The Application of Flow Cytometry for Estimating Genome Size and Ploidy Level in Plants

Aucuba have been cultivated for centuries and are valued as adaptable, broad-leaved, evergreen shrubs that also can have attractive, spotted variegations on the foliage. Improved understanding of the cytogenetics and heritability of specific traits, for specific clones and cultivars, can provide basic information to help facilitate the breeding and improvement of aucuba. The objectives of this study were to determine ploidy level and relative genome size of a diverse collection of species and cultivars of aucuba using flow cytometry and cytology and to make additional observations on heritability of spotted leaf variegation. Chromosome counts were 2n = 2x = 16 for Aucuba chinensis (A. omeiensis), 2n = 4x = 32 for A. japonica ‘Rozannie’, and 2n = 6x = 48 for A. sp. ‘Hosoba’. Relative 2C genome size for the 57 taxa varied from 13.8 pg for A. obcordata to 42.0 pg for A. ‘Hosoba’ and fell within three discrete groups consistent with cytotype. Genome size for diploid taxa (A. chinensis and A. obcordata) ranged from 13.8 to 21.0 pg, tetraploids (A. himalaica var. oblanceolata, A. japonica, and A. japonica var. borealis) ranged from 28.8 to 31.2 pg, and the first-ever reported hexaploids (A. ‘Hosoba’ and A. sp. – Vietnam) ranged from 40.5 to 42.0 pg. Unlike prior reports that indicated inheritance of spotted variegations were extranuclear genes that were maternally inherited, we found that the spotted leaf trait expressed in A. japonica ‘Shilpot’ appears to be a nuclear gene that is inherited in a quantitative fashion and not strictly maternal. These data provide an enhanced foundation for breeding improved aucuba.

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Hidden diversity in wild Beta taxa from Portugal: Insights from genome size and ploidy level estimations using flow cytometry

Plant Science ◽

10.1016/j.plantsci.2013.02.012 ◽

2013 ◽

Vol 207 ◽

pp. 72-78 ◽

Cited By ~ 13

Author(s):

Sílvia Castro ◽

Maria M. Romeiras ◽

Mariana Castro ◽

Maria Cristina Duarte ◽

João Loureiro

Keyword(s):

Flow Cytometry ◽

Genome Size ◽

Ploidy Level

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Flow cytometry as tool in plant sciences, with emphasis on genome size and ploidy level assessment

Genetics & Applications ◽

10.31383/ga.vol2iss2pp1-12 ◽

2018 ◽

Vol 2 (2) ◽

pp. 1 ◽

Cited By ~ 10

Author(s):

Mickael Bourge ◽

Spencer Creig Brown ◽

Sonja Siljak-Yakovlev

Keyword(s):

Flow Cytometry ◽

Genome Size ◽

Ploidy Level ◽

Cell Biology ◽

Base Composition ◽

Good Practice ◽

Plant Cells ◽

Size Measurement ◽

Fast Acquisition ◽

Plant Sciences

Flow cytometry has become the method of choice to measure the DNA content (genome size) in plants. Ease of sample preparation, fast acquisition, and accurate measurements have made the method popular in the domains of plant cell biology, systematics, evolution, genetics and biotechnology. Although the cell wall is a problem when isolating plant cells, cytometry remains a powerful tool in plant sciences. Based on our 30-years’ experience in this field, this review will focus at first on genome size measurement using simply isolated nuclei: the good practice for acquisition, nuclei isolation, appropriate buffers, kind of tissues to use. The second part will briefly review what kind of measurements it is possible to make in plant cytometry, and for what purpose: base composition, ploidy level, cell cycle, endoreplication, seed screening, and nuclei/chromosomes sorting. We will address troubleshooting. The commonly-used mathematic tools will be discussed.

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A new natural hybrid in Saxifraga sect. Porphyrion Tausch (Saxifragaceae)

Phytotaxa ◽

10.11646/phytotaxa.498.1.3 ◽

2021 ◽

Vol 498 (1) ◽

pp. 25-34

Author(s):

DAVID HORÁK ◽

MARTIN HAJMAN ◽

MICHAL HRONEŠ ◽

MOJMÍR PAVELKA

Keyword(s):

Flow Cytometry ◽

Genome Size ◽

Morphometric Analysis ◽

Ploidy Level ◽

Herbarium Specimens ◽

Natural Hybrid ◽

Hybrid Plants

A new natural hybrid Saxifraga ×klimesii Hajman, Horák & Hroneš from Ladakh (NW India) is described and illustrated. This hybrid resulted from cross between Saxifraga meeboldii Engler & Irmscher and Saxifraga pulvinaria Harry Smith. The morphology of the hybrid plants and its parental taxa was evaluated using morphometric analysis of both living plants and herbarium specimens. An artificial cross was also made to compare its morphology with spontaneous hybrids. Ploidy level and relative genome size was established using flow cytometry. Saxifraga ×klimesii is intermediate in morphology and relative genome size between both parents. It differs from S. meeboldii by shorter and wider rosette leaves and lighter yellow, larger and wider petals and from S. pulvinaria by larger rosettes, often more than one pore on leaves and usually yellow coloured petals. A lectotype is selected for S. pulvinaria.

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

Methods in Molecular Biology - Molecular Plant Taxonomy ◽

10.1007/978-1-0716-0997-2_17 ◽

2020 ◽

pp. 325-361

Author(s):

Jaume Pellicer ◽

Robyn F. Powell ◽

Ilia J. Leitch

Keyword(s):

Flow Cytometry ◽

Genome Size ◽

Ploidy Level ◽

Reproductive Modes

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Noninvasive determination of genome size and ploidy level in fishes by flow cytometry: detection of triploidPoecilia formosa

Cytometry ◽

10.1002/(sici)1097-0320(20000201)39:2<91::aid-cyto1>3.0.co;2-4 ◽

2000 ◽

Vol 39 (2) ◽

pp. 91-95 ◽

Cited By ~ 78

Author(s):

D.K. Lamatsch ◽

C. Steinlein ◽

M. Schmid ◽

M. Schartl

Keyword(s):

Flow Cytometry ◽

Genome Size ◽

Ploidy Level ◽

Flow Cytometry Detection

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Identification, Genome Sizes, and Ploidy of Deutzia

Journal of the American Society for Horticultural Science ◽

10.21273/jashs04779-19 ◽

2020 ◽

Vol 145 (2) ◽

pp. 88-94

Author(s):

William G. Hembree ◽

Thomas G. Ranney ◽

Nathan P. Lynch ◽

Brian E. Jackson

Keyword(s):

Flow Cytometry ◽

Genome Size ◽

Ploidy Level ◽

State University ◽

Ploidy Levels ◽

Specific Data ◽

North Carolina State University ◽

The North ◽

Voucher Specimens ◽

First Time

The genus Deutzia, in the Hydrangeaceae family, includes ≈60 species that range in ploidy from diploid (2x) to tetradecaploid (14x). There have been extensive breeding efforts for Deutzia, but this has been limited to a few parental species. Although there have been numerous studies of the cytogenetics of some species of Deutzia, the ploidy level of many species remains unknown, and there are few cytogenetic data available for Deutzia hybrids and cultivars. The purpose of this study was to validate the identification and determine the genome sizes and ploidy of a diverse collection of Deutzia species and hybrids using cytology and flow cytometry. Accessions were identified using the most current taxonomic key and voucher specimens were deposited for each at the North Carolina State University herbarium. Corrected and updated species names are provided for all cultivars and accessions studied. Traditional cytology was performed for roots of representative taxa to calibrate the genome size with the ploidy level. The genome size and estimated ploidy were determined for 43 accessions using flow cytometry. Ploidy levels were reported for the first time for three species of Deutzia including D. calycosa (2n = 4x = 52), D. paniculata (2n = 4x = 52), and D. glauca (2n = 12x = 156). The base and monoploid genome size (1Cx) were somewhat variable and ranged from 1.20 to 2.05 pg. No anisoploid hybrids were documented, suggesting the presence of an interploid block. The information produced from this study are beneficial to future curation, research, development, and improvement of this genus with corrected nomenclature and clone-specific data regarding cytogenetics.

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First estimates of genome size in ribbon worms (phylum Nemertea) using flow cytometry and Feulgen image analysis densitometry

Canadian Journal of Zoology ◽

10.1139/cjz-2014-0068 ◽

2014 ◽

Vol 92 (10) ◽

pp. 847-851 ◽

Cited By ~ 3

Author(s):

Kelly L. Mulligan ◽

Terra C. Hiebert ◽

Nicholas W. Jeffery ◽

T. Ryan Gregory

Keyword(s):

Flow Cytometry ◽

Image Analysis ◽

Body Size ◽

Genome Size ◽

Positive Relationship ◽

Nuclear Dna ◽

Nuclear Dna Content ◽

Size Estimation ◽

Size Diversity ◽

Size Estimates

Ribbon worms (phylum Nemertea) are among several animal groups that have been overlooked in past studies of genome-size diversity. Here, we report genome-size estimates for eight species of nemerteans, including representatives of the major lineages in the phylum. Genome sizes in these species ranged more than fivefold, and there was some indication of a positive relationship with body size. Somatic endopolyploidy also appears to be common in these animals. Importantly, this study demonstrates that both of the most common methods of genome-size estimation (flow cytometry and Feulgen image analysis densitometry) can be used to assess genome size in ribbon worms, thereby facilitating additional efforts to investigate patterns of variability in nuclear DNA content in this phylum.

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