Abstract

The DNA polymorphism diffusely present in the introns of the members of the Eukaryotic beta-tubulin gene families, can be conveniently used to establish a DNA barcoding method, named tubulin-based polymorphism (TBP), that can reliably assign specific genomic fingerprintings to any plant or/and animal species. Similarly, many plant varieties can also be barcoded by TBP. The method is based on a simple cell biology concept that finds a conveniently exploitable molecular basis. It does not depend on DNA sequencing as the most classically established DNA barcode strategies. Successful applications, diversified for the different target sequences or experimental purposes, have been reported in many different plant species and, of late, a new a version applicable to animal species, including fishes, has been developed. Also, the TBP method is currently used for the genetic authentication of plant material and derived food products. Due to the use of a couple of universal primer pairs, specific for plant and animal organisms, respectively, it is effective in metabarcoding a complex matrix allowing an easy and rapid recognition of the different species present in a mixture. A simple, dedicated database made up by the genomic profile of reference materials is also part of the analytical procedure. Here we will provide some example of the TBP application and will discuss its features and uses in comparison with the DNA sequencing-based methods.

Highlights

  • This contribution is inspired by few simple concepts that, applied to the food sector, have found ever increasing and worldwide attention since 1998 when we deposited, for patent assignment, the first version of the tubulin-based polymorphism (TBP) method, developed for plant species and varieties recognition

  • These markers are based on the generation of random fragments and, widely applied for genotyping at subspecies level and QTL mapping, they were not developed for discrimination at species-level

  • The meaning of the word allele in this case is limited to the presence of indels, since nucleotide sequence polymorphism cannot be detected

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Summary

Introduction

This contribution is inspired by few simple concepts that, applied to the food sector, have found ever increasing and worldwide attention since 1998 when we deposited, for patent assignment, the first version of the tubulin-based polymorphism (TBP) method, developed for plant species and varieties recognition. These concepts may be summarized as follows:. We will give an overview of DNA barcoding applications to food in comparison to fingerprinting techniques based on DNA fragment analysis

Barcoding by Sequencing
General Issues about Metabarcoding
Barcoding by Fragment Analysis
Tubulin-Based
Reference Database and Bionformatic Needs
Plant Species Identification and Raw Material Purity
Feed Composition by TBP Metabarcoding
Meat and Fish Food Authentication
Patents
Findings
25 February
Full Text
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