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

2018 ◽  
Vol 30 (1) ◽  
pp. 103-128 ◽  
Author(s):  
Elwira Sliwinska
Keyword(s):  
Flow Cytometry ◽  
Genome Size ◽  
Dna Content ◽  
Plant Material ◽  
Nuclear Dna ◽  
Modern Method ◽  
Hybrid Breeding ◽  
Size Estimation ◽  
Medicinal Species

AbstractFlow 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.

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

2014 ◽  
Vol 92 (10) ◽  
pp. 847-851 ◽  
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.

Nuclear DNA content of the whitefly Bemisia tabaci (Aleyrodidae: Hemiptera) estimated by flow cytometry

2005 ◽  
Vol 95 (4) ◽  
pp. 309-312 ◽  
Author(s):  
J.K. Brown ◽  
G.M. Lambert ◽  
M. Ghanim ◽  
H. Czosnek ◽  
D.W. Galbraith
Keyword(s):  
Flow Cytometry ◽  
Genome Size ◽  
Bemisia Tabaci ◽  
Dna Content ◽  
Blood Cells ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Chromosome Counting ◽  
Male And Female ◽  
Haploid Genome Size

AbstractThe nuclear DNA content of the whitefly Bemisia tabaci (Gennnadius) was estimated using flow cytometry. Male and female nuclei were stained with propidium iodide and their DNA content was estimated using chicken red blood cells and Arabidopsis thaliana L. (Brassicaceae) as external standards. The estimated nuclear DNA content of male and female B. tabaci was 1.04 and 2.06 pg, respectively. These results corroborated previous reports based on chromosome counting, which showed that B. tabaci males are haploid and females are diploid. Conversion between DNA content and genome size (1 pg DNA = 980 Mbp) indicate that the haploid genome size of B. tabaci is 1020 Mbp, which is approximately five times the size of the genome of the fruitfly Drosophila melanogaster Meigen. These results provide an important baseline that will facilitate genomics-based research for the B. tabaci complex.

scholarly journals Genome Sizes in Hepatica Mill: (Ranunculaceae) Show a Loss of DNA, Not a Gain, in Polyploids

2010 ◽  
Vol 2010 ◽  
pp. 1-7 ◽  
Author(s):  
B. J. M. Zonneveld
Keyword(s):  
Flow Cytometry ◽  
Genome Size ◽  
Dna Content ◽  
Close Relationships ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Dna Contents ◽  
Asiatic Species ◽  
Nuclear Dna Contents ◽  
Allotetraploid Species

Genome size (C-value) was applied anew to investigate the relationships within the genus Hepatica (Ranunculaceae). More than 50 samples representing all species (except H. falconeri), from wild and cultivated material, were investigated. Species of Hepatica turn out to be diploid (), tetraploid ( ), and a possible pentaploid. The somatic nuclear DNA contents (2C-value), as measured by flow cytometry with propidium iodide, were shown to range from 33 to 80 pg. The Asiatic and American species, often considered subspecies of H. nobilis, could be clearly distinguished from European H. nobilis. DNA content confirmed the close relationships in the Asiatic species, and these are here considered as subspecies of H. asiatica. Parents for the allotetraploid species could be suggested based on their nuclear DNA content. Contrary to the increase in genome size suggested earlier for Hepatica, a significant (6%–14%) loss of nuclear DNA in the natural allopolyploids was found.

scholarly journals Evolution of Genome Size in Duckweeds (Lemnaceae)

2011 ◽  
Vol 2011 ◽  
pp. 1-9 ◽  
Author(s):  
Wenqin Wang ◽  
Randall A. Kerstetter ◽  
Todd P. Michael
Keyword(s):  
Flow Cytometry ◽  
Genome Size ◽  
Propidium Iodide ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Continuous Increase ◽  
Spirodela Polyrhiza ◽  
Wolffia Arrhiza ◽  
Basic Reference

To extensively estimate the DNA content and to provide a basic reference for duckweed genome sequence research, the nuclear DNA content for 115 different accessions of 23 duckweed species was measured by flow cytometry (FCM) stained with propidium iodide as DNA stain. The 1C-value of DNA content in duckweed family varied nearly thirteen-fold, ranging from 150 megabases (Mbp) in Spirodela polyrhiza to 1,881 Mbp in Wolffia arrhiza. There is a continuous increase of DNA content in Spirodela, Landoltia, Lemna, Wolffiella, and Wolffia that parallels a morphological reduction in size. There is a significant intraspecific variation in the genus Lemna. However, no such variation was found in other studied species with multiple accessions of genera Spirodela, Landoltia, Wolffiella, and Wolffia.

scholarly journals MICROPROPAGATION AND PLOIDY STABILITY OF Lippia lacunosa Mart. & Schauer: AN ENDANGERED BRAZILIAN MEDICINAL PLANT

2019 ◽  
Vol 6 (1) ◽  
pp. 1-7
Author(s):  
Diego Pandeló José ◽  
José Marcello Salabert De Campos ◽  
Lyderson Facio Viccini ◽  
Emilly Ruas Alkimim ◽  
Marcelo De Oliveira Santos
Keyword(s):  
Flow Cytometry ◽  
Medicinal Plant ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Naphthalene Acetic Acid ◽  
Flow Cytometry Analysis ◽  
Ploidy Levels

Lippia lacunosa is a Brazilian savanna plant that belongs to the Verbenaceae family. It has been used in folk medicine as a treatment for different diseases. This species represents an endangered Brazilian medicinal plant, and this is the first report documenting a reliable protocol for the in vitro propagation and regeneration of L. lacunosa. Axenic explants were cultivated in MS medium containing different concentrations of naphthalene acetic acid (NAA) to induce root growth. The mean shoot length and the number of roots were highest with 0.06 mg·L-1 NAA. The highest number of buds in shoot regeneration was induced with 2 mg·L-1 6-benzylaminopurine (BA). To obtain a long-term culture, the dwarf shoots were elongated on MS media containing 0.5 mg·L-1 BA alternated with MS containing 2 mg·L-1 BA every 40 days. In the present protocol, the long-term shoots retained the ability to root even after long periods of BA treatment. In addition, we evaluated the nuclear DNA content and ploidy levels, including the occurrence of endopolyploidy, in long-term micropropagated plant leaves using flow cytometry analysis. The plants propagated in vitro over several years possessed nuclear DNA contents ranging from 2.940 to 3.095 pg, and no differences in DNA content were found among in vitro plants or between these plants and the control (L. lacunosa from a greenhouse with a DNA content of 3.08 pg). The flow cytometry analysis also demonstrated that there was no polyploidization. The present study will be useful for biotechnological approaches and provides the first estimate of the nuclear DNA content of this species using flow cytometry.

Genome size, fluorochrome banding, and karyotype evolution in some Hypochoeris species

Genome ◽  
1995 ◽  
Vol 38 (4) ◽  
pp. 689-695 ◽  
Author(s):  
M. Cerbah ◽  
J. Coulaud ◽  
B. Godelle ◽  
S. Siljak-Yakovlev
Keyword(s):  
Flow Cytometry ◽  
Genome Size ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Nucleolar Organizer ◽  
Phylogenetic Position ◽  
Nucleolar Organizer Regions ◽  
Fluorochrome Banding ◽  
South American

Four South American and two European species of Hypochoeris (Asteraceae) were studied using fluorochrome banding, and genome size was determined by flow cytometry, in order to obtain information about microevolution in this genus and about its primary origin. Fluorochrome banding patterns showed GC-rich repeated sequences, particularly around the nucleolar organizer regions. Few differences appeared among the South American species. Nevertheless, determination of nuclear DNA content and base composition revealed significant differences among these species. The phylogenetic position of Hypochoeris robertia, which has the smallest DNA content, is discussed with regard to chromosome evolution in this genus.Key words: Hypochoeris, Asteraceae, fluorochromes, flow cytometry, nucleolar organizer regions, microevolution.

Genome size differences in Hyalella cryptic species

Genome ◽  
2012 ◽  
Vol 55 (2) ◽  
pp. 134-139 ◽  
Author(s):  
Roland Vergilino ◽  
Kaven Dionne ◽  
Christian Nozais ◽  
France Dufresne ◽  
Claude Belzile
Keyword(s):  
Flow Cytometry ◽  
Cryptic Species ◽  
Dna Barcoding ◽  
Genome Size ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Complex Species ◽  
Numerous Species ◽  
Freshwater Habitats

The Hyalella azteca (Saussure) complex includes numerous amphipod cryptic species in freshwater habitats in America as revealed by DNA barcoding surveys. Two ecomorphs (small and large) have evolved numerous times in this complex. Few phenotypic criteria have been found to differentiate between the numerous species of this complex. The present study aims to explore genome size differences between some species of the H. azteca complex co-occurring in a Canadian boreal lake using flow cytometry. Nuclear DNA content was estimated for 50 individuals belonging to six COI haplotypes corresponding to four provisional species of the H. azteca complex. Species from the large ecomorph had C-values significantly larger than species from the small ecomorph, whereas slight differences were found among species of the small ecomorph. These differences in genome sizes might be linked to ecological and physiological differences among species of the H. azteca complex.

Nuclear DNA content of Pongamia pinnata L. and genome size stability of in vitro-regenerated plantlets

PROTOPLASMA ◽  
2013 ◽  
Vol 251 (3) ◽  
pp. 703-709 ◽  
Author(s):  
Rimjhim Roy Choudhury ◽  
Supriyo Basak ◽  
Aadi Moolam Ramesh ◽  
Latha Rangan
Keyword(s):  
Genome Size ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Pongamia Pinnata ◽  
Size Stability ◽  
Regenerated Plantlets

scholarly journals Estimating Nuclear DNA Content in Peach and Related Diploid Species Using Laser Flow Cytometry and DNA Hybridization

1994 ◽  
Vol 119 (6) ◽  
pp. 1312-1316 ◽  
Author(s):  
W. Vance Baird ◽  
Agnes S. Estager ◽  
John K. Wells
Keyword(s):  
Flow Cytometry ◽  
Chromosome Number ◽  
Genome Size ◽  
Dna Content ◽  
Nuclear Dna ◽  
Diploid Species ◽  
Nuclear Genome ◽  
Nuclear Dna Content ◽  
Gene Copy ◽  
Polyploid Series

Using laser flow cytometry, nuclear DNA amounts were estimated for 12 Prunus species, representing three subgenera [Prunophora (Prunus), Amygdalus, and Cerasus (Lithocerasus)], two interspecific hybrids, four cultivars, and a synthetic polyploid series of peach consisting of haploids, diploids, triploids, and tetraploids (periclinal cytochimeras). Peach nuclear DNA content ranged from 0.30 pg for the haploid nuclei to 1.23 pg for the tetraploid nuclei. The diploid genome of peach is relatively small and was estimated to be 0.60±0.03 pg (or 5.8×108 nucleotide base pairs). The polyploid series represented the expected arithmetic progression, as genome size positively correlated with ploidy level (i.e., DNA content was proportional to chromosome number). The DNA content for the 12 diploid species and two interspecific diploid hybrids ranged from 0.57 to 0.79 pg. Genome size estimates were verified independently by Southern blot analysis, using restriction fragment length polymorphism clones as gene-copy equivalents. Thus, a relatively small and stable nuclear genome typifies the Prunus species investigated, consistent with their low, basic chromosome number (× = 8).

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