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Electrofusion for the production of somatic hybrid plants of Solanum melongena L. and Solanum khasianum C.B. Clark

Identifieur interne : 000283 ( France/Analysis ); précédent : 000282; suivant : 000284

Electrofusion for the production of somatic hybrid plants of Solanum melongena L. and Solanum khasianum C.B. Clark

Auteurs : Darasinh Sihachakr [France] ; Robert Haicour [France] ; Isabelle Serraf [France] ; Edilberto Barrientos [France] ; Catherine Herbreteau [France] ; Georges Ducreux [France] ; Line Rossignol [France] ; Vongsay Souvannavong [France]

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RBID : ISTEX:260FE9E74444621F27876810D92A955F8770CCBD

Abstract

Electrofusion has successfully been used for the production of somatic hybrid plants of Solanum melongena (eggplant) and S. khasianum. This fusion was carried out in a movable multi-electrode (2 mm apart) fusion chamber (500–700 μl capacity) containing a mixture (1:1) of mesophyll protoplasts of both species. Following an alignment of protoplasts induced by an A.C. fields of 125 V/cm and 1 Mhz, fusion was initiated by an exposure of the protoplast samples to a train of 3–4 D.C. pulses of 1.2 kV/cm, each 20 μs. The fusion rate was estimated at 30–40%, at least 30% of which were binary fusions. The mixture of fused protoplasts cultured in KM8p medium containing 0.2 mg/l 2,4-D, 0.5 mg/l zeatin, 1 mg/l NAA and 6.5% (w/v) glucose produced abundant calli, some of which gave rise to shoots on regeneration medium. Although no selection methods have been used, a total of 83 somatic hybrid plants were recovered from 83 individual calli in 3 fusion experiments. They accounted for 40–50% of all the regenerated plants. Several traits of the hybrids were intermediate to those of the parents. All the hybrid plants flowered preciously. The pollen viability averaged 12%, but none of them had set fruits. A random sample of the hybrids gave chromosome numbers ranging from 46 to 48. These numbers approximated to the expected tetraploid level (2n = 4x = 48 chromosomes) The hybridity was confirmed by the banding patterns ofperoxidase activities whcih were composed of the bands of both parents.

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DOI: 10.1016/0168-9452(88)90127-6


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ISTEX:260FE9E74444621F27876810D92A955F8770CCBD

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<div type="abstract" xml:lang="en">Electrofusion has successfully been used for the production of somatic hybrid plants of Solanum melongena (eggplant) and S. khasianum. This fusion was carried out in a movable multi-electrode (2 mm apart) fusion chamber (500–700 μl capacity) containing a mixture (1:1) of mesophyll protoplasts of both species. Following an alignment of protoplasts induced by an A.C. fields of 125 V/cm and 1 Mhz, fusion was initiated by an exposure of the protoplast samples to a train of 3–4 D.C. pulses of 1.2 kV/cm, each 20 μs. The fusion rate was estimated at 30–40%, at least 30% of which were binary fusions. The mixture of fused protoplasts cultured in KM8p medium containing 0.2 mg/l 2,4-D, 0.5 mg/l zeatin, 1 mg/l NAA and 6.5% (w/v) glucose produced abundant calli, some of which gave rise to shoots on regeneration medium. Although no selection methods have been used, a total of 83 somatic hybrid plants were recovered from 83 individual calli in 3 fusion experiments. They accounted for 40–50% of all the regenerated plants. Several traits of the hybrids were intermediate to those of the parents. All the hybrid plants flowered preciously. The pollen viability averaged 12%, but none of them had set fruits. A random sample of the hybrids gave chromosome numbers ranging from 46 to 48. These numbers approximated to the expected tetraploid level (2n = 4x = 48 chromosomes) The hybridity was confirmed by the banding patterns ofperoxidase activities whcih were composed of the bands of both parents.</div>
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