Skip to main content
SpringerLink
Log in

Identification of barriers to interspecific crosses in the genus Trifolium

  • Published:
Euphytica Aims and scope Submit manuscript

Summary

A study of pre- and post-fertilisation barriers after interspecific crosses of diploid and tetraploid Trifolium pratense L. and wild species T. alpestre L., T. medium L. and T. sarosiense Hazsl. was aimed at finding of a promising cross combination for obtaining hybrids. The growth of pollen tubes was arrested in interspecific crosses mainly when T. pratense was at a diploid level. To investigate the post-fertilisation barriers in detail, the hybrid embryo viability was traced by two clearing treatments of immature seeds: (1) using chloral hydrate (which proved to be most appropriate); and (2) a mixture of benzyl benzoate and dibutly phthalate. In interspecific combinations T. pratense (4×) × either T. alpestre or T. sarosiense, enlargement of immature seeds occurred, but no hybrid embryo was traced. Of the wild species used as a male parent for crosses, T. medium was the only exception from the point of view of fertilisation. Globular, heart and the early torpedo stages of hybrid embryos were observed 7 days after pollination (DAP) but only when T. pratense was at a tetraploid level. When T. pratense (2×, 4×) was used as a male parent for interspecific crosses with T. alpestre, T. medium and T. sarosiense, strong defects in various stages of embryogenesis were observed, particularly wrinkled and narrowing embryo sacs caused by an expansion of endothelial cells. We conclude with the following finding: (1) to make crosses only in one direction with T. pratense as a female parent and T. medium as a male; (2) to use tetraploid plants of T. pratense; (3) and to excise hybrid embryos at an early torpedo stage, about 7 DAP.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Aida, M., T. Ishida, H. Fukaki, H. Fujisawa & M. Tasaka, 1997. Genes involved in organ separation in Arabidopsis: analysis of the cup-shaped cotyledon mutant. Plant Cell 9: 841–857.

    Article  PubMed  CAS  Google Scholar 

  • Algan, G. & H.N. Bakar, 1996. Light and electron microscopic examination of the embryo and endosperm development in the natural tetraploid Trifolium pratense L. Israel J Plant Sci 44: 273–288.

    Google Scholar 

  • Bharathi, M., P.B. Kirti, U.R. Murty & N.G.P. Rao, 1982. Occurrence of atypical embryo-sac like structures in Arachis glabrata Benth. Curr Sci 51: 612–612.

    Google Scholar 

  • Christiansen, P., J.M. Gibson, A. Moore, C. Pedersen, L. Tabe & P.J. Larkin, 2000. Transgenic Trifolium repens with foliage accumulating the high sulphur protein, sunflower seed albumin. Transgen Res 9: 103–113.

    Article  CAS  Google Scholar 

  • Collins, G.B., N.L. Taylor & G.C. Phillips, 1981. Successful hybridization of red clover with perennial Trifolium species via embryo rescue. In: Proc XIV Int Grassl Congr. Lexington, pp. 168–170.

  • Fehr, W.R., 1987. Principles of cultivar development. Vol. 1 Theory and technique. McGraw Hill, New York.

  • George, G.P., R.A. George & J.M. Herr Jr., 1979. A comparative study of ovule and megagametophyte development in field-grown and greenhouse-grown plants of Glycine max and Phaseolus aureus (Papilionaceae). Amer J Bot 66: 1033–1043.

    Article  Google Scholar 

  • Hendrych, R., 1990. The third series of complement on Trifolium – monograph by Zohary and Heller (Plantae hybridae). Preslia, Praha 62: 43–60.

    Google Scholar 

  • Herr, J.M., 1982. An analysis of the methods for permanently mounting ovules cleared in four-and-half type clearing fluids. Stain Technology 57: 161–169.

    PubMed  Google Scholar 

  • Herr, J.M., 1999. Endosperm development in Arabidopsis thaliana (L.) Heynh. Acta Biologica Cracoviensia 41: 103–109.

    Google Scholar 

  • Hoshino, Y., E. Nishino & M. Mii, 2000. Isolation of embryo sacs from Dianthus ovules by enzymatic treatment and microdissection. Plant Cell Rep 19: 443–447.

    Article  CAS  Google Scholar 

  • Hughes, J.W., 1986. Tissue culture derived crossing barriers. Amer J Bot 73: 323–329.

    Article  Google Scholar 

  • Huyghe, C., 1987. A cytological and physiological study of haploid-diploid polyembryony in flax (Linum-usitatissimum L.). Agronomie 7: 567–573.

    Google Scholar 

  • Kazimierska, E.M., 1978. Embryological studies of cross compatibility in the genus Trifolium L. I. Hybridization of T. pratense L. with some species in the subgenus Lagopus Bernh. Genet Pol 19: 1–12.

    Google Scholar 

  • Kazimierska, E.M., 1980. Embryological studies of cross compatibility of species within the genus Trifolium. L. III. Development of the embryo and endosperm in crossing T. repens L. with T. hybridum L. and T. fragiferum L. Genet Pol 21: 37–61.

    Google Scholar 

  • Kazimierski, T., E.M. Kazimierska & Cz. Strzyzewska, 1972. Species crossing in the genus Trifolium. Genet Pol 13: 11–32.

    Google Scholar 

  • Lee, R.W.H., J. Strommer, D. Hodgins, P.E. Shewen, Y. Niu & R. Lo, 2001. Towards development of an edible vaccine against bovine pneumonic pasteurellosis using transgenic white clover expressing a Mannheimia haemolytica A1 leukotoxin 50 fusion protein. Infect Immun 69: 5786–5793.

    Article  PubMed  CAS  Google Scholar 

  • Mackiewicz, T., 1965. Low seed-setting in tetraploid red clover (Trifolium pratense L.) in the light of cytoembryologic analyses. Genet Pol 6: 5–39.

    Google Scholar 

  • Mayer, U., G. Buettnet & G. Jurgens, 1993. Apical-basal pattern formation in the Arabidopsis embryo studies on the role of the gnom gene. Development 117: 149–162.

    Google Scholar 

  • Merker, A., 1984. Hybrids between Trifolium medium and T. pratense. Hereditas 101: 267–268.

    Google Scholar 

  • Merker, A., 1988. Amphidiploids between Trifolium alpestre and T. pratense. Hereditas 108: 267.

    Google Scholar 

  • Morris, J.B. & S.L. Greene, 2001. Defining a multiple-use germplasm collection for the genus Trifolium. Crop Sci 41: 893–901.

    Article  Google Scholar 

  • Nedbalkova, B., J. Repkova & L. Bartosova, 1995. Germplasm TBZP1, TBZP2, TBZP3, TBZP4 of interspecific Trifolium hybrids. Scientific Studies – Research Institute for Fodder Plants, Ltd. Troubsko 13: 129–131.

    Google Scholar 

  • Phillips, G.C., G.B. Collins & N.L. Taylor, 1982. Interspecific hybridization of red clover (T. pratense L.) with T. sarosiense Hazsl. Using in vitro embryo rescue. Theor Appl Genet 62: 17–24.

    Google Scholar 

  • Phillips, G.C., J.W. Grosser, S. Buger, N.L. Taylor & G.B. Collins, 1992. Interspecific hybridization between red clover and Trifolium alpestre using in vitro embryo rescue. Crop Sci 32: 1113–1115.

    Article  Google Scholar 

  • Pittock, C., J.J. Weinman & B.G. Rolfe, 1997. The activity of a tobacco basic chitinase promotor in transgenic white clover provides insights into plant development and symbiosis. Aust J Plant Physiol 24: 555–561.

    Article  CAS  Google Scholar 

  • Quesenberry, K.H., D.S. Wofford, R.L. Smith, P.A. Krottje & F. Tcacenco, 1996. Production of red clover transgenic for neomycin phosphotransferase II using Agrobacterium. Crop Sci 36: 1045–1048.

    Article  Google Scholar 

  • Ramsay, G., & B. Pickersgill, 1986. Interspecific hybridization between Vicia faba and other species of Vicia – approaches to delaying embryo abortion. Biol Zbl 105: 171–179.

    Google Scholar 

  • Sawai, A. & S. Ueda, 1987. Embryo development of the hybrid of Trifolium medium L. × 4 x Trifolium pratense L. J Jpn Soc Grassl Sci 33: 157–162.

    Google Scholar 

  • Sawai, A., S. Ueda, M. Gau & K. Uchiyama, 1990. Interspecific hybrids of Trifolium medium L. × 4x T. pratense L. obtained through embryo culture. J Jpn Soc Grassl Sci 35: 267–272.

    Google Scholar 

  • Schwer, J.F. & R.W. Cleveland, 1972. Tetraploid and triploid interspecific hybrids of Trifolium pratense, T. diffusum and some telated species. Crop Sci 12: 419–422.

    Article  Google Scholar 

  • Scott, A., D. Woodfield & W.R. White, 1998. Allelic composition and genetic background effects on transgene expression and inheritance in white clover. Mol Breed 4: 479–490.

    Article  CAS  Google Scholar 

  • Sharma, S.B., K.R. Hancock, P.M. Ealing & D.W.R. White, 1998. Expression of a sulfur-rich maize seed storage protein, delta-zein, in white clover (Trifolium repens) to improve forage quality. Mol Breed 4: 435–448.

    Article  CAS  Google Scholar 

  • Stelly, D.M. & S.J. Peloquin, 1983. Methods to detect 2n female gametophytes in Solanums. Amer Potato J 60: 821–821.

    Google Scholar 

  • Taylor, N.L., R.F. Quarles & M.K. Anderson, 1980. Methods of overcoming interpecific barriers in Trifolium. Euphytica 29: 441–450.

    Article  Google Scholar 

  • Taylor, N.L. & K.H. Quesenberry, 1996. Red clover science. Kluwer Acad Publ, Dordrecht Hardbound, pp. 170–187.

  • Vogt, M. & W. Schweiger, 1983. Study of the methodology of embryo culture in Trifolium. Arch Zücht-Forsch, Berlin 13: 273–283.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Masaryk University, Brno, Czech Republic

    Jana Repkova, Barbara Jungmannova & Hana Jakesova

Authors
  1. Jana Repkova
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Barbara Jungmannova
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hana Jakesova
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jana Repkova.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Repkova, J., Jungmannova, B. & Jakesova, H. Identification of barriers to interspecific crosses in the genus Trifolium . Euphytica 151, 39–48 (2006). https://doi.org/10.1007/s10681-006-9126-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10681-006-9126-3

Keywords

Search

Navigation