scholarly journals The Effect of Storage in Vitro on the Dna Content of Bull Spermatozoa

1972 ◽  
Vol 25 (1) ◽  
pp. 175 ◽  
Author(s):  
AW Blackshaw ◽  
GW Salisbury
Keyword(s):  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Feulgen Staining ◽  
Linear Measurements ◽  
Fast Green ◽  
Orange Fluorescence ◽  
Nuclear Histone ◽  
Photometric Measurements

Bull spermatozoa were aged in vitro for periods of up to 9 days. Microspectro. photometric measurements were made of the nuclear DNA content of the sperm heads using ultraviolet absorption, acridine orange fluorescence, and Feulgen staining and of the nuclear histone content using fast green staining. The surface area of the sperm heads was calculated from linear measurements of head length and width.

Assessment of the cellular DNA content of whole mounted mouse and human oocytes and of blastomeres containing single or multiple nuclei

Zygote ◽  
1993 ◽  
Vol 1 (1) ◽  
pp. 17-25 ◽  
Author(s):  
Nicola J. Winston ◽  
Martin H. Johnson ◽  
Peter R. Braude
Keyword(s):  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Structural Features ◽  
Chromosomal Dna ◽  
Individual Values ◽  
Mean Values ◽  
Mouse Oocytes ◽  
Cellular Dna

SummaryThe nuclear DNA content of intact, live or fixed, human and mouse oocytes and blastomeres has been measured rapidly and reliably. Chromosomal DNA has been stained with DAPI, the fluorescent emission from which has been measured photocytometrically.In vitrofertilised mouse oocytes and embryos at various stages of development were assessed for their DNA content. The mean values of 1C, 2C and 4C DNA content were clearly different, and it was possible to assign correctly individual values for DNA content to each class with 92%, 61% and 81% confidence respectively. Maintaining the cells as whole mounts allowed other morphological and structural features to be examined. When formation of multiple micronuclei was induced in mouse oocytes by their insemination in the presence of nocodazole, the additive signal from all the micronuclei in one zygote was equivalent to the expected DNA content. Application to early human blastomeres of this photocytometric technique for measurement of the total cellular DNA content revealed that multinucleated blastomeres contained 2C to 4C DNA levels, consistent with a diploid DNA content.

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.

Nuclear DNA content and isozyme variation in relation to morphogenic potential of strawberry (Fragaria × ananassa) callus cultures

1991 ◽  
Vol 69 (2) ◽  
pp. 239-244 ◽  
Author(s):  
Narender S. Nehra ◽  
Kutty K. Kartha ◽  
Cecil Stushnoff
Keyword(s):  
Dna Content ◽  
Nuclear Dna ◽  
Flow Cytometric Analysis ◽  
Nuclear Dna Content ◽  
Leaf Explants ◽  
Callus Cultures ◽  
Fragaria Ananassa ◽  
Flow Cytometric ◽  
Ploidy Changes

Callus cultures of strawberry cv. Redcoat (2n = 8x = 56) initiated from greenhouse and in vitro leaf explants were examined at various culture periods for morphogenic response, changes in nuclear DNA content, and isozyme banding patterns of four enzymes. The flow cytometric analysis of nuclear DNA content revealed the occurrence of polyploid and aneuploid changes as a function of ageing of callus cultures. The calli initiated from in vitro leaf explants were more prone to such changes than those initiated from greenhouse leaf explants. The in vitro morphogenic ability of callus cultures was affected by the ploidy changes, but the latter were not the only cause for loss in regeneration potential of long-term callus cultures. The isozyme phenotypes of esterase, phosphoglucomutase, phosphoglucoisomerase, and leucine aminopeptidase did not change with the chromosomal variation in callus cultures. Key words: strawberry, Fragaria × ananassa, callus culture, flow cytometry, nuclear DNA content, isozyme.

Nuclear DNA content in different plant materials of Plantago asiatica L. cultured in vitro

2008 ◽  
Vol 94 (1) ◽  
pp. 65-71 ◽  
Author(s):  
Joanna Makowczyńska ◽  
Emilia Andrzejewska-Golec ◽  
Elwira Sliwinska
Keyword(s):  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Plant Materials ◽  
Plantago Asiatica

scholarly journals In Vitro Propagation and Nuclear DNA Content of Tripleurospermum Insularum (Asteraceae) – A Critically Endangered Insular Endemic Species From Turkey

2021 ◽  
Author(s):  
Huseyin Inceer ◽  
Mustafa Cuce ◽  
Kemal Vehbi Imamoglu ◽  
Tugba Ergin ◽  
Ali Omer Ucler
Keyword(s):  
Natural Population ◽  
Endemic Species ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Critically Endangered ◽  
Ex Situ ◽  
Nodal Segments ◽  
Node Number

Abstract Tripleurospermum insularum Inceer & Hayirlioglu-Ayaz (Asteraceae) is a critically endangered endemic species in Turkey that is face the risk of extinction as a result of the fragmentation of its habitat as well as overgrazing and trampling of its natural population. However, the protocol for micropropagation of this threatened species has not been developed yet. Here, its regeneration ability on MS media supplemented with different plant growth regulators were evaluated using nodal segments. The higher number and length of shoot per explant was achieved with the addition 4.6 µM ZEA and 0.5 µM IAA to the culture medium. Besides, the highest node number of shoot per explant was obtained from MS medium supplemented with 4.6 µM ZEA and 0.5 µM IBA. Flow cytometric analysis also revealed that most of the in vitro developed shoots of T. insularum possessed similar nuclear DNA content as well as ploidy level as initial material and plants from natural population. In vitro rooting of shoots was achieved at 100 % efficiency containing 2.9 µM IAA. Rooted and well-developed plantlets were initially acclimatized under greenhouse conditions and then moved to the botanical garden, where they matured and flowered. Finally, 76% and 74% survivals were achieved during the acclimatization process, respectively. This is the first report of a successfully developed micropropagation protocol of threatened T. insularum for its ex situ conservation.

In vitro culture of Aloe barbadensis Mill.: Morphogenetic ability and nuclear DNA content

Plant Science ◽  
1988 ◽  
Vol 55 (1) ◽  
pp. 53-59 ◽  
Author(s):  
I. Castorena Sanchez ◽  
L. Natali ◽  
A. Cavallin
Keyword(s):  
In Vitro Culture ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Aloe Barbadensis

scholarly journals Nuclear DNA content as an indicator of inflorescence colour stability of in vitro propagated solid and chimera mutants of chrysanthemum

2020 ◽  
Vol 143 (2) ◽  
pp. 421-430 ◽  
Author(s):  
Natalia Miler ◽  
Dariusz Kulus ◽  
Elwira Sliwinska
Keyword(s):  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Γ Irradiation ◽  
Periclinal Chimera ◽  
Single Node ◽  
Periclinal Chimeras ◽  
Fcm Analysis ◽  
The Stability

Abstract In chrysanthemum, breeders seek for desirable characteristics of the inflorescence, which can first be established once the plant is mature. The present study aims to determine whether measurement of DNA content can be useful in the detection of somaclonal variants and/or separation of chimera components in chrysanthemum at the early in vitro multiplication stage. Eleven Chrysanthemum × morifolium (Ramat.) Hemsl. cultivars of the Lady group (a mother cultivar and ten of its radiomutants obtained by X-ray- or γ-irradiation; solid and periclinal chimeras) were propagated in vitro. Single-node explants were cultured in Murashige and Skoog (MS) medium, either without plant growth regulators (PGRs) or supplemented with 6-benzyladenine (BA) and indole-3-acetic acid (IAA). The nuclear DNA content was measured by flow cytometry (FCM) in the shoots produced in vitro. After acclimatization and growth of the plants in a glasshouse, inflorescence colour was recorded. The addition of PGRs to the medium almost doubled the mean number of shoots produced in vitro per explant, but caused a change in inflorescence colour of all (‘Lady Apricot’; periclinal chimera) or part of the plants (‘Lady Amber’; solid mutant and ‘Lady Salmon’; periclinal chimera). All radiomutants contained less DNA than the mother cultivar ‘Richmond’. There were significant differences in DNA content between plants of the same cultivar grown in media with or without PGRs for ‘Lady Apricot’ and ‘Lady Salmon’, but no phenotype alternation occurred in chrysanthemums produced in PGR-free medium compared to the original cultivars. Conversely, in medium with PGRs, chimeras produced flowers different from the original colour. In all except one cultivar (‘Lady Amber’; solid mutant) a lack of differences in genome size between plants grown in either medium coincided with a stable inflorescence colour. The occurrence of some plants of ‘Lady Amber’ with different inflorescence colour may be due to small DNA changes, undetectable by FCM. It can be concluded that FCM analysis of DNA content in young plantlets can be indicative of the stability of inflorescence colour in chrysanthemum, especially chimeric cultivars, and for mutant detection.

scholarly journals Nuclear DNA Content and Chromatin Pattern of Rat Rhabdomyosarcoma Cell Sublines with Different Metastatic Potentials

2000 ◽  
Vol 20 (1) ◽  
pp. 41-48 ◽  
Author(s):  
Jean Dufer ◽  
Marie-France Poupon ◽  
Sonia Yatouji
Keyword(s):  
Tumor Cells ◽  
Cell Lines ◽  
Dna Content ◽  
Nuclear Dna ◽  
Chromatin Condensation ◽  
Nuclear Dna Content ◽  
Image Cytometry ◽  
Chromatin Pattern ◽  
Rhabdomyosarcoma Cell

There is a constant need of features able to characterize potentially metastatic cells among the heterogeneous cell subpopulations which constitute a tumor. Image cytometry of metastatic tumor cells give rise to variable results, partly because of a heterogeneous origin of cells, or potential drug effects. The aim of this work was to characterize nuclear changes observed in metastatic cell clones issuedin vitrofrom the same parental cell population The nuclear phenotypes of 6 cell sublines isolated from a rat rhabdomyosarcoma cell line and differing in their metastatic ability were evaluated by image cytometry on Feulgen‐stained preparations. Densitometric [5], geometric [3] and textural [9] features were computed from each nuclear image. For each cell subline, a metastatic score, ranging from 0 to 10, was calculated on the basis ofin vitroinvasivity data, by measuring the number of pulmonary metastases observed after s.c. graft of tumor cells in rats. Data obtained were compared to karyotype, growth characteristics, and oncogene expressions of cell lines. The nuclear DNA content, the chromosome numbers, the cell sublines doubling times, and the distribution of cells within the cell cycle appear unrelated with this score. On the contrary, increase in metastatic ability is accompanied by changes in chromatin pattern as assessed by textural features. Progressive increase in chromatin condensation can be observed in cell sublines with increasing metastatic score. These results were confirmed by an unsupervised multivariate partitioning of rhabdomyosarcoma cells which identified two separate subsets whose distributions within the analyzed cell lines correlate with their metastatic ability. These data suggest that, in rat rhabdomyosarcoma cell sublines, metastatic ability could be associated with nuclear morphological changes at the level of chromatin texture.

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