Universität Tübingen Botanisches Institut
Lehrstuhl Physiologische Ökologie der Pflanzen
Naton B, Ecke M, Hampp R (1992) Production of fertile hybrids by electrofusion of vacuolated and evacuolated tobacco mesophyll protoplasts. Plant Science 85: 197-208. The aim of our experiments was to produce somatic hybrids without using mutants as fusion partners and an expensive cell sorter. Vacuolated mesophyll protoplasts of Nicotiana tabacum L. cv. Xanthi were electrically fused with evacuolated protoplasts of the same species or genus (N. rustica L.) in a mass fusion chamber. Fusion yields of 6-11% were obtained. After density gradient centrifugation the fraction enriched in fusion products (90-100%) was embedded in low melting point agarose lenses fixed to the bottom of plastic Petri dishes and cultured in the presence of feeder cells. The development of individual fusion products could be followed for several weeks. Plating efficiencies were estimated to be 10-39% for heterokaryocytes. Hybrid character was tested by chromosome counting, isozyme analysis and comparison of morphological properties. About 86% of the regenerants showed hybrid character and 4% were putative cybrids. This study demonstrates that somatic hybrid plants may be selected after electrofusion of mesophyll protoplasts from cultivates lacking specific markers and without using micro-methods or a cell sorter.
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