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Interspecific hybridization of Trifolium alexandrinum with T. constantinopolitanum using embryo rescue

  • Genetic Transformation and Hybridization
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Abstract

The embryo rescue technique was successfully used to raise hybrids between Trifolium alexandrinum and T. constantinopolitanum. As a result of its narrow genetic base, genetic improvement in Egyptian clover (syn. Berseem; T. alexandrinum), an important fodder crop in tropical and subtropical countries, is hampered, thereby making it imperative to introduce alien genes from related species. In a conventional interspecific hybridization program, hybrids could not be raised due to post-fertilization barriers. Of the several combinations tried, pollination 2 days after emasculation was found to be the best. Globular embryos were observed 5–7 days after pollination (DAP), followed by heart-shaped embryos 10–12 DAP. Embryos excised at the heart-shaped stage responded well to EC3 culture medium. Of 612 crosses, 33 healthy embryos could be excised and cultured on EC3 medium. The plumule emerged 8–12 days following inoculation. The embryo-rescued plants were hardened, inoculated with Rhizobium and transferred to the field. The hybrids showed intermediate morphological features with reduced pollen fertility (55–65%) and a chromosomal complement of 2n=16. Biochemical characterization using isozymes confirmed hybridity.

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Fig. 1 a, b
Fig. 2

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Abbreviations

ACP::

Acid phosphatase

IAA::

Indole-3-acetic acid

BA::

6-Benzyl adenine

DAP::

Days after pollination

Est::

Esterase

NAA::

α-Naphthaleneacetic acid

SOD::

Superoxide dismutase

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Acknowledgements

The authors are grateful to the Director of the institute and Head, Crop Improvement Division for use of the facilities and his encouragement. Financial support from the Indian Council of Agricultural Research (project AP CESS) is also acknowledged.

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Authors and Affiliations

  1. Crop Improvement Division, Indian Grassland and Fodder Research Institute, 284003, Jhansi, India

    A. K. Roy, D. R. Malaviya, P. Kaushal, B. Kumar & A. Tiwari

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  1. A. K. Roy
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  2. D. R. Malaviya
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  3. P. Kaushal
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  4. B. Kumar
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  5. A. Tiwari
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Correspondence to A. K. Roy.

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Communicated by P.P. Kumar

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Roy, A.K., Malaviya, D.R., Kaushal, P. et al. Interspecific hybridization of Trifolium alexandrinum with T. constantinopolitanum using embryo rescue. Plant Cell Rep 22, 705–710 (2004). https://doi.org/10.1007/s00299-004-0759-1

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  • DOI: https://doi.org/10.1007/s00299-004-0759-1

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