Cereal barcoding by assessing intron length polymorphism of γ-tubulin genes

DOI: https://doi.org/10.7124/FEEO.v34.1625
Keywords: wheat, aegilops, barley, DNA barcoding, γ-tubulin

Abstract

Aim. The aim of the study was to obtain typical DNA profiles for common cereal plants using a method of assessing γ-tubulin intron length polymorphism and to evaluate the possibility of using this gene for plant species barcoding. Methods. Cereals, in particular wheat, aegilops, barley, and rice, have been studied. The method of γ-tubulin intron length polymorphism assessment was used. Amplified DNA fragments were separated via polyacrylamide gel electrophoresis under non-denaturating conditions and were visualized via silver nitrate staining. Results. The analyzed cereal species-specific DNA profiles of amplicons of the γ-tubulin gene introns were obtained, which made it possible to differentiate the species. Fingerprinting data was used for cluster analysis and dendrogram construction. Conclusions. The feasibility of using the proposed cereal barcoding method has been established. The studied plant DNA profiles can be effectively used in assessing the quality of food products such as flour.

References

Dinu M., Whittaker A., Pagliai, G., Benedettelli, S., Sofi F. Ancient wheat species and human health: Biochemical and clinical implications. J. Nutr. Biochem. 2018. Vol. 52. P. 1–9. doi: 10.1016/j.jnutbio.2017.09.001.

Li H., Xiao W., Tong T., Li Yo., Zhang M., Lin X., Zou X., Wu Q., Guo X. The specific DNA barcodes based on chloroplast genes for species identification of Orchidaceae plants. Scientific Reports. 2021. Vol. 11. P. 1424. doi: 10.1038/s41598-021-81087-w.

Techen N., Parveen I., Pan Z., Khan I. A. DNA barcoding of medicinal plant material for identification. Curr. Opin. Biotech. 2014. Vol. 25. P. 103–110. doi: 10.1016/j.copbio.2013.09.010.

Coissac E., Hollingsworth P. M., Lavergne S. From barcodes to genomes: Extending the concept of DNA barcoding. Mol. Ecol. 2016. Vol. 25. P. 423–1428. doi: 10.1111/mec.13549.

Ponzoni E., Morello L., Gianì S., Breviario D. Traceback identification of plant components in commercial compound feed through an oligonucleotide microarray based on tubulin intron polymorphism. Food Chemistry. 2014. Vol. 162. P. 72–80. doi: 10.1016/j.foodchem.2014.04.021.

Casazza A. P., Morcia C., Ponzoni E., Gavazzi F., Benedettelli S., Breviario D. A reliable assay for the detection of soft wheat adulteration in Italian pasta is based on the use of new DNA molecular markers capable of discriminating between Triticum aestivum and Triticum durum. J. Cereal Sci. 2012. Vol. 56. P. 733–740. doi: 10.1016/j.jcs.2012.08.015.

Braglia L., Gavazzi F., Giani S., Morello L., Breviario D. Tubulin-Based Polymorphism (TBP) in Plant Genotyping. Plant Genotyping: Methods and Protocols, Methods in Molecular Biology. Science+Business Media. 2023. Vol. 2638. doi: 10.1007/978-1-0716-3024-2_28.

Pirko Ya. V., Buy D. D., Postovoytova A. S., Rabokon A. M., Kalafat L. O., Blume Ya. B. Intron length polymorphism of γ-tubulin genes as a new approach to plant genotyping. Reports of the National Academy of Sciences of Ukraine. 2018. Vol. 12. P. 87–92. [in Ukrainian]

Green M. R., Sambrook J. Molecular cloning. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press, 2012. 1890 p.

Benbouza H., Jacquemin J-M., Baudoin J-P., Mergeai G. Optimization of a reliable, fast, cheap and sensitive silver staining method to detect SSR markers in polyacrylamide gel. Biotechnol. Agron. Soc. Environ. 2006. Vol. 10 (2). P. 77–81.

Herrera T. G., Duque D. P., Almeida I. P., Nunez G. T. et al. Assessment of genetic diversity in Venezuelan rice cultivars using simple sequence repeats markers. Electron. J. Biotechnol. 2008. Vol. 11 (5). P. 3–4. doi: 10.4067/S0717-34582008000500003.