Analysis of Hypericum accessions by DNA fingerprinting and flow cytometry

Hypericum perforatum, H. umbellatum, H. maculatum, and H. hircinum accessions originating from botanical gardens across Europe were examined by flow cytometry and molecular markers. 2C DNA content of 17 Hypericum perforatum accessions (Hp) and the H. perforatum cultivar Topaz amounted to between 1.56 pg and 1.62 pg. In four Hp accessions some individual plants were found with a DNA content corresponding to 6Cx (2.34 - 2.39 pg). All plants of accession Hp8 showed a DNA content of 6Cx (2.41 pg). In root tips of Hp plants with an average DNA amount of 1.58 pg, 32 chromosomes were detected, corresponding to 2n = 4x. This is the first ploidy and/or DNA content report for H. umbellatum, H. maculatum and H. hircinum. H. umbellatum and H. maculatum, each contained 0.76 pg DNA and 16 chromosomes were counted. The 2C DNA content of H. hircinum was 1.00 pg with the best metaphase plate revealing 32 chromosomes. Additionally, a combined marker analysis, based on inter-simple sequence repeats (ISSR) and sequence related amplified polymorphism (SRAP), was conducted to gain a better understanding of diversity especially within the accessions of H. perforatum. A total of 27 (11 ISSR and 16 SRAP) primer combinations were screened, showing 699 bands, of which 661 were polymorphic. UPGMA clustering revealed that accessions from the same geographic area tended to be more closely related, while H. maculatum was grouped separately from all H. perforatum accessions. Both methods have shown similar sensitivities in detecting the genetic diversity of the analyzed genotypes. Our results may be useful for Hypericum breeding programs and the development of effective conservation strategies.

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Chromosome Number, Ploidy Level, and Nuclear DNA Content in 23 Species of Echeveria (Crassulaceae)

Genes ◽

10.3390/genes12121950 ◽

2021 ◽

Vol 12 (12) ◽

pp. 1950

Author(s):

Guadalupe Palomino ◽

Javier Martínez-Ramón ◽

Verónica Cepeda-Cornejo ◽

Miriam Ladd-Otero ◽

Patricia Romero ◽

...

Keyword(s):

Flow Cytometry ◽

Chromosome Number ◽

Ploidy Level ◽

Dna Content ◽

Chromosome Numbers ◽

Nuclear Dna ◽

Nuclear Dna Content ◽

Root Tips ◽

Ploidy Levels ◽

Dna Amounts

Echeveria is a polyploid genus with a wide diversity of species and morphologies. The number of species registered for Echeveria is approximately 170; many of them are native to Mexico. This genus is of special interest in cytogenetic research because it has a variety of chromosome numbers and ploidy levels. Additionally, there are no studies concerning nuclear DNA content and the extent of endopolyploidy. This work aims to investigate the cytogenetic characteristics of 23 species of Echeveria collected in 9 states of Mexico, analyzing 2n chromosome numbers, ploidy level, nuclear DNA content, and endopolyploidy levels. Chromosome numbers were obtained from root tips. DNA content was obtained from the leaf parenchyma, which was processed according to the two-step protocol with Otto solutions and propidium iodide as fluorochrome, and then analyzed by flow cytometry. From the 23 species of Echeveria analyzed, 16 species lacked previous reports of 2n chromosome numbers. The 2n chromosome numbers found and analyzed in this research for Echeveria species ranged from 24 to 270. The range of 2C nuclear DNA amounts ranged from 1.26 pg in E. catorce to 7.70 pg in E. roseiflora, while the 1C values were 616 Mbp and 753 Mbp, respectively, for the same species. However, differences in the level of endopolyploidy nuclei were found, corresponding to 4 endocycles (8C, 16C, 32C and 64C) in E. olivacea, E. catorce, E. juarezensis and E. perezcalixii. In contrast, E. longiflora presented 3 endocycles (8C, 16C and 32C) and E. roseiflora presented 2 endocycles (8C and 16C). It has been suggested that polyploidization and diploidization processes, together with the presence of endopolyploidy, allowed Echeveria species to adapt and colonize new adverse environments.

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NUCLEAR DNA CONTENT IN BLUEBERRY DETERMINED BY FLOW CYTOMETRY

HortScience ◽

10.21273/hortsci.27.6.580e ◽

1992 ◽

Vol 27 (6) ◽

pp. 580e-580

Author(s):

Rodomiro Ortiz ◽

D.E. Costich ◽

T.P. Meagher ◽

N. Vorsa

Keyword(s):

Flow Cytometry ◽

Dna Content ◽

Nuclear Dna ◽

Nuclear Dna Content ◽

Haploid Genome ◽

Dna Flow Cytometry ◽

Dna Amount ◽

Ploidy Levels ◽

Polyploid Evolution ◽

Haploid Genome Size

DNA flow cytometry was used to determine nuclear DNA content in diploid blueberry species, and 3x, 4x, 5x, and 6x ploidy levels. Relative fluorescence intensity of stained nuclei measured by flow cytometry was a function of the number of chromosome sets (X): Y = 3.7X – 2.3 (r2 = 95.1%). DNA flow cytometry should be useful for ploidy level determination in the seedling stage. A significant linear relationship was established between nuclear DNA content and number of chromosomes (x); DNA (pg) = 0.52 x1 (r2 = 99.8%). Based on this equation the haploid genome DNA amount (1C) was calculated as 0.62 ± 0.08 pg, with an approximate haploid genome size of 602 Mbp/1C. The results indicate that conventional polyploid evolution occured in the section Cyanococcus, genus Vaccinium: the increase in DNA was concurrent with increase in chromosome number. DNA content differences among 2x species were correlated with Nei's genetic distance estimates based on 20 isozyme markers. Most of the variation was among species (49%), with 26% between populations within species, and 25% within populations.

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Karyotype characterization and nuclear DNA content measurement in Bromeliaceae: State of the art and future perspectives

Anais da Academia Brasileira de Ciências ◽

10.1590/0001-3765201420140224 ◽

2014 ◽

Vol 86 (4) ◽

pp. 1849-1862 ◽

Cited By ~ 1

Author(s):

ANDREI C.P. NUNES ◽

WELLINGTON R. CLARINDO

Keyword(s):

Flow Cytometry ◽

Chromosome Number ◽

Dna Content ◽

Nuclear Dna ◽

State Of The Art ◽

Nuclear Dna Content ◽

Basic Chromosome Number ◽

Basic Chromosome ◽

Content Measurement ◽

2C Dna Content

In Bromeliaceae, cytogenetic and flow cytometry analyses have been performed to clarify systematic and evolutionary aspects. Karyotyping approaches have shown the relatively high chromosome number, similar morphology and small size of the chromosomes. These facts have prevented a correct chromosome counting and characterization. Authors have established a basic chromosome number of x = 25 for Bromeliaceae. Recently, one karyomorphological analysis revealed that x = 25 is no longer the basic chromosome number, whose genome may have a polyploid origin. Besides cytogenetic characterization, the 2C DNA content of bromeliads has been measured. Nuclear DNA content has varied from 2C = 0.60 to 2C = 3.34 picograms. Thus, in relation to most angiosperms, the 2C DNA content of Bromeliaceae species as well as their chromosome size can be considered relatively small. In spite of some advances, cytogenetic and flow cytometry data are extremely scarce in this group. In this context, this review reports the state of the art in karyotype characterization and nuclear DNA content measurement in Bromeliaceae, emphasizing the main problems and suggesting prospective solutions and ideas for future research.

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Determination of DNA content and relative 2C genome sizes of some promising commercial varieties of sugarcane using flow cytometer

Current Botany ◽

10.19071/cb.2017.v8.3207 ◽

2017 ◽

Vol 8 ◽

Author(s):

A. Mondal S.K. Ghosal ◽

T. Pal Kalyan Kumar De

Keyword(s):

Flow Cytometry ◽

Genome Size ◽

Dna Content ◽

Nuclear Dna ◽

Nuclear Dna Content ◽

Cane Yield ◽

Sugarcane Varieties ◽

Yield Information ◽

A Genome ◽

2C Dna Content

In the present study, 2C DNA content and the genome sizes (in picograms-pg and megabase pairs-Mbp respectively) of 19 promising commercial varieties of sugarcane, the derivatives of man-made interspecific hybrids between cultivated and wild species were analyzed using flow cytometry. In this work, 2C nuclear DNA content was determined. Knowing the 2C nuclear DNA content, the unknown chromosome numbers of the varieties could be predicted. Large differences (65 % variation) in DNA content (2C) of 19 varieties were detected, ranging, from 3.80 pg to 10.96 pg, which corresponds to a genome size ranging from 3724.00 Mbp to 10740.80 Mbp due to the variation of ploidy level and are considered the most complex genomes among crop plants. However, the relationship between chromosome number and genome size was highly significant (P < 0.001). In the present study, internode diameter, Sugar juice content and cane yield/ha are also positively correlated with DNA content. The estimated genome sizes would also yield information critical for sugarcane breeding and genome sequencing programs. Keywords: Genome size, Sugarcane varieties, Flow cytometry, DNA content.

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DNA content estimation of Fig and Black Mulberry using flow cytometry

Revista Brasileira de Fruticultura ◽

10.1590/0100-29452018568 ◽

2018 ◽

Vol 40 (2) ◽

Author(s):

Zeynel Dalkiliç ◽

Gonca Günver Dalkiliç

Keyword(s):

Flow Cytometry ◽

Genome Size ◽

Dna Content ◽

Dna Contents ◽

2C Dna Content ◽

Dapi Fluorescence

Abstract In this study, fig and black mulberry DNA contents were estimated using DAPI fluorescence stain in flow cytometry. The 2C DNA contents of the fig and black mulberry were found as 0.82 pg and 8.34 pg, respectively. The calculated 1C value of genome size of fig is 401.8 Mbp and that of black mulberry is 4086.6 Mbp. The ratio of 2C DNA content and 1C genome of the black mulberry was 10.17 times that of the fig although fig is diploid and black mulberry is decosaploid.

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Genome size in 21 Artemisia L. species (Asteraceae, Anthemideae): Systematic, evolutionary, and ecological implications

Genome ◽

10.1139/g01-004 ◽

2001 ◽

Vol 44 (2) ◽

pp. 231-238 ◽

Cited By ~ 27

Author(s):

Montserrat Torrell ◽

Joan Vallès

Keyword(s):

Flow Cytometry ◽

Genome Size ◽

Dna Content ◽

Nuclear Dna ◽

Nuclear Dna Content ◽

Life Forms ◽

Dna Amount ◽

Ploidy Levels ◽

Ecological Selection ◽

Ecological Implications

Genome size was estimated by flow cytometry in 24 populations belonging to 22 Artemisia taxa (21 species, 1 with two subspecies), which represent the distinct subgenera, life forms, basic chromosome numbers, and ploidy levels in the genus. 2C nuclear DNA content values range from 3.5 to 25.65 pg, which represents a more than sevenfold variation. DNA content per haploid genome ranges from 1.75 to 5.76 pg. DNA amount is very well correlated with karyotype length and ploidy level. Some variations in genome size have systematic and evolutionary implications, whereas others are linked to ecological selection pressures.Key words: Artemisia, Asteraceae, flow cytometry, genome size, nuclear DNA amount variation, systematics, evolution, ecology.

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Nuclear DNA content of commercially important Eucalyptus species and hybrids

Canadian Journal of Forest Research ◽

10.1139/x94-142 ◽

1994 ◽

Vol 24 (5) ◽

pp. 1074-1078 ◽

Cited By ~ 47

Author(s):

D. Grattapaglia ◽

H.D. Bradshaw Jr.

Keyword(s):

Flow Cytometry ◽

Dna Content ◽

Positional Cloning ◽

Nuclear Dna ◽

Nuclear Dna Content ◽

Eucalyptus Species ◽

Ploidy Levels ◽

2C Dna Content ◽

Haploid Genome Size ◽

Commercially Important

This paper reports the nuclear DNA content estimates obtained by flow cytometry for a group of twelve Eucalyptus species and five fast-growing hybrids that includes those most widely planted throughout the world. Estimates of nuclear (2C) DNA content for the species surveyed ranged from 0.77 pg/2C for Eucalyptuscitriodora Hook. (subgenus Corymbia) to 1.47 pg/2C for Eucalyptussaligna Smith (subgenus Symphyomyrtus). This range corresponds to a haploid genome size range of 370–700 megabase pairs. The average physical equivalent of a 1 cM distance could be as low as 200 kilobase pairs in Eucalyptus, an attractive feature for positional cloning efforts in woody plants. The closer the species were in phylogenetic relationship the more similar were their nuclear DNA content values. All the interspecific hybrids surveyed displayed a nuclear DNA content in the expected intermediate range between the respective parental species, with the exception of one originating from Rio Claro, Brazil, whose exact parentage is unknown. No evidence of polyploidy was observed in any of the hybrids. The flow cytometry procedure employed in this study is an efficient method for investigating ploidy levels of high yielding hybrids of Eucalyptus.

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Ploidy Level and DNA Content of Perennial Ryegrass Germplasm as Determined by Flow Cytometry

HortScience ◽

10.21273/hortsci.44.7.2049 ◽

2009 ◽

Vol 44 (7) ◽

pp. 2049-2052 ◽

Cited By ~ 20

Author(s):

Ying Wang ◽

Cale A. Bigelow ◽

Yiwei Jiang

Keyword(s):

Flow Cytometry ◽

Perennial Ryegrass ◽

Ploidy Level ◽

Dna Content ◽

Cool Season ◽

Ploidy Levels ◽

Seedling Plant ◽

2C Dna Content ◽

National Plant Germplasm System ◽

Seedling Leaf

Perennial ryegrass (Lolium perenne L.) is a widely used cool-season turfgrass species. The exact ploidy levels of the worldwide perennial ryegrass accessions in the USDA National Plant Germplasm System (NPGS) are unknown, which could complicate future use and breeding efforts. The objective of this study was to determine the ploidy level and DNA content of the 194 USDA NPGS perennial ryegrass accessions and six commercial cultivars (Brightstar SLT, Catalina II, Divine, Inspire, Manhattan 4, Silver Dollar) using flow cytometry. Among the 200 accessions, 194 diploids and six tetraploids were identified. Three tetraploids originated from Canada with the remaining from Ireland, Japan, and The Netherlands. The average DNA content was 5.60 pg/2C for the diploid and 11.45 pg/2C for the tetraploid. The 2C DNA content was positively correlated (r = 0.23, P < 0.01) with seedling plant height but not seedling leaf width. This ploidy data provide important information for future marker trait analysis and cultivar improvement.

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