In the formt provided y the uthors nd unedited. Genetic divergence of erly song discrimintion etween two young songird species Dvid Whetcroft* nd Ann Qvrnström SUPPLEMENTARY INFORMATION VOLUME: 1 ARTICLE NUMBER: 0192 NATURE ECOLOGY & EVOLUTION DOI: 10.1038/s41559-017-0192 www.nture.com/ntecolevol 1
NATURE ECOLOGY & EVOLUTION DOI: 10.1038/s41559-017-0192 www.nture.com/ntecolevol 2
Supplementry Figure 1. Song plycks to nestling collred nd pied flyctchers.. Proportions of nestlings looking up t the nest ox opening in response to song plycks.. Proportions of nestlings moving in response to song plycks. For. nd., proportions of nestlings were first tken within ech nest nd then summrized y ox plots. For presenttion purposes, the dt is presented without ccounting for rndom effects (yer, song recording, nest, nd nestling identity) nd dditionl significnt fixed effects (ehvior efore song plyck, verge condition of nestlings, nd clutch size) tht were included during the modeling procedure. Box plots give the verge numer of egging clls produced y the nestlings oserved within nests in response to collred (white) or pied flyctcher (lck) songs. Lels ove ox plots ( Con, Het ) denote whether the song plyck ws conspecific or heterospecific reltive to the species of the nestlings. Boxes give 1 st nd 3 rd qurtiles, thick lines give medins, nd whiskers give 95 th percentiles. Numers ove oxes give P-vlues for post-hoc tests of differences in generl liner mixed models, which included dditionl fixed nd rndom effects (Supplementry Tles 2-3 nd Supplementry Tles 5-6). NATURE ECOLOGY & EVOLUTION DOI: 10.1038/s41559-017-0192 www.nture.com/ntecolevol 3
NATURE ECOLOGY & EVOLUTION DOI: 10.1038/s41559-017-0192 www.nture.com/ntecolevol 4
Supplementry Figure 2. Song plycks to nestling hyrid flyctchers.. Proportions of nestlings looking up t the nest ox opening in response to song plycks.. Proportions of nestlings moving in response to song plycks. For. nd., proportions of nestlings were first tken within ech nest nd then summrized y ox plots. For presenttion purposes, the dt is presented without ccounting for rndom effects (yer, song recording, nest, nd nestling identity) nd dditionl significnt fixed effects (ehvior efore song plyck, verge condition of nestlings, nd clutch size) tht were included during the modeling procedure. Box plots give the verge numer of egging clls produced y the nestlings oserved within nests in response to collred (white) or pied flyctcher (lck) songs. Boxes give 1 st nd 3 rd qurtiles, thick lines give medins, nd whiskers give 95 th percentiles. Numers ove oxes give P-vlues for post-hoc tests of differences in generl liner mixed models, which included dditionl fixed nd rndom effects (Supplementry Tles 8-9). NATURE ECOLOGY & EVOLUTION DOI: 10.1038/s41559-017-0192 www.nture.com/ntecolevol 5
Supplementry Tle 1. Begging clls produced during song plycks y nestlings rised y conspecific prents model formul, log-lik χ (d.f.) P null -1014.2 - - egging efore plyck -995.1 38.2 (1) < 0.001 egging efore plyck + order -991.4 17.9 (1) < 0.001 egging efore plyck + order + condition -990.0 3.9 (1) 0.089 egging efore plyck + order + clutch size -990.9 1.1 (1) 0.304 egging efore plyck + order + song -983.9 15.1 (1) < 0.001 tretment egging efore plyck + order + song tretment species -982.7 2.4 (2) 0.296 Bold indictes model with lowest log-likelihood in likelihood rtio test. Models with dditionl terms evluted step-wise. NATURE ECOLOGY & EVOLUTION DOI: 10.1038/s41559-017-0192 www.nture.com/ntecolevol 6
Supplementry Tle 2. Looking during song plycks y nestlings rised y conspecific prents model formul, log-lik χ (d.f.) P null -410.1 - - looking efore plyck -386.3 47.7 (1) < 0.001 looking efore plyck + order -385.8 1.0 (1) 0.329 looking efore plyck + condition -386.1 0.4 (1) 0.528 looking efore plyck + clutch size -384.5 3.4 (1) 0.064 looking efore plyck + song tretment -378.7 15.1 (1) < 0.001 looking efore plyck + song tretment species -378.7 0.1 (2) 0.980 Bold indictes model with lowest log-likelihood in likelihood rtio test. Models with dditionl terms evluted step-wise. NATURE ECOLOGY & EVOLUTION DOI: 10.1038/s41559-017-0192 www.nture.com/ntecolevol 7
Supplementry Tle 3. Moving during song plycks y nestlings rised y conspecific prents model formul, log-lik χ (d.f.) P null -591.0 - - moving efore plyck -568.2 45.5 (1) < 0.001 moving efore plyck + order -567.9 0.7 (1) 0.406 moving efore plyck + condition -564.4 7.7 (1) 0.006 moving efore plyck + condition + clutch size -560.6 7.6 (1) 0.006 moving efore plyck + condition + clutch -550.3 20.5 (1) < 0.001 size + song tretment moving efore plyck + condition + clutch size + song tretment species -549.4 1.7 (2) 0.419 Bold indictes model with lowest log-likelihood in likelihood rtio test. Models with dditionl terms evluted step-wise. NATURE ECOLOGY & EVOLUTION DOI: 10.1038/s41559-017-0192 www.nture.com/ntecolevol 8
Supplementry Tle 4. Begging clls produced during song plycks y nestlings rised y heterospecific prents model formul, log-lik χ (d.f.) P null -347.0 - - egging efore plyck -347.0 0.0 (1) 0.985 order -340.9 12.3 (1) < 0.001 order + condition -340.6 0.5 (1) 0.497 order + clutch size -336.8 8.1 (1) 0.005 order + clutch size + song tretment -331.9 9.9 (1) 0.002 order + clutch size + song tretment species -330.9 2.0 (2) 0.376 Bold indictes model with lowest log-likelihood in likelihood rtio test. Models with dditionl terms evluted step-wise. NATURE ECOLOGY & EVOLUTION DOI: 10.1038/s41559-017-0192 www.nture.com/ntecolevol 9
Supplementry Tle 5. Looking during song plycks y nestlings rised y heterospecific prents model formul, log-lik χ (d.f.) P null -115.1 - - looking efore plyck -101.2 27.7 (1) < 0.001 looking efore plyck + order -99.4 3.6 (1) 0.059 looking efore plyck + condition -100.7 1.1 (1) 0.301 looking efore plyck + clutch size -96.8 8.7 (1) 0.003 looking efore plyck + clutch size + -87.5 18.8 (1) < 0.001 song tretment looking efore plyck + clutch size + song tretment species -85.2 4.6 (2) 0.101 Bold indictes model with lowest log-likelihood in likelihood rtio test. Models with dditionl terms evluted step-wise. NATURE ECOLOGY & EVOLUTION DOI: 10.1038/s41559-017-0192 www.nture.com/ntecolevol 10
Supplementry Tle 6. Moving during song plycks y nestlings rised y heterospecific prents model formul, log-lik χ (d.f.) P null -131.9 - - moving efore plyck -124.2 15.5 (1) < 0.001 moving efore plyck + order -124.1 0.1 (1) 0.828 moving efore plyck + condition -124.1 0.0 (1) 0.835 moving efore plyck + clutch size -123.8 0.8 (1) 0.388 moving efore plyck + song tretment -123.7 0.9 (1) 0.355 moving efore plyck + song tretment species -122.2 3.9 (3) 0.268 Bold indictes model with lowest log-likelihood in likelihood rtio test. Models with dditionl terms evluted step-wise. NATURE ECOLOGY & EVOLUTION DOI: 10.1038/s41559-017-0192 www.nture.com/ntecolevol 11
Supplementry Tle 7. Begging clls produced during song plycks y hyrid nestlings hypothesis tested model formul AICc AICc mternl effects egging efore plyck + song plyck 429.5 - species mternl species egging efore plyck + song plyck 455.9 26.5 species egging efore plyck + mternl 591.6 162.1 species song egging efore plyck + clutch size 632.5 203.0 egging efore plyck 632.7 203.2 egging efore plyck + order 634.7 205.2 null 657.0 227.6 sex-linkge egging efore plyck + order + song 213.7 - plyck species mternl species egging efore plyck + order+ song 216.9 3.2 plyck species mternl species sex egging efore plyck + order + song 230.9 17.2 plyck species sex egging efore plyck + order + song 231.2 17.5 plyck species egging efore plyck + order 358.3 144.6 egging efore plyck + order + clutch 359.2 145.5 size egging efore plyck 364.7 151.0 null 376.9 163.2 Models included ll clutches of hyrid nestlings. Models included those clutches of hyrid nestlings where the sex of ll individul nestlings ws known. NATURE ECOLOGY & EVOLUTION DOI: 10.1038/s41559-017-0192 www.nture.com/ntecolevol 12
Supplementry Tle 8. Looking during song plycks y hyrid nestlings hypothesis tested model formul AICc AICc mternl effects looking efore plyck + song plyck 179.8 - species mternl species looking efore plyck + song plyck 186.2 6.5 species looking efore plyck + mternl 189.8 10.0 species song looking efore plyck + order 192.1 12.3 looking efore plyck 193.8 14.0 looking efore plyck + clutch size 194.2 14.4 null 211.1 31.3 sex-linkge looking efore plyck + order + song 126.8 - plyck species mternl species looking efore plyck + order + song 130.9 4.1 plyck species sex looking efore plyck + order + song 132.0 5.2 plyck species mternl species sex looking efore plyck + order + clutch 132.0 5.2 size looking efore plyck + order 132.0 5.2 looking efore plyck + order + song 132.2 5.4 plyck species looking efore plyck 135.8 9.0 null 148.3 21.5 Models included ll clutches of hyrid nestlings. Models included those clutches of hyrid nestlings where the sex of ll individul nestlings ws known. NATURE ECOLOGY & EVOLUTION DOI: 10.1038/s41559-017-0192 www.nture.com/ntecolevol 13
Supplementry Tle 9. Moving during song plycks y hyrid nestlings hypothesis tested model formul AICc AICc mternl effects moving efore plyck + song plyck 202.8 - species moving efore plyck 204.0 1.2 moving efore plyck + order 204.9 2.1 moving efore plyck + mternl 205.5 2.7 species song moving efore plyck + clutch size 205.9 3.1 moving efore plyck + song plyck 206.4 3.6 species mternl species null 233.9 31.1 sex-linkge moving efore plyck 133.0 - moving efore plyck + order 133.1 0.1 moving efore plyck + clutch 134.4 1.4 moving efore plyck + song plyck 134.6 1.7 species moving efore plyck + song plyck 135.9 2.9 species sex moving efore plyck + song plyck 138.6 5.6 species mternl species moving efore plyck + song plyck 143.1 10.1 species mternl species sex null 162.9 29.9 Models included ll clutches of hyrid nestlings. Models included those clutches of hyrid nestlings where the sex of ll individul nestlings ws known. NATURE ECOLOGY & EVOLUTION DOI: 10.1038/s41559-017-0192 www.nture.com/ntecolevol 14
Supplementry Tle 10. Song responses of pure species nd hyrid nestlings egs - model formul, log-lik χ (d.f.) P null -2399.8 - - egging efore plyck -2349.9 99.72 (1) < 0.001 egging efore plyck + order -2342.6 14.7 (1) < 0.001 egging efore plyck + order + clutch -2340.8 3.5 (1) 0.062 egging efore plyck + order + song plyck species species -2177.4 330.4 (5) < 0.001 looking - model formul, log-lik χ (d.f.) P null -681.4 - - looking efore plyck -635.6 91.5 (1) < 0.001 looking efore plyck + order -635.6 0.0 (1) 0.958 looking efore plyck + clutch -633.0 5.2 (1) 0.023 looking efore plyck + clutch + song plyck species species -611.5 43.1 (5) < 0.001 moving - model formul, log-lik χ (d.f.) P null -885.2 - - moving efore plyck -845.1 80.2 (1) < 0.001 moving efore plyck + order -844.4 1.5 (1) 0.229 moving efore plyck + clutch -844.2 1.7 (1) 0.196 moving efore plyck + song plyck species species -829.1 31.9 (5) < 0.001 Bold indictes model with lowest log-likelihood. Models with dditionl terms evluted step-wise. NATURE ECOLOGY & EVOLUTION DOI: 10.1038/s41559-017-0192 www.nture.com/ntecolevol 15
Supplementry Tle 11. List of song recordings used for ech experiment (spredsheet) Supplementry Tle 12. Rw ehviorl mesurements extrcted from video recordings (spredsheet) NATURE ECOLOGY & EVOLUTION DOI: 10.1038/s41559-017-0192 www.nture.com/ntecolevol 16