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Annals of Botany 64: 613-624, 1989
© 1989 Annals of Botany Company

RESEARCH-ARTICLE

Interspecific Hybridization of Trifolium repens with T. hybridum Using In Ovulo Embryo and Embryo Culture

L. PRZYWARA*,{dagger}, D. W. R. WHITE{dagger}, P. M. SANDERS{dagger} and D. MAHER{dagger}

*Jagiellonian University Krakow, Poland
{dagger}Grasslands Division, DSIR Palmerston North, New Zealand

For correspondence

Accepted: 6 January 1989   

In ovulo embryo culture followed by culture of excised immature embryos produced interspecific hybrids between Trifolium repens L. (white clover) and autotetraploid T. hybridum L. (alsike clover).

Ovules containing hybrid embryos were excised 12–14 d after pollination and cultured on Nitsch (1951) medium supplemented with 15% young cucumber juice for 5–6 d. Embryos were subsequently excised and transferred to hormone-free EG medium, a medium suitable for the culture of immature embryos.

A total of 118 hybrid seedlings were obtained from 1978 reciprocal pollinations. All seedlings produced showed various chlorophyll deficiencies, either totally albino or albino with green sectors. Transmission electron microscope studies were carried out to investigate plastid development in embryos and seedlings. Some embryos produced only callus. Plants were regenerated from seven calli. Two semi-albino plants survived transfer to soil, and one plant produced flowers. Backcrosses to T. repens produced one green plant.

Hybridity is supported by analysis of morphological characters, karyotype and the gel electrophoretic separation of leaf isozymes.

Pollen irradiated with 40 Gy of gamma rays was also used for pollinations. Results indicate that in certain cases ionizing radiation might be useful in overcoming hybrid inviability.

Trifolium repens, Trifolium hybridum, clover, interspecific hybridization, in ovulo embryo culture, irradiation


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