Flow Cytometry in Plant Research: A Success Story

Vrána, Jan; Cápal, Petr; Bednářová, Martina; Doležel, Jaroslav

doi:10.1007/978-3-642-41787-0_13

Cited by 24 publications

(17 citation statements)

References 222 publications

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“…Our protocol should also be applicable in these cases. For example, nuclei isolated from leaf tissues are the most common sample type in plant flow cytometry [Vrána et al, 2014]. Flow cytometric analysis has also been done with nuclei isolated from seeds [Matzk et al, 2000;Śliwinska et al, 2005], fruit tissues [Cheniclet et al, 2005], flower petals [Kudo 5.…”

Section: Resultsmentioning

confidence: 99%

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Proteomic Analysis of Barley Cell Nuclei Purified by Flow Sorting

Petrovská

Jeřábková

Chamrád

et al. 2014

Cytogenet Genome Res

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Many proteins are present in the nucleus; some are involved with its structural and functional organization, some with gene expression, and some with cell division. The plant nuclear proteome has not been well explored. Its characterization requires extraction methods which minimize both the artifactual alteration of the proteins and the extent of contamination with non-nuclear proteins. The conventional multi-step fractionation procedure is both laborious and prone to contamination. Here, we describe a single-step method based on flow sorting. The method allows the separation of G1, S and G2 phase nuclei and minimizes the risk of contamination by non-nuclear proteins. Preliminary results obtained using G1 phase cell nuclei from barley root tips indicate that flow sorting coupled with a protein/peptide separation and mass spectrometry will permit a comprehensive characterization of the plant nuclear proteome.

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Section: Resultsmentioning

confidence: 99%

“…Flow cytometry is particularly suited to identify nuclei which underwent one or more endoreduplication cycles [Vrána et al, 2014]. Thus, our method provides an attractive opportunity to study proteins of nuclei in differentiated endopolyploid cells at different levels of endoreduplication.…”

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confidence: 99%

Proteomic Analysis of Barley Cell Nuclei Purified by Flow Sorting

Petrovská

Jeřábková

Chamrád

et al. 2014

Cytogenet Genome Res

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“…Flow cytometry (FCM), commonly applied in multiparametric analysis in medicine, is most frequently used in plant sciences to establish only one parameter, the nuclear DNA content. Because of its advantages, such as high throughput, accuracy and resolution, negligible destructiveness to plants, as well as low operating cost per sample, it has replaced other methods of ploidy and/or genome size (GS) estimation in a large number of laboratories (Vrána et al, 2014). A few years ago, Bennett and Leitch (2011) reported that the proportion of all plant GS estimates using FCM was 85%, and, taking into account the recent popularity of this method, it is likely that this figure is significantly higher today.…”

Section: Introductionmentioning

confidence: 99%

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

Śliwińska

2018

Folia Horticulturae

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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.

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“…However, cytological counting of willow chromosome numbers is very difficult because of their high chromosome numbers and small chromosome size2829. Nowadays, flow cytometry (FCM) is widely adopted for determining the ploidy level of organisms30. Many studies have demonstrated FCM efficiency for ploidy level estimation for different plant species, including Salix species29.…”

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confidence: 99%

“…Many studies have demonstrated FCM efficiency for ploidy level estimation for different plant species, including Salix species29. However, for FCM analysis, sample preparation is complicated and laborious because of plant cell wall rigidity30; thus, it is not suitable for large-scale analyses.…”

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An analytical toolkit for polyploid willow discrimination

Guo

Hou

Yin

et al. 2016

Sci Rep

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Polyploid breeding is an important means for creating elite willow cultivars, and therefore provokes an active demand for discriminating the ploidy levels of natural willow stands. In this study, we established an analytical toolkit for polyploid willow identification by combining molecular markers and flow cytometry (FCM). A total of 10 single-copy fully informative SSRs were chosen for marker-aided selection based on a segregation test with a full-sib willow pedigree and a mutability test with a collection of natural willow stands. Aided by these molecular markers, we performed polyploid selection in two tree species and two shrub species of the genus Salix. The ploidy levels of the investigated samples were further examined using a flow cytometer. It was previously shown that results from marker-aided selection were consistent with those from FCM measurements. Based on ploidy level assessment in different willow species, it was found that tree willows were dominantly tetraploid, whereas shrub willows were most frequently diploid. With this analytical toolkit, polyploids can be rapidly screened from a large number of natural stands; thereafter, the exact ploidy levels of the polyploid candidates can be efficiently confirmed by FCM. This analytical toolkit will greatly enhance polyploid breeding programs for willows.

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Flow Cytometry in Plant Research: A Success Story

Cited by 24 publications

References 222 publications

Proteomic Analysis of Barley Cell Nuclei Purified by Flow Sorting

Proteomic Analysis of Barley Cell Nuclei Purified by Flow Sorting

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

An analytical toolkit for polyploid willow discrimination

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