The Application of Flow Cytometry for Estimating Genome Size, Ploidy Level Endopolyploidy, and Reproductive Modes in Plants

Pellicer, Jaume; Powell, Robyn F.; Leitch, Ilia J.

doi:10.1007/978-1-0716-0997-2_17

Cited by 93 publications

(33 citation statements)

References 75 publications

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“…In total, 43 individuals were sampled, representing two populations of each species from Ports Massif, and one population of C. podospermifolia from Cardó Massif. We collected one leaf per individual, since approximately only 1 cm 2 of leaf material is necessary to carry out genome size assessments by flow cytometry [ 35 ]. Herbarium vouchers from previous collections in the same area by López-Pujol et al [ 1 ] are deposited in herbarium BC (Botanical Institute of Barcelona).…”

Section: Methodsmentioning

confidence: 99%

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Detecting Introgressed Populations in the Iberian Endemic Centaurea podospermifolia through Genome Size

Pellicer

López‐Pujol

Aixarch

et al. 2021

Plants

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Based on results from previous studies, populations of the Iberian endemic Centaurea podospermifolia north of the Ebro River are considered genetically pure, while those southward are introgressed, with genetic input from C. cephalariifolia. This phenomenon is particularly relevant, especially given both the endangered and protected status for the species, which can have consequences in how to best apply conservation strategies to maintain genetic resources in the species. The main goal of this study was to evaluate whether genome size assessments using flow cytometry can help distinguishing between pure, hybrid and introgressed populations, and hence become a powerful and cost-effective tool to complement comprehensive population genetic surveys. The results indicate that there are significant genome size differences between populations of C. podospermifolia, which are coincident with previous considerations of pure and introgressed populations. Given the simplicity and reproducibility of this technique, flow cytometry could become an effective tool for monitoring pure populations of this species and, indeed, become an integral part of the management plans that are mandatory for listed taxa.

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Section: Methodsmentioning

confidence: 99%

“…Fuss. ‘Champion Moss Curled’ (2C = 4.50 pg [ 38 ]) as calibration standard and the general purpose buffer GPB [ 39 ] supplemented with 3% PVP-40 [ 35 ]. The number of individuals analysed per population is depicted in Table 1 .…”

Section: Methodsmentioning

confidence: 99%

Detecting Introgressed Populations in the Iberian Endemic Centaurea podospermifolia through Genome Size

Pellicer

López‐Pujol

Aixarch

et al. 2021

Plants

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“…Genome size measurements. Nuclear DNA content of Euphrasia samples was estimated by flow cytometry using propidium iodide (PI) stained nuclei, following the one step method (see Pellicer et al, 2021). Briefly, for each Euphrasia accession, two small leaves (c. 1-2 cm) were chopped together with the internal standard Oryza sativa 'IR36' (1C = 0.5 pg; Bennett & Smith, 1991) using a new razor blade, in a petri dish containing 1 mL of 'general purpose isolation buffer' (GPB; Loureiro et al, 2007), supplemented with 3% PVP-40 and 0.4…”

Section: Methodsmentioning

confidence: 99%

“…The above types of genetic variation can be subsumed under the term presence/absence variants (PAVs), a type of structural genomic variation, and may be detectable by methods for estimating GS, such as flow cytometry. Modern protocols using flow cytometry with appropriate reference standards, and following best practice approaches, can be accurate and highly precise (Greilhuber et al, 2007;Pellicer et al, 2021) and reveal genuine intraspecific variation that can be confirmed by genome sequencing. Such sequencing has also been used to reveal that repeat differences can be useful genetic markers, including microsatellites and AFLPs.…”

Section: Introductionmentioning

confidence: 99%

The nature of intraspecific genome size variation in taxonomically complex eyebrights

Becher

et al. 2021

Preprint

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Genome size (GS) is a key trait related to morphology, life history, and evolvability. Although GS is, by definition, affected by presence/absence variants (PAVs), which are ubiquitous in population sequencing studies, GS is often treated as an intrinsic property of a species. Here, we studied intra- and interspecific GS variation in taxonomically complex British eyebrights (Euphrasia). We generated GS data for 192 individuals of diploid and tetraploid Euphrasia and analysed GS variation in relation to ploidy, taxonomy, population affiliation, and geography. We further compared the genomic repeat content of 30 samples. We found considerable genuine intraspecific GS variation, and observed isolation-by-distance for GS in outcrossing diploids. Tetraploid Euphrasia showed contrasting patterns, with GS increasing with latitude in outcrossing Euphrasia arctica, but little GS variation in the highly selfing Euphrasia micrantha. Interspecific differences in GS genomic repeat percentages were small. We show the utility of treating GS as the outcome of polygenic variation. Like other types of genetic variation, such as single nucleotide polymorphisms, GS variation may be increased through hybridisation and population subdivision. In addition to selection on associated traits, GS is predicted to be affected indirectly by selection due to pleiotropy of the underlying PAVs.

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“…Flow cytometry is a rapid and efficient method for nuclear DNA content estimation in thousands of cells and has been widely used for the rapid screening of ploidy levels in regenerated plants [13,14]. Additionally, flow cytometry was used for several different purposes, including the determination of the amount of species-specific DNA, the analysis of cell cycle activity in different tissues, and the measurement of endopolyploidization levels [15][16][17].…”

Section: Introductionmentioning

confidence: 99%

Genome Estimation and Phytochemical Compound Identification in the Leaves and Callus of Abrus precatorius: A Locally Endangered Plant from the Flora of Saudi Arabia

Al‐Qurainy

Tarroum

Khan

et al. 2022

Plants

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Abrus precatorius is considered to be a valuable source of natural products for the development of drugs against various diseases. Herein, the genome size and phytochemical compounds in the leaves and callus of A. precatorius were evaluated. The endangered A. precatorius was collected from the Al-Baha mountains, Saudi Arabia and identified based on the phylogenetic analysis of a DNA sequence amplified by ITS1 and ITS4 primers. The callus was induced by the culture of stem explants onto Murashige and Skoog medium (MS) supplemented with various combinations of 2,4-dichlorophenoxyacetic acid (2,4D) and 6-Benzylaminopurine (BAP). The callus with the highest fresh weight (2.03 g) was obtained in the medium containing 0.5µM BA and 5 µM 2,4-D after 8 weeks of culture; thus, the callus of this combination was selected for the genome estimation and phytochemical compound extraction. The genetic stability of the leaves from the donor as well as in the regenerated callus was analyzed by flow cytometry with optimized tomato (2C = 1.96 pg) as an external reference standard. The 2C DNA content was estimated to 1.810 pg ± 0.008 and 1.813 pg ± 0.004 for the leaves and callus, respectively. Then, the total phenol and total flavonoid contents in the methanol extract of the callus and leaves were measured using a spectrophotometer and the High-performance liquid chromatography (HPLC ) methods. The results showed that the methanolic extract of the leaves was higher in total phenols and total flavonoids than the callus extract. Finally, the extracts of callus and leaves were analyzed for phytochemical compound through the Gas chromatography and Mass spectroscopy (GC-MS). A total of 22 and 28 compounds were detected in the callus and leaves, respectively. The comparative analysis showed that 12 compounds of the secondary metabolites were present in both extracts.

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The Application of Flow Cytometry for Estimating Genome Size, Ploidy Level Endopolyploidy, and Reproductive Modes in Plants

Cited by 93 publications

References 75 publications

Detecting Introgressed Populations in the Iberian Endemic Centaurea podospermifolia through Genome Size

Detecting Introgressed Populations in the Iberian Endemic Centaurea podospermifolia through Genome Size

The nature of intraspecific genome size variation in taxonomically complex eyebrights

Genome Estimation and Phytochemical Compound Identification in the Leaves and Callus of Abrus precatorius: A Locally Endangered Plant from the Flora of Saudi Arabia

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