Production and analysis of plants that are somatic hybrids of barley (Hordeum vulgare L.) and carrot (Daucus carota L.) |
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Authors: | H Kisaka M Kisaka A Kanno T Kameya |
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Institution: | (1) Institute of Genetic Ecology, Tohoku University, Sendai, 980-77, Japan, JP |
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Abstract: | In order to obtain plants that were somatic hybrids of barley (Hordeum vulgare L.) and carrot (Daucus carota L.), we fused protoplasts that had been isolated from 6-month-old suspension cultures of carrot cells with protoplasts isolated
from barley mesophyll by electrofusion. After culture for 1 month at 25°C , the cells were cultured for 5 weeks at 4°C , and
were then returned to 25°C for culture on a shoot-inducing medium. Three plants (nos. 1, 2 and 3) were regenerated from the
cells. The morphology of the regenerated plants closely resembled that of the parental carrot plants. A cytological analysis
of callus cultures induced from these plants indicated that most of the cells had about 24 chromosomes, fewer than the sum
of the numbers of parent chromosomes which was 32. Southern hybridization analysis with fragments of the rgp1 gene used as probe showed that the regenerated plants contained both barley and carrot genomic DNA. Chloroplast (ct) and
mitochondrial (mt) DNAs were also analyzed with several probes. The ctDNA of the regenerated plants yielded hybridization
bands specific for both barley and carrot when one fragment of rice ctDNA was used as probe. Furthermore, the regenerated
plants yielded a barley specific band and a novel band with another fragment of rice ct DNA as a probe. One of the regenerated
plants (no. 1) yielded a novel pattern of hybridized bands of mt DNA (with an atp6 probe) that was not detected with either of the parents. These results indicated that the regenerated plants were somatic
hybrids of barley and carrot and that recombination of both the chloroplast genomes and the mitochondrial genomes might have
occurred.
Received: 28 May 1996 / Accepted: 2 August 1996 |
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Keywords: | Hordeum vulgare L Daucus carota L Interfamilial somatic hybrid Low temperature Recombination |
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