Optimized flow cytometric protocol and genome size estimation of Sabah snake grass (Clinacanthus nutans)

The Malva genus contains species that reveal therapeutic properties and are mostly important in medicine and the functional food industry. Its breeding, cultivation, and utilization are based on proper germplasm/plant identification, which is difficult using morphological features. For this reason, we applied flow cytometry and inter simple sequence repeat polymerase chain reaction (ISSR-PCR) for fast and accurate species identification. Genome size estimation by flow cytometry was proposed as the first-choice method for quick accession screening. Out of the 12 tested accessions, it was possible to identify six genotypes based on genome size estimation, whereas all species and varieties were identified using ISSR markers. Flow cytometric analyses revealed that Malva species possessed very small (1.45–2.77 pg/2C), small (2.81–3.80 pg/2C), and intermediate (11.06 pg/2C) genomes, but the majority of accessions possessed very small genomes. Additionally, this is the first report on genome size assessment for eight of the accessions. The relationships between the investigated accessions showed the presence of two clusters representing malvoid and lavateroid group of species. Flow cytometry and ISSR molecular markers can be effectively used in the identification and genetic characterization of Malva species.

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Karyomorphology, ploidy analysis, and flow cytometric genome size estimation of Medicago monantha populations

TURKISH JOURNAL OF BOTANY ◽

10.3906/bot-2105-22 ◽

2021 ◽

Keyword(s):

Genome Size ◽

Size Estimation ◽

Ploidy Analysis ◽

Flow Cytometric

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Novel nuclei isolation buffer for flow cytometric genome size estimation of Zingiberaceae: a comparison with common isolation buffers

Annals of Botany ◽

10.1093/aob/mcw173 ◽

2016 ◽

Vol 118 (6) ◽

pp. 1057-1070 ◽

Cited By ~ 6

Author(s):

Abhishek Sadhu ◽

Sreetama Bhadra ◽

Maumita Bandyopadhyay

Keyword(s):

Genome Size ◽

Size Estimation ◽

Nuclei Isolation ◽

Flow Cytometric ◽

Isolation Buffer

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Flow cytometric analysis using SYBR Green I for genome size estimation in coffee

Acta Histochemica ◽

10.1016/j.acthis.2009.10.005 ◽

2011 ◽

Vol 113 (2) ◽

pp. 221-225 ◽

Cited By ~ 9

Author(s):

Wellington Ronildo Clarindo ◽

Carlos Roberto Carvalho

Keyword(s):

Genome Size ◽

Flow Cytometric Analysis ◽

Sybr Green ◽

Sybr Green I ◽

Size Estimation ◽

Flow Cytometric

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Leaves and Seeds as Materials for Flow Cytometric Estimation of the Genome Size of 11 Rosaceae Woody Species Containing DNA-Staining Inhibitors

Journal of Botany ◽

10.1155/2010/930895 ◽

2010 ◽

Vol 2010 ◽

pp. 1-9 ◽

Cited By ~ 23

Author(s):

Iwona Jedrzejczyk ◽

Elwira Sliwinska

Keyword(s):

Genome Size ◽

Dna Content ◽

Nuclear Dna ◽

Nuclear Dna Content ◽

Woody Species ◽

Size Estimation ◽

Flow Cytometric ◽

Cell Cytosol ◽

High Concentrations ◽

First Time

The presence of some secondary metabolites in the cell cytosol can cause a stoichiometric error in the flow cytometric estimation of nuclear DNA content. There is no fully reliable method to completely eliminate the effect of these compounds on nuclei fluorescence, and therefore using plant organs/parts free of staining inhibitors is recommended. Eleven species of Rosaceae with high concentrations of propidium-iodide-staining inhibitors were studied to check the possibility of using seeds instead of leaves for genome size estimation. Despite optimizing the concentration and composition of antioxidants in nuclei-isolation buffer for each species, the effect of cytosolic compounds present in the leaves could not be avoided entirely. None of the seeds of the studied species contained inhibitors, and they produced histograms of good quality. The genome size of the studied species ranged from 1.15 to 3.17 pg/2C; for 10 species the DNA content was estimated for the first time.

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

Genes ◽

10.3390/genes12040563 ◽

2021 ◽

Vol 12 (4) ◽

pp. 563

Author(s):

Monika Rewers ◽

Iwona Jedrzejczyk ◽

Agnieszka Rewicz ◽

Anna Jakubska-Busse

Keyword(s):

Genome Size ◽

Species Identification ◽

Dna Content ◽

Size Estimation ◽

Orchid Species ◽

Infraspecific Taxon ◽

2C Dna Content ◽

Plant Families ◽

Liparis Loeselii ◽

Identification And Characterization

Orchidaceae is one of the largest and the most widespread plant families with many species threatened with extinction. However, only about 1.5% of orchids’ genome sizes have been known so far. The aim of this study was to estimate the genome size of 15 species and one infraspecific taxon of endangered and protected orchids growing wild in Poland to assess their variability and develop additional criterion useful in orchid species identification and characterization. Flow cytometric genome size estimation revealed that investigated orchid species possessed intermediate, large, and very large genomes. The smallest 2C DNA content possessed Liparis loeselii (14.15 pg), while the largest Cypripedium calceolus (82.10 pg). It was confirmed that the genome size is characteristic to the subfamily. Additionally, for four species Epipactis albensis, Ophrys insectifera, Orchis mascula, Orchis militaris and one infraspecific taxon, Epipactis purpurata f. chlorophylla the 2C DNA content has been estimated for the first time. Genome size estimation by flow cytometry proved to be a useful auxiliary method for quick orchid species identification and characterization.

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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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