TRACKING BREEDING SUCCESS OF DEPLOYED OYSTERS THROUGH MICROSATELLITE VARIATION Carlsson J., S.K....
-
Upload
cynthia-rice -
Category
Documents
-
view
280 -
download
0
Transcript of TRACKING BREEDING SUCCESS OF DEPLOYED OYSTERS THROUGH MICROSATELLITE VARIATION Carlsson J., S.K....
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
The problemRESTORATION EFFORTS HAVE BEEN FOCUSED ON CONSTRUCTING REEFS
SEEDED WITH DOMESTICATED AQUACULTURE STOCKS (MOSTLY DISEASE-RESISTANCE SELCTED STOCKS-DEBY AND CROSBREED LINEs)
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
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
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
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
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)
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)
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
Rappahannock wild – yellow
Deployed spat-on-shell - blue
Example Rappahannock River, Example Rappahannock River, Drumming GroundDrumming Ground
Factorial Correspondence Analysis (Genetix)
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)
RappannockF1 hybrids Deployed
Hybrid partitioning of simulated hybrid progeny based microsatellite multilocus genotypes (8 loci)
Hybrid partitioning (Structure)
Rappannock F1 hybridsDeployed
Hybrid partitioning (BAPS)
Hybrid partitioning of simulated hybrid progeny based microsatellite multilocus genotypes (8 loci)
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)
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