Estimation of the relative DNA content in species of the genus Spiraea, sections Chamaedryon and Glomerati by flow cytometry

The relative DNA content was studied in seven species of the genus Spiraea L., section Chamaedryon Ser., and in two species, section Glomerati Nakai, from 28 natural populations growing in Asian Russia. The cell nuclei were isolated from a leaf tissue. The relative intensity of fluorescence was measured using flow cytometry of propidium iodide-stained nuclei. The analysis was performed using a CyFlowSpace device (Germany, Sysmex Partec) with a laser radiation source of 532 nm. Fresh leaves of Solanum lycopersicum cv. ‘Stupice’ were used as an internal standard. Data on the relative DNA content are presented for the first time for S. flexuosa Fisch ex Cambess. (0.42–0.47 pg), S. ussuriensis Pojark. (0.49–0.52; 0.85 pg), S. alpina-Pall. (0.49–0.51 pg), S. media Schmidt. (0.45; 0.98–1.01 pg), S. trilobata L. (0.46 pg), S. hypericifolia L. (0.49–0.52 pg) and S. aquilegifolia Pall. (0.48–0.51 pg). Mesophytic species of the genus Spiraea growing in the forest zone (S. chamaedryfolia L. and S. media) exhibit a 2-fold higher relative DNA content (C-value) compared to xerophytic species (S. hypericifolia, S. crenata L., S. aquilegifolia and S. trilobata) and species confined only to mountain areas (S. alpinа, S. trilobata and S. aquilegifolia).

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Flow Cytometrically Determined DNA Content of Breast Carcinoma and Benign Lesions: Correlations with Histopathological Parameters

Tumori Journal ◽

10.1177/030089168607200209 ◽

1986 ◽

Vol 72 (2) ◽

pp. 171-177 ◽

Cited By ~ 10

Author(s):

Raffaella Uccelli ◽

Alberto Calugi ◽

Donato Forte ◽

Francesco Mauro ◽

Paolo Polonio-Balbi ◽

...

Keyword(s):

Flow Cytometry ◽

Dna Content ◽

Cell Subpopulation ◽

Aneuploid Cell ◽

Breast Carcinomas ◽

Benign Lesions ◽

Dna Index ◽

Tissue Samples ◽

Normal Tissues ◽

Relative Dna Content

The relative DNA content of cellular samples from 54 patients affected by breast carcinomas and 20 affected by benign breast lesions (including 11 fibroadenomas) was measured by flow cytometry. All normal tissue samples and 17/20 (85%) specimens from benign lesions exhibited a cytometrically diploid DNA distribution, 3/20 (15%) benign lesions an abnormal DNA content, and 35/54 (65%) carcinomas at least one aneuploid cell subpopulation. Furthermore, 9/54 (17%) tumors were characterized by the presence of more than one aneuploid cell subpopulation. The results also indicate that flow cytometry can be used to recognize lymph nodes infiltrated by aneuploid cells. Statistically significant correlations were evidenced between the occurrence of aneuploidy or the ploidy level measured as DNA index and the nodal infiltration status. The percentage of S cells can also be extracted from DNA content distribution histograms. Statistically significant differences (p < 0.01) were also observed for the percentage of S cells between normal tissues (6.2±3.2 SD) and benign lesions (11.1±6.6 SD), normal tissues (6.2 ± 3.2 SD) and aneuploid tumors (19.7 ± 10.3 SD), benign lesions (11.1 ± 6.6 SD) and aneuploid tumors (19.7 ± 10.3 SD), and diploid (7.9 ± 4.0 SD) and aneuploid tumors (19.7 ± 10.3 SD).

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NUCLEAR DNA CONTENT OF DENDROBIUM ORCHID SPECIES AS DETERMINED BY LASER FLOW CYTOMETRY

HortScience ◽

10.21273/hortsci.31.3.322c ◽

1996 ◽

Vol 31 (3) ◽

pp. 322c-322

Author(s):

W.E. Jones ◽

A.R. Kuehnle ◽

K. Arumuganathan

Keyword(s):

Flow Cytometry ◽

Dna Content ◽

Nuclear Dna ◽

Evolutionary Relationship ◽

Internal Standard ◽

Nuclear Dna Content ◽

Orchid Species ◽

Group Assignment ◽

Taxonomic Groups

Flow cytometry (FC) has proven to be an efficient and reliable method to estimate nuclear DNA content (genome size) in quantifiable units useful for genetic and molecular biology studies. This method also makes possible determination of the variation in nuclear DNA content between related taxa, which gives insights into the process of speciation. In this study, DNA content was determined in nuclei isolated from leaves of 21 Dendrobium species representing each of the major taxonomic groups used in the Univ. of Hawaii breeding program. Nuclei were mechanically isolated, stained with the nucleic acid-specific fluorochrom propidium iodide, and DNA content determined using a Coulter Epics 753 laser flow cytometer. Chicken erythrocyte nuclei (2C = 2.33 pg DNA) were used as an internal standard for direct comparative measurement. The mean diploid genome (2C) values for Dendrobium species ranged from 3.36 to 5.06 pg. Genome sizes were evaluated for possible use as discrete characters for taxonomic group assignment and compared to previous data on breeding compatibility and evolutionary relationship between species.

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Variation of nuclear DNA content in seeds of Nitraria schoberi L.

BIO Web of Conferences ◽

10.1051/bioconf/20181100046 ◽

2018 ◽

Vol 11 ◽

pp. 00046

Author(s):

Maria Voronkova ◽

Evgeniy Banaev ◽

Maria Tomoshevich ◽

Taigana Ak-Lama

Keyword(s):

Flow Cytometry ◽

Dna Content ◽

Nuclear Dna ◽

Natural Populations ◽

Nuclear Dna Content ◽

Relative Content ◽

Average Value ◽

The Crimea ◽

First Time

For the first time are search of nuclear DNA relative content in Nitraria schoberi L. seeds from 15 natural populations of Siberia, the Crimea, Kazakhstan, and Tajikistan has been conducted by the method of flow cytometry. High intra-and interpopulational variations of nuclear DNA content - 2,93-3,39 pg, at average value - 3,22±0,108 pg is revealed.

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Genome size in Arachis duranensis: a critical study

Genome ◽

10.1139/g01-081 ◽

2001 ◽

Vol 44 (5) ◽

pp. 826-830 ◽

Cited By ~ 16

Author(s):

Eva M Temsch ◽

Johann Greilhuber

Keyword(s):

Flow Cytometry ◽

Genome Size ◽

Sea Level ◽

Dna Content ◽

Internal Standard ◽

Critical Study ◽

Dna Flow Cytometry ◽

Technical Problems ◽

Arachis Duranensis ◽

Collection Sites

Arachis duranensis is a diploid wild relative of the tetraploid cultivated peanut Arachis hypogaea. The literature indicates two 2C genomic DNA mean values (genome size) for A. duranensis, 4.92 and 5.64 pg, and intraspecific variation of up to 11% negatively correlated with altitude above sea level of the collection sites has been reported. Our recent investigations of Arachis species have shown that unrecognized technical problems with peanut material may have influenced previous genome-size data and rendered them open to critical comments. In the present study, 20 accessions of A. duranensis were investigated by means of DNA flow cytometry (propidium iodide staining) and several of these also by Feulgen DNA image analysis. Pisum sativum was used as the internal standard (2C = 8.84 pg). 2C values in A. duranensis were about half those described previously and varied between 2.49 and 2.87 pg (flow cytometry). This variation was statistically significant and reproducible. There was a negative correlation of genome size with latitude and altitude above sea level of the collection sites. Such a correlation had been already found in one of the previous studies. However, the incongruences between the absolute DNA content values obtained in the present investigation and those in the literature point to the importance of carrying out methodological studies on best practice in DNA-content determinations in plants.Key words: Arachis duranensis, genome size, flow cytometry, Feulgen densitometry.

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Flow cytometric determination of DNA content in isolated nuclei of cereals

Genome ◽

10.1139/g88-095 ◽

1988 ◽

Vol 30 (4) ◽

pp. 565-569 ◽

Cited By ~ 12

Author(s):

E. Hülgenhof ◽

R. A. Weidhase ◽

R. Schlegel ◽

A. Tewes

Keyword(s):

Flow Cytometry ◽

Dna Content ◽

Internal Standard ◽

Aegilops Speltoides ◽

Isolated Nuclei ◽

Triticum Timopheevi ◽

Flow Cytometric ◽

Nuclear Isolation ◽

Cereal Plants

Isolated nuclei from cereal plants were used for quantitative determination of DNA per nucleus by flow cytometry. The technique is based on enhanced fluorescence of ethidium bromide and olivomycin when they bind to DNA. Nuclei were isolated from protoplasts derived from leaves of seedlings. The diploid Hordeum vulgare cv. Trumpf was used as an internal standard. Analysis of nuclei from several cultivars of hexaploid wheat (Triticum aestivum), Triticum durum var. hordeiforme, T. durum var. alexandrinum, Triticum araraticum, Triticum timopheevi, Triticum monococcum, Aegilops speltoides, Secale cereale, and one hexaploid and one octoploid triticale revealed significant intra- and inter-specific differences in DNA values per 2C nucleus. The advantages of the procedure are discussed along with its utilization for quantitative DNA measurement in Gramineae.Key words: nuclear isolation, flow cytometry, DNA content, cereals.

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Determination of Genome Size Differentiation and Ploidy Levels in Some Citrus Rootstock Populations

10.21203/rs.3.rs-1106171/v1 ◽

2021 ◽

Author(s):

Sefa POLATÖZ ◽

Murat Seker ◽

Çağlar KAYA

Keyword(s):

Flow Cytometry ◽

Leaf Tissue ◽

Soil Conditions ◽

Citrus Species ◽

Cell Nuclei ◽

Isolated Cells ◽

Ploidy Levels ◽

Citrus Rootstock ◽

Leaf Tissues

Abstract In plants, knowing the ploidy level of plant material used in breeding studies, and especially for biotechnology applications, carries great importance. The presence of a rapid variety of dynamics in citrus fruits allows their use as rootstock and varieties ensuring adaptability to various climate and soil conditions with different breeding methods. A variety of appropriate rootstocks are used for commercial citrus species. This study investigated the genome sizes and ploidy levels in citrus rootstocks commonly used around the world with flow cytometry in seedling populations. The study used Gou-Tou, C-35, Troyer Citrange, Taiwanica, Citremon, Yuzu, Sunki mandarin, Flying Dragon, Yuma Citrange, Macrophylla and Chinese orange rootstocks. Fresh leaf tissues were mixed with the triploid Tahiti lime leaf tissue, used as standard species, and cell nuclei were isolated. Cells stained with propidium iodide were read with flow cytometry and histograms and cytograms were obtained. According to the obtained results, all seedlings of species had diploid genome volumes. In terms of genome volume, there were differences found between species. Yuzu seedlings were determined to be the species with largest genome volume (0.808 pg/2C), while Flying Dragon trifoliate had smallest genome volume (0.700 pg/2C).

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Nuclear DNA Content and Ploidy Levels in the Genus Ipomoea

Journal of the American Society for Horticultural Science ◽

10.21273/jashs.119.1.110 ◽

1994 ◽

Vol 119 (1) ◽

pp. 110-115 ◽

Cited By ~ 42

Author(s):

Peggy Ozias-Akins ◽

Robert L. Jarret

Keyword(s):

Flow Cytometry ◽

Dna Content ◽

Large Scale ◽

Nuclear Dna ◽

Internal Standard ◽

Nuclear Dna Content ◽

Ploidy Levels ◽

Genetic Structure Of Populations ◽

Chicken Erythrocytes ◽

Nuclei Ploidy Level

The nuclear DNA content of 53 accessions from 24 Ipomoea (Convolvulaceae) species, including four sweetpotato cultivars, was determined by flow cytometry of DAPI-stained nuclei. Ploidy level and DNA content were significantly correlated within the genus, but more highly so within species that contained multiple cytotypes. DNA content of cultivated Z. batatas (L.) Lam. (4.8 to 5.3 pg/2C nucleus) and feral tetraploid I. batatas (3.0 to 3.5 pg/2C nucleus) was estimated from the known DNA content of chicken erythrocytes (2.33 pg), which were used as an internal standard. Tetraploid forms of Z. cordato-triloba Dennstedt also were identified. Ploidy analysis using flow cytometry is rapid and suitable for large-scale experiments such as studying the genetic structure of populations of Z. batatas and related species. Chemical name used: 4′,6-diamidino-2-phenylindole (DAPI).

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Nuclear DNA content of 11 fungal species in Glomales

Genome ◽

10.1139/g98-038 ◽

1998 ◽

Vol 41 (3) ◽

pp. 422-428 ◽

Cited By ~ 48

Author(s):

Michel Hosny ◽

Vivienne Gianinazzi-Pearson ◽

Hubert Dulieu

Keyword(s):

Flow Cytometry ◽

Dna Content ◽

Nuclear Dna ◽

Internal Standard ◽

Nuclear Dna Content ◽

Fungal Species ◽

Dapi Staining ◽

Interspecific Variations ◽

Propidium Iodide Staining ◽

The Difference

The nuclear DNA content of 11 species of Glomales was evaluated by flow cytometry after DAPI staining relative to Gigaspora margarita, which was used as internal standard. The nuclear DNA content of this species was calibrated by propidium iodide staining relative to chicken red blood cells. A correction was applied when the difference in AT content of the DNA was significant between a sample and the standard. A single unimodal peak of fluorescence was observed for nuclei from the quiescent spores of the 11 fungal species studied. It was considered that this peak corresponded to the amount of DNA in the genome of each species. Important interspecific variations in DNA content per nucleus (1- to 8-fold) were observed among four species of the genus Scutellospora.Key words: nucleus, DNA content, flow cytometry, spore, Glomales.

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