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Simple verification of in vitro - grown clones of the genus Drosera L. using its molecular markers

100%

Fratrikova M., Bauer M., Jopcik M., Libantova J.

Acta Scientiarum Polonorum. Hortorum Cultus

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2018

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tom 17

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nr 1

Drosera L. is a genus of carnivorous plants that comprises approximately 250 species, although this number is probably not complete. Some of these taxa exhibit only small differences in morphological traits that can be partly influenced if the taxa are propagated in vitro. Here, we focus on the verification of putative clones of Drosera spathulata Labill., Drosera rotundifolia L. and Drosera binata var. Dichotoma species cultivated in vitro using molecular markers covering the internal transcribed spacer (ITS) region of 45S ribosomal DNA (rDNA). Following the polymerase chain reaction (PCR) amplification of ~360-bp DNA fragments and sequencing, the sequences were aligned with corresponding sequences in the National Center for Biotechnology Information (NCBI) database. In addition, each of tested PCR amplicons had a specific restriction profile that predominantly enables the differentiation of D. rotundifolia and D. spathulata; the shape of the leaves does not have to be a clear morphologically distinguishable trait.

2

Application of Arabidopsis tissue-specific CRUC promoter in the Cre/loxP self-excision strategy for generation of marker-free oilseed rape: potential advantages and drawbacks

100%

Boszaradova E., Libantova J., Matusikova I., Moravcikova J.

Acta Physiologiae Plantarum

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2014

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tom 36

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nr 06

The aim of the present study was to explore the usability of Arabidopsis cruciferin C (CRUC) promoter in the Cre/loxP self-excision strategy with the minimal rate of an ectopic expression of the cre recombinase. For this, a plant transformation vector containing the gus reporter gene driven by the light-sensitive Lhca3.St1 instead of double dCaMV 35S promoter and one pair loxP sites flanking the cre and the nptII genes was prepared. Agrobacterium-mediated transformations of three economically important oilseed rape (Brassica napus L.) cultivars Campino, Hunter and Topas as well as tobacco (Nicotiana tabacum L.) as a reference system were performed. The integration of the T-DNA into genome of all Brassica cultivars was accompanied by DNA deletions/rearrangements on the both T-DNA ends. The disruption of the Cre/ loxP recombination system in oilseed rape was observed. On the contrary, no such events were detected in tobacco. The applied strategy did restrict an ectopic CRUC activity to some extent and a set of transgenic tobacco T₀ plants without premature excision event was obtained. Our data showed that a choice of the promoter can limit undesired activation of the Cre/loxP cassette.

3

Beta-1,3-glucanase accumulation in roots of maize and soybean exposed to lead, cadmium and arsenic

100%

Pirselova B., Libantova J., Moravcikova J., Matusikova I.

Acta Biologica Cracoviensia. Series Botanica. Supplement

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2009

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tom 51

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nr 2

4

Expression pattern of Arabidopsis thaliana pollen- and embryo-specific promoter in transgenic tobacco plants

100%

Jopcik M., Matusikova I., Moravcikova J., Libantova J.

Acta Biologica Cracoviensia. Series Botanica

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2014

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tom 56

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nr 1

5

Development of embryo-like structures in the suspension cultures of flax coincides with secretion of chitinase-like proteins

88%

Petrovska B., Salaj T., Moravcikova J., Libantova J., Salaj J.

Acta Physiologiae Plantarum

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2010

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tom 32

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nr 4

Flax suspension cultures have been established from the callus induced from the cotyledons, hypocotyls, and immature zygotic embryos (iZE). The growth of flax suspension culture (expressed as a sedimented cell volume) was compared in both conditioned (by liquid from embryogenic Pinus nigra suspension culture) and nonconditioned media. Conditioning of media significantly increased the growth of the cell lines of hypocotyl and iZE origin; however, it had no promotive effect on embryogenic response of these flax liquid cultures. Formation of embryo-like structures (ELS), confirmed also histologically, has only been found in the cell line derived from iZE and cultivated in non-conditioned MS medium supplemented with 1 mg l⁻¹ 2,4-D. The process of ELS formation in this cell line was accompanied by the expression of the protein(s) with chitinolytic activity and molecular weight approximately of 25 kDa. The relationship between the formation of ELS and secretion of chitinase(s) is discussed.

6

Cre-mediated marker gene removal for production of biosafe commercial oilseed rape

88%

Boszoradova E., Matuskova I., Libantova J., Zimova M., Moravcikova J.

Acta Physiologiae Plantarum

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2019

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tom 41

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nr 06

The Cre/loxP-based self-excision represents a promising strategy for removal of hazardous transgene(s) from genomes of targeted crops prior to their introduction into environment. Here, we applied the Cre/loxP self-excision strategy in which the cre recombinase gene is driven by the embryo-specific CRUC promoter from Arabidopsis thaliana. Besides, the Cre/loxP cassette, the T-DNA also consisted of the β-glucuronidase gene, controlled by the double dCaMV 35S promoter and the selectable neomycin phosphotransferase gene; the latter was aimed to be removed from the transgenic genome. The Cre/loxP self-excision cassette was introduced into genomes of four commercial oilseed rape cultivars Haydn, Heros, Hunter and Topas (Brassica napus L.) via Agrobacterium tumefaciens. Transgenic T₀ plants were regenerated from all cultivars. Their detailed molecular analyses revealed premature activation of the CRUC promoter that resulted in partial excision of the nptII gene. Progenies of four self-pollinated T₀ lines were further analysed, and the data confirmed complete excision event in 5 out of 105 (4.8%) of T₁ transgenic oilseed rape plants. The excision efficiency does not seem to depend on the target cultivar. However, the poor transformation efficiency of rapeseed and the limited specificity of the CRUC promoter are clearly the bottleneck of this approach, and the feasibility of (tissue-specific) self-excision of selectable marker gene from genomes of each commercial rapeseed variety adds to their perspective to cope the increasing negative impacts of climate changes.

7

SDS-PAGE protein profiles of developing seeds of Abies concolor

76%

Kormutak A., Libiakova G., Salaj J., Libantova J., Moravcikova J., Vookova B.

Acta Biologica Cracoviensia. Series Botanica. Supplement

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2005

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tom 47

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nr 1

8

Stress-induced expression of cucumber chitinase and Nicotiana plumbaginifoliabeta-1,3-glucanase genes in transgenic potato plants

64%

Moravcikova J., Libantova J., Hedlak J., Salaj J., Bauer M., Matusikova I., Galova Z., Mlynarova L.

Acta Physiologiae Plantarum

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2007

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tom 29

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nr 2

The genes encoding for a cucumber class III chitinase and Nicotiana plumbaginifolia class I glucanase were co-introduced into Slovak potato (Solanum tuberosum L.) cultivar ETA using Agrobacterium tumefaciens. Expression of both genes was driven by wound-inducible polyubiquitin promoter isolated from Slovak potato breeding line 116/86. Analyses showed inducible, peel-specific expression of both transgenes under stress conditions. The effect of transgene expression on fungal susceptibility of transformants was evaluated in vitro and in vivo. Experiments with crude protein extracts isolated from transgenic microtubers showed growth inhibition of Rhizoctonia solani hyphae in the range from 7.3 to 14.2%. In contrast, experiments performed in growth chamber conditions revealed that the polyubiquitin promoter driven transgene expression did not ensure any obvious increase of transgenic potato resistance against Rhizoctonia solani.

9

Hybridization processes in putative hybrid swarms of scots pine and mountain dwarf pine as revealed by chloroplast DNA

64%

Kormutak A., Brana M., Manka P., Galgoci M., Libantova J., Camek V., Bolecek P., Gomory D.

Acta Biologica Cracoviensia. Series Botanica

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2014

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tom 56

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nr 2

Ograniczanie wyników

Simple verification of in vitro - grown clones of the genus Drosera L. using its molecular markers

Application of Arabidopsis tissue-specific CRUC promoter in the Cre/loxP self-excision strategy for generation of marker-free oilseed rape: potential advantages and drawbacks

Beta-1,3-glucanase accumulation in roots of maize and soybean exposed to lead, cadmium and arsenic

Expression pattern of Arabidopsis thaliana pollen- and embryo-specific promoter in transgenic tobacco plants

Development of embryo-like structures in the suspension cultures of flax coincides with secretion of chitinase-like proteins

Cre-mediated marker gene removal for production of biosafe commercial oilseed rape

SDS-PAGE protein profiles of developing seeds of Abies concolor

Stress-induced expression of cucumber chitinase and Nicotiana plumbaginifoliabeta-1,3-glucanase genes in transgenic potato plants

Hybridization processes in putative hybrid swarms of scots pine and mountain dwarf pine as revealed by chloroplast DNA