tracking breeding success of deployed oysters through microsatellite variation carlsson j., s.k....
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TRACKING BREEDING SUCCESS OF DEPLOYED TRACKING BREEDING SUCCESS OF DEPLOYED OYSTERS THROUGH MICROSATELLITE VARIATION OYSTERS THROUGH MICROSATELLITE VARIATION
Carlsson J., S.K. Allen Jr, and K.S. Reece
Supported by grants from:NOAA Oyster Restoration Program through the Chesapeake Bay Program Office
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The problemRESTORATION EFFORTS HAVE BEEN FOCUSED ON CONSTRUCTING REEFS
SEEDED WITH DOMESTICATED AQUACULTURE STOCKS (MOSTLY DISEASE-RESISTANCE SELCTED STOCKS-DEBY AND CROSBREED LINEs)
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ARE THE DEPLOYED DOEMSTICATED OYSTERS CONTRIBUTE SIGNIFICANTLY TO SPAT PRODUCTION?
DEBY DEPLOYMENT – USE mtDNA RFLP (note this also works for Gulf of Mexico strains)
The Question
PRIOR TO DEPLOYMENT
DEPLOYED DEBY
PRODUCED SPAT
Rare
BB
AA
Data from Great Wicomico 2002-2006
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CROSBREED DEPLOYMENT – USE?
The Problem 1
Hinf I digest of mtDNA coIII in natural population
Hinf I digest of mtDNA coIII in Crosbreed strain
The coI and coIII mitochondrial genes do not allow for discriminating among non-DEBY or non-Gulf of Mexico strains
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The frequency of the BB haplotypes is not 100% in the DEBY line or Gulf strains
An oyster showing the AA haplotype could still be a DEBY or Gulf of Mexico oyster
An oyster showing the BB haplotype has a high probability of being a DEBY or Gulf of Mexico oyster
The Problem 2
DEBY
mtDNA is good for assigning proportion of DEBY/Gulf of Mexico strains but not for assigning individuals to strains
WILD
Rare
BB
AA
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The Solution?Microsatellites
+High power
+Highly variable
+High throughput
+Nuclear
-Time consuming to develop
-Expensive Today the most widely used genetic marker for population genetic studies
ATCTATATATATATATATATATATCGTGG
TCGATATATATATATATATATATAGCACC
ATCTATATATATATATATATCGTGG
TCGATATATATATATATATAGCACC
Chromosome from
♀ (TA)10
Chromosome from
♂ (TA)8
Microsatellite
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Example 1Example 1
Wild pop1 – blueWild pop2 – whiteDEBY – yellow
Test pop – greyNote!
4 microsatellites4 microsatellites
2 mtDNA genes2 mtDNA genes
Over 85% did Over 85% did significantlysignificantly not assign to DEBY not assign to DEBY
1 individual assigned to DEBY1 individual assigned to DEBY
Factorial Correspondence Analysis (Genetix)
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Example 2Example 2
Wild pop 1 – blue
Test pop A – pinkDEBY – yellow
Wild pop 2 - white
Test pop B - grey
4 microsatellites4 microsatellites
2 mtDNA genes2 mtDNA genes
Over 90% did Over 90% did significantlysignificantly not assign to DEBY not assign to DEBY
No individuals assigned to DEBYNo individuals assigned to DEBY
Factorial Correspondence Analysis (Genetix)
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Microsatellite Work in ProgressMicrosatellite Work in Progress
Spat-on-shell – deployment of domesticated oysters
Six sites in the Chesapeake Bay
Six samples of wild oysters collected prior to deployment (n=100/site)
Sampling of deployed spat on shell (n=50 per deployment, 3 deployments/site)
Sampling of 8 domesticated aquaculture lines (n=50/line)
Analyze samples with a suite of 8 microsatellite loci
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Rappahannock wild – yellow
Deployed spat-on-shell - blue
Example Rappahannock River, Example Rappahannock River, Drumming GroundDrumming Ground
Factorial Correspondence Analysis (Genetix)
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Example Rappahannock River, Example Rappahannock River, Drumming GroundDrumming Ground
Hybrids - white
Deployed spat-on-shell - blue
Rappahannock wild – yellow
Factorial Correspondence Analysis of simulated hybrid progeny (Genetix and Hybridlab)
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RappannockF1 hybrids Deployed
Hybrid partitioning of simulated hybrid progeny based microsatellite multilocus genotypes (8 loci)
Hybrid partitioning (Structure)
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Rappannock F1 hybridsDeployed
Hybrid partitioning (BAPS)
Hybrid partitioning of simulated hybrid progeny based microsatellite multilocus genotypes (8 loci)
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Concluding remarksConcluding remarks
-mtDNA RFLP has provided and will continue to provide essential information about the contribution of DEBYs on a group level
-Microsatellites can clearly discriminate between wild and domesticated oysters on a group and on an individual level
-Microsatellites can also be utilized to detect hybrids and to determine the amount of genetic introgression between wild and domesticated oysters (both current and future)
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Thanks!Thanks!
Jim WessonJim Wesson
Tommy LeggettTommy Leggett
Wes HeydeckWes Heydeck
LABORATORYLABORATORY
Georgeta ConstantinGeorgeta Constantin
Elizabeth FrancisElizabeth FrancisMissy SouthworthMissy Southworth
P.G. RossP.G. Ross
Mark LuckenbachMark LuckenbachSharon FurinessSharon Furiness
SAMPLINGSAMPLING
Mike CongroveMike Congrove