supporting information...2015/07/21 · dominican republic iac-nhm; es & rg 105 195 0.0135 0.06...

Supporting InformationSherratt et al. 10.1073/pnas.1506516112

Fig. S1. Toepad morphology of fore- and hindfeet among recent (representing five ecomorphs) and fossil anoles. Lamella numbers for the modern species aregiven beside digits III and IV. The amber fossils are colored by ecomorph according to DFA results if assigned with a probability greater than 0.90 (Table S3).Modern specimens from Museum of Comparative Zoology Reptile collection, Harvard University (prefix MCZ R).

Sherratt et al. www.pnas.org/cgi/content/short/1506516112 1 of 12

www.pnas.org/cgi/content/short/1506516112

Twig

(A

nolis

inso

litus

)Tr

unk

(A. b

revi

rost

ris)

Gra

ss-b

ush

(A. d

olic

ocep

halu

s)Tr

unk-

crow

n (A

. chl

oroc

yanu

s)Tr

unk-

grou

nd

(A. c

ybot

es)

O

K

J

F

MCZ R-59788T-0845

MCZ R-124846MCZ R-74865

MCZ R-69274MCZ R-155836

MCZ R-186752T-0339

MCZ R-107016 MCZ R-107014

Fig. S2. Cranial shape of five recent anoles representing five ecomorphs compared with four amber fossils. Crania of modern juveniles (Left) and adults(Center), as well as amber fossil skulls (Right) are shown in lateral and dorsal views. The amber fossils are colored by ecomorph if assigned with a probabilitygreater than 0.90 (Table S3). Modern specimens from Museum of Comparative Zoology Reptile collection, Harvard University (prefix MCZ R), or Field Tag forT.J. Sanger (prefix T).



Fig. S3. Variation among the fossils and how it corresponds with variation among the ecomorphs for all 10 cranial measurements. Size-corrected residuals areplotted for the amber fossils labeled by letters (A–T) and modern specimens (colored dots by ecomorph). Amber fossils are colored according to the DFA results(detailed in Table S3) when assigned with a probability greater than 0.90. A plot of the first two canonical variables of the DFA using all 10 variables (LowerRight) illustrates how well the cranial measurements discriminate the ecomorphs (modern specimens only).



Leng

th (r

esid

uals

)-0.2

-0.15

-0.1

-0.05

0

0.05

0.1

0.15

AMB

J

Pelvis width

-0.1

-0.05

0

0.05

0.1

LA E

RH

M

Sacrum width

-0.25

-0.2

-0.15

-0.1

-0.05

0

0.05

0.1

A

RMJH

Pubis length

-0.25

-0.2

-0.15

-0.1

-0.05

0

0.05

0.1

0.15

0.2

A J

Sternum width

-0.15

-0.1

-0.05

0

0.05

0.1

O

A JC

Humerus

-0.2

-0.15

-0.1

-0.05

0

0.05

0.1

0.15

OA RCF JH IG

Ulna

-0.2

-0.15

-0.1

-0.05

0

0.05

0.1

L

MEC GAJFIDB H

Femur

-0.3

-0.25

-0.2

-0.15

-0.1

-0.05

0

0.05

0.1

LE MC JA F TB IGH

D

Tibia

-0.3

-0.2

-0.1

0

0.1

LE MA JC TH IGB FD

Metatarsal IV

-0.3

-0.25

-0.2

-0.15

-0.1

-0.05

0

0.05

0.1

MEC H IJ TB DA GF

Hindtoe IV

-0.25

-0.2

-0.15

-0.1

-0.05

0

0.05

0.1

0.15

LME H JC ID FA

Hindtoe III

-0.25

-0.2

-0.15

-0.1

-0.05

0

0.05

0.1

0.15

C H JO

Foretoe III

-0.25

-0.2

-0.15

-0.1

-0.05

0

0.05

0.1

CG

H J O

Foretoe IV

Leng

th (r

esid

uals

)Le

ngth

(res

idua

ls)

Leng

th (r

esid

uals

)Le

ngth

(res

idua

ls)

EcomorphGB TR TC TG TW

Amber EcomorphGB TR TC TG TW


Amber




Amber




Amber




Amber


Amber-10

-8

-6

-4

-2

0

2

4

6

Can

onic

al 2

-10 -5 0 5 10Canonical 1

DFA, All postcranial variables, modern only

Fig. S4. Variation among the fossils and how it corresponds with variation among the ecomorphs for all 13 postcranial measurements. Size-corrected residualsare plotted for the amber (A–T) and modern specimens. Amber fossils are colored according to the discriminant function analysis (DFA) results (Table S3) whenassigned with a probability greater than 0.90. A plot of the first two canonical variables of the DFA using all 13 variables (Lower Right) illustrates how well thepostcranial measurements discriminate the ecomorphs (modern specimens only).



Fig. S5. Summary tree showing the phylogenetic relationships of the amber fossils inferred by parsimony analyses. Names of Anolis subclades are given on theright. The amber fossils are represented by letters corresponding to those in Fig. 2, and their relationships are indicated by yellow-orange branches; brokenbranches indicate the prune and graft results of alternative placements. DFA classifications for the fossils are indicated by pie charts, colored by ecomorph (onlyprobabilities greater than 0.1 are shown).



foretoe IIIforetoe IV

hindtoe IV

hindtoe III

frontal w

max l

prf w

skull w

jugal horbit d

mand lmand w

snout l

pmx w

2mm

tibia l

femur l

fused

sacrum w

metatarsal IV

hindtoe IV

humerus l

radius l

Postcranial

A. chlorocyanus (MCZ R-187268)47mm SVL (trunk-crown)

Cranial

Lamella Number

fused

hindtoe III

foretoe IIIforetoe IV

2mmA. baleatus (MCZ R-186663)49mm SVL (crown-giant)

unfused

unfusedb

a

A. chlorocyanus (MCZ R-59788)

A. coelestinus (MCZ R- 187469)

A. coelestinus (MCZ R- 187469)

Fig. S6. Schematic of the morphometric measurements and meristic counts taken from fossil and recent anole specimens shown on an adult trunk-crownanole (A). A juvenile crown-giant anole (B) of approximately the same SVL. Red arrows point to where there are clearly unfused (B) and fused (A) epiphyses anddiaphyses of fore and hindlimb bones, which allow the identification of juveniles vs. adults. Specimens from Museum of Comparative Zoology Reptile col-lection, Harvard University (prefix MCZ R).



Table

S1.

The20

fossils

exam

ined

inthis

study

Specim

enSp

ecim

enID

Elem

ents

preserved

Specim

enlocation

Notesonspecim

enLo

calityoforigin

Scan

ned

wherean

dbywhom

kVμA

Voxe

lsize

(mm)

Rotation

step

(o)

Exposure

(ms)

AAMNH

DRSH

1W

hole

body

American

Museum

ofNaturalH

istory,

New

York

CuratorDav

idGrimaldi

Dominican

Rep

ublic

Harva

rdCen

trefor

Nan

oscaleSy

stem

sMaterialSynthesis

Facility(CNS);

EmmaSh

erratt

(ES)

&Jasm

ine

Casart

7012

00.02

670.11

1,00

0

BI

Whole

body

Privateco

llection

Owned

byMarily

nHalonen

and

Michae

lCusanovich

of

Tuscon,Arizo

na

Dominican

Rep

ublic

CNS;

D.Lu

keMah

ler

5518

50.04

520.25

—

CII

Whole

body

Privateco

llection

Owned

byJim

Work

ofCen

tral

Point,

Oregon

Dominican

Rep

ublic

CNS;

ES65

195

0.06

090.11

1,00

0

DIII

Whole

body

Privateco

llection

Jose

Calbetoof

Guay

nab

o,Pu

erto

Rico

Dominican

Rep

ublic

CNS;

ES55

165

0.01

490.11

1,41

5

EM-106

2Pa

rt(hindlim

bsan

dtail)

Privateco

llection

Owned

byEttore

MoroneofTo

rino,

Italy

Dominican

Rep

ublic

Imag

ingan

dAnalysisCen

tre,

NaturalHistory

Museum

London

(IAC-N

HM);ES

&RussellGarwood

(RG)

105

195

0.01

070.06

354

FM-115

3W

hole

body

Privateco

llection

Owned

byEttore

MoroneofTo

rino,

Italy

Dominican

Rep

ublic

IAC-N

HM;ES

&RG

105

195

0.01

350.06

354

GM-341

0W

hole

body

Privateco

llection

Owned

byEttore

MoroneofTo

rino,

Italy

Dominican

Rep

ublic

IAC-N

HM;ES

&RG

105

195

0.03

300.06

354

HM-525

Whole

body

Privateco

llection

Owned

byEttore

MoroneofTo

rino,

Italy

Dominican

Rep

ublic

IAC-N

HM;ES

&RG

105

195

0.02

100.06

500

INMBA

Entom

P52

Whole

body

Naturhistorisches

Museum,Basal

Switze

rlan

d

Obtained

byC.

Baroni-Urban

i,NMBA,Basel

LaTo

camine,

Cordillera

Septentrional,

Dominican

Rep

ublic

Synch

rotron

Rad

iationFa

cility

(ESR

F)in

Grenoble,Fran

ce,

Ren

audBoistel&

AnthonyHerrel

——

0.00

746

——

JOAAAA

Whole

body

Privateco

llection

Supplie

dbyMarco

Greco

(Ambras

Greco

SAS)

of

Mila

n,Italy

Dominican

Rep

ublic

CNS;

ES75

130

0.02

370.11

1,00

0



Table

S1.

Cont.

Specim

enSp

ecim

enID

Elem

ents

preserved

Specim

enlocation

Notesonspecim

enLo

calityoforigin

Scan

ned

wherean

dbywhom

kVμA

Voxe

lsize

(mm)

Rotation

step

(o)

Exposure

(ms)

KSM

NSDo-

4871

-MPa

rt(hea

d)

Staa

tliches

Museum

fürNaturkunde

Stuttgart,

German

y

Donated

byGeo

rgDommel

of

Düsseldorf,

German

yin

1985

Dominican

Rep

ublic

CNS;

ES50

140

0.00

770.11

1,41

5

LSM

NSDo-

4886

-BPa

rt(tail,hindlim

b)

Staa

tliches

Museum

fürNaturkunde

Stuttgart,

German

y

Purchased

in19

85from

ErichBey

na,

Santo

Domingo,

Dominican

Rep

ublic

Dominican

Rep

ublic

CNS;

ES50

180

0.01

240.11

1,00

0

MSM

NSDo-

5566

-XPa

rt(trunk,

forelim

b,

hindlim

bs)

Staa

tliches

Museum

fürNaturkunde

Stuttgart,

German

y

Purchased

1991

from

D.M.Sa

lem,

8950

Sunrise

Lake

Blvd.80

/311

,Su

nrise,FL

3332

2,USA

Dominican

Rep

ublic

CNS;

ES50

130

0.02

560.11

1,41

5

NSM

U74

976

Part

(hea

d)

ShulerMuseum

of

Paleontology,

South

Methodist

University,Dallas

TX

Donated

byWilliam

S.Lo

weof

Granbury

Texa

s

Dominican

Rep

ublic

University

ofTe

xas

High-Resolution

X-ray

CTFa

cility,

RichKetch

am

8020

00.06

800.13

—

OUSN

M58

0060

Part

(hea

d,forelim

b)

Smithsonian

Institution

National

Museum

ofNaturalH

istory,

WashintonDC

Purchased

from

AmbericaWest

LaTo

camine,

Cordillera

Septentrional,

Dominican

Rep

ublic

University

ofTe

xas

High-Resolution

X-ray

CTFa

cility,

Matthew

Colbert

180

133

0.01

270.23

—

““

““

““

CNS;

ES60

165

0.00

510.18

1,00

0P

M-209

6Pa

rt(trunkan

dforelim

bs)

Privateco

llection

Owned

byEttore

MoroneofTo

rino,

Italy

Dominican

Rep

ublic

IAC-N

HM;ES

&RG

105

195

0.01

680.06

354

QRAAA

Part

(forelim

bs)

Privateco

llection

Supplie

dbyMarco

Greco

(Ambras

Greco

SAS)

of

Mila

n,Italy

Dominican

Rep

ublic

CNS;

ES75

130

0.00

890.11

1,00

0

RM-301

6Pa

rt(forelim

b,torso)

Privateco

llection

Owned

byEttore

MoroneofTo

rino,

Italy

Dominican

Rep

ublic

IAC-N

HM;ES

&RG

105

195

0.00

920.06

354

SM-127

3Pa

rt(forelim

bs)

Privateco

llection

Owned

byEttore

MoroneofTo

rino,

Italy

Dominican

Rep

ublic

IAC-N

HM;ES

&RG

105

195

0.00

680.06

354

TSM

NSDo-

5742

Part

(body,

nohea

d)

Staa

tliches

Museum

fürNaturkunde

Stuttgart,

German

y

Donated

byGeo

rgDommel

of

Düsseldorf,

German

yin

1980

Dominican

Rep

ublic

CNS;

ES50

165

0.03

080.11

1,41

5

Inform

ationisprovided

onstateofpreservation(elemen

tspreserved

);cu

rren

tlocationan

dlocalityoforigin

(includingthemine,

wherekn

own);an

dX-ray

microco

mputedtomographyscan

settings,with

inform

ationonwherethescan

was

donean

dbywhom.F

ossilO,o

riginally

scan

ned

atUniversity

ofTe

xas,was

rescan

ned

atHarva

rdat

ahigher

resolutionforthisproject

(datafrom

thehigher

resolutionscan

wereusedforthemorphometrican

dphylogen

etic

analyses).Th

reefossils

previouslywerepublished

:A(18),I(15

),an

dN(19).



Table

S2.

Mea

suredan

destimated

bodysize

(SVL),morphometricva

riab

les(inmm),an

dmeristicco

unts

take

nfrom

each

amber

specim

enan

dusedin

thediscrim

inan

tfunction

analyses

(DFA

s)presentedin

Table

S3

Specim

enA

BC

DE

FG

HI

JK

LM

NO

PQ

RS

T

Mea

suredSV

L23

.830

.052

.830

.225

.723

.225

.423

.1SV

Lestimatefrom

lumbar

length

25.7

27.1

24.4

24.2

31.0

31.0

26.3

SVLestimatefrom

ilium

length

27.6

53.6

17.7

21.8

23.0

21.8

29.6

25.1

SVLestimatefrom

bodylength

24.8

SVLestimatefrom

man

dible

length

28.2

27.7

27.5

24.2

21.1

21.5

20.6

21.2

19.7

SVLestimatefrom

other

(see

notes)

19.4–21

.019

.9–21

.718

.5–20

.025

.2–44

.719

.9–28

.2Man

dible

length

8.63

8.46

8.40

7.40

6.43

6.97

6.57

6.29

6.57

6.00

Prem

axillalength

1.23

1.06

1.29

0.89

0.97

1.05

0.96

1.02

Max

illalength

4.20

5.40

5.27

4.52

4.13

3.91

3.55

3.79

3.64

Skullwidth

4.13

4.94

4.88

3.80

4.23

3.80

3.64

3.63

3.70

Frontalwidth

betwee

norbits

0.64

0.77

0.54

0.51

0.54

0.39

0.35

0.53

Prefrontalwidth

2.89

2.91

2.80

2.46

2.47

2.19

2.58

Orbitdiameter

3.38

3.47

3.50

3.31

2.84

3.04

2.96

2.90

2.90

Snoutlength

3.65

3.81

3.73

3.01

2.59

2.73

2.15

2.51

2.35

Man

dible

width

3.53

6.80

4.12

4.20

3.24

3.38

3.30

3.13

3.15

Jugal

height

1.92

1.81

1.58

1.52

1.68

1.50

1.36

1.54

1.62

Sternum

width

2.29

1.82

Humeruslength

4.38

9.00

3.17

3.20

3.48

4.64

3.40

Ulnalength

3.62

7.80

3.72

2.95

2.72

2.79

2.60

3.03

2.57

4.18

3.13

2.70

ForetoeIV

length

4.83

1.64

2.47

2.16

1.93

2.18

1.77

2.46

1.88

ForetoeIII

length

5.17

1.90

2.83

2.43

2.40

2.33

2.03

2.93

2.18

Pelviswidth

2.38

2.25

2.12

1.47

2.46

Sacrum

width

2.00

1.51

1.51

2.52

1.92

1.62

Pubislength

2.48

1.78

1.70

2.45

1.96

Femurlength

6.05

5.70

12.45

3.23

4.85

6.26

5.95

4.56

4.88

4.74

8.04

7.15

Tibia

length

4.89

5.24

10.98

2.44

4.44

5.35

4.56

4.02

4.38

4.22

6.75

6.21

4.29

MetatarsalIV

length

3.37

3.16

6.40

1.73

2.72

3.04

2.92

2.74

2.83

2.80

3.88

3.80

2.84

HindtoeIV

length

4.72

5.16

9.80

3.10

4.38

4.89

4.63

4.54

4.61

4.13

6.34

4.42

HindtoeIII

length

2.53

5.54

1.66

2.31

2.82

2.56

2.49

2.41

3.65

3.30

HindtoeIV

lamellae

1416

1816

17HindtoeIII

lamellae

2326

1618

2321

1923

17Fo

retoeIV

lamellae

1617

1518

2121

ForetoeIII

lamellae

1313

1620

20

Reg

ressionmodelsofmodernspecieswereusedto

estimateSV

Lofthefossils

from

fourlog-transform

edva

riab

les:thelength

ofalumbar

verteb

ra;length

oftheilium;bodylength;an

dman

dible

length.

Detailsan

deq

uationsaregiven

inMethods.Allsquared

correlationco

efficien

tsare0.96

,excep

tbodylength,w

hichis0.99

.SVLestimates

inbold

arethose

usedin

theDFA

s.W

hen

more

than

oneestimatewas

available,theva

luech

osenwas

decided

acco

rdingto

thepreservationofthefossil.

Rows1–

6providearangeofSV

Lestimates:specim

enA

broke

n,mea

suredSV

Lan

underestimate;

specim

enCinco

mplete,

mea

suredSV

Lan

underestimate;

specim

enD

inco

mplete,ilium

frag

men

tpreserved

;specim

enG

inco

mplete,mea

suredSV

Lan

underestimate;

specim

enH

broke

n,m

easuredSV

Lan

underestimate;

specim

enK,

other

SVLestimated

from

rangeofecomorph-specificslopes

forman

dible

length;specim

enN,o

ther

SVLestimated

from

rangeofecomorph-specificslopes

forman

dible

length;specim

enO,o

ther

SVLestimated

from

rangeofecomorph-specific

slopes

forman

dible

length;specim

enP,

other

SVLestimated

from

rangeofecomorph-specific

slopes

forulnalength

(r2=0.84

);specim

enS,

other

SVLestimated

from

rangeof

ecomorph-specific

slopes

forulnalength

(r2=

0.84

).



Table S3. Results of discriminant function analyses (DFAs) using size-corrected residuals of the morphometric variables (details of thesize correction in Table S2)

Specimen TC TR TG TW GB λ Approximate F (df) Percent misclassified

A 1.00 0.00002 40.50 (84, 227.59) 0B 0.96 0.00099 35.25 (44, 304.19) 0C 1.00 0.00018 50.11 (44, 258.28) 0D 0.99 0.00256 28.33 (36, 260.31) 1.22E 1.00 0.00148 46.78 (28, 257.42) 0F 1.00 0.00004 41.46 (72, 238.29) 0G 0.69 0.31 0.00004 44.46 (68, 241.70) 0H 0.99 0.00002 40.26 (84, 227.59) 0I 0.99 0.00037 45.59 (40, 259.70) 0J 1.00 0.00004 34.31 (88, 243.69) 0K 0.82 0.18 0.01746 18.97 (32, 304.00) 7.4

[0.81] [0.18] 0.02303 16.91 (32, 304.00) 13.8L 0.98 0.00227 49.70 (24, 252.39) 1.22M 1.00 0.00075 36.86 (40, 259.70) 2.4N 0.86 0.12 0.02797 18.21 (28, 300.68) 5.3

[0.52] [0.48] 0.03315 16.88 (28, 300.68) 10.6O 1.00 0.00122 26.99 (48, 275.54) 1.1

[0.99] 0.00183 23.75 (48, 275.54) 1.1P {0.13) {0.49} {0.34} 0.22769 20.81 (8, 152.00) 42.7Q {0.31} {0.13} {0.20} {0.35} 0.22769 20.81 (8, 152.00) 42.7R 0.94 0.00626 43.90 (20, 243.06) 3.7S {0.60} {0.35} 0.22769 20.81 (8, 152.00) 42.7T 0.99 0.04449 43.50 (12, 233.12) 24.2

Probabilities of classification to the five ecomorphs are presented, with a post hoc Wilks’ λ test of the null hypothesis that the means of all of theindependent variables are equal across groups (F statistic and degrees of freedom), and proportion of individuals misclassified in each analysis. Probabilitiesbelow 0.1 are not shown. Each DFA used a different set of variables specific to the fossil (Table S2). For three fossils that comprised an isolated skull (K, N, andO), a second DFA using raw (not size-corrected) variables was conducted, the results of which are given in brackets. For fossils P, Q, and S, only lamella countswere available for the DFA, so these results are given in braces. Bold indicates DFA probabilities above 0.90.



Table

S4.

Resultsofparsimonyan

dBay

esianphylogen

etic

analyses



Movie S1. Photographs and animated 3D renderings of the 20 amber fossils, in order from A to T. For each fossil, a photograph of the specimen is shownfollowed by a 3D reconstructed skeleton and/or soft tissue animation from X-ray micro-CT data. In specimens L and M, there are ant inclusions (genus Azteca)that have been false-colored brown and rendered opaque.

Movie S1


http://movie-usa.glencoesoftware.com/video/10.1073/pnas.1506516112/video-1


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