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Genome modifications in protoplast-derived tobacco plants: Phenotypic evaluation and RFLP Analysis

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Biologia Plantarum

Abstract

Genomic instability of protoplast-derived tobacco plants was studied by means of phenotypic evaluation, karyological analysis, and Southern blot experiments. Of the total number of 91 regenerants belonging to 35 different protoclones 57 plants displayed various morphological and/or functional aberrations, some of them being inherited into the progeny. A karyological study of 20 randomly chosen plants revealed 15 tetraploid and 5 diploid chromosome sets. A Southern blot hybridization analysis of three regenerants displayed some DNA polymorphism (RFLP) and thus confirmed that in such plants alterations in the genome structure could be found and that genotypes of protoplast-derived plants frequently differ from the parental genotype.

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References

  • Brown, P. T. H., Kyozuka, J., Sukekiyo, Y., Kimura, Y., Shimamoto, K., Lörz, H.: Molecular changes in protoplast-derived rice plants. - Mol. gen. Genet.223:324–328, 1990.

    Article  PubMed  CAS  Google Scholar 

  • Cullis, C. A., Cleary, W.: Rapidly varying DNA sequences in flax. - Can. J. Genet. Cytol.28:252–259, 1986.

    CAS  Google Scholar 

  • Dennis, E. S., Brettell, R. I. S., Peacock, W. J.: A tissue culture inducedAdhl mutant of maize results from a single base change. - Mol. gen. Genet.210:181–183, 1987.

    Article  CAS  Google Scholar 

  • Holliday, R.: The inheritance of epigenetic defects. - Science238:163–169, 1987.

    Article  PubMed  CAS  Google Scholar 

  • Karp, A., Bright, S. W. J.: On the causes and origins of somaclonal variation. - Oxford Surv. Plant Mol. Cell Biol.2:199–234, 1985.

    Google Scholar 

  • Kiss, T., Kis, M., Abel, S., Solomosy, F.: Nucleotide sequence of the 17S-25S spacer region from tomato rDNA. - Nucl. Acids Res.16:7179, 1988.

    Article  PubMed  CAS  Google Scholar 

  • Koukalová, B., Reich, J., Matyášek, R., Kuhrová, V., Bezděk, M.: ABamHI family of highly repeated DNA seguences ofNicotiana tabacum, - Theor. appl. Genet.78:77–80, 1989.

    Article  Google Scholar 

  • Kuhrová, V., Bezděk, M., Vyskot, B., Koukalová, B., Fajkus, J.: Isolation and characterization of two middle repetitive DNA sequences of nuclear tobacco genome. - Theor. appl. Genet.81: 740–744, 1991.

    Article  Google Scholar 

  • Larkin, P. J., Scowcroft, W. R.: Somaclonal variation - a novel source of variability from cell cultures for plant improvement. - Theor. appl. Genet.60:197–214, 1981.

    Article  Google Scholar 

  • Li, X. Q., Chetrit, P., Mathieu, C., Vedel, F., De Paepe, R., Remy, R., Ambard-Bretteville, F.: Regeneration of cytoplasmatic male sterile protoclones ofNicotiana sylvestris with mitochondrial variations. - Curr. Genet.13:261–266, 1988.

    Article  CAS  Google Scholar 

  • Lörz, H., Scowcroft, W. R.: Variability among plants and their progeny regenerated from protoplasts ofSu/su heterozygotes ofNicotiana tabacum, - Theor. appl. Genet.66 67–75. 1983.

    Article  Google Scholar 

  • Magnien, E., Dalschaert, X., Faraoni-Sciamanna, P.: Transmission of a cytological heterogeneity from the leaf to the protoplasts in culture. Plant Sci. Lett.25:291–303, 1982.

    Article  CAS  Google Scholar 

  • Müller, E., Brown, P. T. H., Hartke, S., Lörz, H.: RFLP-analysis of rice plants regenerated from tissue cultures. - In: Nijkamp, H. J. J.(et al.) (ed.): Progress in Plant Cellular and Molecular Biology. Pp. 153–156. Kluwer Academic Publ., Dordrecht 1990.

    Google Scholar 

  • Murray, M. G., Thompson, W. F.: Rapid isolation of high molecular weight plant DNA. - Nucl. Acids Res.8:4321, 1983.

    Article  Google Scholar 

  • Peerbolte, R., Ruigrok, P., Wullems, G., Schilperoort, R.: T-DNA rearrangements due to tissue culture: somaclonal variation in crown-gall tissues. - Plant mol. Biol.9:51–57, 1987.

    Article  CAS  Google Scholar 

  • Phillips, R. L., Kaeppler, S. M., Peschke, V. M.: Do we understand somaclonal variation? - Progr. Plant cell. mol. Biol.1990: 131–141, 1990.

    Google Scholar 

  • Prat, D.: Genetic variability induced inNicotiana sylvestris by protoplast culture. - Theor. appl. Genet.64:223–230, 1983.

    Article  Google Scholar 

  • Sugiura, M., Shinoyaki, K., Zaita, N., Kusuda, M., Masanobu, K.: Clone bank of the tobacco(Nicotiana tabacum) chloroplast genome as a set of overlapping restriction endonuclease fragments: mapping of eleven ribosomal protein genes. - Plant Sci.44:211–216, 1986.

    Article  CAS  Google Scholar 

  • Vogt, P.: Potential genetic functions of tandem repeated DNA sequences blocks in the human genome are based on a highly conserved “chromatin folding code“. - Human Genet.84:301–336, 1990.

    CAS  Google Scholar 

  • Vyskot, B., Reich, J., Fajkus, J., Bezděk, M., Soška, J.: Genome modifications in protoplast-derived tobacco plants: contents of repetitive DNA sequences. - Biol. Plant.33: 448–454, 1991.

    CAS  Google Scholar 

  • Walbot, V., Cullis, C. A.: Rapid genomic change in higher plants. - Annu. Rev. Plant Physiol.36:367–396, 1985.

    Article  CAS  Google Scholar 

  • Weisling, K., Kahl, G., Schell, J.: Transfer, structure and expression of foreign genes in plants. - In: Kahl, G. (ed.): Architecture of Eukaryotic Genes. Pp. 57–87. Verlagsgesellschaft mbH, Weinheim 1988.

    Google Scholar 

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Vyskot, B., Fajkus, J., Kuglík, P. et al. Genome modifications in protoplast-derived tobacco plants: Phenotypic evaluation and RFLP Analysis. Biol Plant 33, 455–460 (1991). https://doi.org/10.1007/BF02897719

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  • DOI: https://doi.org/10.1007/BF02897719

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