Chromosomal analysis of Nicotiana asymmetric somatic hybrids by dot blotting and in situ hybridization |
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Authors: | William C Piastuch and George W Bates |
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Institution: | (1) Department of Biological Science and the Institute of Molecular Biophysics, Florida State University, 32306 Tallahassee, FL, USA |
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Abstract: | Summary A species-specific, dispersed repetitive DNA sequence was cloned from Nicotiana plumbaginifolia and used in dot blots and in situ hybridizations to analyze asymmetric somatic hybrids of N. tabacum(+)kanamycin-resistant N. plumbaginifolia. Dot blot hybridization data, using the cloned, species-specific repetitive DNA as a probe, showed that some of the hybrids contain only 1%–5% N. plumbaginifolia DNA, whereas others contain 15%–25%. In situ hybridization of the probe to chromosome spreads showed that the extremely asymmetric hybrids retain a single N. plumbaginifolia chromosome; the hybrids with higher dot blot values were found to have 8 to 12 N. plumbaginifolia chromosomes and chromosome fragments. In situ hybridization also revealed translocations between N. plumbaginifolia and N. tabacum chromosomes in 3 of 8 hybrids studied. RFLP analysis using a 5S gene probe showed the presence of N. plumbaginifolia-specific 5S banding patterns in most hybrids examined, including those that retain only a single N. plumbaginifolia chromosome. |
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Keywords: | Species-specific repetitive DNA Asymmetric somatic hybrids Intergenomic translocations In situ hybridization Tobacco |
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