Abstract
Systematic investigations on the occurrence of differential DNA replication in carrot cultures, expressed at the total genome level, were performed. The genome of Daucus carota L. could be characterized by a pattern of repetitive BstNI fragments that was independent of tissue specificity or cultivar differences. Characterization of the genomic DNA of the secondary phloem of carrot roots, in comparison to the DNA of the induced primary cultures at different growth phases, revealed dramatic differences in the copy number of the repetitive fragments. Highly proliferative tissue showed extensive reduction in the proportion of repetitive sequences in the genome in all of the 37 investigated variants. In contrast, during subsequent transition to stationary growth the repetitive fragments re-amplified. The results suggest that the quantitative genome organisation was involved in the regulation of the growth potential of cells. A hypothesis is discussed suggesting a determining influence of the observed differential DNA replication on cell-cycle rates and the cell program of proliferative tissue by structural and positioning effects on DNA loops. To study the causality of somaclonal variation, research on the relationship between physiological genome variability and the induction of heritable changes is recommended.
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References
Adams RLP (1990) DNA methylation. Biochem J 265:309–320
Allen GC, Hall GE Jr., Childs LC, Weissinger AK, Spiker S, Thompson WF (1993) Scaffold attachment regions increase reporter gene expression in stably transformed plant cells. Plant Cell 5:603–613
Altamura MM, Bassi P, Cavallini A, Cionini G, Cremonini R, Monacelli B, Pasqua G, Sassoli O, Tran Than Van K, Cionini PG (1987) Nuclear DNA changes during plant development and the morphogenetic response in vitro of Nicotiana tabacum tissues. Plant Sci 53:73–79
Arnholdt-Schmitt B (1993a) Rapid changes in amplification and methylation pattern of genomic DNA in cultured carrot root explants (Daucus carota L.). Theor Appl Genet 85:793–800
Arnholdt-Schmitt B (1993b) Hormonsensitivität und genomische DNS-Variabilität in ihrem Bezug zur Ertragsbildung bei Daucus carota L. ssp. sativus. Ein Beitrag zur Zellbiologie pflanzlichen Wachstums. Habilitationsschrift, Giessen
Arnholdt-Schmitt B, Holzapfel B, Schillinger A, Neumann KH (1991) Variable methylation and differential replication of genomic DNA in cultured carrot root explants during growth induction as influenced by hormonal treatments. Theor Appl Genet 82:283–288
Arnholdt-Schmitt B, Herterich S, Neumann K-H (1995) Physiological aspects of genome variability in tissue culture. I: Growth phase-dependent differential DNA methylation of the carrot genome (Daucus carota L.) during primary culture. Theor Appl Genet 91:809–815
Bassi P, Cionini PG, Cremonini R, Seghizzi P (1984) Underrepresentation of nuclear DNA sequences in differentiating root cells of Vicia faba. Protoplasma 123:70–77
Blumenthal AB, Kriegstein HJ, Hogness DS (1974) The units of DNA replication in Drosophila melanogaster chromosomes. Cold Spring Harbor Symp Quant Biol 38:205–223
Bonifer C, Vidai M, Grosveld F, Sippel AE (1990) Tissue-specific and position-independent expression of the complete gene domain for chicken lysozyme in transgenic mice. EMBO J 9: 2843–2848
Bonifer C, Hecht A, Saueressig H, Winter DM, Sippel AE (1991) Dynamic chromatin: the regulatory domain organization of eukaryotic gene loci. J Cell Biochem 47:99–108
Bouchard RA (1982) Moderately repetitive DNA in evolution. Int Rev Cytol 76:113–193
Cavalier-Smith T (1978) Nuclear volume control by nucleoskeletal DNA, selection for cell volume and cell growth rate, and the solution of the DNA C-value paradox. J Cell Sci 34:247–278
Cullis CA (1987) The generation of somatic and heritable variation in response to stress. Am Nat 130:S62-S73
Doolittle WF, Sapienza C (1980) Selfish genes, the phenotype paradigm and genome evolution. Nature 284:601–603
Dührssen E (1983) Untersuchungen zur Charakterisierung der DNS einiger höherer Pflanzen (Reassoziationskinetiken, Interspezieshybridisierungen, Dichtegradienten-zentrifugationen). Dissertation, Giessen
Dührssen E, Neumann KH (1980) Characterization of satellite-DNA of Daucus carota L.. Z Pflanzenphysiol 100:447–454
Dührssen E, Saavedra E, Neumann KH (1980) DNA reassociation kinetics and hybridization in different angiosperms. Plant Syst Evol 136:267–273
Elder JK, Southern EM (1983) Measurement of DNA length by ge electrophoresis II: comparison of methods for relating mobility to fragment length. Anal Biochem 128:227–231
Flavell R (1980) The molecular characterization and organisation of plant chromosomal DNA sequences. Annu Rev Plant Physiol 31:569–96
Flavell RB, Bennett MD, Smith IB, Smith DB (1974) Genome size and the proportion of repeated nucleotide sequence DNA in plants. Biochem Genet 12:257–269
Francis D, Kidd AD, Bennett MD (1985) DNA replication in relation to DNA C values. In: Bryant JA, Francis D (eds) The cell division cycle in plants. Cambridge University Press, pp 61–82
Fujimura T, Komamine A (1982) Molecular aspects of somatic embryogenesis in a synchronous system. In: Fujiwara (ed) Proc 5th Int Cong Plant Tissue and Cell Cult, Tokyo, pp 105–106
Gartenbach-Scharrer U, Habib S, Neumann KH (1990) Sequential synthesis of some proteins in cultured carrot explant (Daucus carota) cells during callus induction. Plant Cell, Tissue and Organ Cult. 22:27–35
Hall G, Allen GC, Loer DS, Thompson WF (1991) Nuclear scaffolds and scaffold-attachment regions in higher plants. Proc Natl Acad Sci USA 88:9320–9324
Harley CB, Reis RJS, Goldstein S (1982) Loss of repetitious DNA in proliferating somatic cells may be due to unequal recombination. J Theor Biol 94:1–12
Havelange A, Jeanny JC (1984) Changes in density of chromatin in the meristematic cells of Sinapis alba during transition to flowering. Protoplasma 122:222–232
Houssa C, Jacqmard A, Bernier G (1990) Activation of replicon origins as a possible target for cytokinins in shoot meristems of Sinapis. Planta 181:324–326
Jacqmard A, Houssa C (1988) DNA fiber replication during a morphogenetic switch in the shoot meristematic cells of a higher plant. Exp Cell Res 179:454–461
Jenns SM, Bryant JA (1978) Correlation between deoxyribonuclease activity and DNA replication in the embryonic axes of germinating peas (Pisum sativum L.). Planta 138:99–103
Kuhrová V, Bezdek M, Vyskot B, Koukalová B, Fajkus J (1991) Isolation and characterization of two middle repetitive DNA sequences of nuclear tobacco genome. Theor Appl Genet 81: 740–744
Larkin PJ, Scowcroft WR (1981) Somaclonal variation — a novel source of variability from cell culture for plant improvement. Theor Appl Genet 60:197–214
Leeton PRJ, Smyth DR (1993) An abundant LINE-like element amplified in the genome of Lilium speciosum. Mol Gen Genet 237:97–104
Liu YG, Ikeda TM, Tsunewaki K (1992) Moderately repeated, dispersed, and highly variable (MRDHV) genomic sequences of common wheat usable for cultivar identification. Theor Appl Genet 84:535–543
Ludérus MEE, Graaf A, Mattia E, Blaauwen JL, Grande MA, Jong L, Driel R (1992) Binding of matrix attachment regions to lamin B1. Cell 70:949–959
Matousek J, Rakousky S, Nemcova B (1987) Occurrence of sugar non-specific nuclease in tobacco callus: changes of nuclease activity during callus growth and plant regeneration. Plant Cell, Tissue Organ Cult 8:61–71
Mellerowicz EJ, Riding RT, Little CHA (1992) Periodicity of cambial activity in Abies balsamea. II. Effects of temperature and photoperiod on the size of the nuclear genome in fusiform cambial cells. Physiol Plant 85:526–530
Mitra J, Mapes MO, Steward FC (1960) Growth and organized development of cultured cells. IV. The behavior of the nucleus. Am J Bot 47:357–368
Murray MG, Thompson WF (1980) Rapid isolation of high-molecular-weight plant DNA. Nucleic Acids Res 8:4321–4325
Nagl W (1976) Zellkern und Zellzyklen. Ulmer, Stuttgart
Nagl W (1979) Differential DNA replication in plants: a critical re-view. Z Pflanzenphysiol 95:283–314
Nagl W (1987) Replication. Progr Bot 49:181–191
Nagl W (1989) Replication. Prog Bot 51:173–180
Nagl W (1990) Gene amplification and related events. Biotechnol agric for 11:15.3–201
Nagl W (1992) Replication. Prog Bot 53:166–180
Nagl W, Jeanjour M, Kling H, Kühner S, Michels I, Müller T, Stein B (1983) Genome and chromatin organisation in higher plants. Biol. Zentralbl 102:129–148
Natali L, Cavallini A, Cremonini R, Bassi P, Cionini PG (1986) Amplification of nuclear DNA sequences during induced plant cell de-differentiation. Cell Differ 18:157–161
Neumann KH (1962) Untersuchungen über den Einfluß essentieller Schwermetalle auf das Wachstum und den Proteinstoffwechsel von Karottengewebekulturen. Dissertation, Giessen
Neumann KH (1966) Wurzelbildung und Nucleinsäuregehalt bei Phloem-Gewebekulturen der Karottenwurzel auf synthetischem Nährmedium verschiedener Hormonkombinationen. Phytohormones et l'Organogenèse 38:95–102
Neumann KH (1968) Über Beziehungen zwischen hormonal gesteuerter Zellteilungsgeschwindigkeit und Differenzierung. Ein Beitrag zur Physiologie der pflanzlichen Ertragsbildung. Habilitationsschrift, Giessen
Neumann KH (1972) Untersuchungen über den Einfluß des Kinetins und des Eisens auf den Nukleinsäure- und Protein-Stoffwechsel von Karottengewebekulturen. Z Pflanzenernaehr Bodenkd 131: 211–220
Neumann KH, Pertzsch C, Moos M, Krömmelbein G (1978) Untersuchungen zur Charakterisierung des Ernährungssystems von Karottengewebekulturen: Photosynthese, Kohlenstoff- und Stickstoff-ernährung, Mineralstoffverbrauch. Z Pflanzenernaehr Bodenkd 141:299–311
Nuti Ronchi V, Bennici A, Martini G (1973) Nuclear fragmentation in de-differentiating cells of Nicotiana glauca pith tissue grown in vitro. Cell Differ 2:77–85
Olszewska MJ, Maszewski J, Bilecka A (1990) DNA replication in relation to S phase duration in six angiosperm annual species. Cytologia 55:31–35
Opstelten RJ, Clement JME, Wanka F (1989) Direct repeats at nuclear matrix-associated DNA regions and their putative control function in the replicating eukaryotic genome. Chromosoma 98:422–427
Orgel LE, Crick HC (1980) Selfish DNA: the ultimate parasite. Nature 284:604–607
Peschke VM, Phillips RL (1992) Genetic implications of somaclonal variation in plants. Adv Genet 30:41–75
Power JB, Davey MR, Freeman JP, Mulligan BJ, Cocking EC (1986) Fusion and transformation of plant protoplasts. Methods Enzymol 118:578–595
Richards GM (1974) Modification of the diphenylamine reaction giving increased sensitivity and simplicity in the estimation of DNA.Anal Biochem 57:369–376
Rogowsky PM, Manning S., Liu JY, Langridge P (1991) The R173 family of rye-specific repetitive DNA sequences: a structural analysis. Genome 34:88–95
Rudnicki K, Rzepecki R, Szopa J (1988) Rearrangement of the nuclease-inhibitor complex components within cells of white bush treated with phytohormones. J Plant Physiol 132:658–663
Schäfer A (1976) Untersuchungen zur Charakterisierung der DNS von Daucus carota L. Dissertation, Giessen
Solano R, Hueros G, Fominaya A, Ferrer E (1992) Organization of repeated sequences in species of the genus Avena. Theor Appl Genet 83:602–607
Southern EM (1979) Measurement of DNA length by gel electrophoresis. Anal Biochem 100:319–323
Steward FC, Caplin SM, Millar FK (1952) Investigations on growth and metabolism of plant cells. I. New techniques for the investigation of metabolism, nutrition and growth in undifferentiated cells. Ann Bot 16(1):57–77
Steward FC, Mapes MO, Kent AE, Hosten RD (1964) Growth and development of cultured plant cells. Science 143:20–27
Stief AD, Winter M, Strätling WH, Sippel AE (1989) A nuclear element mediates elevated and position-independent gene activity. Nature 341:343–345
Szopa J, Wagner KG (1980) Purification and properties of a DNase inhibitor from Nicotiana tabacum cell cultures. Eur J Biochem 111:211–215
Van Driel R, Humbel B, de Jong L (1991) The nucleus: a black box being opened. J Cell Biochem 47:311–316
Van't Hof J (1985) Control points within the cell cycle. In: Bryant JA, Francis D (eds): The cell-division cycle in plants. Cambridge University Press, pp 1–13
Waterborg JH, Shall S (1985) The organisation of replicons. In: Bryant JA, Francis D (eds): The cell-division cycle in plants. Cambridge University Press, pp 15–35
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Communicated by H. R Linskens
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Arnholdt-Schmitt, B. Physiological aspects of genome variability in tissue culture. II. Growth phase-dependent quantitative variability of repetitive BstNI fragments of primary cultures of Daucus carota L.. Theoret. Appl. Genetics 91, 816–823 (1995). https://doi.org/10.1007/BF00220965
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DOI: https://doi.org/10.1007/BF00220965