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Annals of Botany 89: 171-181, 2002
© 2002 Annals of Botany Company

Painting of Parental Chromatin in Beta Hybrids by Multi-colour Fluorescent in situ Hybridization

CHRISTINE DESEL1, RITA JANSEN1, GUE DEDONG2 and THOMAS SCHMIDT*,1

1Plant Molecular Cytogenetics Group, Institute of Crop Science and Plant Breeding, Christian-Albrechts-University of Kiel, Am Botanischen Garten 1–9, D-24098 Kiel, Germany and 2Department of Biotechnology, Heilongjiang University Harbin, China

 * For correspondence. Fax +49 431 880 2566, e-mail Tschmidt{at}plantbreeding.uni-kiel.de

Received: 24 August 2001; Returned for revision: 13 September 2001; Accepted: 20 October 2001.

Sugar beet (Beta vulgaris L.) is a relatively young crop and has a narrow gene pool. In order to introduce genetic variability into the crop, interspecific hybrids, selected from crosses with wild beets of the sections Corollinae and Procumbentes, have been generated. The introgressed B. procumbens chromatin carries resistance genes to beet cyst nematode Heterodera schachtii Schm. These lines are important for breeding of nematode-resistant sugar beet, while Corollinae species are potential donors of tolerance to biotic and abiotic stresses such as drought or saline soils. We have used in situ hybridization of genomic DNA to discriminate the parental chromosomes in these interspecific hybrids. Suppression of cross-hybridization by blocking DNA was not necessary indicating that the investigated Beta genomes contain sufficient species-specific DNA enabling the unequivocal determination of the genomic composition of the hybrids. Interspecific hybrid lines with an additional chromosome (2n = 18 + 1), chromosome fragment (2n = 18 + fragment) or translocation of B. procumbens (2n = 18) were analysed by genomic in situ hybridization (GISH) at mitosis and meiosis. Species-specific satellites and ribosomal genes used in combination with genomic DNA or in rehybridization experiments served as landmark probes for chromosome identification in hybrid genomes. The detection of a B. procumbens translocation of approx. 1 Mbp demonstrated the sensitivity and resolution of GISH and showed that this approach is a powerful method in genome analysis projects of the genus Beta.

Key words: Genomic in situ hybridization (GISH), fluorescence in situ hybridization (FISH), Beta vulgaris, Beta corolliflora, Beta procumbens, sugar beet, repetitive DNA, satellite DNA, nematodes, minichromosome.


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