MOLECULAR TOOLS FOR FINGERPRINTING OF WILD GRASS

Wild grasses represent a source of genetic biodiversity for the introgression of agronomical traits of interest into their cultivated relatives, and contribute a relevant part of livestock diets. Characterization of their genetic material is often difficult due to poor knowledge of genomes, complex taxonomic relationships and difficulty in correct classification due to elevated phenotypic variation. A suitable universal marker for plants is still lacking and genomic barcoding according to CBOL relies on rbcL plus matK gene sequencing. TBP is a PCR-based ILP molecular marker, targeting introns of the beta-tubulin gene family. Amplicons are separated by PAGE or by capillary electrophoresis giving reproducible fingerprinting. This codominant marker is suitable for fingerprinting at the species level, but is also useful for lower taxonomic levels and for elucidating genotype relationships. A case study of genetic diversity among 17 different Phalaris species is shown. CE-TBP has also been successfully applied to the authentication of forage composition within the FILAGRO project. TBP profiles from temporary grasslands (first and second cut) report, in a single analysis, the sum of specie-specific fingerprinting. Analytical results were further confirmed using the trnL intron as a marker.