Tubulin-Based DNA Barcode: Principle and Applications to Complex Food Matrices

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.

Genomic Profiling: The Strengths and Limitations of Chloroplast Genome-Based Plant Variety Authentication

“…Genomic profiling creates a molecular fingerprint of an organism based on its DNA . DNA is stored in each cell, and it contains the unique genetic information on the entire organism .…”

“…Genomic profiling creates a molecular fingerprint of an organism based on its DNA. 1 DNA is stored in each cell, and it contains the unique genetic information on the entire organism. 2 The whole DNA of an organism is defined as its genome, and the closer two organisms are related, the more similar are their genomes.…”

DNA Barcodes for Accurate Identification of Selected Medicinal Plants (Caryophyllales): Toward Barcoding Flowering Plants of the United Arab Emirates

“…Considerably, it is now a well-known fact that the combination of the two plastid markers, ribulose 1,5-bisphosphate carboxylase gene (rbcL) and maturase K (matK), that were accepted as the core barcoding regions [33], do not grant a suitable coverage of plant species. Thus, they must often be implemented along with the other hypervariable sequences, such as nuclear ITS or the plastid interspacer region trnH-psbA [88].…”

Prey partitioning between sympatric wild carnivores revealed by DNA metabarcoding: a case study on wolf (Canis lupus) and coyote (Canis latrans) in northeastern Washington

“…These methods also tend to underestimate the proportion of prey species whose remains are not found (completely digested or not consumed at all) (Deagle, Kirkwood, & Jarman, 2009). Earlier molecular attempts use a conventional genotyping methodology, involving multiplex PCR followed by fragment size determination with capillary electrophoresis and known sized allelic ladders (Morello, Braglia, Gavazzi, Gianì, & Breviario, 2019). Though very affordable, this method does not account for any sequence-based differences between fragments of the same size.…”