PAG-I Plant Genome I Conference

Town & Country Conference Center, San Diego, CA, November, 1992.


PG-I: 81pg1

MOLECULAR ANALYSIS OF HOMOLOGOUS REGIONS OF THE SOYBEAN GENOME FOR HOMEOLOGY AND REPEATED SEQUENCES.

K. M. Polzin 1, E. Calvo 2, and R. C. Shoemaker 1, 1 USDA/ARS/FCR and 2 Dept. of Zoology/Genetics, Iowa State University, Ames, IA 50011.


The modern soybean genome is thought to have been derived by tetraploidization followed by chromosome rearrangements resulting in diploidization. Such events should result in the presence of large and/or small homeologous regions depending on the level of 'scrambling' during diploidization. We have analyzed genomic clones (12-16 kb) of the soybean genome that contain sequences homologous either to pA071, a random genomic Pstl fragment, or pSle, a cDNA of a soybean seed-specific late embryogenic gene, by restriction mapping and cross hybridization for evidence of homeology. Seven pA071- and three pSle-hybridizing regions 11 were assayed. A pair of potentially homeologous clones was identified for each probe. In both cases, a region of >8 kb surrounding the probe hybridizing region showed cross hybridization at high stringency (O.1X SSC, 60C) and conservation of restriction sites, followed by regions which did not cross hybridize even at low stringency (0.5X SSC, 60C) and which showed little or no conservation of restriction sites. The extent of the homologous regions could not be determined since the homology extended to the end of the clone. The remaining clones showed homology only in the probe-hybridizing region suggesting either extensive divergence between homeologs and/or that these clones correspond to other copies of the probe sequences present at different sites in the tetraploid. Since not all pA071-hybridizing fragments of the soybean genome were recovered as EMBL3 clones, the failure to find homeologous partners for the remaining five unpaired pA071 EMBL3 clones may be due to the homeologs being among the uncloned fragments or to the homeologs for these clones having been deleted during diploidization. To further elucidate the structure of the soybean genome we have also analyzed some of the above clones for repeated sequences. Our results show that these regions contain primarily low and middle-repetitive sequences (10-20, and >20 but distinct bands, respectively).


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