Plant Genome II Conference
Town & Country Conference Center, San Diego, CA, January, 1994.
PG-II: USE OF REPRESENTATIONAL DIFFERENCE ANALYSIS FOR GENOME,
REGION-SPECIFIC CLONING IN CEREALS
USE OF REPRESENTATIONAL DIFFERENCE ANALYSIS FOR GENOME,
REGION-SPECIFIC CLONING IN CEREALS.
D.E. Delaney, S.H. Hulbert, B. Friebe and B.S. Gill, Department
of Plant Pathology, Kansas State University, Manhattan, KS
66506.
Genomic subtraction techniques are particularly useful for
obtaining markers closely to genes of agronomic importance.
Representational Difference Analysis (RDA) is a new PCR based
genomic subtraction technique (lisitsyn et al., Science 259:
946-951) that is well suited to organisms with large, complex
genomes. We have used this technique to obtain clones for the
long arm of chromosome 6 in rye (6RL), which carries a gene for
resistance to Hessian fly larvae. After completing three cycles
of RDA, we obtained 43 clones ranging in size from 400-800bp. Of
these 43 clones, 11 were middle repetitive sequences, 11 were low
copy sequences, and 21 clones did not hybridize to wheat or rye
genomic DNA and could be contaminants in the DNA samples that
were amplified during the procedure. All of the eleven low copy
clones mapped to 6RL, and one clone mapped within a one micron
(-70 million bp) fragment of 6RL containing the Hessian fly
resistance gene that is present as an interstitial translocation
in wheat. We are currently repeating this experiment to identify
additional markers on 6RL. We are also constructing a linkage
map of rye chromosome arm 6RL using two mapping populations. One
population is a cross between two wheat-rye translocation stocks
with resistant and susceptible 6RL arms translocated to 6BS of
wheat, and the other is a cross between the resistant rye
cultivar 'Balbo' and the susceptible cultivar 'Imperial'.
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