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Production of somatic hybrids between frost tolerantSolanum commersonii andS. Tuberosum: Protoplast fusion,regeneration and isozyme analysis
Authors:T Cardi  K J Puite  K S Ramulu  F D’Ambrosio  L Frusciante
Institution:1. Research Centre for Vegetable Breeding, CNR, via Università, 133, 80055, Portici, Italy
2. Center for Plant Breeding and Reproduction Research, CPRO, Postbox 16, 6700 AA, Wageningen, The Netherlands
3. Department of Agronomic Sciences and Plant Genetics, University of Naples, via Università, 100, 80055, Portici, Italy
Abstract:Mesophyll protoplasts ofSolanum commersonii, a frost tolerant wild species not crossable with the cultivated potato, were fused with either dihaploid or tetraploid S.tuberosum. Protoplasts were aggregated by means of alternating current (AC) or polyethylene glycol (PEG), and electrofused with three direct current (DC) pulses. The treatments with PEG/DC generally resulted in very low heterofusion frequency and protoplast viabiity. On the other hand, AC/DC fusion conditions were optimized by increasing the fusion density of protoplasts and adding CaCl2 to fusion medium. When a density of 4.8 × 105 protoplasts ml?1 was used in the fusion medium containing 0.2 mM Ca++, AC/DC treated protoplasts showed heterofusion frequencies and plating efficiencies of about 10 and 3%, respectively. Fast growing calli from AC/DC fusion experiments were further cultured for regeneration. Fifty-seven plants were regenerated and clonedin vitro as shoot cultures. Compared to parents they showed heterotic vigor and could be identified as hybrids, based on isozyme analysis.
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