Melissa Gunn, Dave Coltman, Jon Slate, Katie Hartnup, Andrew

Leviston

MicrosatellitesPanel 1 Panel 2

Size (bp)

400

100

Microsatellites• 19 polymorphic m’sats working• ~10 alleles per locus• 200 extra clones available• ~460 individuals genotyped (tissue)

– 2003/2004 Sulphur, Kloo, Agnes all extracted

• 80% of loci scored• Robot re-calibrated, hair extraction kit ready to go

– ~40 days to extract 2500 hairs– All hair samples catalogued– PCR may need re-optimising for hair samples

Parentage Assignment

• Cervus V.2

• Currently assigning 58% sires • known mother genotyped (95% confidence)

– Assigning 54% sires • no mother genotyped

– More sires assigned if all males included • Chase males don’t seem to get all paternities

Assigned Parent Output

Offspring ID Known parent ID KP class0-KP loci mismatching Candidate parent ID

O-CP loci mismatching LOD Delta Confidence

KL36 KL40 Typed 0 SUL18 1 1.02E+01 8.00E+00 *KL68 KL30 Typed 0 KL81 0 9.93E+00 6.27E+00 *KL18 KL10 Typed 0 KL5 0 1.29E+01 7.38E+00 *KL144   Untyped 0 KL53 1 6.49E+00 4.41E+00 *KL145   Untyped 0 KL53 0 7.12E+00 5.66E+00 *KL48 KL11 Typed 0 KL89 0 1.17E+01 3.72E+00 *KL172   Untyped 0 KL9 1 7.08E+00 3.63E+00 *KL71   Untyped 0 KL9 1 6.52E+00 3.98E+00 *SUL28 SUL19 Typed 0 SUL41 0 1.45E+01 1.09E+01 *SUL93 SUL19 Typed 2 SUL7 1 1.13E+01 9.72E+00 *SUL102   Untyped 0 SUL11 3 5.20E+00 2.45E+00 *SUL99   Untyped 0 SUL4 1 6.58E+00 2.35E+00 *SUL154 SUL14 Typed 1 SUL16 2 9.10E+00 7.15E+00 *SUL166 SUL12 Typed 1 SUL41 1 1.32E+01 9.52E+00 *SUL114 SUL17 Typed 2 SUL2 2 1.16E+01 8.61E+00 *SUL167 SUL17 Typed 1 SUL2 1 1.13E+01 6.89E+00 *

Papers• Primer note – “Microsatellites for

Tamiasciurus hudsonicus and their

cross-utility in other rodents”

Sample Thu3 Thu14 Thu21 Thu23 Thu25 Thu29 Thu31 Thu32 Thu33 Thu37 Thu38 Thu40 Thu41 Thu42 Thu49 Thu50

GreySquirrel1 263 263 157 157 186 186 125 129 262 262 159 169 374 395GreySquirrel2 148 148 186 186 187 189 129 141 262 262 141 141 374 374RedSquirrel5 241 241 187 189 102 125 144 149 253 253RedSquirrel6 280 296 137 141 294 296 248 250RedSquirrel11 200 200RedSquirrel12 189 189 200 200 102 102 144 181RedSquirrel30 200 200RedSquirrel49 196 202RedSquirrel50 200 200RedSquirrel53 200 200Microtus agrestis (field vole)Clethrionomys glareolus (Bank vole) 1 121 121 179 179 254 254 178 178Clethrionomys glareolus (Bank vole) 2 178 178Clethrionomys glareolus (Bank vole) 3Clethrionomys glareolus (Bank vole) 4 178 178Clethrionomys glareolus (Bank vole) 5Clethrionomys glareolus (Bank vole) 6Clethrionomys glareolus (Bank vole) 7 121 121

Tamiasciurus hudsonicus hair

H1843 232 252Tamiasciurus hudsonicus muscle 230 232 278 286 156 156 187 187 183 187 125 125 141 141 281 285 149 149 159 174 300 302 382 392 238 264 250 250 198 200 273

Papers & Ideas• Heritability of plasticity of birthdate

– Estimate including maternal effect 0.89±0.6/4.9 (av)– Plasticity measured as birthdate/log cone previous year– Based on 156 mothers with multiple births– Later analyses will include probable fathers– Probable covariates incl. juv. growth rate, weight at

birth, number of maternal breeding events

• Population structure & migration – observed migrations vs. genetic effects

• Productivity vs. genetic diversity – Agnes vs. Kloo? Between-individuals?)

Other Results & Ideas• NeEstimator - Effective pop. size

= ~4.6/56/109 (c.f 456 individuals used for test)– How large an Ne is required to maintain evolvability ?– Compare with Ne from previous years?– Depends on good hair genotyping

• Is selection still favouring plastic females?• More tissue

– mainly new juv’s– Some adults still need sampling

• Comparison with island populations (Ne, birthdate, etc)?

Acknowledgements

Thanks for access to the database, all squirrel sample collectors, and providers of other DNA samples and

primers: Roland Ashford, Chris Faulkes, Chris Barnard, Marie Hale, Amy Plowman, Deborah

Dawson, Terry Burke, Pippa Thomson, Corey Davis