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Use of cellular oncogene probes to identify Morone hybrids
I I Wirgin1, L Maceda, C Mesing
1Institute of Environmental Medicine, New York University Medical Center, Tuxedo 10987.
Abstract:
We tested the ability of cellular oncogene (c-onc) probes to identify F1 hybrids and the lineage of known backcrosses within the fish genus Morone. Total DNA was isolated from five to 14 individuals per North American Morone species (striped bass, white bass, white perch, and yellow bass). The DNA was digested with two restriction enzymes, Eco RI and Hin dIII, Southern blotted, and hybridized to six different c-onc probes including v-abl, v-erb B, c-myc, c-H-ras, c-K-ras, and v-src. We found fixed genotypic differences among the four species for all six probes in single restriction enzyme digests. The heritability of these nuclear DNA genotypes was evaluated in hatchery-produced F1 Morone hybrids (striped bass x white bass and striped bass x white perch) tested with the six informative single probe/restriction enzyme combinations. All F1 individuals exhibited heterozygosity in all diagnostic nuclear DNA fragments, confirming the Mendelian inheritance of these genotypes in these fish. Furthermore, analysis of these nuclear DNA genotypes in hatchery-produced backcrosses of F1 hybrids striped bass x (white bass x striped bass) detected both recombinant and parental genotypes at all six polymorphic c-onc sequences. The lineage of suspected Morone hybrids of unknown descent collected from Lewis Smith Lake, Alabama, and from the Occoquan River, Virginia, was determined using the c-onc probes. Our results suggest that c-onc probes are suitable markers to unequivocally identify F1 hybrids and backcrosses and to quantify introgression in natural populations of fishes. The addition of RFLP analysis of mtDNA provided a complete ancestral history of individual fish.
Insights
Cellular oncogene (c-onc) probes effectively identify fish hybrids and backcrosses. These DNA markers confirm Mendelian inheritance and help trace the lineage of wild Morone hybrids.
Area of Science:
- Genetics
- Molecular Biology
- Ichthyology
Background:
- Accurate identification of fish hybrids and their lineage is crucial for population genetics and conservation.
- Traditional methods may not always provide definitive results for complex hybridizations within the Morone genus.
Purpose of the Study:
- To evaluate the efficacy of cellular oncogene (c-onc) probes for identifying F1 hybrids and backcrosses in the fish genus Morone.
- To assess the heritability of nuclear DNA genotypes derived from c-onc probes.
- To determine the lineage of wild Morone hybrids of unknown ancestry.
Main Methods:
- Isolation of total DNA from four North American Morone species.
- Digestion of DNA with Eco RI and Hin dIII restriction enzymes.
- Southern blotting and hybridization with six different c-onc probes (v-abl, v-erb B, c-myc, c-H-ras, c-K-ras, v-src).
- Analysis of hatchery-produced F1 hybrids and backcrosses.
- Application of probes to wild suspected hybrids from Alabama and Virginia.
- Mitochondrial DNA (mtDNA) RFLP analysis for ancestral history.
Main Results:
- Fixed genotypic differences were observed among the four Morone species using all six c-onc probes and single restriction enzyme digests.
- All F1 hybrids showed heterozygosity in diagnostic nuclear DNA fragments, confirming Mendelian inheritance.
- Backcross analysis revealed both recombinant and parental genotypes at all six polymorphic c-onc sequences.
- c-onc probes successfully identified the lineage of suspected wild Morone hybrids.
Conclusions:
- Cellular oncogene (c-onc) probes are reliable molecular markers for unequivocally identifying F1 hybrids and backcrosses in Morone species.
- These probes facilitate the quantification of introgression in natural fish populations.
- Combined with mtDNA RFLP analysis, c-onc probes provide a comprehensive ancestral history for individual fish.