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The Journal of Heredity 1996:87(4):308-313
© 1996 The American Genetic Association 87:308-313

research-article

Soybean Chromosome Painting: A Strategy for Somatic Cytogenetics

L. Shi, T. Zhu, M. Morgante, J. A. Rafalski, and P. Kelm

Department of Biological Sciences, Northern Arizona University Flagstaff, AZ 86011-5640
Dipartimento di Produzione Vegetale e Tecnologie Agrarte, Universita' di Udine Via delle Scienze 208, 1-33100 Udine, Italy
DuPont Ag Biotechnology Wilmington, Delaware

Corresponding Editor: Reid G. Palmer

Abstract

Cytological identification of soybean mitotic metaphase chromosomes (2n = 40) has been severely limited by their small size and uniform karyomorphology. We have developed fluorescent in situ hybridization (FISH), PCR-primed in situ labeling (PCR-PRINS) procedures, and molecular probes for routine cytological identification and for the physical mapping of soybean somatic chromosomes. Chromosome preparation has been achieved by modifications of previous protocols and through the preparation of root-tip protoplasts prior to chromosome spreading. Initially our probe selection focused on highly repeated DNAs that provide very intense localized hybridization signals. Repetitive gene probes that have proven valuable include the rDNA loci (5S and 45S) which are chromosome specific. We have also developed satellite DNA probes for two different sequence families: the SB92 and the STR120 satellites. Both of these are tandemly arranged at multiple chromosomal loci. By using different cloned examples of each family, we have been able to selectively label unique subsets of soybean chromosomes. Double hybridization with blotin and digoxigenin labeled probes has allowed us to determine the chromosomal overlap between different probes. In addition, we have joined portions of the metaphase chromosome painting patterns with the genetic map by single-copy FISH and PCR-PRINS detection of the RFLP loci G8.15, G17.3, and A199b and A199b. Total genomic DNA in situ hybridization (GISH) patterns were also used to characterize the soybean chromosomes.


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