Flow cytometry – a modern method for exploring genome size and nuclear DNA synthesis in horticultural and medicinal plant species

Abstract

Flow cytometry (FCM) has been used for plant DNA content estimation since the 1980s; however, presently, the number of laboratories equipped with flow cytometers has significantly increased and these are used extensively not only for research but also in plant breeding (especially polyploid and hybrid breeding) and seed production and technology to establish seed maturity, quality and advancement of germination. A broad spectrum of horticultural and medicinal species has been analyzed using this technique, and various FCM applications are presented in the present review. The most common application is genome size and ploidy estimation, but FCM is also very convenient for establishing cell cycle activity and endoreduplication intensity in different plant organs and tissues. It can be used to analyze plant material grown in a greenhouse/field as well asin vitro. Due to somaclonal variation, plant material grown in tissue culture is especially unstable in its DNA content and, therefore, FCM analysis is strongly recommended. Horticultural species are often used as internal standards in genome size estimation and as models for cytometrically studied cytotoxic/anticancer/allelopathic effects of different compounds. With the growing interest in genome modification, increased application of FCM is foreseen.

Various in vitro systems of Ragged Robin (Lychnis flos-cuculi L.): a new potential source of phytoecdysteroids?

“…Methods based on the flow cytometry are applied to ascertain that in vitro culture conditions do not affect the cytogenetic stability of a plant as well as the occurrence of somaclonal variations. DNA content in several micropropagated species was evaluated after growing in in vitro conditions (Thiem and Sliwinska 2003;Thiem et al 2013;Sliwinska 2018). In the presented studies, the stability of genome size (about 5.7-5.8 pg/2C) in different plant materials indicates that in vitro cultures of L. flos-cuculi are a viable, homogenous, alternative, and renewable source of plant material.…”

Assessment of somaclonal variation in indirect morphogenesis-derived plants of Arracacia xanthorrhiza

“…The chromosome number in the somatic cells of the control plant material was 2n=44, which corresponds to the tetraploid chromosomal level (4x) (Figure 2). These results indicate that, despite the general presumption that plant material grown in tissue culture shows especially unstable DNA content (Sliwinska, 2018), arracacha plants proved to be stable and not susceptible to ploidy level changes during in vitro cultivation. These results are similar to those of Viehmannova et al (2014), who studied another Andean root crop, S. sonchifolius, and detected somaclonal variation at the point mutation level in A, inter-simple sequence repeat profile of in vitro regenerants and a control plant using the UBC836 primer, where R1-R15 are 15 randomly chosen regenerants, C is the control plant, L is the ladder, and arrows indicate the position of the polymorphic bands; B, representative histograms showing relative fluorescence intensity of the control plant (on the left) and of a randomly chosen in vitro regenerant (on the right), with the peak of the internal standard, Bellis perennis, indicated with an asterisk; and C, chromosome counts in root tips of arracacha (2n = 4x = 44).…”

Genome Size Diversity in Rare, Endangered, and Protected Orchids in Poland

“…Analysis of nuclear DNA content using flow cytometry is reliable, fast, relatively cheap compared to the molecular methods and an attractive alternative to microspectrophotometry. Moreover, for the analysis only a small amount of tissue is needed, which is important in the case of valuable and/or protected specimens [ 24 , 25 ].…”