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Vol 52, No 4 (2018)

Article

Histochemical Analysis of Tissue-Specific α-Tubulin Acetylation as a Response to Autophagy Induction by Different Stress Factors in Arabidopsis thaliana

Lytvyn D.I., Olenieva V.D., Yemets A.I., Blume Y.B.

Abstract

In order to study the role of posttranslational α-tubulin acetylation in the realization of stressinduced autophagy, levels of α-tubulin acetylation in Arabidopsis thaliana plants subjected to abiotic stress, such as salt and osmotic stresses, starvation, or ultraviolet B irradiation, were determined by Western blot analysis. It was shown a significant increase in the levels of acetylated α-tubulin under autophagy development, moreover, a synergistic action of stressful factors and E-64, an autophagy inhibitor, attenuated this increase. In order to study tissue-specific character of this modification, an immunohistochemical analysis of α-tubulin acetylation in A. thaliana seedlings was conducted. These experiments demonstrated the tissuespecific character of α-tubulin acetylation induced by stress, particularly, it is most markedly manifested in young and meristematic tissues, as well as in root tissues (root cap, epidermis and pericycle cells). Obtained results can serve as a proof of the regulatory role of α-tubulin acetylation in the realization of autophagy as an adaptive response to the influence of stressful stimuli.

Cytology and Genetics. 2018;52(4):245-252
pages 245-252 views

Alterations in the Antioxidant Status of Transgenic Roots of Artemisia spp. Representatives after A. rhizogenes-Mediated Genetic Transformation

Matvieieva N., Shutava H., Shysh S., Drobot K., Ratushnyak Y., Duplij V.

Abstract

The effect of A. rhizogenes-mediated genetic transformation on the antioxidant status of Artemisia tilesii, A. vulgaris, A. dracunculus, and A. annua transgenic roots has been studied. Antioxidant activity (AOA) of aqueous extracts was determined using methods based on the ability to reduce DPPH+ and ABTS+-radicals. The level of AOA (DPPH) in 50% of extracts obtained from transgenic roots was higher than the level of activity possessed by extracts from untransformed roots. An increased ability to reduce the ABTS+ radical was observed in 80% of the extracts. Extracts of A. annua and A. tilesii transgenic roots were the most active, while the lowest antioxidant activity was shown in A. dracunculus extracts. Thus, A. rhizogenes-mediated transformation has led to a change in the antioxidant status of the “hairy” roots of several Artemisia spp. plants (except A. vulgaris). It can be used as a method for the enhancement of the natural antiradical properties of plants belonging to the Artemisia genus.

Cytology and Genetics. 2018;52(4):253-259
pages 253-259 views

Inheritance of Traits Controlled by Odd Chromosomes Using Data on Transmission of Monosomic Addition St Chromosome of the Elymus Sibiricus Genome

Motsnyi I.I., Kulbida M.P.

Abstract

On the basis of the winter bread wheat cultivar Obryi, two independent disomic addition lines BC12F with the chromosome of the E. sibiricus St genome are created. A practical algorithm for determining the probabilities of transmission of the odd chromosome separately through male and female gametes in selfpollination of hemizygous hybrids from the equation p2–(1 + f1f4) × p + f1 = 0 is proposed, where p is the probability of the formation of viable gametes with the considered chromosome and f1 and f4 are the empirical frequencies of the corresponding homozygotes with and without the trait. The probability of transmission of an alien univalent chromosome through pollen (p) is associated with the frequency of its transmission through the egg cell (p) in backcrosses and in self-pollination (1–f4) by the equation p = 1–f4/(1–p). The calculated empirically dependent estimates of the probabilities of transmission of the added chromosome through the egg cell p = 18.7% and through pollen p = 4.3% correspond to the empirical frequencies obtained for backcrosses. The coefficients of the gamete selection V = 0.748 and V = 0.172 are calculated, and the expected segregation for the alien trait controlled by a dominant gene located in the added chromosome is determined—with the trait: without the trait is 0.222: 0.778 in F2; 0.187: 0.813 in equational and 0.043: 0.957 in certational backcrosses.

Cytology and Genetics. 2018;52(4):260-268
pages 260-268 views

Peculiarities of Regeneration and Genetic Variability of Crambe koktebelica and Crambe tataria Plants in vitro

Pushkarova N.O., Lakhneko O.R., Belokurova V.B., Morgun B.V., Kuchuk M.V.

Abstract

The regenerative capability of three types of explants was studied on media with different compositions of growth regulators with the purpose of selecting optimal conditions of fast reproduction of endangered Crambe species that could be used as a relevant source of genetic material for the improvement of industrially valuable plants. PCR-analysis of genotypes of C. koktebelica and C. tataria plants was conducted to identify the influence of in vitro cultivation on the genetic stability of plants. The highest regeneration rates were observed with the use of petiole explants on MS medium with BA and NAA. The absence of somaclonal variability in C. koktebelica and C. tataria in vitro regenerated plants was demonstrated.

Cytology and Genetics. 2018;52(4):269-275
pages 269-275 views

Genotyping of Triticum ssp. Hexaploid Species Samples with ISSR-Markers

Kyrienko A.V., Rozhkov R.V., Parii M.F., Symonenko Y.V.

Abstract

The genetic improvement of soft wheat (Triticum aestivum L.) and other hexaploid wheat species (T. spelta, T. spherococcum, T. petropavlovskyi, and T. compactum) is important. This can be done by transferring genes of interest (resistance to biotic and abiotic factors). For this purpose, phylogenetic connections of the studied genotypes should be investigated. This problem can be solved by using a multilocus system based on ISSR markers. The marker system is highly polymorphic and convenient for analysis. The study presents the results of a comparison of 20 different hexaploid wheat genotypes based on ISSR markers. In addition, the level of polymorphism was determined and a dendrogram reflecting the phylogenetic connections between the studied genotypes was constructed. It was shown that the species located next to each other according to the systematics turned out to be more remote after the ISSR-marker analysis, and, vice versa, the species that were systematically more distant showed a higher level of kinship.

Cytology and Genetics. 2018;52(4):276-282
pages 276-282 views

The Effect of Multipotent Mesenchymal Stromal Cell Derivatives on the Properties of Breast Cancer Cells in vitro and in vivo

Herheliuk T., Perepelytsina O., Ostapchenko L., Sydorenko M.

Abstract

The effects of multipotent mesenchymal stromal cell (MMSC) derivatives on breast cancer cells have been studied in vitro and in vivo. A cytostatic effect of substances produced by human bone marrow MMSCs on the tumor cell population and the inhibition of tumor cell migration into the suspension fraction in vitro were demonstrated. These phenomena were accompanied by upregulation of tumor-associated marker expression: cytokeratin and EpCAM expression was upregulated in 2D and 3D cell cultures and vimentin expression was upregulated in 3D cultures. A single injection of mouse bone marrow MMSC lysate into the experimental animals suppressed tumor growth in thigh muscles. Moreover, this treatment contributed to the preservation of muscle tissue integrity and the normalization of biochemical parameters of the blood in animals that received grafts of Ehrlich adenocarcinoma tumor cells.

Cytology and Genetics. 2018;52(4):283-293
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Novel Mutation in the MECP2 Gene Identified in a Group of Rett Syndrome Patients from Ukraine

Chernushyn S., Gulkovskyi R., Livshits L.

Abstract

Mutations in the MECP2 gene are known to cause Rett syndrome (RTT)—a neurodevelopmental disorder, one of the most common causes of intellectual disability in females, with an incidence of 1 in 10000–15000. We have investigated exons 3 and 4 of the MECP2 gene, that coding MBD and TRD domains of the MeCP2 protein, in 21 RTT patients from Ukraine by PCR-DGGE analysis followed by Sanger sequencing of PCR fragments with abnormal migration profiles. In 13 of 21 (61.9%) patients 7 different mutations were identified one nonsense mutation—c. NC_000023.11:g.154031326G>A (MECP2:c.502C>T) and 4 missense mutation NC_000023.11:g.154031409G>T (MECP2:c.419C>T), NC_000023.11:g.154031355G>A (MECP2:c.473C>T), NC_000023.11:g.154031354A>C (MECP2:c.472A>C), NC_000023.11:g.154031431G>A (MECP2:c.397C>T) located in exon 4, a rare RTT-causing splice site mutation NC_000023.10:g.153296903T>G (MECP2:c.378-2A>C) in intron 3 and deletion NC_000023.10:g.1532 96079_153296122del44 in exon 4. The novel mutation MECP2:c.472A>C identified in our study in patients withclassic RTT phenotype leds to T158P substitution. It is one more confirmation of crucial role that 158 codon in MECP2 protein function.

Cytology and Genetics. 2018;52(4):294-298
pages 294-298 views

Assessment of Nanotoxicity (Cadmium Sulphide and Copper Oxide) Using Cytogenetical Parameters in Coriandrum sativum L. (Apiaceae)

Pramanik A., Datta A.K., Das D., Kumbhakar D.V., Ghosh B., Mandal A., Gupta S., Saha A., Sengupta S.

Abstract

Present investigation deals with cytogenetical consequences (using attributes namely, seed germination, seedling length, mitotic index, mitotic and meiotic abnormalities and pollen grain sterilities) of cadmium sulphide (CdS) and copper oxide (CuO) nanoparticles (NPs) treatment in Coriandrum sativum L. (Family: Apiaceae, spice of commerce). Ethyl methanesulphonate (EMS), a conventional mutagen is used as positive control. Results suggest that both CdS- and CuO-NPs can induce growth inhibition and cause cytological aberrations in both mitotic and meiotic cells in the studied species. EMS also responds similarly as that of NPs. Study highlights that rooted plant species can be effectively used as model for assessment of nanotoxicity considering cytogenetical parameters.

Cytology and Genetics. 2018;52(4):299-308
pages 299-308 views

Morphometric and ISSR-Analysis of Local Populations of Geranium molle L. from the Southern Coast of the Caspian Sea

Esfandani-Bozchaloyi S., Sheidai M., Keshavarzi M., Noormohammadi Z.

Abstract

Geranium molle is known as Dovefoot Geranium or Awnless Geranium. Dovefoot Geranium is a low-growing herb with pink flowers and sharply toothed leaves. Dovefoot Geranium is native to Eurasia and has been introduced to many habitats of the world. This species is very similar to G. robertianum but its palmate-like leaves and bilobed petals show differences. This plant is considered to be anodyne, astringent and vulnerary. We have no information on its population genetic structure, genetic diversity, and morphological variability in Iran. Therefore, due to the importance of these plant species, we performed a combination of morphological and molecular data for this species. For this study, we used 132 randomly collected plants from 18 geographical populations in 4 provinces. Genetic diversity parameters were determined in these populations. STRUCTURE analysis and K-Means clustering identified 14 gene pools in the country and revealed isolation by distance among the studied populations. The Mantel test showed correlation between genetic and geographical distance. AMOVA revealed a significant genetic difference among populations and showed that 40% of total genetic variation was due to within-population diversity. The consensus tree of both molecular and morphological data identified divergent populations. These data may be used in future breeding and conservation of this important medicinal plant in the country.

Cytology and Genetics. 2018;52(4):309-321
pages 309-321 views

Erratum

Erratum to: “Induction of Arabidopsis thaliana Resistance to Pathogenic Bacteria by Lipopolysaccharide and Salicylic Acid”

Shilina J.V., Gushcha M.I., Molozhava O.S., Litvinov S.V., Dmitriev A.P.

Abstract

Page 170, right column, the first sentence of the last paragraph should read:

The relative increase of root length was significantly enhanced in wild-type pretreated with LPS 8614, SA, or the composite preparation (Fig. 3).

Page 171, right column, the second sentence of the last paragraph should read:

For instance, the elicitor chitosan was shown to activate the defense genes by stimulating JA synthesis through the octadecanoid pathway [17].

Cytology and Genetics. 2018;52(4):322-322
pages 322-322 views