Biopolym. Cell. 2013; 29(4):295-310.
Reviews
Evolution of cell populations in vitro: peculiarities, driving forces, mechanisms and consequences
1Kunakh V. A.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680

Abstract

This review outlines the major features and distinctions of cell populations, types and directions of selection in such populations. Population-genetic basis for cell adaptation to growth conditions in vitro is elucidated; in particular, peculiarities of genome evolution in the course of cell dedifferentiation and further cell adaptation to growth conditions in passaged culture are evaluated. Main factors of variation and selection in cell populations in vitro, influence of growth conditions on structure of cell populations and some regularities of cultured cells and regenerated plants are considered. Details of creation of stable cell lines-producers of biologically active substances are presented. Views and suppositions of author resulting from analysis of both literature data and own multiyear studies on cell population genetics are set forth. Among others are substantiated such key statements: cell culture in vitro presents dynamically-heterogeneous biological system, clone population, which is developing (evolving) as a result of major driving factors of evolution – variation, heredity, selection and drift of genes (genotypes); interaction between these processes determines the biological characteristics of each particular cell line grown in specific conditions; in adaptation of cells to growth conditions in vitro one can single out three periods: the initial population of isolated cells, the period of strain (cell line) formation and the established strain. The division into periods is determined by the type, direction and intensity of «natural» selection that acts in cell population. The formed (adapted to growth in vitro) strains are genetically heterogeneous, they are characterized by the presence of physiological and genetic homeostasis, which are mostly caused by the action of stabilizing selection; cultured cells of higher plants are able to synthesize practically all classes of secondary (specialized) compounds (alkaloids, steroids, terpenoids, etc.); any somatic cell with living (functionally active) nucleus during its isolation and further cultivation in tissue culture, as a result of the process of «somaclonal» variability occurring according to the N. I. Vavilov’s law of homologous series in hereditary variability, can restore in it’s descendants, including regenerated plants, the entire genetic polymorphism (or at least a significant part of it) characteristic of the plant’s species and may be even it’s genus. This provides an opportunity to preserve and restore the natural polymorphism in cultured cells and tissues in vitro.
Keywords: genome evolution, cell populations, selection

References

[1] Biotechnology in Agriculture and Forestry / Eds Y. P. S. Bajaj, T. Nagata, J. Kumlehn Berlin etc.: Springer, 1986–2012 p.
[2] Kunakh V. A. Biotechnology of medical plants. Genetic, physiological and biochemical basis Kyiv: Logos, 2005 730 p.
[3] Kunakh V. A. Plant biotechnology for human life improvement Biotechnology (Ukr.) 2008 1, N 1:28–39.
[4] Ignatova S. A. Cell technologies in plant growing, genetics and breeding of crops: tasks, possibilities, development of systems in vitro Odessa: Astroprint, 2011 224 p.
[5] Kunakh V. A., Mozhylevskaya L. P. Human life span and plant biotechnology Molecular and Applied Genetics Minsk, 2013 Vol. 14:56–62.
[6] Mitrofanova I. V. Somatic embryogenesis and organogenesis as a biotech basis of obtaining and preservation of perennial horticultural plants Kyiv: Agrarna nauka, 2011 344 p.
[7] Cherevchenko T. M., Lavrentyeva A. N., Ivannikov R. V. Boitechnology of tropical and subtropical plants in vitro Kyiv: Naukova dumka, 2008 560 p.
[8] Olenov Y. M. Cellular heredity, differentiation of cells and carcinogenesis as problems of evolutionary genetics Leningrad: Nauka, 1967 310 p.
[9] Vakhtin Y. B. Genetic theory of cell populations Leningrad: Nauka, 1980 168 p.
[10] Vakhtin YB, Pinchuk VG, Schvemberger IN, Butenko ZA. Clonal-selectional concept of tumor growth Kiev: Naukova Dumka, 1987 216 p.
[11] Dubrovna O. V., Chugunkova T. V., Bavol A. V., Lal'ko I. I. Biotechnological and cytogenetic basis establishment of plants resistant to stress Kyiv: Logos, 2012 428 p.
[12] Lutova L. A., Ezhova T. A., Dodueva I. E., Osipova M. A. Developmental genetics of plants: for biological specialities of Universities / 2nd issue revised and suppl Saint-Petersburg: Publ. House N-L, 2010 432 p.
[13] Gavrilov V. I. Transplantable cells in virology M.: Meditsina, 1964 264 p.
[14] Mamaeva SE. The patterns of the karyotypic evolution of cells in culture. Tsitologiia. 1996;38(8):787-814.
[15] Dobzhansky Th. Mendelian populations and their evolution Genetics in 20th century / Ed U. C. Dunn New York: Acad. press, 1952:573–589.
[16] Kunakh V. A. Genome plasticity of somatic cells and adaptability of plants Molecular and Applied Genetics Minsk, 2011 Vol. 12:7–14.
[17] Kunakh V. A. Developmental genome plasticity as a basis for plant adaptability. Zhebrak readings III / Institute of Genetics and Cytology of NAS of Belarus / Ed. A. V. Kilchevsky Minsk, 2011 56 p.
[18] Kordium V. A. Our «Shagreen leather» is our problem. We have to solve it ourselves Kyiv: Logos, 2006 264 p.
[19] Solov'yan V. T., Spiridonova K. V., Kunakh V. A. Genome rearrangements in cell culture of Rauwolfia serpentina. Diverse pattern of genome variations Tsitologiya 1994 30, N 2 P. 250–254.
[20] Solov'yan V. T., Spiridonova K. V., Kunakh V. A. Genome rearrangements in cell culture of Rauwolfia serpentina. II Communication with interspecies variability Tsitologiya 1994 30, N 3:399–403.
[21] Solov'yan V. T., Spiridonova K. V., Kunakh V. A. Special features of genomic variation of cell culture of Rauwolfia srpentina Cytol. Genet. 1994 28, N 5:21–25.
[22] Kunakh V. A. Plant genome evolution in cell culture in vitro. Peculiarities, reasons, mechanisms and consiquencies Genetics and breeding in Ukraine at the turn of millenniums / Ed. V.V. Morgun K.: Logos, 2001:53–67.
[23] Kunakh V. A. Mechanisms and some pegularities to somaclonal variability of the plants Bull. of Vavilov Soc. Geneticists and Breeders of Ukr 2003 1, N 1:101–106.
[24] Andreev I. O., Spiridonova K. V., Kunakh V. A. Plant genome rearrangements in cell culture in vitro Biopolym. Cell 2004 20, N 1–2:42–49.
[25] Andreev I. O., Spiridonova K. V., Solovyan V. T., Kunakh V. A. Variability of ribosomal RNA genes in Rauwolfia species: parallelism between tissue culture-induced rearrangements and interspecies polymorphism Cell Biol. Int 2005 29, N 1 P. 21–27.
[26] Kunakh V. A., Andreev I. O., Spiridonova K. V. 18-S and 5-S gene interspecies polymorphism and variability in tissue culture of Rauwolfia serpentina Benth. and Gentiana L. Physiol. Biochem. Cultivated Plants 2006 38, N 2:110–123.
[27] Andreev I. O., Spiridonova K. V., Maidanyuk D. M., Kunakh V. A. Genetic effects of tissue culture on maize Physiol. Biochem. Cultivated Plants 2009 41, N 6:487–495.
[28] Andreev I. O. Plant tissue culture as a model of evolutionary processes Achievements and problems of genetics, breeding and biotechnology / Ed. V. A. Kunakh K.: Logos, 2012 Vol. 4 P. 7–12.
[29] Belyaev D. K. Destabilizing selection The development of the theory of evolution in the USSR: 1917–1970 Leningrad: Nauka, 1983:266–277.
[30] Kunakh V. A. Cytogenetic behaviour of tissue culture of Haplopappus Culture of isolated organs, tissues and cells of plants M.: Nauka, 1970:155–158.
[31] Kunakh V. A. Polyploidy in cell and tissue cultures in vitro and its putative reasons Experimental polyploidy in crops K.: Naukova dumka, 1974:39–56.
[32] Sidorenko PG, Kunakh VA. Character of caryotype variability in cell population of tissue culture of Haplopappus gracilis with long term passaging. Tsitol Genet. 1970; 4(3):235-41.
[33] Sidorenko PG, Kunakh VA. Production of Culture of Haplopappus gracilis and Crepis capillaris and their cytogenetic characteristic. Tsitol Genet. 1972; 6(6):483-86.
[34] Inge-Vechtomov S. G. Genetics with basis of breeding: textbook for high school students / 2nd issue revised and suppl Saint-Petersburg: Publ. House N-L, 2010 720 p.
[35] Severtsov A. N. Principal directions of the evolutionary process. Morphobiological theory of evolution Moscow; Leningrad: Biomedgiz, 1934 150 p.
[36] Kunakh V. A. Genome variability in plant somatic cells. 3. Callus formation in vitro Biopolym. Cell 1997 13, N 5 P. 362–371.
[37] Kunakh V. A. Variability of plant genome in process of dedifferentiation and callus formation in vitro Russ. J. Plant. Physiol 1999 46, N 6:919–929.
[38] Kunakh V. A. Cytogenetic studies on cell populations in culture of isolated plant tissues Ph.D. thesis, 03.00.15 – Genetics K., 1975 16 p.
[39] Twardovska M. O., Drobyk N. M., Mel'nyk V. M., Konvalyuk I. I., Kunakh V. A. Genome variability of some Gentiana L. species in nature and in culture in vitro: RAPD-analysis Biopolym. Cell 2010 26, N 6:499–507.
[40] Bublyk O. M., Andreev I. O., Spiridonova K. V., Kunakh V. A. Inter simple sequence repeat (ISSR) variability in Ungernia victoris tissue culture Topics in experimental evolution of organisms / Ed. V. A. Kunakh K.: Logos, 2010–Vol. 9:8–12.
[41] Bublyk O. M., Andreev I. O., Spiridonova E. V., Kunakh V. A. Variability of PCR-markers based on disease resistance and abiotic stress response genes in Ungernia victoris tissue culture Topics in experimental evolution of organisms / Ed. V. A. Kunakh K.: Logos, 2011 Vol. 11:213–218.
[42] Konvalyuk I. I., Mel'nyk V. M., Drobyk N. M., Kravets N. B., Twardovska M. O., Kunakh V. A. RAPDand ISSR-analysis of genetic variation in Gentiana pneumonanthe L. tissue and organ culture Bull. Vavilov. Soc. Geneticists and Breederes of Ukr 2011 9, N 1:22–31.
[43] Konvalyuk I. I., Drobyk N. M., Mel'nyk V. M., Twardovska M. O., Kravets N. B., Kunakh V. A. RAPDand ISSR-analysis of Gentiana lutea L. tissue and organ culture in various conditions of maintenance Achievements and problems of genetics, breeding and biotechnology / Ed. V. A. Kunakh K.: Logos, 2012 Vol. 4 P. 523–527.
[44] Kunakh VA. Genome variation in plant somatic cells and factors regulating this process. Tsitol Genet. 1980; 14(1):73-81.
[45] Gubar EK, Kunakh VA. Karyotype variability of cultured Crepis cells (Crepis capillaris L. Wallr.). Genetika. 1992; 28(6):51-61.
[46] Kunakh V. A. Somaclonal variation in Rauwolfia Biotechnology in Agriculture and Forestry. Somaclonal variation in crop improvement. II Berlin; Heidelberg; New York: Springer, 1996 Vol. 36:315–332.
[47] Spiridonova KV, Andreev IO, Solov'yan VT, Kunakh VA. Peculiarities of some gene rearrangement in the cell culture of Rauwolfia serpentina Benth. in vitro. Dopovidi Nats Akad Nauk Ukrainy. 2000; (2):165-70.
[48] Spiridonova K. V., Andreev I. O., Solovyan V. T., Kunakh V. A. Molecular and biological characteristics of genomic rearrangements in Rauwolfia serpentina Benth. cell culture in vitro Genetics and breeding in Ukraine at the turn of the millennium / Ed. V.V. Morgun K.: Logos, 2001 Vol. 1:422–427.
[49] Tvardovs'ka MO, Strashniuk NM, Mel'nyk VM, Adonin VI, Kunakh VA. [Chromosome variability in the tissue culture of rare Gentiana species]. Tsitol Genet. 2008;42(4):12-7. Ukrainian.
[50] Bublik EN, Adonin VI, Kunakh VA. Cytogenetic variation of Ungernia victoris cell lines during cultivation on culture media of different composition. Tsitol. Genet. 2008 42(1):29–36.
[51] Bublyk O. M., Andreev I. O., Spiridonova E. V., Kunakh V. A. Variability of Ungernia victoris morphogenic and non-morphogenic tissue culture as results from RAPD-analysis Bull. of Vavilov Soc. Geneticists and Breeders of Ukr 2008 6, N 1:44–51.
[52] Kunakh V. A. Peculiarities of structural mutagenesis in populations of cultured plant cells Advances of modern genetics / Ed. N. P. Dubinin M.: Nauka, 1984 Vol. 12:30–62.
[53] Kunakh VA, Zosimovich VP. Effect of kinetin on the frequency and types of chromosome aberrations in a tissue culture of Haplopappus gracilis. Genetika. 1977; 13(8):1355-65.
[54] Kunakh V. A., Sidorenko P. G., Zossimovich V. P. Kinetin influence on reproduction of cells with varying ploidy Achievements of polyploidy / Ed. V. P. Zossimovich K.: Naukova dumka, 1977:203–215.
[55] Brodskii V. Ya. The forms of variability in a cell population as well as in an organism population. The biology of the development of the cardiac myocytes Ontogenez 1994 25, N 5 P. 29–43.
[56] Braun V. Bacterial genetics M.: Nauka, 1968 446 p.
[57] Kunakh V. A. Genome variability in plant somatic cells. 4. Variability in the process of dedifferentiation and callus formation in vitro Biopolym. Cell 1998 14, N 4:298–319.
[58] Kunakh V. A. Genome variability in the somatic plant cells. 6. Variability and selection in the course of adaptation to in vitro conditions Biopolym. Cell 2000 16, N 3:159–185.
[59] Kunakh V. A. Genome variability in somatic plant cells. 7. Variability of population-genetic parameters in the culture in vitro Biopolym. Cell 2002 18, N 5:377–393.
[60] Kunakh V. A. Genome variability and accumulation of indoline alkaloids in Rauwotfia serpentina Benth. cell culture Biopolym. Cell 1994 10, N 1:3–30.
[61] Kunakh V. A. Twenty five years long stable biosynthesis of ajmaline by related hormone-independed Rauwolfia serpentina cell lines Euromedica-Hannover-2005 (16–17 Juni) International Congress and Exhibition: Programm Abstracts Hannover, 2005:22.
[62] Kunakh V. A., Katsan V. A. Biosynthesis of poppy isoquinoline alkaloids in nature and in the in vitro culture. 1. Opium poppy (Papaver somniferum L.) Ukr. Biokhim. Zh 2003 75, N 5 P. 41–54.
[63] Kunakh V. A., Katsan V. A. Biosynthesis of poppy quinoline alkaloids in nature and in vitro culture. 2. (Papaver bracteatum Lindl.) Ukr. Biokhim. Zh 2004 76, N 5:29–44
[64] Maidanyuk D. M., Andreev I. O., Spiridonova K. V., Kunakh V. A. Genetic polymorphism of the maize somaclonal lines derived from P346 line Biopolym. Cell 2007 23, N 4:324–331.
[65] Maidanyuk D. M., Andreev I. O., Spiridonova K. V., Kunakh V. A. Genomic variability in maize callus cultures of lines P346 and its derivative somaclonal lines Biopolym. Cell 2007 23, N 5:416–424.
[66] Kunakh VA. On relationship between ploidy Crepis capillaris and Haplopappus gracilis strains and spontaneous organogenesis. Tsitol Genet. 1974; 8(4):308–12.
[67] Kunakh V. A. Cytogenetic peculiarities of isolated tissue culture and regenerated plants in connection with their application in breeding New methods for creation and application of original materials for plant breeding K.: Naukova dumka, 1979 P. 186–193.
[68] Kunakh V. A. Cytogenetic variability of cell populations in culture of isolated plant tissues Tissue and cell cultures in plant breeding M.: Kolos, 1979:38–51.
[69] Kunakh VA, Alkhimova EG, Voityuk LI. Variability of the chromosome number in callus tissues and pea regenerants. Tsitol Genet. 1984; 18(1):20-5.
[70] Kuchko A. A., Oliynyk T. M. Somaclonal variation in potatoes Kyiv: Dovira, 1998 192 p.
[71] Bublyk O. M., Andreev I. O., Spiridonova K. V., Kunakh V. A. Genetic variability in regenerated plants of Ungernia victoris Biologia plantarum 2012 56, N 2:395–400.
[72] Poronnyk O. O., Kunakh V. A. Biosynthesis of naphthoquinoine pigments in plants from Boraginaceae family in nature and in vitro culture Ukr. Biokhim. Zh 2005 77, N 6. P. 24–36.
[73] Strashniuk N. M., Les'kova O. M., Mel'nyk V. M., Twardovska M. O., Konvalyuk I. I., Kunakh V. A. Biologically active substances of Gentiana L. genus. 3. Flavonoid contents in tissue culture Phytotherapy 2008 N 3:82–87.
[74] Poronnik O. O., Shablij V. A., Kunakh V. A. Generation of Echium plantagineum L. tissue culture which is producent of shikonin pigments Biotechnology (Ukr.) 2008 1, N 3 P. 56–63.
[75] Poronnik O. O., Kuchma M. D., Kunakh V. A. Generation of Echium plantagineum L. callus culture – producent of shikonin Bull. of Vavilov Soc. Geneticists and Breeders of Ukr 2008 6, N 3:282–286.
[76] Konvalyuk I. I., Hrytsak L. R., Mel'nyk V. M., Drobyk N. M., Kunakh V. A. Obtaining and characterization of isolated root culture from plants of genus Gentiana Biotechnology (Ukr.) 2011 4, N 3. P. 29–35.
[77] Miryuta N. Ju., Parnikoza I. Ju., Ammouri Ju., Kunakh V. A. The application of thermodynamic approach for population cell dynamic ivestigation in vitro for Rauwolfia serpentina Benth. tissue culture as indole alkaloids producent Biotechnology (Moscow) 2006 N 2:78–95.
[78] Miryuta N. Yu., Kunakh V. A. The dynamics of cell systems in vitro. I. Temporal organization and stability of Rauwolfia serpentina culture tissues at circadian level Biotechnology (Ukr.) 2011 4, N 5:25–38.
[79] Miryuta N. Yu., Kunakh V. A. Dynamic of cell population systems in vitro. II. Temporal organization and robustness of Rauwolfia serpentina Benth culture tissues system at passage level Biotechnology (Ukr.) 2011 4, N 6:18–30.
[80] Miryuta N. Yu., Kunakh V. A. Dynamic of cell population systems in vitro. III. Hypothesis of cell differential process self-control and it's phenomenology realization by the example of Rauwolfia serpentina Benth tissues culture Biotechnology (Ukr.) 2012 5, N 3:40–51.