Study of chromosomes, DNA amount, and in vitro growth in different species of Luzula

Genome ◽  
1990 ◽  
Vol 33 (1) ◽  
pp. 143-147 ◽  
Author(s):  
Jayanti Sen ◽  
Sumona Mukherjee ◽  
A. K. Sharma

Cells of six different species of Luzula were subjected to in vitro culture and examined for callus formation. The cells of calli from the different species were analyzed for variation in chromosome number. The cells of different species showed a differential response in culture with respect to callus growth. Chromosome number variation in vitro, including both hypo- and hyper-diploidy, was recorded in all species except L. pediformis and L. luzuloides. The chromosome fragments survived as a result of a nonlocalized centromere. With prolonged culture, normal diploid cells were frequent. The nuclear DNA content of cells of these species was also measured, both in vivo and in vitro. The DNA values ranged from 6.05 to 7.03 pg per 4C nucleus. No marked change in DNA value was noted in cells even with high chromosome numbers, thereby confirming their origin through fragmentation of chromosomes. In cells of callus, the DNA amount was observed to be higher than in normal cells, indicating fragmentation and duplication of individual chromosomes. Their importance in the origin of new genotypes has been indicated.Key words: Luzula spp., in vitro growth, chromosomes, nuclear DNA content.

2016 ◽  
Vol 58 (1) ◽  
pp. 81-93 ◽  
Author(s):  
Małgorzata Podwyszyńska ◽  
Dorota Kruczyńska ◽  
Aleksandra Machlańska ◽  
Barbara Dyki ◽  
Iwona Sowik

Abstract Apple species and cultivars differ in nuclear (2C) DNA content and ploidy level. The majority of these genotypes are diploids, but there are some triploids and a few tetraploids. Nuclear DNA content is a specific feature and its flow cytometric evaluation can be helpful in differentiating taxa. For many apple genotypes – including all the Polish ones, these characteristics are not known. 2C DNA was evaluated in relation to leaf, flower, fruit, pollen grain and stomata sizes as well as to the flowering time for seventy genotypes (including 46 Polish cultivars) gathered in the gene bank of the Research Institute of Horticulture, Skierniewice, Poland. For standard cultivars with the known chromosome number, 2C value was 1.71 pg for diploid cultivar ‘Alwa’ (2n=2x=34), 2.55 pg for triploid ‘Boskoop’ (3x=51), and 3.37 pg for tetraploid genome (4x=68) of mixoploid ‘McIntosh 2x+4x’. In 61 cultivars (including 41 Polish ones), the nuclear DNA content ranged from 1.58 to 1.78 pg indicating their diploid chromosome number. Five cultivars were identified as triploids (‘Bursztówka Polska’, ‘Pagacz’, ‘Rapa Zielona’, ‘Rarytas Śląski’ and ‘Witos’) owing to their nuclear DNA amount ranging between 2.42 and 2.58 pg. Leaf, flower, fruit, stomata and pollen grain sizes were on average significantly larger in triploids. Thus, in 3x plants the mean leaf surface was 49.1 cm2, flower diameter – 52.4 mm, fruit weight – 204.7 g, stomata length – 32.1 μm and pollen grain diameter – 33.7 μm, whereas in diploids – 36.0 cm2, 46.1 mm, 162.7 g, 28.4 μm and 30.7 μm, respectively. Pollen grain viability was on average significantly higher in diploids (75.6%), compared to triploids (22%). These results confirm that in apple, as in many other plant species, the higher ploidy level of triploids is generally associated with increased sizes of pollen grains, stomata, flowers, fruits and leaves but decreased pollen viability. No clear correlation between ploidy level and flowering time was found. In the case of mixoploid apple genotypes possessing diploid and tetraploid genomes, some phenotype observation is helpful in describing the ploidy level of the histogenic layers, L1 and L2. Small stomata sizes (similar to diploid) indicate diploid L1 and larger leaf sizes, compared to diploid counterparts, show tetraploid L2. The results will be used for breeding, in which it is important to determine maternal and paternal genotypes as well as the direction of the crossing that is of great importance in obtaining seeds and materials for further selection.


2017 ◽  
Vol 17 (4) ◽  
pp. 320-326 ◽  
Author(s):  
Luis Felipe Lima e Silva ◽  
Vânia Helena Techio ◽  
Luciane Vilela Resende ◽  
Guilherme Tomaz Braz ◽  
Kátia Ferreira Marques de Resende ◽  
...  

Zygote ◽  
1993 ◽  
Vol 1 (1) ◽  
pp. 17-25 ◽  
Author(s):  
Nicola J. Winston ◽  
Martin H. Johnson ◽  
Peter R. Braude

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.


Genome ◽  
1990 ◽  
Vol 33 (5) ◽  
pp. 659-662 ◽  
Author(s):  
H. L. Porter ◽  
A. Lane Rayburn

The B-chromosome and C-band numbers were determined in 12 Arizona Indian maize populations. These populations were originally collected from altitudes ranging from 100 to 5300 ft (1 ft = 0.3048 m). In addition, the total nuclear DNA amounts of these populations have been observed to vary by as much as 20%. The number of B-chromosomes was not significantly correlated with altitude, C-band number, or nuclear DNA amount. C-band number was significantly correlated with both altitude and genome size. It does not appear that the amount of C-band variation can account for the large nuclear DNA variation observed in these accessions. Additional A-chromosomal DNA sequences may be involved in the nuclear DNA content variation that exists among these accessions.Key words: heterochromatin, DNA content, evolution, repeated DNA.


Genes ◽  
2021 ◽  
Vol 12 (12) ◽  
pp. 1950
Author(s):  
Guadalupe Palomino ◽  
Javier Martínez-Ramón ◽  
Verónica Cepeda-Cornejo ◽  
Miriam Ladd-Otero ◽  
Patricia Romero ◽  
...  

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.


HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 580e-580
Author(s):  
Rodomiro Ortiz ◽  
D.E. Costich ◽  
T.P. Meagher ◽  
N. Vorsa

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.


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

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.


1991 ◽  
Vol 69 (2) ◽  
pp. 239-244 ◽  
Author(s):  
Narender S. Nehra ◽  
Kutty K. Kartha ◽  
Cecil Stushnoff

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.


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