ISHS Acta Horticulturae 528: VII International Symposium on Grapevine Genetics and Breeding
SOMATIC HYBRIDIZATION OF GRAPEVINE PROTOPLASTS |
| Authors: | A. Matt, R. Blaich, G.M. Reustle |
| Keywords: | Grapevine, protoplast fusion, electrofusion (electrical parameters), hybrid selection, iodoacetamide |
| Abstract:
An electrofusion protocol for the somatic hybridization of grapevine protoplasts has been developed. The highest yield of heterokaryons was obtained by subjecting the protoplast mixture to an AC field of 1 Mhz and 120 V/cm for 25 sec followed by three pulses of 1000 V/cm for 40 μsec. The fusion medium was 0.65 M mannitol containing 1 mM CaCl2. The addition of 10 % PEG (polyethylene glycol, MW 3350) and 10 μM MOPS (3-morpholino-propanesulfonic-acid) to the fusion solution did not increase the fusion frequency. Different pH-values and increasing Ca-concentrations in the fusion medium have been tested, too. Somatic hybrids could be selected due to the inactivation of embryogenic callus protoplasts of Vitis sp. cv. Seyval blanc by iodoacetamide (IOA) and the regeneration inability of Vitis vinifera cvs. Orion, Silvaner, Trollinger, Riesling, Vitis sp. cv. Chancellor and V. rotundifolia protoplasts. | |
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