From repetitive DNA sequences to single-copy gene mapping in plant chromosomes by in situ hybridization
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Plant Physiology and Biochemistry v. 27, no. 2, p. 161-168609964Summary: Haploid leaf segments of Nicotiana plumbaginifolia were transformed with Agrobacterium rhizogenes and actively growing haploid root cultures established.Using protoplasts prepared from these cell populations, the single copy, 1.6 kb Adh (alcohol dehydrogenase) gene and repetitive ribosomal DNA sequences were tentatively mapped by in situ hybridization to chromosomes 1 or 5, and 1B, 6B and 5D respectively in metaphase preparations. The addition of sodium dodecylsulfate (SDS) and polyethylene glycol (PEG) to the probe mixture reduced background noise and allowed longer autoradiographic exposures (12-15 weeks)
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Haploid leaf segments of Nicotiana plumbaginifolia were transformed with Agrobacterium rhizogenes and actively growing haploid root cultures established.Using protoplasts prepared from these cell populations, the single copy, 1.6 kb Adh (alcohol dehydrogenase) gene and repetitive ribosomal DNA sequences were tentatively mapped by in situ hybridization to chromosomes 1 or 5, and 1B, 6B and 5D respectively in metaphase preparations. The addition of sodium dodecylsulfate (SDS) and polyethylene glycol (PEG) to the probe mixture reduced background noise and allowed longer autoradiographic exposures (12-15 weeks)
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Elsevier
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