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Restriction endonuclease studies on the chloroplast and mitochondrial DNAs of alfalfa (Medicago sativa L.) protoclones

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Summary

Alfalfa protoclones were regenerated from the mesophyll protoplasts of two cloned source plants (parents), RS-K1 and RS-K2, initiated from Regen S seed. Because of the high frequency of karyotypic upset previously observed in these plants, chloroplast DNAs (cpDNA) from 23 protoclones and mitochondrial DNAs (mtDNA) from 20 protoclones were examined by restriction endonuclease analysis in order to assess recombination in their cytoplasmic genomes. Seven and four endonucleases were separately used for cpDNA and mtDNA analysis, respectively. Data were consistent with no, or a low frequency of, major sequence rearrangements in either the chloroplast or the mitochondrial genomes as a result of protocloning. However, two types of cpDNA were detected in the 23 protoclones, with only one protoclone possessing the cpDNA type of the cloned parental populations sampled. Possible explanations include a preferential selection during protocloning for one of two parental cpDNA types, an in planta sorting out of cpDNA types in the parental material or both.

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References

  1. Akada S, Hirai A: Studies on the mode of separation of chloroplast genomes in parasexual hybrid calli. II. Heterogenous distribution of two kinds of chloroplast genomes in hybrid callus. Plant Sci Lett 32: 95–100, 1983.

    Google Scholar 

  2. Bayliss MW: Chromosomal variation in plant tissues in culture. Int Rev Cytol Suppl 11A: 113–144, 1980.

    Google Scholar 

  3. Belliard G, Pelletier G, Vedel F, Quetier F: Morphological characteristics and chloroplast DNA distribution in different cytoplasmic parasexual hybrids of Nicotiana tabacum. Molec Gen Genet 165: 231–237, 1978.

    Google Scholar 

  4. Bingham ET, Hurley LV, Kaatz DM, Saunders JW: Breeding alfalfa which regenerates from callus tissue in culture. Crop Sci 15: 719–721, 1975.

    Google Scholar 

  5. Birky CWJr: Relaxed cellular controls and organelle heredity. Science 222: 468–475, 1983.

    Google Scholar 

  6. Bohnert HJ: Organisation and expression of plastid genomes. In: Parthier B, Boulter D (eds) Nucleic Acids and Proteins in Plants II. Encyclopedia of Plant Physiology, N.S. 14B, 1982, pp 475–530.

  7. Chen K, Wildman SG, Smith HH: Chloroplast DNA distribution in parasexual hybrids as shown by polypeptide composition of fraction I protein. Proc Natl Acad Sci (U.S.A.) 74: 5109–5112, 1977.

    Google Scholar 

  8. Chu NM, Tewari KK: Arrangement of the ribosomal RNA genes in chloroplast DNA of Leguminosae. Molec Gen Genet 186: 23–32, 1982.

    Google Scholar 

  9. Douglas GC, Wetter LR, Keller WA, Setterfield G: Somatic hybridization between Nicotiana rustica and N. tabacum IV. Analysis of nuclear and chloroplast genome expression in somatic hybrids. Can J Bot 59: 1509–1513, 1981.

    Google Scholar 

  10. Evans DA, Sharp WR: Single gene mutations in tomato plants regenerated from tissue culture. Science 221: 949–951, 1983.

    Google Scholar 

  11. Flick CE, Evans DA: Evaluation of cytoplasmic segregation in somatic hybrids of Nicotiana: tentoxin sensitivity. J Hered 73: 264–266, 1982.

    Google Scholar 

  12. Fluhr R: The segregation of organelles and cytoplasmic traits in higher plant somatic fusion hybrids. In: Potrykus I, Harms CT, Hinnen A, Hutter R, King PJ, Shillito RD (eds) Protoplasts 1983, Lecture Proceedings, Birkhauser Press, Basel, 1983, pp 85–92.

    Google Scholar 

  13. Fluhr R, Aviv D, Edelman M, Galun E: Cybrids containing mixed and sorted-out chloroplasts following interspecific somatic fusions in Nicotiana. Theor Appl Genet 65: 289–294, 1983.

    Google Scholar 

  14. Fluhr R, Aviv D, Galun E, Edelman M: Generation of heteroplastidic Nicotiana cybrids by protoplast fusion: analysis for plastid recombinant types. Theor Appl Genet 67: 491–497, 1984.

    Google Scholar 

  15. Gengenbach BG, Connelly JA, Pring DR, Conde MF: Mitochondrial DNA variation in maize plants regenerated during tissue culture selection. Theor Appl Genet 59: 161–167, 1981.

    Google Scholar 

  16. Glimelius K, Chen K, Bonnett HT: Somatic hybridization in Nicotiana: Segregation of organellar traits among hybrid and cybrid plants. Planta 153: 504–510, 1981.

    Google Scholar 

  17. Iwai S, Nakata K, Nagao T, Kawashima N, Matsuyama S: Detection of the Nicotiana rustica chloroplast genome coding for the large subunit of Fraction I protein in a somatic hybrid in which only the N. tabacum chloroplast genome appeared to have been expressed. Planta 152: 478–480, 1981.

    Google Scholar 

  18. Johnson LB, Stuteville DB, Higgins RK, Skinner DZ: Regeneration of alfalfa plants from protoplasts of selected Regen S clones. Plant Sci Lett 20: 297–304, 1981.

    Google Scholar 

  19. Johnson LB, Stuteville DL, Schlarbaum SE, Skinner DZ: Variation in phenotype and chromosome number in alfal protoclones regenerated from nonmutagenized calli. Crop Sci 24: 948–951, 1984.

    Google Scholar 

  20. Kemble RJ, Flavell RB, Brettell RIS: Mitochondrial DNA analyses of fertile and sterile maize plants derived from tissue culture with the Texas male sterile cytoplasm. Theor Appl Genet 62: 213–217, 1982.

    Google Scholar 

  21. Kemble RJ, Gun RE, Flavell RB: Classification of normal and male-sterile cytoplasms in maize. II. Electrophoretic analysis of DNA species in mitochondria. Genetics 95: 451–458, 1980.

    Google Scholar 

  22. Kemble RJ, Shepard JF: Cytoplasmic DNA variation in a potato protoclonal population. Theor Appl Genet 69: 211–216, 1984.

    Google Scholar 

  23. Kumar A, Cocking EC, Bovenberg WA, Kool AJ: Restriction endonuclease analysis of chloroplast DNA in interspecies somatic hybrids of Petunia. Theor Appl Genet 62: 377–383, 1982.

    Google Scholar 

  24. Larkin PJ, Scowcroft WR: Somaclonal variation—a novel source of variability from cell cultures for plant improvement. Theor Appl Genet 60: 197–214, 1981.

    Google Scholar 

  25. Leaver CJ, Gray MW: Mitochondrial genome organization and expression in higher plants. Ann Rev Plant Physiol 33: 373–402, 1982.

    Google Scholar 

  26. Melchers G, Sacristan MD, Holder AA: Somatic hybrid plants of potato and tomato regenerated from fused protoplasts. Carlsberg Res Commun 43: 203–218, 1978.

    Google Scholar 

  27. Nikiforova ID, Negruk VI: Comparative electrophoretical analysis of plasmid-like mitochondrial DNAs in Vicia faba and in some other legumes. Planta 157: 81–84, 1983.

    Google Scholar 

  28. Palmer JD: Physical and gene mapping of chloroplast DNA from Atriplex triangularis and Cucumis sativa. Nucleic Acids Res 10: 1593–1605, 1982.

    Google Scholar 

  29. Palmer JD, Thompson WF: Chloroplast DNA rearrangements are more frequent when a large inverted repeat sequence is lost. Cell 29: 537–550, 1982.

    Google Scholar 

  30. Schiller B, Herrmann RG, Melchers G: Restriction endonuclease analysis of plastid DNA from tomato, potato and some of their somatic hybrids. Mol Gen Genet 186: 453–459, 1982.

    Google Scholar 

  31. Scowcroft WR, Larkin PJ: Chloroplast DNA assorts randomly in intraspecific somatic hybrids of Nicotiana debneyi. Theor Appl Genet 60: 179–184, 1981.

    Google Scholar 

  32. Scowcroft WR, Larkin PJ: Somaclonal variation: a new option for plant improvement. In: Vasil IK, Scowcroft WR, Frey KJ (eds) Plant improvement and somatic cell genetics. Academic Press, New York, 1982, pp 159–178.

    Google Scholar 

  33. Shepard JF, Bidney D, Shahin E: Potato protoplasts in crop improvement. Science 208: 17–24, 1980.

    Google Scholar 

  34. Shepard JF, Bidney D, Barsby T, Kemble R: Genetic transfer in plants through interspecific protoplast fusion. Science 219: 683–688, 1983.

    Google Scholar 

  35. Smith SE, Bingham ET, Fulton RW: Transmission of chlorophyll deficiencies in Medicago sativa. Evidence for biparental inheritance of plastids. J Hered 77: (in press), 1986.

  36. Solignac M, Monnerot M, Mounolou J-C: Mitochondrial heteroplasmy in Drosophillia mauritiana. Proc Natl Acad Sci (U.S.A.) 80: 6942–6946, 1983.

    Google Scholar 

  37. Tilney-Bassett RAE, Abdel-Wahab OAL: Irregular segregation at the Pr locus controlling plastid inheritance in Pelargonium: Gametophytic lethal or incompatability system? Theor Appl Genet 62: 185–191, 1982.

    Google Scholar 

  38. Umbeck PF, Gengenbach BG: Reversion of male-sterile T-cytoplasm maize to male fertility in tissue culture. Crop Sci 23: 584–588, 1983.

    Google Scholar 

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Author notes

  1. R. J. Rose

    Present address: Department of Biological Sciences, The University of Newcastle, 2308, New South Wales, Australia

  2. R. J. Kemble

    Present address: Department of Plant Biology, Allelix, Inc., 6850 Goreway Drive, L4V 1P1, Mississauga, Ont., Canada

Authors and Affiliations

  1. Department of Plant Pathology, Kansas State University, 66506, Manhattan, KS, U.S.A.

    R. J. Rose, Lowell B. Johnson & R. J. Kemble

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  2. Lowell B. Johnson
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  3. R. J. Kemble
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Rose, R.J., Johnson, L.B. & Kemble, R.J. Restriction endonuclease studies on the chloroplast and mitochondrial DNAs of alfalfa (Medicago sativa L.) protoclones. Plant Mol Biol 6, 331–338 (1986). https://doi.org/10.1007/BF00034940

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  • DOI: https://doi.org/10.1007/BF00034940

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