Utilization of plant profilins as DNA markers

Authors

  • Simona Čerteková
  • Adam Kováčik
  • Lucia Klongová
  • Jana Žiarovská SPU v Nitre

Keywords:

profilins, PCR, length polymorphism, DNA marker

Abstract

This study aims to explore the possibility of utilizing markers derived from profilin sequences for the genomic fingerprinting of plant organisms. Profilins are a category of small actin-binding proteins that are present in all eukaryotic cells. Despite profilins being ubiquitous, some forms are also clinically relevant because of their ability to induce allergic responses in sensitized individuals. We conducted a PCR analysis on DNA samples obtained from 11 vegetable species (Brassica oleracea L. in 4 varieties) using two types of primers: non-degenerate and degenerate. In the case of degenerate primers, a total of 51 amplification products of different lengths were recorded, while their average amount was in the range of 7-8 amplicons for one species. The most frequently occurring product was the product with a length of 249 bp. A lower degree of polymorphism was noted when non-degenerate primers were used. The total number of different products created by amplification using non-degenerate primers was 33 and there was an average of 5 amplicons in one sample. As indicated by the findings, implementation of degenerate primers was more suitable for genomic fingerprinting based on profilin sequences in vegetable species, as it led to a higher level of variability in the amplification profiles of distinct species. It can be stated that amplification based on profilin sequences proved to be sufficient in its versatility and efficiency in generating variable-length polymorphism of PCR products.

References

Breiteneder, H., & Ebner, C. (2000). Molecular and biochemical classification of plant-derived food allergens. Journal of Allergy and Clinical Immunology, 106(1), 27–36. https://doi.org/10.1067/mai.2000.106929

Carlsson, L. et al. (1977). Actin polymerizability is influenced by profilin, a low molecular weight protein in non-muscle cells. Journal of Molecular Biology, 115(3), 465–483.

Collard, B. C. Y., & Mackill, D. J. (2009). Conserved DNA-Derived Polymorphism (CDDP): A Simple and Novel Method for Generating DNA Markers in Plants. Plant Molecular Biology Reporter, 27(4), 558–562. https://doi.org/10.1007/s11105-009-0118-z

Davey, R. J., & Moens, P. D. (2020). Profilin: Many facets of a small protein. Biophysical Reviews, 12(4), 827–849. https://doi.org/10.1007/s12551-020-00723-3

Harris, C. R. et al. (2020). Array programming with NumPy. Nature, 585(7825), Art. 7825. https://doi.org/10.1038/s41586-020-2649-2

Hasan, N. et al. (2021). Recent advancements in molecular marker-assisted selection and applications in plant breeding programmes. Journal of Genetic Engineering and Biotechnology, 19(1), 128. https://doi.org/10.1186/s43141-021-00231-1

Huang, S. et al. (1996). The Arabidopsis profilin gene family. Evidence for an ancient split between constitutive and pollen-specific profilin genes. Plant Physiology, 111(1), 115–126.

Kandasamy, M.K. et al. (2002). Plant profilin isovariants are distinctly regulated in vegetative and reproductive tissues. Cell motility. https://doi.org/10.1002/cm.10029

Klongová, L. et al. (2021). Utilization of specific primers in legume allergens based polymorphism screening. Science, Technology and Innovation, 13(2), Art. 2. https://doi.org/10.5604/01.3001.0015.5431

Kwok, S. et al. (1994). A guide to the design and use of mismatched and degenerate primers. Genome Research, 3(4), S39–S47. https://genome.cshlp.org/content/3/4/S39.long

Linhart, C., & Shamir, R. (2005). The degenerate primer design problem: Theory and applications. Journal of Computational Biology: A Journal of Computational Molecular Cell Biology, 12(4), 431–456. https://doi.org/10.1089/cmb.2005.12.431

Maeda, K. et al. (2020). Actin Filament Disruption Alters Phragmoplast Microtubule Dynamics during the Initial Phase of Plant Cytokinesis. Plant and Cell Physiology, 61(3), 445–456. https://doi.org/10.1093/pcp/pcaa003

Mahoney, N. M. et al. (1997). Structure of the profilin-poly-L-proline complex involved in morphogenesis and cytoskeletal regulation. Nature Structural & Molecular Biology, 4(11), 953–960. https://doi.org/10.1038/nsb1197-953

Mahoney, N. M. et al. (1999). Profilin binds proline-rich ligands in two distinct amide backbone orientations. Nature Structural Biology, 6(7), Art. 7. https://doi.org/10.1038/10722

Masojć, P. (2002). The application of molecular markers in the process of selection. Cellular & Molecular Biology Letters, 7(2A), 499–509.

McKinney, W. (2010). Data Structures for Statistical Computing in Python. 56–61. https://doi.org/10.25080/Majora-92bf1922-00a

Mills, E. N. C., & Shewry, P. R. (Ed.). (2004). Plant food allergens. Blackwell Science.

Qu, X. et al. (2015). Organization and regulation of the actin cytoskeleton in the pollen tube. Frontiers in Plant Science, 5. https://doi.org/10.3389/fpls.2014.00786

Pandey, D.K. and Chaudhary, B. (2020). Evolution of Functional Diversity Among Actin-Binding Profilin Genes in Land Plants. Frontiers in Cell and Developmental Biology. https://doi.org/10.3389/fcell.2020.588689

Ramachandran, S. et al. (2000). Profilin plays a role in cell elongation, cell shape maintenance, and flowering in Arabidopsis. Plant Physiology, 124, 1637–1647. https://doi.org/10.1104/pp.124.4.1637

Rodríguez del Río, P. et al. (2018). Profilin, a Change in the Paradigm. Journal of Investigational Allergology and Clinical Immunology, 28(1), 1–12. https://doi.org/10.18176/jiaci.0193

Roldán-Ruiz, I. et al. (2000). AFLP markers reveal high polymorphic rates in ryegrasses (Lolium spp.). Molecular Breeding, 6(2), 125–134. https://doi.org/10.1023/A:1009680614564

Ruiz‐Hornillos, J. et al. (2020). Profilin is a marker of severity in allergic respiratory diseases. Allergy, 75(4), 853–861. https://doi.org/10.1111/all.14140

Sankian, M. et al. (2005). Sequence homology: A poor predictive value for profilins cross-reactivity. Clinical and molecular allergy : CMA, 3, 13. https://doi.org/10.1186/1476-7961-3-13

Schlüter, K., Jockusch, B. M., & Rothkegel, M. (1997). Profilins as regulators of actin dynamics. Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, 1359(2), 97–109. https://doi.org/10.1016/S0167-4889(97)00100-6

Sokal, R., & Michener, C. (1902). A statistical method for evaluating systematic relationships. http://archive.org/details/cbarchive_33927_astatisticalmethodforevaluatin1902

Tessier, C. et al. (1999). Optimization of the choice of molecular markers for varietal identification in Vitis vinifera L. Theoretical and Applied Genetics, 98, 171–177. https://doi.org/10.1007/s001220051054

Tordesillas, L. et al. (2009). Characterization of IgE epitopes of Cuc m 2, the major melon allergen, and their role in cross-reactivity with pollen profilins: B epitopes of melon profiling. Clinical & Experimental Allergy, 40(1), 174–181. https://doi.org/10.1111/j.1365-2222.2009.03401.x

Valenta, R. et al. (1991). Identification of Profilin as a Novel Pollen Allergen; IgE Autoreactivity in Sensitized Individuals. Science, 253(5019), 557–560. https://doi.org/10.1126/science.1857985

Valenta, R. et al. (1993). Identification of profilin as an actin-binding protein in higher plants. Journal of Biological Chemistry, 268(30), 22777–22781. https://doi.org/10.1016/S0021-9258(18)41594-3

Virtanen, P. et al. (2020). SciPy 1.0: Fundamental algorithms for scientific computing in Python. Nature Methods, 17(3), 261–272. https://doi.org/10.1038/s41592-019-0686-2

Wallner, M. et al. (2017). The Concept of Pollen Panallergens: Profilins and Polcalcins. V J. Kleine-Tebbe & T. Jakob (Ed.), Molecular Allergy Diagnostics: Innovation for a Better Patient Management (s. 43–56). Springer International Publishing. https://doi.org/10.1007/978-3-319-42499-6_3

Waskom, M. L. (2021). seaborn: Statistical data visualization. Journal of Open Source Software, 6(60), 3021. https://doi.org/10.21105/joss.03021

Yamashita, H. et al. (2011). Chloroplast actin filaments organize meshwork on the photorelocated chloroplasts in the moss Physcomitrella patens. Planta, 233(2), 357–368. https://doi.org/10.1007/s00425-010-1299-2

Zamiesková, L., & Žiarovská, J. (2021). Variability of DNA based amplicon profiles generated by Bet v 1 homologous among different vegetable species. Acta Fytotechnica et Zootechnica, 24. https://doi.org/10.15414/afz.2021.24.mi-apa.1-6

Žiarovská, J., & Urbanová, L. (2022). Utilization of Bet v 1 homologs based amplified profile (BBAP) variability in allergenic plants fingerprinting. Biologia, 77(2), 517–523. https://doi.org/10.1007/s11756-021-00943-2

Downloads

Published

2023-10-16

Issue

Section

Plant Science