Summary
Intergeneric symmetric and asymmetric somatic hybrids have been obtained by fusion of metabolically inactivated protoplasts from embryogenic suspension cultures of tall fescue (Festuca arundinacea Schreb.) and unirradiated or 10-500 Gy-irradiated protoplasts from non-morphogenic cell suspensions of Italian ryegrass (Lolium multiflorum Lam.). Genotypically and phenotypically different somatic hybrid Festulolium mature flowering plants were regenerated.
Species-specific sequences from F. arundinacea and L. multiflorum being dispersed and evenly-represented in the corresponding genomes were isolated and used for the molecular characterization of the nuclear make-up of the intergeneric, somatic Festulolium plants recovered. The irradiation of Italian ryegrass protoplasts with ≤ 250 Gy X-rays prior to fusogenic treatment favoured the unidirectional elimination of most or part of the donor chromosomes. Irradiation of L. multiflorum protoplasts with 500 Gy produced highly asymmetric (over 80% donor genome elimination) nuclear hybrids and clones showing a complete loss of donor chromosomes.
The RFLP analysis of the organellar composition in symmetric and asymmetric tall fescue (+) Italian ryegrass regenerants confirmed their somatic hybrid character and revealed a bias towards recipient-type organelles when extensive donor nuclear genome elimination had occurred.
Approaches aimed at improving persistence of ryegrasses based on asymmetric somatic hybridization with largely sexually-incompatible grass species (F. rubra and Alopecurus pratensis), and at transferring the cytoplasmic male sterility trait by intra- and inter-specific hybridization in L. multiflorum and L. perenne, have been undertaken.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
Abbreviations
- cpDNA:
-
chloroplast DNA
- CMS:
-
cytoplasmic male sterility
- 2,4-D:
-
2,4-dichlorophenoxy-acetic acid
- IOA:
-
iodoacetamide
- mtDNA:
-
mitochondrial DNA
- RFLP:
-
restriction fragment length polymorphism
References
Akagi, H., M. Sakamoto, T. Negishi & T. Fujimura, 1989. Construction of rice cybrid plants. Mol. Gen. Genet. 215: 501–506.
Creemers-Molenaar, J., R.D. Hall & F.A. Krens, 1992. Asymmetric protoplast fusion aimed at intraspecific transfer of cytoplasmic male sterility (CMS) inLolium perenneL. Theor. Appl. Genet. 84: 763–770.
Crowder, L.V., 1953. Interspecific and intergeneric hybrids ofFes-tucaandLolium. J. Hered. 44: 195–203.
de Lautour, G. & B.M. Cooper, 1971. Cold and chemical pretreat-ments to aid chromosome counts in a grass leaf squash technique incorporating hot pectinase maceration. Stain. Technol. 46: 305– 310.
Eizenga, G.C. & R.C. Buckner, 1986. Cytological and isozyme eval-uation of tall fescue x Italian ryegrass hybrids. Plant Breeding 97: 340–344.
Feinberg, A.P. & B. Vogelstein, 1983. A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 132: 6–13.
Gieba, Y.Y., S. Hinnisdaels, V.A. Sidorov, V.A. Kaleda, A.S. Parokonny, N.V. Boryshuk, N.N. Cherup, I. Negrutiu & M. Jacobs, 1988. Intergeneric asymmetric hybrids between Nicotiana plumbaginifoliaand Atropa belladonnaobtained by ‘gamma-fusion’. Theor. Appl. Genet. 76: 760–766.
Hayashi, Y., J. Kyozuka & K. Shimamoto, 1988. Hybrids of rice (Oryza sativaL.) and wildOryzaspecies obtained by cell fusion. Mol. Gen. Genet. 246: 6–10.
Kyozuka, J., Y. Hayashi & K. Shimamoto, 1987. High frequency plant regeneration from rice protoplasts by novel nurse culture methods. Mol. Gen. Genet. 206: 408–413.
Kyozuka, J., T. Kaneda & K. Shimamoto, 1989. Production of cytoplasmic male sterile rice(Oryza sativaL.) by cell fusion. Bio/Technology 7: 1171–1174.
Lichtenstein, C. & J. Draper, 1985. Genetic engineering of plants. p. 67–119. In: D.M. Glover (Ed). DNA Cloning, Vol. II. IRL Press, Oxford Washington.
Müller, A.J. & R. Grafe, 1978. Isolation and characterization of cell lines ofNicotiana tabacumlacking nitrate reductase. Mol. Gen. Genet. 161:67–76.
Murashige, T. & F. Skoog, 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant. 15: 473–497.
Perez-Vicente, R., L. Petris, M. Osusky, I. Potrykus & G. Span-genberg, 1992. Molecular and cytogenetic characterization of repetitive DNA sequences fromLoliumandFestuca: applica-tions in the analysis ofFestuloliumhybrids. Theor. Appl. Genet. 84: 145–154.
Sambrook, J., E.F. Fritsch & T. Maniatis, 1989. Molecular cloning -a laboratory manual. Second edition. Cold Spring Harbor Lab-oratory Press. CSH.
Sanger, F., A.R. Coulson, B.G. Barrel, A.J.H. Smith & B.A. Roe, 1980. Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J. Mol. Biol. 143: 161–178.
Saul, M.W. & I. Potrykus, 1984. Species-specific repetitive DNA used to identify interspecific somatic hybrids. Plant Cell Rep. 3: 65–67.
Shillito, R.D., J. Paszkowski, M. Müller & I. Potrykus, 1983. Agarose plating and a bead type culture technique enable and stimulate development of protoplast-derived colonies in a num-ber of plant species. Plant Cell Rep. 2: 244–247.
Spangenberg, G., M.P. Vallés, Z.Y. Wang, P. Montavon, J. Nagel & I. Potrykus, 1994a. Asymmetric somatic hybridization between tall fescue(Festuca arundinaceaSchreb.) and irradiated Italian rye-grass(Lolium multiflorumLam.) protoplasts. Theor. Appl. Genet. 88: 509–519.
Spangenberg, G., Z.Y. Wang, J. Nagel & I. Potrykus, 1994b. Pro-toplast culture and generation of transgenic plants in red fescue (Festuca rubraL.). Plant Sci. 97: 83–94.
Tabaeizadeh, Z., R.J. Ferl & I.K. Vasil, 1986. Somatic hybridization in the Gramineae:Saccharum officinarumL. (Sugarcane) +Pen-nisetum americanum(L.) K. Schum. (Pearl millet). Proc. Natl. Acad. Sci. USA 83:5616–5619.
Takamizo, T. & G. Spangenberg, 1994. Somatic hybridization inFes-tucaandLolium. p. 112–131. In: Y.P.S. Bajaj (Ed). Biotechnolo-gy in Agriculture and Forestry, Vol. 27: Somatic hybridization in crop improvement. Springer Verlag, Berlin, Heidelberg.
Takamizo, T., G. Spangenberg, K. Suginobu & I. Potrykus, 1991. Intergeneric somatic hybridization in Gramineae: somatic hybrid plants between tall fescue(Festuca arundinaceaSchreb.) and Italian ryegrass(Lolium multiflorumLam.). Mol. Gen. Genet. 231: 1–6.
Takamizo, T., K. Suginobu & R. Ohsugi, 1990. Plant regeneration from suspension culture derived protoplasts of tall fescue(Festuca arundinaceaSchreb.) of a single genotype. Plant Sci. 72: 125– 131.
Terada, R., J. Kyozuka, S. Nishibayashi & K. Shimamoto, 1987. Plantlet regeneration from somatic hybrids of rice(Oryza sativaL.) and barnyard grass(Echinochloa oryzicolaVasing). Mol. Gen. Genet. 210: 39–43.
Vasil, V., R.J. Ferl & I.K. Vasil, 1988. Somatic hybridization in the Gramineae:Triticum monococcumL. (Einkorn) +Pennisetum americanum(L.) K. Schum. (Pearl millet). J. Plant. Physiol. 132: 160–163.
Wang, Z.Y., T. Takamizo, V.A. Iglesias, M. Osusky, J. Nagel, I. Potrykus & G. Spangenberg, 1992. Transgenic plants of tall fes-cue(Festuca arundinaceaSchreb.) obtained by direct gene trans-fer to protoplasts. Bio/Technology 10: 691–696.
Wang, Z.Y., M.P. Vallés, P. Montavon, I. Potrykus & G. Span-genberg, 1993a. Fertile plant regeneration from protoplasts of meadow fescue(Festuca pratensisHuds.). Plant Cell Rep. 12: 95–100.
Wang, Z.Y., J. Nagel, I. Potrykus & G. Spangenberg, 1993b. Plants from cell suspension-derived protoplasts inLoliumspecies. Plant Sci. 94: 179–193.
Yamashita, Y., R. Terada, S. Nishibayashi & K. Shimamoto, 1989. Asymmetric somatic hybrids of Brassica: partial transfer ofB. campestrisgenome intoB. oleraceaby cell fusion. Theor. Appl. Genet. 77: 189–194.
Yang, Z.Q., T. Shikanai, K. Mori & Y. Yamada, 1989. Plant regeneration from cytoplasmic hybrids of rice(Oryza sativaL.). Theor. Appl. Genet. 77: 305–310.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1995 Springer Science+Business Media New York
About this chapter
Cite this chapter
Spangenberg, G. et al. (1995). Intergeneric symmetric and asymmetric somatic hybridization in Festuca and Lolium . In: Cassells, A.C., Jones, P.W. (eds) The Methodology of Plant Genetic Manipulation: Criteria for Decision Making. Developments in Plant Breeding, vol 3. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-0357-2_28
Download citation
DOI: https://doi.org/10.1007/978-94-011-0357-2_28
Publisher Name: Springer, Dordrecht
Print ISBN: 978-94-010-4159-1
Online ISBN: 978-94-011-0357-2
eBook Packages: Springer Book Archive