Optimization of sample preparation technique and the possibility of using different types of plant material of two species of the genus Colchicum (Colchicaceae) in flow cytometry

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Abstract

The article presents the results of testing and optimization of the sample preparation technique for fl ow cytometric analysis of two species that have not been previously studied in this aspect: Colchicum bulbocodium subsp. versicolor and C. laetum. We are using optimal LB01 buff er with added sodium thiosulfate, as internal standards we take Petroselinum crispum and Secale cereale. For the fi rst time, the DNA content in living samples of C. bulbocodium subsp. versicolor (2C = 8.049 pg) and C. laetum (2C = 5.412 pg) from natural habitats in the territory of the Russian Federation was determined. We have proven that not only living material (sprouts), but also seeds can be used as material for studying the genome size by fl ow cytometry of two species of the genus Colchicum. The main thing is to take into account their feature – the number of events corresponding to the embryo is much less than the number of endosperm cells. We do not recommend using herbarium samples of Colchicum as material for DNA content analysis, as their values were much overestimated in comparison with sprouts. The data we obtained complement and expand our understanding of the DNA level within the genus Colchicum.

About the authors

Alena Sergeevna Parhomenko

Saratov State University

83, Astrakhanskaya str., Saratov, 410012, Russia

Kristina S. Pavlenko

Saratov State University

83, Astrakhanskaya str., Saratov, 410012, Russia

Vladimir S. Epifanov

Saratov State University

ORCID iD: 0009-0007-5229-8151
83, Astrakhanskaya str., Saratov, 410012, Russia

Savelii F. Efimenko

Saratov State University

ORCID iD: 0009-0007-7828-9499
83, Astrakhanskaya str., Saratov, 410012, Russia

Lyudmila V. Grebenyuk

Saratov State University

83, Astrakhanskaya str., Saratov, 410012, Russia

Mikhail V. Skaptsov

Altai State University, South-Siberian Botanical Garden

61 Lenina Pr., Barnaul 656049, Russia

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