scholarly journals УРОВНИ ПЛОИДНОСТИ И ОТНОСИТЕЛЬНОГО СОДЕРЖАНИЯ ДНК В КУЛЬТУРЕ КЛЕТОК И ТКАНЕЙ РАСТЕНИЙ IN VITRO

Author(s):  
M. V. Skaptsov ◽  
M. A. Krasnoborodkina ◽  
M. G. Kutsev ◽  
S. V. Smirnov ◽  
A. I. Shmakov ◽  
...  

<p>We presented results of variations in the ploidy level and the genome size of the <em>R. acetosa</em> regenerants. These regenerants was obtained by indirect and direct morphogenesis in in vitro culture. Explants were prepared from seedlings on the three-leaf stage of plant development. More than 100 explants were used to stimulate the indirect and direct morphogenesis. Mesophilic explants were cultured on the MS nutrient medium containing auxin to callus proliferation (2 mg/L NAA, 1 mg/L BA). Cultivation of the callus was maintained for 4 weeks followed by an indirect morphogenes. Indirect morphogenesis stimulated on the MS medium with cytokinin and gibberellic acid predominance (0.5 mg/L BA, 0.2 mg/L GA3). Direct stimulate morphogenesis from the apical meristem of seedlings on nutrient media with a predominance of cytokinins (1 mg/L BA, 0.25 mg/L NAA). Rhizogenesis have stimulated by transferring of the regenerants to the ½MS medium supplemented with 0.2 mg/L of NAA. Research of a ploidy level and genome size was performed by flow cytometry used propidium iodide staining with <em>Vicia faba</em> cv “Innovec” (2C=26.90 pg) as internal DNA standard. We calculated the relative DNA content (2C) for <em>R. acetosa</em> equal to 6,98 pg. Cytogenetical analisis showed that the maximum genome size variation recorded for regenerants obtained through the indirect morphogenesis. Variations in the genome size of the regenerants obtained by direct morphogenesis deviates from the control group to 0.30 pg (2С=7.28 pg) and after indirect morphogenesis to 1.04 pg (2С=8.2 pg). Cytogenetical analysis of the regenerated plants showed the presence of different somatic chromosome numbers ranging from 2n = 14 to 2n = 28. The relative DNA content of tetraploid forms was 11.87 pg. In our study was shown, that the most effective method of plant conservation in the <em>in vitro</em> culture is a direct morphogenesis. Analysis of the relative nuclear DNA content and chromosome counts of regenerants obtained by indirect morphogenesis from the callus cultures showed significant variations in the DNA content, as well as the appearance of polyploid forms. Therefore, long-term cultivation of callus cultures increases the probability of genomic aberrations, which reduces the stability of the plant genome.</p>

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.


PROTOPLASMA ◽  
2013 ◽  
Vol 251 (3) ◽  
pp. 703-709 ◽  
Author(s):  
Rimjhim Roy Choudhury ◽  
Supriyo Basak ◽  
Aadi Moolam Ramesh ◽  
Latha Rangan

2020 ◽  
Vol 1 (40) ◽  
pp. 28-38
Author(s):  
Trang Phuong Nguyen Thi ◽  
Quang Minh Bui ◽  
Hai Duc Le ◽  
Linh Quoc Nguyen

Blepharis maderaspatensis (L.) Heyne ex. Roth is a short-term plant which contains many important secondarycompounds with high medicinal value. Currently, most of the researches focus on chemical composition and pharmacological activity, but the source of raw materials is very limited. In this study, the first step is transferring the samples from nature into in vitro culture conditions to understand the effects of the factors related to shooting and callus morphogenesis was performed, the first node from shoots apical meristem was isolated and sterilized with 1.5% NaOCl for 20 minutes to achieve high efficiency with 86.11% sterile samples and 85.56% shoot growth rate after 2 weeks of culture on MS medium. The shoot generation from axillary shoots was continued to be investigated with the highest number of shoots formed on MS medium supplemented with BA (1 mg / l) showed 1.53 shoots/implant which the height and the number of leavesare 3.65cm and 6.67, respectively. Besides, the formation of callus from leaves of MS medium supplemented with 2.4 - D (0.25 mg / l) achieved the rate of 66.67% of cultured samples, forming good callus after 4 weeks of culture. The results of the study not only contribute importantly to understanding morphogenesis for micropropagation purposes but also serve as the scientific database for further studies at the cellular and molecular levels of this plant.


2021 ◽  
Vol 21 (2) ◽  
Author(s):  
Asmaa A.A. Abdel- Kareem ◽  
H.A. El- Shamy ◽  
A.K. Dawh ◽  
S.G. Gwiefel

The present work was conducted in order to investigate the effect of auxin type (2,4-D and NAA) and concentration (0.00, 0.25, 0.50, 1.00 and 2.00 mg/l) on Balanites aegyptiaca callus cultures growth and production of secondary metabolites. Obtained results demonstrated that supplementation MS medium with 2,4-D at 2.0 mg/l could enhanced and recorded the ultimate values of callus fresh weight, antioxidant activity (%), total flavonoids, total phenolic compounds and total saponins contents and yields of Balanites aegyptiaca L. callus.


2015 ◽  
Vol 57 (1) ◽  
pp. 104-113
Author(s):  
Sandra Cichorz ◽  
Maria Gośka ◽  
Monika Rewers

AbstractSinceM. sinensisAnderss.,M. sacchariflorus(Maxim.) Hack. andM. ×giganteusJ.M.Greef & Deuter ex Hodk. and Renvoize have considerably the highest potential for biomass production amongMiscanthusAnderss. species, there is an urgent need to broaden the knowledge about cytological characteristics required for their improvement. In this study our objectives were to assess the genome size variation among eighteenMiscanthusaccessions, as well as estimation of the monoploid genome size (2C and Cx) of theM. sinensiscultivars, which have not been analyzed yet. The characterization of threeMiscanthusspecies was performed with the use of flow cytometry and analysis of the stomatal length. The triploid (2n = 3x = 57)M. sinensis‘Goliath’ andM. ×giganteusclones possessed the highest 2C DNA content (8.34 pg and 7.43 pg, respectively). The intermediate 2C-values were found in the nuclei of the diploid (2n = 2x = 38)M. sinensisaccessions (5.52–5.72 pg), whereas they were the lowest in the diploid (2n = 2x = 38)M. sacchariflorusecotypes (4.58–4.59 pg). The presented study revealed interspecific variation of nuclear DNA content (P<0.01) and therefore allowed for recognition of particular taxa, inter- and intraspecific hybrids and prediction of potential parental components. Moreover, intraspecific genome size variation (P<0.01) was observed inM. sinensiscultivars at 3.62%. The values of the stomatal size obtained for the triploidM. ×giganteus‘Great Britain’ (mean 30.70 μm) or ‘Canada’ (mean 29.67 μm) and diploidM. sinensis‘Graziella’ (mean 29.96 μm) did not differ significantly, therefore this parameter is not recommended for ploidy estimation.


2003 ◽  
Vol 83 (4) ◽  
pp. 873-876 ◽  
Author(s):  
A. N. Aziz ◽  
R. J. Sauvé ◽  
S. Zhou

Daylily (Hemerocallis sp. ‘Stella de Oro’) callus cultures initiated from ovules were bombarded with gold particles coated with plasmid harboring Basta® resistance gene. Resulting putative transgenic calli were selected after 3 wk on semi-solid Murashige and Skoog’s (MS) basal medium supplemented with 10 mg L-1 1-naphthaleneacetic acid, 2 mg L-1 6-benzylaminopurine and 3 mg L-1 phosphinothricin (PPT). Surviving calli regenerated shoots after 2 mo on semi-solid MS medium supplemented with 2 mg L-1 thiadiazuron and 1 mg L-1 PPT. Polymerase chain reaction and Southern blotting were used to confirm independent transformation events. Key words: Basta® resistance, in vitro, Hemerocallis


Genome ◽  
1989 ◽  
Vol 32 (5) ◽  
pp. 834-839 ◽  
Author(s):  
C. Juan ◽  
E. Petitpierre

The relative amount of C-banded heterochromatin varies strikingly in seven species of tenebrionid beetles, from 25 to 58%, but most species show procentric bands only. Nevertheless, Gonocephalum patruele exhibits an almost completely heterochromatic X chromosome. The nuclear DNA content of Feulgen-stained spermatids has yielded up to a threefold difference, from 0.27 to 0.86 pg, which is not completely in accordance with the amount of C-banded heterochromatin. However, the genome sizes correlate significantly with the total chromosome areas at metaphase I and with the spermatid areas. Furthermore, the genome sizes agree with the subfamilial taxonomic groupings of these tenebrionids.Key words: Tenebrionidae, genome size, C-banding.


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