Ewa Grzebelus

Ewa Grzebelus
University of Agriculture in Krakow | UR · Department of Plant Biology and Biotechnology

Ph.D.

About

44
Publications
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891
Citations
Citations since 2016
21 Research Items
739 Citations
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2016201720182019202020212022050100150
2016201720182019202020212022050100150
2016201720182019202020212022050100150

Publications

Publications (44)
Article
Full-text available
Background The genus Daucus (Apiaceae) comprises about 40 wild species and the cultivated carrot, a crop of great economic and nutritional importance. The rich genetic diversity of wild Daucus species makes them a valuable gene pool for carrot improvement breeding programs. Therefore, it is essential to have good knowledge of the genome structure a...
Article
Full-text available
Key message We have developed and validated an efficient protocol for producing gene-edited carrot plants that do not result in the stable incorporation of foreign DNA in the edited plant’s genome. Abstract We report here a method for producing transgene-free, gene-edited carrot (Daucus carota subs. sativus) plants. With this approach, PEG-mediate...
Article
Full-text available
Background: In the course of evolution, chromosomes undergo evolutionary changes; thus, karyotypes may differ considerably among groups of organisms, even within closely related taxa. The genus Daucus seems to be a promising model for exploring the dynamics of karyotype evolution. It comprises some 40 wild species and the cultivated carrot, a crop...
Article
Full-text available
The genus Lachenalia J. Jacq. ex Js. Murray (Asparagaceae) comprises 136 species endemic to South Africa and Namibia. These bulbous geophytes are considered to have excellent horticultural potential due to their morphological and genetic diversity, and thus have been chosen for the initiation of a breeding program to develop new cultivars that suit...
Article
Full-text available
In this study we report the development of effective in vitro systems for a medicinal plant Nigella damascena L. comprising: (1) callus induction, (2) somatic embryogenesis in callus cultures with subsequent plant regeneration, and (3) isolation and regeneration of callus-derived protoplasts. Callus development was achieved on 83–100% of hypocotyl...
Article
Full-text available
Somatic hybridisation in the carrot, as in other plant species, enables the development of novel plants with unique characteristics. This process can be induced by the application of electric current to isolated protoplasts, but such electrofusion requires an effective hybrid cell identification method. This paper describes the non-toxic fluorescen...
Article
Full-text available
The CRISPR/Cas9 system enables precise genome editing and is a useful tool for functional genomic studies. Here we report a detailed protocol for targeted genome editing in the model grass Brachypodium distachyon and its allotetraploid relative B. hybridum, describing gRNA design, a transient protoplast assay to test gRNA efficiency, Agrobacterium¬...
Chapter
Light microscopy with a bright field mode offers an easy and fast examination of plant specimen for carotenoid presence in its cells. Using basic techniques such as hand sectioned or squashed preparations, carotenoid-rich chromoplasts can be identified without applying any staining procedure and their localization within the cell, their shape and n...
Article
Full-text available
Phytosulfokine-α (PSK), a peptidyl plant growth factor, has been recognized as a promising intercellular signaling molecule involved in cellular proliferation and dedifferentiation. It was shown that PSK stimulated and enhanced cell divisions in protoplast cultures of several species leading to callus and proembryogenic mass formation. Since PSK ha...
Article
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Background: Clearing methods allow relatively quick processing of plant material and examination of cellular structures by rendering tissues and organs translucent. They have been adapted for plant embryology, primarily to study ovule development, megasporogenesis, megagametogenesis and embryogenesis. Such clearing methods overcome several disadvan...
Chapter
In this chapter, we review the contribution of cytogenetics to our understanding of the genome organization of the carrot (Daucus carota subsp. sativus) and its wild Daucus relatives. The genus Daucus includes about 40, mainly diploid, species with basic chromosome numbers ranging from n = 8 to n = 11. Early studies have suffered the difficulty to...
Article
Full-text available
This is the first study to generate carrot plants for enhanced salinity tolerance using a single-cell in vitro system. Protoplasts of three carrot accessions were exposed to treatment by seven different concentrations of NaCl (10–400 mM). Salt concentrations higher than 50 mM decreased plating efficiency and those of 200–400 mM of NaCl completely a...
Article
Intriguingly, we have observed that carotenoid microcrystals, extracted from cells of carrot roots and built from 95% achiral β‐carotene, exhibit a very intense chiroptical (ECD and ROA) signal. This serendipitous discovery of preferential chirality of crystalline aggregates, composed mostly from achiral building blocks is a newly observed phenomen...
Article
Intriguingly, we have observed that carotenoid microcrystals, extracted from cells of carrot roots and built from 95% achiral β‐carotene, exhibit a very intense chiroptical (ECD and ROA) signal. This serendipitous discovery of preferential chirality of crystalline aggregates, composed mostly from achiral building blocks is a newly observed phenomen...
Article
Three non-destructive and complementary techniques, Raman imaging, Atomic Force Microscopy and Scanning Near-field Optical Microscopy were used simultaneously to show for the first time chemical and structural differences of carotenoid crystals. Spectroscopic and microscopic scanning probe measurements were applied to the released crystals or to cr...
Article
Full-text available
Carrot is one of the most important vegetables worldwide, owing to its capability to develop fleshy, highly nutritious storage roots. It was domesticated ca. 1,100 years ago in Central Asia. No systematic knowledge about the molecular mechanisms involved in the domestication syndrome in carrot are available, however, the ability to form a storage r...
Article
Full-text available
We report a high-quality chromosome-scale assembly and analysis of the carrot (Daucus carota) genome, the first sequenced genome to include a comparative evolutionary analysis among members of the euasterid II clade. We characterized two new polyploidization events, both occurring after the divergence of carrot from members of the Asterales order,...
Article
Full-text available
Carrot (Daucus carota L.) chromosomes are small and uniform in shape and length. Here, mitotic chromosomes were subjected to multicolour fluorescence in situ hybridization (mFISH) with probes derived from conserved plant repetitive DNA (18-25S and 5S rDNA, telomeres), a carrot-specific centromeric repeat (Cent-Dc), carrot-specific repetitive elemen...
Data
##Assembly-Data-START## Sequencing Technology :: Sanger dideoxy sequencing ##Assembly-Data-END##
Article
Full-text available
Protoplasts of three carrot cultivars were isolated from in vitro-grown plantlets by overnight incubation in an enzyme mixture composed of 1% (w/v) cellulase Onozuka R-10 and 0.1% (w/v) pectolyase Y-23. After cell immobilization in modified thin alginate layers, three types of β-lactam antibiotics (cefotaxime, carbenicillin, or timentin) at five di...
Article
Full-text available
Protoplasts of four Daucus carota subspecies and three wild Daucus species were isolated from 2-weekold shoot cultures during overnight incubation in an enzyme mixture composed of 1 % (w/v) cellulase Onozuka R-10 and 0.1 % (w/v) pectolyase Y-23. Before the culture, they were embedded in autoclave- or filter-sterilized alginate solution. Modified th...
Article
Full-text available
Protoplasts isolated from three accessions of cultivated carrot and 5-day-old protoplast-derived aggregates were subjected to selection to identify somaclonal variants with enhanced tolerance to the fungal disease black rot incited by Alternaria radicina. Different concentrations [1, 2, 3.5, 5, 10, 20, 35 and 50 % (v/v)] of a fungal culture filtrat...
Article
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We investigated nine families of Stowaway-like miniature inverted-repeat transposable elements (MITEs) in the carrot genome, named DcSto1 to DcSto9. All of them were AT-rich and shared a highly conserved 6 bp-long TIR typical for Stowaways. The copy number of DcSto1 elements was estimated as ca. 5,000 per diploid genome. We observed preference for...
Article
One BC1 [('Kutnowska' × A. roylei) × 'Kutnowska'] plant was obtained after hand-pollinations of F1 A. cepa × A. roylei hybrids with Allium cepa pollen and application of embryo rescue technique in in vitro cultures. Genomic in situ hybridization (GISH) revealed that the BC1 plant had 16 chromosomes, from which 8 displayed blue fluorescence, which p...
Article
Full-text available
The aim of this work was to improve plating efficiency of sugar beet mesophyll protoplast cultures. Preliminary experiments showed that cultures of good quality, viable protoplasts were obtained in rich media based on the Kao and Michayluk formulation and with the calcium alginate as an embedding matrix. Nevertheless, in these cultures cell divisio...
Article
Full-text available
An easy and effective regeneration system from leaf- and hypocotyl-derived protoplasts was established for carrot. The protoplast isolation efficiency after preplasmolytic treatment and digestion of source material in enzyme mixture consisted of 1% cellulase Onozuka R-10 and 0.1% pectolyase Y-23 reached on average 3×106 and 106 protoplasts per g of...
Article
Carrot (Daucus carota L.) chromosomes are small and poorly differentiated in size and morphology. Here we demonstrate that fluorescent in situ hybridization (FISH) signals derived from arbitrary PCR probes can be used for chromosome identification in carrot. To prepare probes, we searched for nonpolymorphic products abundantly amplified with arbitr...
Article
It is the first report of direct, in situ detection of carotenoids at the subcellular level by using Raman microspectroscopy. Single crystals sequestered in a carrot cell were measured using FT-Raman spectrometer equipped with a microscope and 40× objective. The observed characteristic bands centered at 1518cm−1 and 1156cm−1 proved the crystals wer...
Article
Full-text available
Genomic in situ hybridization (GISH) has been proved to be the most effective and accurate technique for confirmation of hybrid character. The objective of our study was to adapt and optimize a GISH protocol for identification of donor chromatin in hybrids obtained by interspecific crosses between five Rhododendron taxa (R. aureum, R. brachycarpum,...
Article
We examined the toxicity of two antibiotics belonging to the betalactam group, to carrot (Daucus carota L.) protoplasts. Leaf protoplasts were cultured in the presence of cefotaxime or carbenicillin applied in five concentrations in the range from 0.1 to 0.5 mg ml-1. Cell viability, division frequency, and regeneration capacity were assessed to det...
Article
Lachenalia Jacq. f. ex Murray, belonging to the Hyacinthaceae family, is a bulbous plant endemic to South Africa and Namibia (Kleynhans, 2007). As a pot plant, it is a relatively new product on the international flower market. The aim of the present study was to examine the level of incompatibility after self- and cross-pollination using four culti...
Article
Karyotype analysis provides insights into genome organization at the chromosome level and into chromosome evolution. Chromosomes were marked for comparative karyotype analysis using FISH localization of rDNA genes for the first time in Apioideae species including taxa of economic importance and several wild Daucus relatives. Interestingly, Daucus s...
Article
The experiment was carried out in attempt to develop an efficient and simple protocol for diploidization of gynogenic onion (Allium cepa L.) embryos. Well-developed embryos derived from flower bud cultures of different onion materials were used. Doubling took place in vitro, on two regeneration media supplemented with one of four anti-mitotic agent...
Article
During the two years of the studies (1997-1998) 32802 flower buds of thirty Polish onion genotypes were plated on different media. In total 310 embryos (0.9 %) were obtained, out of which more than a half underwent regeneration. lt was established that 3.5-4.5 mm long flower buds were the most responsive to gynogenesis. Out of 6 studied media combi...

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Projects

Projects (3)
Project
The main goal of this project is to answer the following research questions: (1) How have the evolutionary relationships within Daucus species been shaped on the chromosomal level? (2) What were the directions of evolutionary changes in their karyotypes? (3) How important was the role of the repetitive sequences in this process? (4) Are the morpho-anatomical data of pollen and seeds/fruits useful in inferring the evolutionary relationships between the Daucus species?
Project
Submit your manuscript to Special Issue "In #Vitro-Based Systems for #Horticultural and #Floricultural Crop Improvement" of Agronomy (IF 2.603) Edited by Dr. Ewa Grzebelus and Dr. Agnieszka Kiełkowska Uniwersytet Rolniczy im. Hugona Kołłątaja w Krakowie. Deadline: 15 May 2021 https://lnkd.in/e4ki2Pz
Project
development of somatic hybrids with CMS in parsley and onion by wide hybridization; development of somatic hybrids in garlic