Abstract
The DNA relative content in nuclei from several Solanum species, which were used as partners for somatic hybridization, were determined using a flow cytometry method. The nuclei were isolated mechanically or via protoplasts from leaves of in vitro grown plants. In the case of S. nigrum as well as S. tuberosum cv. Bzura and dihaploid clone H8105, the nuclei were also obtained from suspension cultured cells by lysis of protoplasts. The source and the method of nuclei isolation affected the pattern of nuclear DNA in the genotypes studied. The mesophyll nuclei showed two distinct peaks on the DNA histograms, whereas the DNA peaks produced by cell suspension nuclei were broad and less distinct. The DNA content in the nuclei, calculated from the DNA histograms of the samples and a DNA standard historgam (Trout Red Blood Cells, having DNA content of 5.05 pg per nucleus), were much lower in mesophyll nuclei than in those obtained from the cell suspension for the same genotypes. The results are discussed in respect of the genetic instability of Solanum genotypes. The usefulness of a flow cytometry approach in somatic hybridization research is also discussed.
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Avan M.F., Maciejewska U., Kleczkowski K., and Wielgat B. 1997. Differential responses of potato cell suspensions to a culture filtrate of Phytophthora infestans. Z. Naturforsch. 52c: 333–338.
Doležel J., Binarova P., and Lucretti S. 1989. Analysis of nuclear DNA content in plant cells by flow cytometry. Biol. Plant. 31: 113–120
Doležel J. 1991. Flow cytometry analysis of nuclear DNA content in higher plants. Phytochem. Anal. 2: 143–154
Galun E. 1981. Plant protoplasts as physiological tools. Ann. Rev. Plant. Physiol. 32: 237–266
Haberlach G.T., Cohen B.A., Reichert N.A., Baer M.A., Towill L.E. and Helgeson J.P. 1985. Isolation, culture and regeneration of protoplasts from potato and several related Solanum species. Plant Science 39: 67–74.
Hanisch ten Cate Ch.H., and Ramulu K.S. 1987. Callus growth, tumor development and polyploidization in the tetraploid potato cultivar Bintje. Plant Sci. 49: 209–216.
Jakuczun H., Strzelczyk-Żyta D., Narkiewicz M. 1997. Zastosowanie pośrednich metod oznaczania poziomu ploidalności ziemniaka. Biul. Inst. Ziemniaka 48: 91–98.
Menczel L., Nagy F., Kiss Zs.R., and Maliga P. 1981. Streptomycin resistant and sensitive somatic hybrids of Nicotiana tabacum + Nicotiana knightiana: Correlation of resistance to N. tabacum plastids. Theor. Appl. Genet. 59: 191–195.
Menke U., Schilde-Rentschler L., Ruoss B., Zanke C., Hemleben V., Ninnemann H. 1996. Somatic hybrids between the cultivated potato Solanum tuberosum L. and the 1EBN wild species Solanum pinnatisectum Dun.: morphological and molecular characterization. Theor. Appl. Genet., 92: 617–626.
Murashige T., and Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant 15: 473–497.
Nagy J.I., and Maliga P. 1976. Callus induction and plant regeneration from mesophyll protoplasts of Nicotiana sylvestris. Z. Pflanzenphysiol. 78: 453–455.2
Nicoloso F.T., Val J., van der Keur M., van Iren F., and Kijne J.W. 1994. Flow-cytometric cell counting and DNA estimation for the study of plant cell population dynamics. Plant Cell, Tissue and Organ Cult. 39: 251–259.
Ramulu K.S., Dijkhuis P., Hanisch ten Cate Ch.H., and Groot B. De. 1985. Patterns of DNA and chromosome variation during in vitro growth in various genotypes of potato. Plant Sci. 41: 69–78.
Ramulu K.S., Dijkhuis P. 1986. Flow cytometric analysis of polysomaty and in vitro genetic instability in potato. Plant Cell Rep. 3: 234–237
Ramulu K.S., Dijkhuis P., and Roest S. 1989. Patterns of phenotypic and chromosome variation in plants derived from protoplast cultures of monohaploid, dihaploid and diploid genotypes and in somatic hybrids of potato. Plant Sci. 60: 101–110
Sgorbati S., Levi M., Sparvoli E., Trezzi F. and Lucchini G. 1986. Cytometry and flow cytometry of 4′, 6-diamidino-2-phenylindole (DAPI) -stained suspensions of nuclei released from fresh and fixed tissues of plants. Physiol.Plant. 68: 471–476.
Ulrich I. and Ulrich W. 1991. High-resolution flow cytometry of nuclear DNA in higher plants. Protoplasma 165: 212–215.
Valkonen J.P.T. 1994. Nuclear DNA content of the Solanum spp. in the series Etuberosa as determined by laser flow cytometry. Ann. Appl. Biol. 125: 589–600.
Valkonen J.P.T., Watanabe K.N. and Pehu E. 1994. Analysis of correlation between nuclear diploid Solanum brevidens and (di)haploid S. tuberosum. Jpn J. Genet. 69: 525–536.
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Maciejewska, U., Skierski, J.S. & Szczerbakowa, A. Nuclear DNA content of Solanum species grown in vitro, as determined by flow cytometry. Acta Physiol Plant 21, 37–43 (1999). https://doi.org/10.1007/s11738-999-0025-9
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DOI: https://doi.org/10.1007/s11738-999-0025-9