Yakiv Ivanovych Ratushnyak
Contents |
M.Sc., Ph.D
Date & Place of birth
April 5, 1961; Studena, Vinnytsya distr., Ukraine
Contacts
phone: | +380 (44) 266-70-53 |
fax: | +380 (44) 252-17-86 |
Education & Degrees
1981-1986 | M.Sc. in Genetics | Thesis: “Genetic complementation of tomates in somatic and sexual hybridization” Kyiv State University, Kyiv, Ukraine |
1986-1991 | Ph.D. in Genetics. | Thesis: “Transgenomic plants in Lycopersicon obtained by γ-hybridization of protoplasts” Institute of Cell Biology & Genetic Engineering Academy of Sciences of the Ukraine, Kyiv, Ukraine |
Positions Held
1997- present | Senior Researcher | Laboratory of Cytophysiology Institute of Cell Biology & Genetic Engineering National Academy of Sciences of Ukraine, Kyiv, Ukraine |
1997-1998 | Postdoctoral Fellow | Laboratory of Prof. Dr. Peter Meyer University of Leeds, Leeds, United Kingdom |
1991-1996 | Researcher | Laboratory of Cytophysiology & Cell Engineering Institute of Cell Biology & Genetic Engineering National Academy of Sciences of Ukraine, Kyiv, Ukraine |
1989-1991 | Research Assistant | Laboratory of Cytophysiology & Cell Engineering Institute of Cell Biology & Genetic Engineering National Academy of Sciences of Ukraine, Kyiv, Ukraine |
1986-1989 | Research Assistant | Laboratory of Cytophysiology & Cell Engineering Institute of Botany National Academy of Sciences of Ukraine, Kyiv, Ukraine |
1983-1986 | Research Technician | Laboratory of Cytophysiology & Cell Engineering Institute of Botany National Academy of Sciences of Ukraine, Kyiv, Ukraine |
Teaching experience
1987-1990 | Teaching Assistant | Principles of Tomato Cell Engineering | Institute of Cell Biology & Genetic Engineering Academy of Sciences of Ukraine, Kyiv, Ukraine |
1992-1997 | Teacher | Plant Tissue Culture and Micropropagation | Department of Cell Biology & Genetic Engineering, National University of Kyiv, Kyiv, Ukraine |
Grants
1993 | Individual Grant from the International Scientific Fund, USA |
1994 | Travel Grant from the International Scientific Fund, USA |
1997 | Posdoctoral Fellowship from the Royal Society/NATO, UK |
Summary of Research Activities
As a MSc student, I worked on protoplast culture and plant regeneration of Lycium barbarum (woody bush).
My PhD thesis was devoted to obtaining and analysing transgenomic plants of: L. esculentum with L. peruvianum var. dentatum, L. pennellii, L. hirsutum var. glabratum, Solanum rickii and Lycium barbarum. The results obtained and perspective of future investigations of fertile asymmetric somatic hybrids L. esculentum + L. peruvianum var. dentatum were announced in Rice Biotechnology Quarterly (Gamma Fusion of Tomato Protoplasts, 1991, V 8, p 38-39).
My postdoctoral work started with studies of tomato cybrid plants with chloroplasts of L. peruvianum var. dentatum and reconstructed mitochondria with phenotypic traits of nucleo-cytoplasmic incompatibility.
Then cybrid plants of L. peruvianum var. dentatum were obtained containing tomato chloroplasts and L. peruvianum var. dentatum mitochondria that along with cybrid tomato/peruvianum could serve as an excellent model for studies of nuclear-cytoplasmic and chloroplast-mitochondrial interactions.
Also, by restriction analysis of chloroplast DNA for asymmetric somatic hybrids between L. esculentum and L. peruvianum var. dentatum the presence of the new nonparental patterns was shown. Blot-hybridization of chloroplast DNA with Nicotiana tabacum chloroplast DNA fragments revealed rearrangements which might be interpreted as recombination of chloroplast DNA.
At the same time, for fertile asymmetric somatic hybrids L. esculentum + L. peruvianum var. dentatum a considerable genetic variability was shown. Morphological, cytogenetic, isozyme, ribosomal DNA and random amplified polymorphism DNA analyses of F1 and F2 progenies obtained after self-pollination of somatic hybrids revealed new nonparental patterns of DNA and enzymes, chromosome elimination and variation of leaf morphology.
Also, in collaboration with Prof. C.M.Rick (Tomato Stock Center, Davis, USA) we studied protoplast culture of other species of Petota section S. ochrantum, S. juglandifolium, as well as possibilities of their somatic hybridization with tomato.
Then I worked on genetic transformation of Arabidopsis by leaf-disk and root co-cultivation with Agrobacterium in a project named “Identification, isolation and analysis of stress-induced plants genes in Arabidopsis thaliana by T-DNA tagging and gene fusion” funded by the INTAS.
Also, I worked on obtaining haploid plants from Petunia cultivars and Nicotiana species (using anther and pollen culture). These are used in activation tagging experiments aimed to achieve genetic transformation by co-cultivation of haploid protoplasts with Agrobacterium. Co-cultivation allowed the selection some PEG- and salt-resistant clones of N. tabacum. These clones might become useful material to isolate salt- and PEG-resistant plants, and to identify genes responsible for salt- and PEG-resistant. I also study possibilities of the use of diploid protoplasts of Arabidopsis thaliana in these experiments. The title of the project is “Activation tagging Arabidopsis thaliana and Petunia hybrida” and it is funded by the Royal Society and NATO.
At present, I am studying possibilities for successful integration of foreing sequences into plastome of some Solanaceae species via PEG-mediated protoplast transformation.
Scientific Interests
- organelle DNA recombination and nucleo-cytoplasmic interactions under somatic hybridization in higher plants,
- genetic variability of fertile asymmetric somatic hybrids,
- plant genetic transformation.
Experience
Plant cell tissue and protoplast culture, mutagenesis and mutant selection, haploid production, plant genetic transformation (Agrobacterium-mediated procedure, direct gene transfer with and without electroporation, chloroplast transformation), somatic hybridization, biochemical and molecular analyses (DNA-fragment amplification by PCR, isozyme analysis).
Selected Publications
- Ratushnyak YI, Piven NM, Rudas VA (1989) Protoplast culture and plant regeneration in Lycium barbarum L. Plant Cell Tissue Organ Cult 17:183-190</P>
- Ratushnyak YI, Piven NM, Rudas VA (1990) Regeneration of Lycium barbarum L. plants from leaf tissue, callus culture and callus protoplasts. Plant Cell Rep 9:84-87</P>
- Ratushnyak YI, Latypov SA, Samoylov AM, Piven NM, Gleba YY (1991) Introgressive hybrydization of tomatoes by "gamma-fusion" of the Lycopersicon esculentum Mill. and Lycopersicon peruvianum var. dentatum Dun. protoplasts. Plant Sci 73:65-78</P>
- Ratushnyak YI, Piven NM, Latypov SA, Samoylov AM, Zavgorodnyaya AV, Gleba YY (1991) Changes of the genetic constitution of the tomato plants by 'gamma-fusion' of Lycopersicon esculentum Mill. and Lycopersicon peruvianum var. dentatum Dun. protoplasts. Biopolymers and Cell (Rus) 7:82-91</P>
- Ratushnyak YI (1991) Gamma fusion of tomato protoplasts. Rice Biotechnology Quarterly 8:38-39</P>
- Ratushnyak YI, Rudas VA, Piven NM (1993) Regeneration of plants from protoplasts of Lycium barbarum L. (Wolfberry). In: Bajaj YPS (ed) Biotechology in Agriculture and Forestry, Vol 22, Plant Protoplasts and Genetic Engineering III, Springer, New York, pp 79-86</P>
- Ratushnyak YI, Cherep NN, Zavgorodnyaya AV, Latypov SA, Borozenko IV, Rachkovskaya RI, Gleba YY (1993) Fertile asymmetric somatic hybrids between Lycopersicon esculentum Mill. and Lycopersicon peruvianum var. dentatum Dun. Mol Gen Genet, 236: 427-432</P>
- Ratushnyak YI, Cherep NN, Zavgorodnyaya AV, Latypov SA, Borozenko IV, Rachkovskaya RI, Gleba YY (1993) Cell engineering synthesis of tomato fertile asymmetric somatic hybrids by "fusion" of protoplasts. Genetics (Russia), 29:164-176</P>
- Ratushnyak YI, Cherep NN, Latypov SA (1994) Somatic hybridization of Lycopersicon esculentum Mill. + Lycopersicon peruvianum var. dentatum Dun. In: Bajaj YPS (ed) Biotechology in Agriculture and Forestry, Vol. 27 Somatic Hybridization in Crop Improvement, Springer, New York, pp 224-243</P>
- Ratushnyak YI, Kochevenko AS, Cherep NN, Zavgorodnyaya AV, Latypov SA, Gleba YY (1995) Alloplasmatic incompatibility in cybrid plants possessing Lycopersicon esculentum Mill. genome and Lycopersicon peruvianum var. dentatum Dun. plasmagenes. Russian Journal of Genetics, 31:565-571</P>
- Kochevenko AS, Ratushnyak YI, Gleba YY (1996) Protoplast culture and somaclonal variability of species of series Juglandifolia. Plant Cell Tissue Organ Cult, 44:103-110 </P>
- Kochevenko AS, Rudas VA, Ratushnyak YI (1999) Analysis chlorophyll-deficiency mutation of Lycopersicon peruvianum var. dentatum by obtain the cytoplasmic hybrids. Russian Journal of Genetics 35:1-8</P>
- Kochevenko AS, Ratushnyak YI, Korneev DU, Stasik OO, Shevchenko VV, Kochubey SM, Gleba YY (1999) Photosynthetic apparatus in cytoplasmic hybrid of cultured tomato carrying nuclear-cytoplasmic incompatibility traits. Russian Journal of Plant Physiology, 46:474-481 </P>