scholarly journals Deschampsia antarctica E. Desv. plants with different chromosome number cultivated in vitro. Plants length and flavonoids in in vitro culture and in nature

Author(s):  
O. O. Poronnik ◽  
I. Yu. Parnikoza ◽  
N. Yu. Miryuta ◽  
G. Yu. Myryuta ◽  
V. P. Grakhov ◽  
...  

Aim. The aim of this work is to research two adaptability indices (leaf length and flavonoid content and spectrum) in eleven genotypes revealed in seeds germ of Deschampsia antarctica E. Desv. in standardized cultivation condition in vitro. Then these adaptability indices would compare with the same for native populations. Methods. We have used biochemical method for determination of flavonoid contents in plant leaves, HPLC method for research of flavonoid spectrum in plant leaves. Results. Heterogeneity of leaf length and flavonoid content has been shown. Both parameters difference have been detected in comparison with native populations. Flavonoid spectrum has no difference in comparison with native populations. Conclusions. Deschampsia antarctica genotypes under standardized cultivation condition in vitro have had longer leaves than plants in nature populations. Length of the leaves was varied depending on karyotype of corresponding genotype. Cultivated plants have had flavonoid spectrum similar to the same plants in nature. This fact allows to assume plants in vitro possible producent of secondary metabolites. Keywords: Deschampsia antarctica, leaf length, flavonoids, chromosome number.

Author(s):  
I. Yu. Parnikoza ◽  
N. Yu. Miryuta ◽  
M. Roiek ◽  
A. A. Betiekhtin ◽  
O. O. Poronnik ◽  
...  

Aim. We have researched probabilistic relations between genome size and two adaptability indices in eleven genotypes revealed in seeds germ of Deschampsia antarctica E. Desv. which has different genome size under standardized cultivation condition in vitro. Methods. We have used biochemical method for determination of flavonoid contents in plant leaves and flow cytometry analysis for determination of genome size in plant leaves. Results. Eleven genotypes of Deschampsia antarctica with different genome size have been researched by three data rows. Probabilistic scheme have been created and analyzed for all researched genotypes. Conclusions. The Deschampsia antarctica genome size or karyotype changes have been shown to have effect on its probabilistic relations with flavonoids content and leaf length. Mechanisms of such relations is required a detailed research. Keywords: Deschampsia antarctica, genome size, leaf length, flavonoids.


Author(s):  
N. Yu. Miryuta ◽  
I. Yu. Parnikoza ◽  
O. O. Poronnik ◽  
G. Yu. Myryuta ◽  
V. A. Kunakh

Aim. This work deals with probabilistic relations between genome size and adaptability indices research in 11 genotypes plants revealed among seeds of Deschampsia antarctica E. Desv. which has different genome size under standardized cultivation condition in vitro. Methods. We have used electrophoretic method for determination of protein relative contents in plant leaves, and statistical analysis for probabilistic relations determination. Results. Protein densitometric profiles for 11 D. antarctica genotypes plants were analyzed. Probabilistic scheme have been created for all researched plants genotypes for all previously analyzed parameters. Three probabilistic schemes of genotypes plants received from seeds collected in the same loci and season have been shown and analyzed. Conclusions. Protective protein densitometric profiles were similar for 11 genotypes of D. antarctica plants under standardized cultivation condition in vitro but quantitative part of protective protein has been changed. Genome size had an influence on its probabilistic relations with flavonoids content, leaf length and some protein relative contents in general pool. Probabilistic relations between genome size and some protein relative content and between the last and flavonoids content and leaf length is assumed to express underlying in this plants adaptation process to uniqueness nature condition in dynamic hereditary memory special information in certain degree. Mechanisms of probabilistic relations changes are required a detailed research.Keywords: Deschampsia antarctica, plants in vitro, probabilistic relations between genome size and adaptability indices, plant adaptability.


2021 ◽  
pp. 56-81
Author(s):  
N. Miryuta ◽  
◽  
I. Parnikoza ◽  
O. Poronnik ◽  
G. Myryuta ◽  
...  

The research was to develop and describe in detail the algorithm for calculating the United Quality Latent Index (UQLI, Iq ) of plant adaptability from the collection of Deschampsia antarctica Ė. Desv. genotypes obtained from seeds collected at different sites in the Argentine Islands region, the maritime Antarctic, and grown in vitro at the laboratory conditions. Genome size and genetic distances by ISSR and IRAP markers according to data from published articles were used as basic indices of initial genetic heterogeneity for analyzed plant genotypes. To assess individual adaptability indices for eleven D. antarctica genotypes, we used measurement of the leaf length morphometric index and determination of the flavonoids content by rutin and the content of photosynthetic pigments. The spectra of reserve and protective proteins in leaves were investigated by polyacrylamide gel electrophoresis. To obtain the United Quality Latent Index of Adaptability (Iqi, UQLI), the method of extreme grouping was used. The estimation of Iqi (UQLI) was performed using pairwise comparisons of indices from differences sets for each pair of genotypes. We developed and described in detail the algorithm for Iqi estimation for eleven D. antarctica genotypes. As an example of application, correlation models of probability relations of the indices are presented. To evaluate the complex adaptability for eleven D. antarctica genotypes grown in vitro we used developed algorithm for the UQLI calculation. The individuality of the adaptive portrait for all studied genotypes under in vitro cultivation conditions was shown. The influence of basic genetic characteristics (genome size and genetic distances) on auxin metabolism-related indices of leaf length and flavonoid content was shown. Such effect may be carried out by genetic characteristics both individually and together, probably via auxin metabolism. Among the eight genotypes researched, we distinguish four different variants by correlation models and two (positive and negative) by the general Iqi value. Thus the Iqi (UQLI) is proposed to describe a large number of source data at different organization levels which characterize sample genotypes by reducing the dimensions to one dimensionless number. This genotypes’ individuality and the peculiarities of their grouping by Iqi should be taken into account when doing experimental studies using these genotypes as model plants, especially in experiments studying the regulation of productivity and the effect of the various exogenous factors, etc.


2016 ◽  
Vol 14 (1) ◽  
pp. 36-43 ◽  
Author(s):  
V. A. Kunakh ◽  
D. O. Navrotska ◽  
M. O. Twardovska ◽  
I. O. Andreev

Aim. To clarify the details of chromosome variation in calli derived from D. antarctica plants in the initial passages of the culture in vitro. Methods. Induction of callus from root explants of plants, which were grown from seeds, and consequent subcultivation of tissue culture. Cytogenetic analysis of squashed slides stained by acetic-orcein and counting the number of chromosomes in mitotic metaphase plates. Results. There were analyzed the cultured tissues derived from D. antarctica plants with different chromosome numbers: diploid plants (2n=26), mixoploid plant with B-chromosomes (2n=26+1-3B), and mixoploid plant with near-triploid modal class (2n=36, 38). Analysis of callus tissues of all plants at 2-4 passages revealed mixoploidy, presence of polyploid and aneuploid cells. The modal class in all studied calli was composed of diploid and aneuploid cells with near-diploid chromosome number. The cytogenetic structure of cell population of cultured tissues was found to vary with characteristics of the karyotype of donor plant. The largest range of variation in the number of chromosomes (from 18 to 63 chromosomes) was found in tissue culture of diploid plant (2n=26) from the Galindez Island, and the highest frequencies of polyploid (47 %) and aneuploid cells were in the culture of mixoploid plant with near-triploid modal class from the Big Yalour Island. Conclusions. In different D. antarctica cultured tissues at the early stages of the culture, the modal class was composed of diploid cells and cells with near-diploid chromosome number irrespective of karyotype of donor plant (diploid, mixoploid poliploid).Key words: Deschampsia antarctica Desv., plant tissue culture, chromosomal variability in vitro, mixoploidy.


2015 ◽  
Vol 10 (3) ◽  
pp. 1934578X1501000 ◽  
Author(s):  
Agnieszka Kicel ◽  
Monika Anna Olszewska

This study was designed to evaluate the flavonoid, saponin (TSC) and phenolic (TPC) contents and in vitro antioxidant activity of the crude (CME) and dry extracts and fractions of Medicago lupulina L. aerial parts. A validated RP-HPLC method led to quantitation of flavonols (kaempferol, quercetin and myricetin) and flavones (apigenin and luteolin) in the hydrolyzed extract. TSC and TPC were assayed spectrophotometrically at 560 and 760 nm, respectively. The antioxidant activity of the CME and the dry fractions were followed in vitro by DPPH free radical and ferric reducing antioxidant power (FRAP) methods. The flavonoid content of CME was 1.27 mg/g dw. The prevailing flavonoids were luteolin and myricetin, at concentrations of 0.37 and 0.36 mg/g dw, respectively. TSC and TPC were detected in CME at the level of 90.4 mg ESE/g dw, and 12.9 mg GAE/g dw, respectively. In the DPPH and FRAP tests, the CME exhibited antioxidant capacity with TEAA and FRAP values of 45.4 μmol Trolox®/g dw and 0.2 mmol Fe2+/g dw, respectively. The diethyl ether dry fraction was the most valuable one, showing the highest antioxidant activity (TEAA = 726.1 μmol Trolox®/g dw, FRAP = 2349.4 μmol Fe2+/g dw) that was in accordance with its high TPC (162.4 mg/g dw).


2021 ◽  
Vol 74 ◽  
Author(s):  
Nataliia Nuzhyna ◽  
Viktor Kunakh ◽  
Oksana Poronnik ◽  
Ivan Parnikoza

Abstract We studied the anatomical features of the leaf structure of Deschampsia antarctica É. Desv. (Poaceae) obtained from seeds collected from different habitats of the maritime Antarctic. These plants have been maintained in culture in vitro for more than 7 years. The plants include stable chromosome forms comprising diploids (2 n = 26), a diploid plant with B-chromosomes (2 n = 26 + 0–3B), a triploid with rearranged chromosome morphology, and myxoploids with different ratios of diploid and nondiploid cells in the root meristem. The D. antarctica plants that were studied generally had a similar anatomical structure in culture in vitro and in nature. At the same time, plants of different cultivated genotypes also displayed their own leaf structure features. In particular, qualitative features included asymmetric four-ribs and five-rib leaves instead of typical three-rib leaves for some genotypes, some individuals characterized by the presence of unicellular nonglandular pointed trichomes on an adaxial side, differences in vascular bundle sheath, and other features. No clear dependence of the anatomical structural features on the chromosomal status of the studied genotypes was evident. At the same time, differences were evident in traits that included the number of stomata, thickness of the outer cell walls of the epidermis, bundles sheath state, number of leaf ribs, and the persistent presence of trichomes under prolonged in vitro cloning. The findings indicate that D. antarctica plants collected from different locations of the maritime Antarctic for long-term cultivation in vitro under standard cultivation conditions retain the anatomical characteristics of the wild plants. The polymorphism of in vitro-cultivated plants is not related to the polymorphism of their karyotype, but is apparently due to the heterogeneity of the original plants and possibly the epigenetic fixation of a number of anatomical features produced in the natural microhabitats of D. antarctica populations.


2020 ◽  
Vol 26 ◽  
pp. 276-281
Author(s):  
M. O. Twardovska ◽  
I. I. Konvalyuk ◽  
K. V. Lystvan ◽  
I. O. Andreev ◽  
V. A. Kunakh

Aim. The aim of the study was a comparative assessment of total phenolic content and total flavonoid content in in vitro plants, regenerated plants, plants grown in a growth chamber, and tissue culture of several genotypes of Deschampsia antarctica. Methods. In vitro culture, Folin-Ciocalteu method, spectrophotometry, high-performance liquid chromatography. Results. The total content of phenolic compounds and total flavonoid content was determined in the samples of three D. antarctica genotypes: G/D12-2a (2n=26), DAR12 (2n=26+0–3B) and Y66 (2n=36–39). The content of these biologically active compounds was the highest in in vitro plants and it was higher than in plants grown in a growth chamber. The highest content of BAC was found in DAR12 in vitro plants (16.50 mg of ferulic acid equivalent and 21.26 mg of rutin equivalent per g of dry weight, respectively). The regenerated plants did not differ significantly in the content of BAC from the original in vitro plants. In tissue culture, the content of BAC was lesser. One- and two-year-old tissue cultures did not differ significantly in the content of phenolic compounds and flavonoids. Conclusions. The relatively high content of phenolic compounds and flavonoids in in vitro plants and in regenerated plants indicates that in vitro cultivated D. antarctica plants can be a promising raw material for production of valuable BACs. Keywords: Deschampsia antarctica E. Desv., in vitro plants, plant tissue culture, phenolic compounds, flavonoids.


2020 ◽  
Vol 16 ◽  
Author(s):  
M. Alarjah

Background: Prodrugs principle is widely used to improve the pharmacological and pharmacokinetic properties of some active drugs. Much effort was made to develop metronidazole prodrugs to enhance antibacterial activity and or to improve pharmacokinetic properties of the molecule or to lower the adverse effects of metronidazole. Objective: In this work, the pharmacokinetic properties of some of monoterpenes and eugenol pro metronidazole molecules that were developed earlier were evaluated in-vitro. The kinetic hydrolysis rate constants and half-life time estimation of the new metronidazole derivatives were calculated using the validated RP-HPLC method. Method: Chromatographic analysis was done using Zorbbax Eclipse eXtra Dense Bonding (XDB)-C18 column of dimensions (250 mm, 4.6 mm, 5 μm), at ambient column temperature. The mobile phase was a mixture of sodium dihydrogen phosphate buffer of pH 4.5 and methanol in gradient elution, at 1ml/min flow rate. The method was fully validated according to the International Council for Harmonization (ICH) guidelines. The hydrolysis process carried out in an acidic buffer pH 1.2 and in an alkaline buffer pH 7.4 in a thermostatic bath at 37ºC. Results: The results followed pseudo-first-order kinetics. All metronidazole prodrugs were stable in the acidic pH, while they were hydrolysed in the alkaline buffer within a few hours (6-8 hr). The rate constant and half-life values were calculated, and their values were found to be 0.082- 0.117 hr-1 and 5.9- 8.5 hr., respectively. Conclusion: The developed method was accurate, sensitive, and selective for the prodrugs. For most of the prodrugs, the hydrolysis followed pseudo-first-order kinetics; the method might be utilised to conduct an in-vivo study for the metronidazole derivatives with monoterpenes and eugenol.


Pharmaceutics ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 158
Author(s):  
Snežana Ilić-Stojanović ◽  
Ljubiša Nikolić ◽  
Vesna Nikolić ◽  
Slobodan Petrović ◽  
Violeta Oro ◽  
...  

In this study, poly(N-isopropylacrylamide-co-2-hydroxypropyl methacrylate) hydrogels were synthesized using free radical initiated copolymerization method. Four hydrogels with different cross-linker concentrations were prepared. Semi-crystalline, cross-linked copolymer networks were confirmed by FTIR, SEM and XRD analysis. Variation of swelling behaviour was monitored gravimetrically and thermo-responsiveness has been noticed. An application of synthesized thermo-responsive hydrogels as carriers for the modulated release of anti-inflammatory model drug was investigated. Moreover, naproxen loading into these hydrogels was also determined using FTIR, SEM and XRD techniques and release was analyzed using HPLC method at simulated physiological conditions. Swelling kinetic and mechanism of water transport, as well as diffusion of naproxen through the hydrogels were analyzed. Thus, the aim of this work was to study various compositions of obtained hydrogels and their possibility of application as a thermo-responsive carrier for prolonged naproxen release in order to evaluate as a potential candidate for drug carrier in future pharmaceutical applications.


Author(s):  
Aakriti Bhandari ◽  
Harminder Singh ◽  
Amber Srivastava ◽  
Puneet Kumar ◽  
G. S. Panwar ◽  
...  

Abstract Background Sophora mollis Royle (family Fabaceae, subfamily-Papilionaceae) is a multipurpose legume distributed in plains and foothills of the North-West Himalaya to Nepal and is facing high risk of extinction due to habitat loss and exploitation by the local people for its fuel and fodder values. Therefore, the present study was conducted to standardize a micropropagation protocol for Sophora mollis by using shoot tip explants and to study the meiotic chromosome count in the species. Results Multiple shoots were induced in shoot tip explants of Sophora mollis in Murashige and Skoog medium supplemented with different concentrations of cytokinins alone (BAP, TDZ, and Kinetin) and in combination with varying concentrations of NAA. MS medium supplemented with BAP (8.9 μM) was observed to be the optimal medium for multiple shoot induction and maximum 25.32 shoots per explant was obtained with average length of 4.5 ± 0.8 cm. In vitro developed shoots were transferred onto rooting media supplemented with different concentrations of auxin (IAA, IBA, and NAA). Maximum 86% rooting was observed in half-strength MS medium supplemented with 21.20 μM NAA with an average of 21.26 roots per culture. In vitro raised plantlets were adapted to greenhouse for better acclimatization and 60% plants were successfully transferred to the open environment. Based on the chromosome counts available from the literature and the current study, the species tend to show a basic chromosome number of x = 9. Conclusion The micropropagation protocol standardized can be helpful for the ex situ mass multiplication and germplasm conservation of the endangered species. Moreover, the ex situ conservation approach will be helpful in actively bridging the gap between ex situ and in situ approaches through the reintroduction of species in the wild. The cytological studies revealed the basic chromosome number x = 9 of the species.


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