Di3A), University of Catania, Via Santa Sofia 100, 95123, Catania, Italy; AgriUniTech, spin-off of the University of Catania, Via Valdisavoia 5, 95123, Catania, ItalyDepartment of Agriculture, Food and Environment (Di3A), University of Catania, Via Santa Sofia 100, 95123, Catania, Italy; Corresponding author.Centro de Citricultura y Producción Vegetal, Instituto Valenciano de Investigaciones Agrarias (IVIA), Valencia, 46113, SpainDepartment of Agriculture, Food and Environment (Di3A), University of Catania, Via Santa Sofia 100, 95123, Catania, Italy; Corresponding author.Department of Agriculture, Food and Environment (Di3A), University of Catania, Via Santa Sofia 100, 95123, Catania, ItalyDepartment of Agriculture, Food and Environment (Di3A), University of Catania, Via Santa Sofia 100, 95123, Catania, ItalyThe economic relevance of sweet orange [Citrus sinensis (L.) Osbeck] raised the interest for the set-up of robust and scalable methods to assess the origin of plants and derived products. Molecular markers, such as single-nucleotide polymorphisms (SNPs), represent a robust tool for the univocal identification of the genetic origin of a specific cultivar (or group of accessions) with relevant application for traceability.In this work, the whole-genome sequencing of 29 accessions representing the four varietal groups of sweet orange (i.e.: Common, Navel, Acidless and Pigmented) allowed the in silico detection of unique SNPs to discriminate each group (group-specific SNPs) and ten pigmented orange accessions (genotype-specific SNPs).A subset of the SNPs detected in silico was then validated through a high-resolution melting assay (HRM) on additional 81 genotypes held in three ex-situ collections in Italy and Spain and on several matrices: leaves, hand‐squeezed and commercial fresh-squeezed juices, processed orange juices, and beverages. The HRM assay allowed the identification of 13 group-specific and 30 genotype-specific SNPs showing consistent results across the whole germplasm and the different food matrices tested. The HRM assay proved its efficacy also for the identification of juice blending with other sweet orange varieties (with a detection limit of 5 %). The identified unique SNPs represent a valuable tool to trace the varietal correspondence of plants and fruit-derived products. Their implementation with a scalable and robust HRM assay could be readily employed by growers and the food processing industry to ensure traceability along the chain.http://www.sciencedirect.com/science/article/pii/S2666154325001267Citrus sinensisSingle-nucleotide polymorphism (SNP)HRMMislabellingWhole-genome sequencingJuice blending
spellingShingle Sebastiano Seminara
Stefania Bennici
Francesco Scollo
Mario Di Guardo
Pablo Aleza
Stefano La Malfa
Alessandra Gentile
Gaetano Distefano
High Resolution Melting DNA analysis for the traceability of plants and juices of blond and pigmented sweet orange
Journal of Agriculture and Food Research
Citrus sinensis
Single-nucleotide polymorphism (SNP)
HRM
Mislabelling
Whole-genome sequencing
Juice blending
title High Resolution Melting DNA analysis for the traceability of plants and juices of blond and pigmented sweet orange
title_full High Resolution Melting DNA analysis for the traceability of plants and juices of blond and pigmented sweet orange
title_fullStr High Resolution Melting DNA analysis for the traceability of plants and juices of blond and pigmented sweet orange
title_full_unstemmed High Resolution Melting DNA analysis for the traceability of plants and juices of blond and pigmented sweet orange
title_short High Resolution Melting DNA analysis for the traceability of plants and juices of blond and pigmented sweet orange
title_sort high resolution melting dna analysis for the traceability of plants and juices of blond and pigmented sweet orange
topic Citrus sinensis
Single-nucleotide polymorphism (SNP)
HRM
Mislabelling
Whole-genome sequencing
Juice blending
url http://www.sciencedirect.com/science/article/pii/S2666154325001267
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