Severely distorted Feulgen-DNA amounts in Pinus (Coniferophytina) after nonadditive fixations as a result of meristematic self-tanning with vacuole contents

1986 ◽  
Vol 28 (3) ◽  
pp. 409-415 ◽  
Author(s):  
Johann Greilhuber
Keyword(s):  
Acetic Acid ◽  
Allium Cepa ◽  
Dna Content ◽  
Nuclear Dna ◽  
Large Fraction ◽  
Internal Standard ◽  
Feulgen Reaction ◽  
Root Tips ◽  
Meristematic Tissue ◽  
Dna Amounts

Highly divergent nuclear DNA amounts were obtained in Pinus mugo and Pinus cembra when meristematic tissue from root tips was fixed either with neutral formaldehyde or various nonadditive agents as methanol – acetic acid, ethanol – acetic acid, alcohols alone, Carnoy's fluid, acetone, or was directly hydrolyzed with 5 M HCl. After formaldehyde fixation, the 1C values in P. mugo and P. cembra amount to 20.16 and 24.16 pg, respectively, when calibrated against Allium cepa as internal standard, but 1C values after application of nonadditive fixatives are strongly reduced to 25–41% of the former values. This phenomenon is explained by the observation that in Pinus a large fraction of the meristematic cells contains a considerably large vacuome, whose content (probably condensed tannins) becomes immobilized after formaldehyde fixation and further on does not interfere with the Feulgen reaction, whereas after nonadditive fixations the vacuole contents extravasate and strongly tan the whole meristem and especially the nuclei. The Feulgen reaction is impaired. The tint and absorbance spectra are different in pine and Allium cepa nuclei. Pinus Feulgen-DNA values obtained after fixations other than formaldehyde must be regarded as highly distorted from the true genomic DNA content. Self-tanning as a source of methodical error in DNA content determinations by cytochemical techniques may be widespread because of the frequent occurrence of tannins in the plant kingdom.Key words: Feulgen reaction, DNA contents, tannins, Pinus, conifers.

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

Genes ◽  
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.

Nuclear DNA amount in pure species and hybrid willows (Salix): a flow cytometric investigation

1998 ◽  
Vol 76 (1) ◽  
pp. 157-165 ◽  
Author(s):  
Jérôme Thibault
Keyword(s):  
Flow Cytometry ◽  
Ploidy Level ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Natural Hybridization ◽  
Ploidy Levels ◽  
Key Words Flow Cytometry ◽  
Dna Values ◽  
Dna Amounts

Flow cytometry (FCM) has been used to estimate the nuclear DNA content of 11 Salix species and 5 hybrids. One hundred and sixty nine individuals were studied including 159 individuals from a sequence of 32 communities along a stretch of river in France and 10 individuals from French and English collections for comparison. Isolated nuclei were stained with propidium iodide. FCM was a significantly more practical and rapid technique than that of establishing the karyotype to survey many samples of Salix for variation in ploidy. The 2C DNA amounts for diploid species ranged from 0.76 to 0.98 pg, and tetraploid values ranged from 1.62 to 1.80 pg. The DNA values were consistent with the known ploidy levels. With the exception of a doubtful Salix xquercifolia, ploidy levels and DNA amounts of hybrids were intermediate compared with those of their parents. Intraspecific variation of nuclear DNA values including instrumental variation was low (i.e., 6-11% at the same ploidy level). FCM appeared to be an accurate tool for determination of Salix triploid hybrids. However, it remains limited concerning hybrids from crosses between species of the same ploidy level. Results suggest that natural hybridization might not be frequent in the communities studied, although they have been subject to disturbance. Previous overestimates of hybridization frequency in willows were probably due to misinterpretation of the effects of the environment on Salix spp. morphology; however, the extent and mechanisms of introgression in the genus remain to be further investigated. Key words: flow cytometry, Salix, hybridization, nuclear DNA content, riparian vegetation, disturbance.

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

HortScience ◽  
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.

Associated chromosomal DNA changes in polyploids

Genome ◽  
1994 ◽  
Vol 37 (4) ◽  
pp. 560-564 ◽  
Author(s):  
S. N. Raina ◽  
A. Parida ◽  
K. K. Koul ◽  
S. S. Salimath ◽  
M. S. Bisht ◽  
...  
Keyword(s):  
Genome Size ◽  
Dna Content ◽  
Nuclear Dna ◽  
Seed Set ◽  
Diploid Species ◽  
Expected Value ◽  
Chromosomal Dna ◽  
Seed Fertility ◽  
Dna Loss ◽  
Dna Amounts

The 2C and 4C nuclear DNA amounts were estimated in eight diploid species, belonging to three diverse genera (Vicia, Tephrosia, and Phlox) and their corresponding colchitetraploids. In P. drummondii, T. purpurea, and T. oxygona tetraploids the deviation from the expectation was highly significant. The DNA in P. drummondii was further discarded in subsequent (C1, C2) generations, thus attaining an overall reduction of about 25%. The DNA content in the subsequent generations was the same as that of C2. It is concluded that rapid DNA loss in the first and subsequent generations was not only associated with the substantial increase (30–66%) in the seed set, but it also helped in the establishment and stabilization of the tetraploid. The possible relationship between such a nucleotypic change and success of polyploids is discussed. The DNA change from the expected value in the P. drummondii tetraploid was achieved by equal decrement to each chromosome independent of size, i.e., small chromosomes loose the same amount of DNA as the large chromosomes.Key words: colchitetraploid, genome size, DNA loss, seed fertility, stability, DNA distribution.

scholarly journals RIBOSOMAL DNA AMOUNTS IN PISUM SATIVUM

Genetics ◽  
1975 ◽  
Vol 81 (3) ◽  
pp. 485-492
Author(s):  
C A Cullis ◽  
D Roy Davies
Keyword(s):  
Pisum Sativum ◽  
Ribosomal Dna ◽  
Dna Content ◽  
Leaf Size ◽  
Rrna Genes ◽  
Root Tips ◽  
The One ◽  
Cotyledon Cells ◽  
Dna Amounts

ABSTRACT Different varieties of peas have different proportions of rDNA in their genomes; there is no obvious correlation between the proportions and seed or leaf size. The rDNA proportions in root tips, seedlings, leaves and in the cotyledon cells of high DNA content, were compared in four varieties. In three, there was no difference between tissues; the fourth showed an amplification of rRNA genes in the cells of high DNA content of the seed cotyledon, and also in the cells of young but not of older leaves. The fourth variety was the one that had the lowest proportion of rDNA of all those examined. Studies of the tissues of hybrids between genotypes with "low" and "high" proportions of rDNA showed that heterozygotes had the "high" value, showing again the occurrence of an amplification phenomenon.

Méthode d'étude des quantités d'ADN métaphasique après écrasement de méristèmes radiculaires

Genome ◽  
1992 ◽  
Vol 35 (4) ◽  
pp. 706-708
Author(s):  
C. Le Coq ◽  
C. Guervin ◽  
M. Esclapez ◽  
J. Moret
Keyword(s):  
Dna Content ◽  
Deoxyribonucleic Acid ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Feulgen Reaction ◽  
Root Cells ◽  
Meristematic Root

A method is described for standardized preparation of meristematic root cells treated with colchicine for cytophotométrie analysis of metaphase DNA. Deoxyribonucleic acid has been stained by the Feulgen reaction prior to crushing of the cells.Key words: cytophotometry, Ornithogalum, nuclear DNA content.

Effects of Spent Pot Liner on mitotic activity and nuclear DNA content in meristematic cells of Allium cepa

2012 ◽  
Vol 107 ◽  
pp. 140-146 ◽  
Author(s):  
Larissa Fonseca Andrade-Vieira ◽  
José Marcello Salabert de Campos ◽  
Lisete Chamma Davide
Keyword(s):  
Mitotic Activity ◽  
Allium Cepa ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Meristematic Cells

Nuclear DNA content and base composition in 28 taxa of Musa

Genome ◽  
2001 ◽  
Vol 44 (4) ◽  
pp. 622-627 ◽  
Author(s):  
K Kamaté ◽  
S Brown ◽  
P Durand ◽  
J -M Bureau ◽  
D De Nay ◽  
...  
Keyword(s):  
Dna Content ◽  
Petunia Hybrida ◽  
Base Composition ◽  
Nuclear Dna ◽  
Internal Standard ◽  
Nuclear Dna Content ◽  
Leaf Sheath ◽  
Musa Balbisiana ◽  
A Genome ◽  
Aa Genome

The nuclear DNA content of 28 taxa of Musa was assessed by flow cytometry, using line PxPC6 of Petunia hybrida as an internal standard. The 2C DNA value of Musa balbisiana (BB genome) was 1.16 pg, whereas Musa acuminata (AA genome) had an average 2C DNA value of 1.27 pg, with a difference of 11% between its subspecies. The two haploid (1C) genomes, A and B, comprising most of the edible bananas, are therefore of similar size, 0.63 pg (610 million bp) and 0.58 pg (560 million bp), respectively. The genome of diploid Musa is thus threefold that of Arabidopsis thaliana. The genome sizes in a set of triploid Musa cultivars or clones were quite different, with 2C DNA values ranging from 1.61 to 2.23 pg. Likewise, the genome sizes of tetraploid cultivars ranged from 1.94 to 2.37 pg (2C). Apparently, tetraploids (for instance, accession I.C.2) can have a genome size that falls within the range of triploid genome sizes, and vice versa (as in the case of accession Simili Radjah). The 2C values estimated for organs such as leaf, leaf sheath, rhizome, and flower were consistent, whereas root material gave atypical results, owing to browning. The genomic base composition of these Musa taxa had a median value of 40.8% GC (SD = 0.43%).Key words: Musa, cytometry, banana.

scholarly journals Nuclear DNA Content and Ploidy Levels in the Genus Ipomoea

1994 ◽  
Vol 119 (1) ◽  
pp. 110-115 ◽  
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).

scholarly journals DNA endoreplication level in endosperm during seed development in three monocotyledonous species

2014 ◽  
Vol 62 (3-4) ◽  
pp. 143-147
Author(s):  
Kazimierz Marciniak
Keyword(s):  
Plant Species ◽  
Dna Content ◽  
Species Differences ◽  
Nuclear Dna ◽  
Endosperm Development ◽  
Asparagus Officinalis ◽  
Feulgen Reaction ◽  
Monocotyledonous Plant ◽  
2C Dna Content ◽  
Dna Endoreplication

The DNA content after the Feulgen reaction in the endosperm of three monocotyledonous plant species (<i>Asparagus officinalis</i>, <i>Muscari comosom</i>, <i>Haemanthus kurharinae</i>) differing in their 2C DNA content, was cytophotometrically measured. During endosperm development 1-6 endoreplication cycles take place, depending on the species. Differences in nuclear DNA endoreplication dynamics in the tested species are similar to those occurring in root parenchyma, but the endoreplication level in the endosperm is higher.

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