Microsatellite Markers in Olives (Olea europaea L.): Utility in the Cataloging of Germplasm, Food Authenticity and Traceability Studies

The olive fruit, a symbol of Mediterranean diets, is a rich source of antioxidants and oleic acid (55–83%). Olive genetic resources, including cultivated olives (cultivars), wild olives as well as related subspecies, are distributed widely across the Mediterranean region and other countries. Certain cultivars have a high commercial demand and economical value due to the differentiating organoleptic characteristics. This might result in economically motivated fraudulent practices and adulteration. Hence, tools to ensure the authenticity of constituent olive cultivars are crucial, and this can be achieved accurately through DNA-based methods. The present review outlines the applications of microsatellite markers, one of the most extensively used types of molecular markers in olive species, particularly referring to the use of these DNA-based markers in cataloging the vast olive germplasm, leading to identification and authentication of the cultivars. Emphasis has been given on the need to adopt a uniform platform where global molecular information pertaining to the details of available markers, cultivar-specific genotyping profiles (their synonyms or homonyms) and the comparative profiles of oil and reference leaf samples is accessible to researchers. The challenges of working with microsatellite markers and efforts underway, mainly advancements in genotyping methods which can be effectively incorporated in olive oil varietal testing, are also provided. Such efforts will pave the way for the development of more robust microsatellite marker-based olive agri-food authentication platforms

How to choose a good marker to analyze the olive germplasm (Olea europaea l.) and derived products

The olive tree (Olea europaea L.) is one of the most cultivated crops in the Mediterranean basin. Its economic importance is mainly due to the intense production of table olives and oil. Cultivated varieties are characterized by high morphological and genetic variability and present a large number of synonyms and homonyms. This necessitates the introduction of a rapid and accurate system for varietal identification. In the past, the recognition of olive cultivars was based solely on analysis of the morphological traits, however, these are highly influenced by environmental conditions. Therefore, over the years, several methods based on DNA analysis were developed, allowing a more accurate and reliable varietal identification. This review aims to investigate the evolving history of olive tree characterization approaches, starting from the earlier morphological methods to the latest technologies based on molecular markers, focusing on the main applications of each approach. Furthermore, we discuss the impact of the advent of next generation sequencing and the recent sequencing of the olive genome on the strategies used for the development of new molecular markers

Autenticación de aceite de oliva mediante análisis de ADN

Olive oil, which has been produced mainly in the Mediterranean area since the ancient times, has a high nutritional value linked to many health benefits. Extra virgin, which is the purest form of olive oil, has excellent quality and premium prices. Many cases of adulteration and fraud necessitate the development of reliable and accurate methods for olive oil authentication. DNA-based methods analyze the residual DNA extracted from olive oil and use molecular markers for genetic identification of different species, subspecies or cultivars because these markers act as signs which reflect distinct genetic profiles. This study reviews the process by which DNA from olive oil is extracted and analyzed by the most recently used markers in the authentication of olive oil, such as Simple Sequence Repeats (SSR) or microsatellites and the single nucleotide polymorphisms (SNPs). Methods of analysis such as qPCR and digital PCR are also discussed with a special emphasis placed on the method of High-Resolution Melting (HRM), a post-polymerase chain reaction method, which enables rapid, high performing identification of genetic variants in the DNA regions of interest without sequencing, and may differentiate very similar cultivars which differ in only one nucleotide in a specific locus.El aceite de oliva, producido principal­mente en el área mediterránea desde la antigüedad, tiene un alto valor nutricional vinculado a muchos benefi­cios para la salud. El aceite de oliva virgen extra, que es la forma más pura de aceite de oliva, tiene una excelente calidad y precios premium. Muchos casos de adulteraciones y fraudes requieren el desarrollo de métodos fiables y precisos para la autenticación del aceite de oliva. Los métodos basados en el ADN analizan el ADN residual extraído del aceite de oliva y usan marcadores moleculares para la identificación genética de diferentes espe­cies, subespecies o cultivares, porque estos marcadores actúan como signos que producen perfiles genéticos distintos. Este estudio revisa el proceso mediante el cual el ADN del aceite de oliva es extraído y analizado por los marcadores utilizados más recientemente en la autenticación del aceite de oliva, como las repeticiones de secuencia simple (SSR) o los micro satélites y los polimorfismos de un solo nucleótido (SNP). Los métodos de análisis como qPCR y PCR digital también se analizan haciendo especial énfasis en el método de fusión de alta resolución (HRM), un método de reacción en cadena posterior a la polimerasa, que permite la identificación rápida y con alto rendimiento de variantes genéticas en regiones del ADN de interés sin secuenciación, y pueden diferenciar cultivares muy similares, que difieren en un solo nucleótido, en un lugar específico

DNA-Based Approaches for Traceability and Authentication of Olive Oil

Authentication and traceability of extra virgin olive oil is a challenging research task due to the complexity of fraudulent practices. Various chemical and biochemical techniques have been developed for determining the authenticity of olive oil and in recent years non-conventional methods based on DNA analysis have gained attention, due to high specificity, sensitivity and reliability. DNA analyses have very high discriminating power because ultimately the unique identity of a variety or species is to a great extent genetically dependent. Polymorphisms are genetic variations which refer to the variation in populations or species. Molecular markers provide information on genetic variations and are valuable tools to determine olive oil authenticity. Recently several DNA-based methods have been developed to authenticate olive oil, since analysis of the residual oil DNA with the use of molecular markers can lead to the identification of the variety or the plant species from which it was extracted. The aim of this chapter is to provide an overview of the current trends and critical issues on DNA-targeted approaches used for traceability and authenticity of olive oil. This is considered a rapidly expanding field with significant challenges and prospects which shall be discussed thoroughly

Recovery, assessment, and molecular characterization of minor olive genotypes in Tunisia

Olive is one of the oldest cultivated species in the Mediterranean Basin, including Tunisia, where it has a wide diversity, with more than 200 cultivars, of both wild and feral forms. Many minor cultivars are still present in marginal areas of Tunisia, where they are maintained by farmers in small local groves, but they are poorly characterized and evaluated. In order to recover this neglected germplasm, surveys were conducted in different areas, and 31 genotypes were collected, molecularly characterized with 12 nuclear microsatellite (simple sequence repeat (SSR)) markers, and compared with 26 reference cultivars present in the Tunisian National Olive collection. The analysis revealed an overall high genetic diversity of this olive’s germplasm, but also discovered the presence of synonymies and homonymies among the commercialized varieties. The structure analysis showed the presence of different gene pools in the analyzed germplasm. In particular, the marginal germplasm from Ras Jbal and Azmour is characterized by gene pools not present in commercial (Nurseries) varieties, pointing out the very narrow genetic base of the commercialized olive material in Tunisia, and the need to broaden it to avoid the risk of genetic erosion of this species in this country