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Theoretical and Applied Genetics

, Volume 70, Issue 3, pp 227–233 | Cite as

Culture of plant somatic hybrids following electrical fusion

  • G. W. Bates
  • C. A. Hasenkampf
Article

Summary

Electrically-induced protoplast fusion has been used to produce somatic hybrids between Nicotiana plumbaginifolia and Nicotiana tabacum. Following fusion of suspension culture protoplasts (N. plumbaginifolia) with mesophyll protoplasts (N. tabacum) heterokaryons were identified visually and their development was followed in culture. Because electrical fusion is a microtechnique, procedures were developed for culturing the heterokaryons in small numbers and at low density. The fusion and culture procedures described are rapid, uncomplicated and repeatable. Good cell viabilities indicate that the fusion procedure is not cytotoxic. Fused material was cultured 1–2 days at high density in modified K3 medium (Nagy and Maliga 1976). The heterokaryons were isolated manually and grown, at low density in conditioned media. Calli have been regenerated. Esterase isozyme patterns confirm the hybrid character of calli and clonally-derived plantlets recovered from these fusions.

Key words

Electrical fusion Protoplast regeneration Isozymes Nicotiana 

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Copyright information

© Springer-Verlag 1985

Authors and Affiliations

  • G. W. Bates
    • 1
  • C. A. Hasenkampf
    • 1
  1. 1.Department of Biological ScienceFlorida State UniversityTallahasseeUSA

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