Soybean chromosome painting: A strategy for somatic cytogenetics

L. Shi, T. Zhu, M. Morgante, J. A. Rafalski, Paul S Keim

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Cytological identification of soybean mitotic metaphase chromosomes (2n = 40) has been severely limited by their small size end 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 biotin 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 A199a and A199b. Total genomic DNA in situ hybridization (GISH) patterns were also used to characterize the soybean chromosomes.

Original languageEnglish (US)
Pages (from-to)308-313
Number of pages6
JournalJournal of Heredity
Volume87
Issue number4
StatePublished - Jul 1996

Fingerprint

Chromosome Painting
Soybeans
cytogenetics
Cytogenetics
Chromosomes
soybeans
chromosomes
Primed In Situ Labeling
Metaphase
fluorescence in situ hybridization
Fluorescence In Situ Hybridization
metaphase
loci
hybridization
Satellite DNA
Digoxigenin
satellite DNA
Molecular Probes
Polymerase Chain Reaction
digoxigenin

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Genetics
  • Genetics(clinical)

Cite this

Shi, L., Zhu, T., Morgante, M., Rafalski, J. A., & Keim, P. S. (1996). Soybean chromosome painting: A strategy for somatic cytogenetics. Journal of Heredity, 87(4), 308-313.

Soybean chromosome painting : A strategy for somatic cytogenetics. / Shi, L.; Zhu, T.; Morgante, M.; Rafalski, J. A.; Keim, Paul S.

In: Journal of Heredity, Vol. 87, No. 4, 07.1996, p. 308-313.

Research output: Contribution to journalArticle

Shi, L, Zhu, T, Morgante, M, Rafalski, JA & Keim, PS 1996, 'Soybean chromosome painting: A strategy for somatic cytogenetics', Journal of Heredity, vol. 87, no. 4, pp. 308-313.
Shi L, Zhu T, Morgante M, Rafalski JA, Keim PS. Soybean chromosome painting: A strategy for somatic cytogenetics. Journal of Heredity. 1996 Jul;87(4):308-313.
Shi, L. ; Zhu, T. ; Morgante, M. ; Rafalski, J. A. ; Keim, Paul S. / Soybean chromosome painting : A strategy for somatic cytogenetics. In: Journal of Heredity. 1996 ; Vol. 87, No. 4. pp. 308-313.
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