Systematics of the Polymorphic North American Rat Snake (Elaphe Obsoleta)

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2000 Systematics of the Polymorphic North American Rat Snake (Elaphe Obsoleta). Frank Thomas Burbrink Louisiana State University and Agricultural & Mechanical College

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. SYSTEMATTCS OF THE POLYMORPHIC NORTH AMERICAN RAT SNAKE 0ELAPHE OBSOLETA )

A Dissertation

Submitted to the Graduate Faculty of the Louisiana State University and Agricultural and Mechanical College in partial fulfillment of the requirement for the degree of Doctor of Philosophy

The Department of Biological Sciences

by Frank T. Burbrink B.S., University of Illinois, 1993 M.S., University of Illinois, 1995 May 2000

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Acknowledgments

I would like to thank my parents, Charolet and Glen Burbrink, and my gal,

Angela Cinquemano, for all of their support. I owe a debt of gratitude to my committee

members: J. McGuire, M, Fitzsimons, M. Hafner, D. Foltz, F. Sheldon and D. Rossman.

J. Slowinski and R. Lawson must be acknowledged for help their kind with Chapter

one. K. Naoki allowed me use of his computer during his absence in the summer of

1999. I am very grateful to the following persons who provided help in obtaining

samples used in this study: P. Moler, A. Meier, A. Meier, Jr., G. Burbrink, C. Burbrink,

K. Krysko, A. Smythe, A. Bass, G. Clark, C. Arm ant, G. Watkins, A. Cinquemano, D.

Rossman, S. Rossman, E. Censky, W. Gibbons, T. Tuberville, D. Wills, M. Braun, B.

Payst, J. Cole, P. Frank, J. Decker, L. Giodano, H. Dessauer, E. Liner, W. Shoop, S.

Cardiff, J. McLean, W. Sanderson, R. Vaeth, S. Secor, R. Axtel, C. Smith, H. Dowling,

R. Knight, G. Schafer, C. Guyer, D. Hartman, A. Wilson, R. Brandon, F. Scott, M.

Sabaj, M. Waters, S. Trauth, J. Babin, D. Dittmann, V. Remsen, F. Sheldon, C. Phillips,

J. Petzing, M. Dreslik, C. Sheil, J. Collins, T. Taggart, J. Boundy, T. Majors and J.

Demastes. A special debt o f gratitude is extended to C. Elliger and E. Visser for help

with laboratory procedures and sequence alignment. I am grateful for financial

assistance from the California Academy of Sciences In-House Research Fund, the

Theodore Roosevelt Memorial Fund, Ernst Mayr Award, and Charles Stems Grants-in-

Aid. I would also like to express my gratitude to M. Hafner, J. McGuire, and D. Reed

for reviewing various manuscripts related to this study.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Finally, I wish to thank the following individuals and institutions for loaning

numerous specimens required for the completion of this study:

AMNH: American Museum of Natural History: New York: Linda Ford, Darrel Frost

ANSP: Academy of Natural Sciences, Philadelphia: Ted Daeschler, Ned Gilmore

ASUMZ: Arkansas State University, Fayetteville: Stanley Trauth

AUM: Auburn University Museum, Auburn: Craig Guyer, Robert Reed

BCB: Bryce C. Brown Collection, Baylor University, Waco: Frederich Gehlbach, David

Lintz

CAS: California Academy of Sciences, San Francisco: Jens Vindum, Joseph Slowinski

CR: Charleston Museum, Charleston: Albert Sanders

CM: Carnegie Museum of Natural History, Pittsburgh: Ellen Censky, John Wiens,

Steve Rogers

FMNH: Field Museum of Natural History, Chicago: Alan Resetar, Harold Voris

INHS: Illinois Natural History Survey, Champaign: Chris Phillips, John Petzing

KU: University of Kansas Museum of Natural History, Lawrence: William Duellman,

John Simmons, Chris Sheil

LSUMZ: Louisiana State University Museum of Natural Science, Baton Rouge:

Douglas Rossman, Frederick Sheldon, and Donna Dittman

MCZ: Museum of Comparative Zoology, Cambridge: John Cadle, Jose Rosado

MPM: Milwaukee Public Museum, Milwaukee: Robert Henderson, Gary Casper

NCSM: North Carolina State Museum, Raleigh: Alvin Braswell, Jeffrey Beane

OMNH: Oklahoma Museum of Natural History, Norman: Laurie Vitt

SM: Strecker Museum, Baylor University, Waco: Frederick Gehlbach, David Lintz

iii

permission of the copyright owner. Further reproduction prohibited without permission. SREL: Savannah River Ecology Laboratory, Aiken: Whit Gibbons, Tracy Tuberville

TCWC: Texas Cooperative Wildlife Collection, College Station: Lee Fitzgerald,

Kathryn Vaughan

TNHC: Texas Natural History Collection, Austin: David Cannatella

UF: Florida State Museum, Gainesville: David Auth, Kenney Krysko

UGAMNH: University of Georgia Museum of Natural History, Athens: Elizabeth

McGhee

UIMNH: University of Illinois Museum of Natural History, Urbana: Chris Phillips,

John Petzing

UNSM: University of Nebraska Science Museum, Lincoln: John Lynch

UTA: University of Texas, Arlington: Jonathon Campbell

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table of Contents

ACKNOWLEDGEMENTS...... ii

LIST OF TABLES...... vi

LIST OF FIGURES...... viii

ABSTRACT...... xii

INTRODUCTION...... 1

CHAPTER I MITOCHONDRIAL DNA PHYLOGEOGRAPHY OF THE POLYTYPIC NORTH AMERICAN RAT SNAKE {ELAPHE OBSOLETA): A CRITIQUE OF THE SUBSPECIES CONCEPT...... 3 Introduction ...... 3 Methods and Materials ...... 7 Results...... 11 Discussion ...... 29

CHAPTER 2 MULTIVARIATE STATISTICAL ANALYSIS OF MORPHOLOGICAL VARAITION IN THE ELAPHE OBSOLETA COMPLEX...... 34 Introduction ...... 34 Methods and Materials ...... 39 Results...... 57 Discussion ...... 124

SUMMARY AND CONCLUSION...... 163

LITERATURE CITED...... 165

APPENDIX A: TISSUE EXAMINED...... 173

APPENDIX B: OLIGONUCLEOTIDE PRIMERS USED FOR AMPLIFICATION AND SEQUENCING OF ELAPHE VULPINA, E. BAIRDI, AND E. OBSOLETA CYTOCHROME B AND CONTROL REGIONS...... 176

APPENDIX C: MORPHOLOGICAL SPECIMENS EXAMINED...... 178

VITA...... 187

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. List of Tables

Pace

2.1 List of morphological characters used in this study ...... 41

2.2 Results of ANOVAs performed on raw variables for males for the four clades of rat snakes ...... 58

2.3 Results of ANOVAs performed on raw variables for females for the four clades of rat snakes ...... 66

2.4 Canonical discriminant functions based on all Case 1 variables for Males...... 106

2.5 Canonical discriminant functions based on all Case 2 variables for males...... 107

2.6 Canonical discriminant functions based on all Case 3 variables for males...... 108

2.7 Canonical discriminant functions based on all Case 4 variables for males...... 109

2.8 Canonical discriminant functions based on all Case 1 variables for females...... 110

2.9 Canonical discriminant functions based on all Case 2 variables for females...... I l l

2.10 Canonical discriminant functions based on all Case 3 variables for females...... 112

2.11 Canonical discriminant functions based on all Case 4 variables for females...... 113

2.12 Canonical discriminant functions based on all Case I variables for combined male and female data ...... 114

2.13 Canonical discriminant functions based on all Case 2 variables for combined male and female data ...... 115

2.14 Canonical discriminant functions based on all Case 3 variables for combined male and female data ...... 116

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2.15 Canonical discriminant functions based on all Case 4 variables for combined male and female data ...... 117

2.16 Principal component loadings for all Case 2 variables for males ...... 118

2.17 Principal component loadings for all Case 2 variables for females ...... 119

2.18 Principal component loadings for all Case 2 variables for combined male and female data ...... 120

2.19 Discriminant factor classification matrix for males only ...... 121

2.20 Discriminant factor classification matrix for females only ...... 122

2.21 Discriminant factor classification matrix for combined male and female data...... 123

2.22 Discriminant factor classification matrix by subspecies for males only ...... 125

2.23 Discriminant factor classification matrix by subspecies for females only ...... 126

2.24 Discriminant factor classification matrix by subspecies for combined male and female data ...... 127

2.25 Mean and standard deviation of mensural ratio measurements for males...... 128

2.26 Mean and standard deviation of mensural ratio measurements for females...... 133

vii

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. List of Figures

Pace

1.1 Geographic distribution of all Elaphe obsoleta subspecies and Elaphe bairdi ...... 6

1.2 Localities of specimens sampled molecularly in this study ...... 8

1.3 Bootstrap, strict consensus tree for 18,479 shortest trees of length 424 steps...... 15

1.4 Map showing range extension of Elaphe obsoleta mitochondrial clades from southern refugia following glacial retreat ...... 17

1.5 Pair-wise genetic distance regressed against pair-wise geographic distance for all eastern clade samples against all eastern clade samples ...... 20

1.6 Pair-wise genetic distance regressed against pair-wise geographic distance for all eastern clade samples against all central clade samples ...... 21

1.7 Pair-wise genetic distance regressed against pair-wise geographic distance for all eastern clade samples against all western clade samples...... 22

1.8 Pair-wise genetic distance regressed against pair-wise geographic distance for all central clade samples against all central clade samples ...... 23

1.9 Pair-wise genetic distance regressed against pair-wise geographic distance for all central clade samples against all western clade samples...... 24

1.10 Pair-wise genetic distance regressed against pair-wise geographic distance for all western clade samples against all western clade samples...... 25

1.11 Pair-wise genetic distance regressed against pair-wise geographic distance for all eastern clade samples against all E. bairdi clade samples...... 26

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 1.12 Pair-wise genetic distance regressed against pair-wise geographic distance for all central clade samples against all E. bairdi clade samples...... 27

1.13 Pair-wise genetic distance regressed against pair-wise geographic distance for all western clade samples against all E. bairdi clade samples...... 28

2.1 Map of the US showing the general location of specimens examined in this study...... 40

2.2 Illustration of selected characters measured on the head and dorsal head scales Elaphe obsoleta and E. bairdi ...... 49

2.3 Illustration of selected characters measured on the lateral and ventral head scales of Elaphe obsoleta and E. bairdi ...... 50

2.4 Illustration of selected color pattern and body scale measurements made on Elaphe obsoleta and E. bairdi ...... 51

2.5 Illustration of dorsal body stripe measurements made on Elaphe obsoleta and E. bairdi ...... 52

2.6 Graph of HL/SV against S V for all samples of Elaphe obsoleta...... 54

2.7 Three dimensional plot of the first three discriminant function scores based on all Case I variables for males ...... 75

2.8 Three dimensional plot of the first three discriminant function scores based on all Case 2 variables for males ...... 76

2.9 Three dimensional plot of the first three discriminant function scores based on all Case 3 variables for males ...... 77

2.10 Three dimensional plot of the first three discriminant function scores based on all Case 4 variables for males ...... 78

2.11 Three dimensional plot of the first three discriminant function scores based on all Case 1 variables for females...... 79

2.12 Three dimensional plot of the first three discriminant function scores based on all Case 2 variables for females ...... 80

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2.13 Three dimensional plot of the first three discriminant function scores based on all Case 3 variables for females ...... 81

2.14 Three dimensional plot of the first three discriminant function scores based on all Case 4 variables for females ...... 82

2.15 Three dimensional plot of the first three discriminant function scores based on all Case 1 variables for combined male and female data ...... 83

2.16 Three dimensional plot of the first three discriminant function scores based on all Case 2 variables for combined male and female data ...... 84

2.17 Three dimensional plot of the first three discriminant function scores based on all Case 3 variables for combined male and female data ...... 85

2.18 Three dimensional plot of the first three discriminant function scores based on all Case 4 variables for combined male and female data ...... 86

2.19 Notched box plots of the first two discriminant functions based on all characters included in Case 1 for males ...... 87

2.20 Notched box plots of the first two discriminant functions based on all characters included in Case 2 for males ...... 88

2.21 Notched box plots of the first two discriminant functions based on all characters included in Case 3 for males...... 89

2.22 Notched box plots of the first two discriminant functions based on all characters included in Case 4 for males ...... 90

2.23 Notched box plots of the first two discriminant functions based on all characters included in Case 1 for females ...... 91

2.24 Notched box plots of the first two discriminant functions based on all characters included in Case 2 for females ...... 92

2.25 Notched box plots of the first two discriminant functions based on all characters included in Case 3 for females ...... 93

2.26 Notched box plots of the first two discriminant functions based on all characters included in Case 4 for females ...... 94

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 2.27 Notched box plots o f the first two discriminant functions based on all characters included in Case 1 for combined female and male data ...... 95

2.28 Notched box plots o f the first two discriminant functions based on all characters included in Case 2 for combined female and male data...... 96

2.29 Notched box plots o f the first two discriminant functions based on all characters included in Case 3 for combined female and male data ...... 97

2.30 Notched box plots of the first two discriminant functions based on all characters included in Case 4 for combined female and male data ...... 98

2.31 Three dimensional plot of the first three principal component scores based on all Case 2 variables for males only ...... 99

2.32 Three dimensional plot of the first three principal component scores based on all Case 2 variables for females only ...... 100

2.33 Three dimensional plot of the first three principal component scores based on all Case 2 variables for combined male and female data ...... 101

2.34 Notched box plots of the first three principal component scores based characters included in Case 2 for males ...... 102

2.35 Notched box plots of the first three principal component scores based characters included in Case 2 for females ...... 103

2.36 Notched box plots of the first three principal component scores based characters included in Case 2 for combined male and female data ...... 104

2.37 Geographic distribution of E. alleghaniensis ...... 145

2.38 Geographic distribution of E. spiloides...... 151

2.39 Geographic distribution of E. osboleta...... 156

2.40 Geographic distribution of E. bairdi ...... 160

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Abstract

The North American rat snake (Elaphe obsoleta) complex is composed o f seven

uniquely colored subspecies of E. obsoleta and the sister species E. bairdi. Maximum

parsimony (MP) and maximum likelihood (ML) analyses of two mitochondrial DNA

genes from 73 E. obsoleta and E. bairdi, produced well-supported clades that do not

conform to the currently accepted subspecies. Both ML and MP trees were

significantly shorter than trees in which each subspecies was constrained to be

monophyletic. The subspecies of E. obsoleta do not represent genetic lineages.

Instead, molecular evidence revealed the existence of four well-supported mtDNA

clades confined to particular geographic areas in North America. Lack of genetic

variability in each of the clades of E. obsoleta may indicate that the lineages expanded

northward from southern glacial refiigia recently.

Univariate and multivariate analysis of 67 morphological characters scored from

1006 specimens provided additional statistical support for the existence of the same four

evolutionary lineages identified in the molecular phylogeographic study. Specimens

could be classified using canonical discriminant function analysis into the four

molecular clades better than they could be partitioned into subspecific categories.

Moreover, the identification of these subspecies proved difficult when using the

traditional characters ascribed to them.

In light of the corroborating molecular and morphological evidence, it is

suggested that the recognition of the subspecies of Elaphe obsoleta be discontinued.

Instead, the four molecular clades should be recognized as four species: E.

xii

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. alleghaniensis, E. spiloides, E. obsoleta, and E. bairdi. This research demonstrates that

the recognition of subspecies from one or two characters may be detrimental to

understanding evolutionary history.

xiii

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Introduction

The North American rat snake (Elaphe obsoleta) is a large polymorphic colubrid

commonly found in the forested regions of the eastern United States. This species is

known to have eight colored subspecies. The subspecies may be unpatterned, blotched,

or striped and have black, grey, brown, yellow, or orange ground coloration. To the

casual observer, they might appear to be different species. Several subspecies cross

geographical barriers without any known interruption in color pattern. It is not clear if

these different races represent independent evolutionary lineages (i.e., species).

In Chapter One, two complete mitochondrial genes were sequenced from 73

individual specimens sampled throughout the range of this Elaphe obsoleta and it

putative sister taxon E. bairdi. Members of all seven subspecies o f E. obsoleta

represented in this molecular data set. These data were analyzed phylogeographically

by using maximum parsimony and maximum likelihood to determine if any of the color

patterns reflect independently evolving lineages. Patterns of sequence divergence

between individuals also were examined to see if these molecular data support a

hypothesis of recent range expansion following the retreat o f the last North American

glacier at the end of the Pleistocene.

Chapter Two focuses on morphology in light of molecular patterns discovered

for Elaphe obsoleta in Chapter One. Sixty-seven morphometric measurements were

obtained from 1006 preserved specimens o f Elaphe obsoleta. This morphological data

set was examined univariately by using ANOVA and raultivariately by using canonical

discriminant function analysis and principal components analysis. Classification

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. matrices based on discriminant function analyses scores were constructed to determine

how well the groups could be classified according to the molecular data set. Taxonomic

conclusions for this polymorphic group were based upon the independence of evolving

lineages as determined by concordance among molecular and morphological data sets.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Chapter I

Mitochondrial DNA Phylogeography of the Polytypic North American Rat Snake (Elaphe obsoleta)'. A Critique of the Subspecies Concept

INTRODUCTION

The validity of the subspecies rank has received criticism for over 50 years from

(Mayr, 1942; Wilson and Brown, 1953; Mayr, 1982; Cracraft, 1983; McKitrick and

Zink, 1988, and Frost et al., 1992; Frost and Kluge, 1994). Commenting on the use of

subspecies, Mayr (1982) stated that “ the subspecies was not a concept of evolutionary

biology but simply a handle of convenience for the clerical work of the museum

curator.” Moreover, subspecies that represent only arbitrary sections of a cline should

not be considered as evolutionarily distinct entities (Wilson and Brown, 1953; Mayr,

1982; Cracraft, 1983; Frost and Hillis, 1990). Alternatively, subspecies may be used to

designate isolated sublineages that may evolve into full species (incipient species;

Mayr, 1942). The suggestion that some subspecies may evolve into full species requires

unobtainable knowledge of the future. Furthermore, incipient species cannot be

differentiated from real species phylogenetically (Frost et al., 1992). Therefore,

susbspecies have no real taxonomic meaning if they are used to represent arbitrary

pattern classes or incipient species.

It is likely that there are species masquerading as subspecies and only further

studies can reveal their true status. That is, many subspecies may actually represent

independently evolving lineages. In fact, Frost et al. (1992) suggest that in the

curatorial operations of a museum, it is prudent to retain trinomials for taxa with

diagnosable characters having para pa trie or allopatric ranges in case future work may

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. find them to be distinct species. In this paper we examine the phylogeography of a

textbook example of a polytypic species with numerous subspecies: the North American

rat snake ( Elaphe obsoleta) (Futuyma, 1998).

The North American Rat Snake is a common reptile in most forested areas of

the central and eastern United States. Over the past 176 years, the following eight

subspecies of Elaphe obsoleta have been described based on adult color pattern; E. o.

obsoleta. E. o. quadrivittata, E. o. lindheimeri, E. o. spiloides, E. o. bairdi, E. o.

deckerti, E. o. williamsi, and E o. rossalleni. Another distinct form, E. o. parallela,

was described by Barbour and Engels (1942), but has since been considered an

intergrade between E. o. quadrivittata and E. o. obsoleta (Neill, 1949).

The subspecies are defined by the following key characters: Elaphe obsoleta

obsoleta (Say, 1823) has a dark brown or black dorsum with little evidence of any

pattern (Wright and Wright, 1957; Conant and Collins, 1991); E. o. quadrivittata

(Holbrook, 1842) has four dark dorsal stripes on a ground color of yellow or tan in

Florida to gray in North Carolina (Conant and Collins, 1991; Schultz, 1996); E. o.

lindheimeri (Baird and Girard, 1853) has 25-35 large dorsal blotches on a brown,

yellow, or orange ground color. This subspecies is subject to considerable variation in

color pattern (Conant and Collins, 1991; Schultz, 1996); E. o. spiloides (Dumeril,

Bibron, and Dumeril, 1854) resembles E. o. lindheimeri. They both retain the blotches

evident in the juvenile color pattern in all of E. obsoleta. The distinguishing character

is that the dorsal ground color in E. o. spiloides is gray or grayish-white rather than the

brown, yellow, or orange as in E. o. lindheim eri (Schultz, 1996);E. o. deckerti (Brady,

1932) has four brown longitudinal stripes on an orange, yellow, or tan ground color,

while still retaining the dorsal blotches found in the juveniles (Neil, 1949; Wright and

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Wright, 1957); E. o. williamsi (Barbour and Carr, 1940) exhibits both blotches and

stripes on a ground color o f white or gray (Wright and Wright, 1957; Schultz 1996); E.

o. rossalleni (Neil, 1949) has an orange, orange-yellow, or orange-brown ground color

with four poorly defined longitudinal stripes (Wright and Wright, 1957). E„ bairdi

(Yarrow, 1880 in Cope, 1888) often has four poorly defined longitudinal stripes and

numerous dorsal blotches on a ground color of brown to grey with a wash o f yellow or

orange at the edge of each scale (Conant and Collins, 1991). This subspecies was

elevated to species status by Olson (1977); Ranges of these subspecies are illustrated in

Fig. 1.1.

Intergrades are assumed to occur where the ranges of these subspecies meet

(Neill 1949; Schultz, 1996). However, there are no intergrades between Elaphe

obsoleta lindheimeri and E. bairdi (Lawson and Lieb, 1990). Neill (1949) and Dowling

(1951) examined the range of color variation in E. obsoleta. Each relied on color

pattern to describe distinct geographic subspecies. Neill (1949), with no mention of

actual specimens examined, believed that E. o. obsoleta, E. o. spliloides (referred to in

his paper asE. o. confmis), E. o. williamsi, E. o quadrivittata, E. o. rossalleni, E. o.

deckerti , and E. o. lindheimeri were all valid subspecies. However, Dowling (1951,

1952) recognized only four subspecies: E. o. obsoleta, E. o. spiloides, E. o.

quadrivittata, and E. o. bairdi. Elaphe o. rossalleni, E. o. deckerti, and E. o. williamsi

were placed in synonomy with E. o. quadrivittata. Elaphe bairdi was considered a

subspecies o f E. obsoleta because it was assumed that it intergrades with E. o.

lindheimeri. Schwartz (1996) reviewed the taxonomic history of the group.

To examine the relationships and phylogeography o f Elaphe obsoleta, I

sequenced the complete cytochrome b and control region 1 mitochondrial genes from

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.

E. E. o. obsoleta E, E, o. deckerti E. E. o. rossalleni E. E. o quadrivittata

a Elaphe bairdi. williamsi spiloides subspecies and lindheimeriE. bairdi lindheimeriE. Elaphe obsoleta X - Fig. Fig. 1.1. Geographic distribution ofall

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. specimens of E. obsoleta and E. bairdi sampled from throughout its range (Fig. 1.2 and

Appendix A). I documented patterns of variation in the nucleotide sequences of these

two genes within and among subspecies and reconstructed the phytogeny of these

mtDNA genes. I used this gene tree to assess whether the currently recognized

subspecies form natural groups. I maintain that if these eight subspecies form

independently evolving lineages (species), then their distinct histories should be

apparent in the mtDNA phylogeny, barring incomplete lineage sorting. In addition, I

present a hypothesis that we believe best accounts for the current geographical

distribution of the sampled mtDNA haplotypes within E. obsoleta and E. bairdi.

MATERIALS AND METHODS

Sequencing

As a source of total genomic DNA we used shed skin, liver tissue, or, in a few

instances, blood drawn directly from the caudal vein of living snakes. Tissues were

digested for three to four hours at 65 C with constant motion in 2ml of lysis buffer

(Tris HCl lOOmM at pH 8.0, EDTA 50mM at pH 8.0, NaCl lOmM, SDS 0.5%)

containing 60ug of proteinase K per ml. Digestion was followed by extraction twice

with phenol/CHCl3 at pH 7.3 and once with CHCI 3 . DNA was precipitated from the

aqueous layer with 2.5 volumes of pure ethanol and the usually spooled DNA was then

washed in 80% ethanol, dried and redisolved in TE buffer (Tris lOmM, EDTA 1 mM,

pH 8.0). Template DNA for the polymerase chain reaction (PCR) was prepared by

diluting stock DNA with TE buffer to give spectrophotometric readings at A260

between 0.2A and 0.7A. Mitochondrial DNA was amplified from template DNA in

lOOul reactions using a hot start method in a thermal cycler with a 7 minute denaturing

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Fig. 1,2. Localities of specimens sampled genetically in this study. Numbers point to specific localities listed in Appendix A. step at 94° C followed by 40 cycles of denaturing for 40 sec at 94 C, primer annealing

for 30 sec at 46° C and elongation for one min at 72 C with a final 7 min elongation

step at 72° C. PCR products were purified using the Promega Wizard® PCR Preps

DNA Purification System according to manufacturers instructions.

Cycle sequencing was performed on the PCR products by using the Perkin-

Elmer Big Dye® reaction premix for 50 cycles of 96 C, 10 sec, 45 C, 5 sec and 60 C

for four min. The nucleotide sequence was determined by using an ABI model 310

Genetic Analyzer. Oligonucleotide primers for amplification and sequencing were

taken from the literature or designed for this project and are listed in Appendix 2. The

entire cytochrome b gene was amplified by using primers L14910 and H16064. The

sequence of single stranded DNA was obtained using primers LI 5324, LI 5584 and

HI 5399 for cycle sequencing. If ambiguous sites were found, both strands were

sequenced by using the primers L14919 and H16064 in addition to those listed above.

The entire control region was obtained by first amplifying with LI6090 and H690

followed by cycle sequencing with all four control region primers (Appendix B). The

nucleotide sequence of gene segments was aligned by using the program Xesee©

version 3.2 (Cabot, 1998). The nucleotide sequence of the control region of the colubrid

snake Dinodon semicarinatus (Kumazawa et al., 1998) was used as an aid in alignment

of the Elaphe control region sequence segments. AH sequences for both the

cytochrome b and control region genes will be deposited in Genbank.

Phylogenetic Analyses

The aligned sequences resulted in 2151 characters (1117 cytochrome b positions

and 1034 control region positions). The 70 Elaphe obsoleta, two E. bairdi and one E.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. vulpm a sequences were analyzed by using maximum parsimony and maximum

likelihood optimality criteria implemented with PAUP* 4.0 (SwofFord, 1999). The

maximum parsimony (MP) analysis was performed with all sites weighted equally. For

MP, 100 heuristic searches were performed with the starting trees obtained by random

stepwise addition, followed by TBR branch-swapping. The reliability of clades on the

shortest trees was assessed by using nonparametric bootstrapping (Felsenstein, 1985)

performed with 100 replicates, each executed as a heuristic search as above. Skewness

(gl; Hi 11 is, 1991) was estimated from a random sample of 10,000 trees generated by

PAUP* 4.0. Elaphe vuipina was used to root the tree, as it appears to be the sister

taxon to the E. obsoleta -E. bairdi clade (Robsin Lawson, unpubl. data).

For the maximum likelihood (ML) searches, it was necessary to determine an

appropriate model of sequence evolution (Swofford et al., 1996). This was

accomplished by using likelihood-ratio tests (Huelsenbeck and Rannala, 1997) to

compare the following nested and successively more parameter-rich models using

PAUP 4.0* (Swofford, et al., 1996): the F81 (Felsenstein, 1981), HKY85 (Hasegawa et

al., 1985), and the GTR (General time-reversible) (Lanave et al., 1984; Tavare, 1986;

Rodriguez et al., 1990) models. Because the three codon sites in protein-coding genes

experience different substitution rates, rate heterogeneity was assumed and rate

variation was modeled as a discrete gamma distribution (Yang, 1993, 1996) with four

rate categories. The three models were tested by using the MP tree. The gamma

HKY85 model with a -In value of 5688.85 was shown to perform significantly better

than the gamma F81 model with a-ln value of 5937.68 (X^=497.65, df=l, P«0.001).

The gamma GTR model with a -In value of 5685.07 did not perform significantly better

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. than the HKY85 (X^=5.56, df=4, P=0.4). Thus, the gamma HKY85 is considered the

appropriate model for these data.

Different tree topologies were examined statistically using Templeton

(Templeton, 1983), winning sites (Prager and Wilson, 1988), and Kishino-Hasegawa

(Kishino and Hasegawa, 1989) tests. With these tests it will be possible to determine if

different tree topologies produced by MP or ML are significantly different. In addition,

trees that constrain subspecies into monophyletic groups can be statistically compared

to the shortest MP and ML trees without topological constraints.

RESULTS

Sequence Characteristics

Cytochrome b

Because the amplification primers are located within highly conserved transfer

RNA genes which flank the cytochrome b gene and because our sequences contain an

open reading frame without stop codons, it may be inferred that the sequences represent

the functional cytochrome b gene rather than nuclear pseudogenes. Additionally, the

translated amino acid sequence is sufficiently similar to that of other snakes to support

this contention.

Characteristics of the complete snake cytochrome b nucleotide sequences were

discussed by Campbell (1997) and Slowinski and Keogh (1999). In the rat snakes, as in

other advanced snakes, the cytochrome b sequence commences with the start codon

ATG coding for the amino acid methionine. In the Elaphe examined in this study, the

cytochrome b gene is always 1116 bp long (372 amino acids). Instead of a stop codon,

the gene ends with a thymine which is post-transcriptionally polyadenylated to form a

functional stop codon (UAA), a common mechanism in snakes (Campbell, 1997;

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Kumazawa et al., 1998; Slowinski and Keogh, in press). Additionally, as in other

advanced snakes, 10 amino acid positions are deleted relative to the cytochrome b

molecule of most other vertebrates; nine codon deletions occur near the 5' end and one

near the 3' end of the gene (Campbell, 1997; Slowinski and Keogh, 1999). These

deletions are in the matrix domain of the cytochrome b molecule (Degli Esposti, 1993),

which because of functional differences between domains has been shown to be the

least conserved of the three domains comprising the molecule (Griffiths, 1997). Of the

372 amino acids coded in the snake cytochrome b gene there are two fixed differences

between the ingroup and outgroup taxa and 12 positions that show variability among the

ingroup taxa.

For the cytochrome b gene, nucleotide compositional bias exists mainly among

the three codon sites, rather than as bias among the sampled sequences (all cytochrome

b sites: = 14.35, 216 df, P = 1.00). The frequencies o f A, C, G, and T at the first

position were: 0.350, 0.230,0.178, and 0.241, respectively. For the second positions,

they were: 0.198, 0.280, 0.111, and 0.410. For the third positions, they were: 0.451,

0.269, 0.036, and 0.245. This pattern of bias, with A’s dominating at first sites, T’s at

second sites, and A’s at third sites, is similar to the pattern found by Campbell (1997)

and Slowinski and Keogh (1999) in other snakes and is generally similar to the pattern

in other vertebrates (e.g., mammals: Irwin et al., 1991).

Control Region

There are few descriptions of the control region in snakes. Here we compare the

control region sequence for the rat snakes used in this study with that published for

Dinodon semicarinatus, an Asian colubrine snake (Kumazawa e t al., 1998). The

control region in vertebrates is composed of three domains, a 5r domain that contains

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. one or more termination-associated sequences (TAS), a central conserved domain and a

3' domain that contains the site of initiation for heavy-strand replication, as well as two

to three conserved sequence blocks (CSB) (Brown et al., 1986; Saccone et al., 1991;

Taberlet, 1996). The displacement loop is part of the control region from the

termination associated sequences and spanning the site of initiation of heavy-strand

replication (Clayton, 1982, 1991; Taberlet, 1996). In vertebrates, sites o f increased

variability vary among species, but generally are found in the 5’ and 3’ domains where

insertions, deletions and base substitutions occur more commonly (Taberlet, 1996).

In Dinodon semicarinatus, the entire control region has been duplicated, with

the duplicate sequence inserted within the IQM tRNA gene cluster (Kumazawa et al'.,

1996, 1998). As this duplication has been found in coiubrids, boids and viperids, but not

in Leptotyphlops, its presence in all advanced snakes seems probable (Kumazawa et al.,

1996). These two control regions have been designated control region 1 for the original,

located between tRNA-proiine and tRNA-phenylalanine, and control region 2 for the

duplicate (Kumazawa et al., 1998). To ensure that only control region 1 was amplified,

primers located in tRNA-Thr and a highly conserved segment of 12S ribosomal DNA

flanking this region were used.

The control region 1 of the three species of Elaphe sampled show a high level of

sequence similarity with that ofDinodon. Near the S' end of the left domain is a region

known as the C- rich region, which in Dinodon consists of nine C’s followed by TA and

then an additional nine C’s. In all three Elaphe species sampled, the C-region is similar

to that of Dinodon, with nine Cs in the 5' block but only eight in the 3' block. Unique to

theElaphe vulpina examined was a sequence of ten nucleotides inserted within the 5' C

block. Of three left domain and one right domain hairpin-like secondary structures in

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. theDinodon control region, hairpins one and two are nearly identical in Elaphe, hairpin

three is identical and hairpin four is variously identical or nearly so among different E.

obsoleta specimens, E. bairdi and the single E. vulpina. Likewise, TAS 1 and TAS 2 are

identical between Dinodon and the three Elaphe species. CSB 1,2 and 3 are very

similar in sequence between Dinodon and Elaphe, but do show some variability among

E. obsoleta individuals. Finally, a singly repeated 20 bp segment and a doubly repeated

49 bp segment found in Dinodon also occur in the three species of Elaphe, where they

have a high degree of sequence similarity to Dinodon.

In Dinodon, there are six nucleotide sites that are absent from E. vulpina and

eight that are absent from E. obsoleta and E. bairdi. In E. vulpina, there are, in addition

to the ten nucleotide segment inserted within the C-rich region mentioned above, four

nucleotide sites not found in Dinodon. Among E. bairdi and E. obsoleta individuals,

there are two to six sites absent from Dinodon. With the present data it is not possible to

tell whether these indels are insertions or deletions.

TheDinodon control region is 1018 bp (Kumazawa et al., 1996), E. vulpina is

1032 bp and E. obsoleta is variably between 1020 and 1022 bp.

Phylogenetic Analyses

The parsimony search found 18,479 shortest trees of length 424 steps

(uninformative sites excluded; Cl = 0.644, R1 = 0.950). The gl statistic for these data

was -0.607, indicating highly structured data. Results of the non parametric bootstrap

analysis are spresented in Fig. 1.3. Very high support for three major mtDNA clades of

Elaphe obsoleta was obtained from the bootstrap analysis: 1) an eastern clade

comprised of rat snakes east of the Apalachicola River and the

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Co. n (i.o) Co.. SC (10. Q) ChadiamCo. GA (12. Q) {. Waha Co.. NC (9.0) Panpamans Co.. NC (8.0 ) IM taaCo.CT1l2.OI I M t a B Co.. CT 2 (3.0) Levy C o . H. 1 (22. W) Lavy Co.. FL (23. W) Taylor Co.. FL (24. S) Ltarty Co.. FL (28. S) Co.. FL (19. Q) Co.. FL (20.0) Co.. R. (15. R) Co.FL(16.R> Monro* Co.. FL (17. D) M om * Co.. FL (18.0) Puhiam Co.. FL (13.0) I lamandp C o. FL (21. Q) Alachua Co.. FL (14.0) Fredrick Co.. MD (5.0) Rodongham Co. VA (7.0) Orangt C o. NY (4.0) JaaparC o. S C (ll.Q ) Loudon C o. VA (6.0 ) Fonaat Co.. MS (30. L) BaMari Co.. AL (29. S) Hinds C o. MS (31. L) Otlaana P ar. LA (32. L) E. Btaon Rouga Par. LA 1 (40. L) E. Faiaana P ar. LA (30. L) C rd g ia ri C o. AR (58. L) * Dacriur C o. TN (64. L) GaM xi Co.. IL (62. L) Johnaon Co. t. (63. L) I | SL Tammany Par. LA 1 (33. L) a. Tamreny Par. LA 2 (34. L) Tan g p aho* Par.. LA 2. (35. L) Co.. FL (28. S) Co.. FL (27. S) Wafcuka C o. FL (25. S) La* C o . AL (68. L) Dataware Co.. OH (70.0) PAts C o . L (60.0) Madaon Co.. AL (88.0) Wood C o. WV (72.0) SL CtaraCo.. L (61. L) TaBadaga Co.. AL (67. L) Knox C o. TN (88.0) Grundy C o. TN (65.0) Stark C o. OH (71.0)

E. Baton Rouga Par. LA 2 (41. L) ' Tanppahoa Par. LA l 0 6 . L) * SL LanOy P ar. LA (43. L) E vaigaria Par. LA (44. L) MatrhAomai P ar. LA (46. L) 2* Tanahotma P ar. LA 1 0 7 . L) b a n ria P ar. LA (42. L) Camaron Par. LA (45. L) Tanabonna P ar. LA 2 (38. L) Marina Co.. TX (48. L) Corrta C o. TX (47. L) Karr Co.. TX (48. L) Palo Pink) C o. TX (52. L) G avy C o. KS (56.0) Graana Co. MO (56.0) Surmar Co.. KS (54. L) Ctamland C o. OK (53. L) Garland Co.. AR (58.0) Madaon Co.. AR (57.0) JaHaraon Oavs Co. TX 1 (50. B) JaRanon Davis Co.. TX 2 (51. B) Elaphe vulpsia

Fig. 1.3. Bootstrap, strict consensus tree for 18,479 shortest trees of length 424 steps. Numbers in parentheses correspond to listed localities in Fig. 1.2. and Appendix A. Letters in parentheses correspond to subspecies of Elaphe obsoleta, where: O=obsoleta, Q=quadrivittata, D=deckerti, S=spiloides, R=rossalleni, W=williamsi, L=lindheimeri, and B =Elaphe bairdi. Specimens found outside of the geographic area represented by their clade are marked with an*.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appalachian Mountains (98% bootstrap support), 2) a central clade located west of the

Apalachicola River and the Appalachian Mountains and east of the Mississippi River

(99% bootstrap support), and 3) a clade west of the Mississippi River (79% bootstrap

support). A monophyletic Elaphe bairdi mtDNA clade (100% bootstrap support) is

placed as the sister taxon to the western clade. The geographic areas occupied by these

groups are illustrated in Fig. 1.4.

The ML analysis provided a tree with -InL of 5676.90. This value was

compared to the MP tree’s -InL value o f5679.52 using the Kishino-Hasegawa test,

which did not show a significant difference (p=0.375). The parsimony tree length for

the ML topology was 428 steps (CI=0.638 and RI=0.949), which was not significantly

different from the length of the MP tree according to the Templeton (P=0.157),

winning sites (P=0.375), and Kishino-Hasegawa (P=0.158) tests.

To test the hypothesis that the subspecies form natural groups, a parsimony

analysis was undertaken in which subspecies were constrained to be monophyletic.

This generated 10,811 equally parsimonious unconstrained trees o f695 steps (CI=0.393

and RI=0.861). This tree is 271 steps greater than the most parsimonious trees (424),

which is significant according to Kishino-Hasegawa (p<0.0001; Kishino and Hasegawa,

1989), Templeton (PO.OOOl; Templeton, 1983), and winningsites (p<0.001; Prager,

and Wilson, 1988). Constraining subspecies monophyly also produced a ML tree with a

significantly lower likelihood score than the unconstrained ML estinmate to the

Kishino-Hasegawa test (diff-InL 1083.51, P < 0.0001).

Haplotypes and pairwise genetic comparisons

In the 72 ingroup specimens examined in this study, a total of 59 distinct

haplotypes were discovered. The cytochrome b gene alone produced 56 distinct

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.

Elapkeobsoleta Western Haplotypes r t * Central Haplotypes A Haplotypes Eastern V Mississippi River Apalachicola River Fig. Fig. Map showing 1.4. northern dispersal patternsof Elaphe obsoleta mitochondrial clades from southern refugia following glacial retreat.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. haplotypes for 72 ingroup specimens, whereas the control region yielded 26 distinct

haplotypes for 68 ingroup specimens. Members of the following groups of taxa share

the same haplotypes: a) Johnson Co., IL-Gallatin Co., IL-Craighead Co., AR-Decatur

Co., TN; b) Delaware Co., OH-Stark Co., OH-Pike Co., IL; c) Madison Co., AR-

Garland Co., AR; d) Orleans Par., LA-East Baton Rouge Par., LA 2; e) Pinellas Co., FL-

Broward, Co., FL; f) Monroe Co. FL 1- Monroe Co., FL 2. Both the control region and

the cytochrome b gene were sequenced in all 73 specimens except for the following:

Sumner Co., KS; Greene Co., MO; Cleveland Co., OK; and St. Landry Par., LA. Only

the control region sequences were determined for those snakes. These four specimens

shared the same control region haplotypes with several other specimens found within

the western clade: a) Sumner Co., KS-Greene Co., MO-CIeveland Co., OK-Geary Co.

KS; b) Cameron Co., LA-St. Landry Par., LA; c) Natchitoches Par., LA-St Landry Par.,

LA; d) Terrebone Par., LA I-St. Landry Par., LA; e) Terre bone Par. LA 2-St. Landry

Par., LA.

To compare genetic distance and geographic distance, all pair-wise genetic

distances of individuals with both complete gene sequences corrected for multiple

substitutions by the gamma HKY85 model with invariable sites estimated from the data

were regressed against estimated pair-wise geographic distance between collecting

sites. Straight-line distances between pairs of samples were measured on a map in

order to obtain the geographic distances used in this study. Pair-wise geographic and

genetic distances are categorized as intra-clade comparisons if all of the comparisons

were made between members found only within the eastern clade, central clade, or

western clade. They are categorized as inter-clade comparisons if all the comparisons

were made between individuals from different clades (i.e, eastern and central clades,

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. eastern and western clades, central and western clades, eastern and E. bairdi clades,

central and E. bairdi clades, and western and E. bairdi clades). These inter-and intra-

clade comparisons are shown in Fig. 1.5-1.13. All o f the inter-clade and intra-clade

comparisons produced regressions that were inconsistent with isolation by distance (a

2 slope close to zero with low r values; no points on the regression are non-

independent). The lowest inter-clade pairwise genetic distance values (eastern clade x

central clade = 0.016 substitutions/site; eastern clade x western clade=0.086

substitutions/site; central clade x western clade = 0.090 substitutions/site) are greater

than all intra-clade genetic distance values (eastern clade x eastern clade = 0.014

substitutions/site, central clade x central clade = 0.0085 substitutions/site; western clade

x western clade = 0.012 substitutions/site). This can be used as evidence to support the

notion that these mtDNA clades represent independent evolutionary lineages and that

members within a clade are more closely related than they are to members between

clades (Avise, 1994).

Because the inter-clade genetic distances are much greater than the intra-clade

distances, a correlation between genetic distance and geographic distance between

members of different clades is not expected. However, it is surprising that the intra-

clade comparisons failed to demonstrate that pair-wise genetic distance increases with

pair-wise geographic distance. This may indicate that the populations within each clade

have recently expanded outward from a smaller area and there has not been enough

time for these genes to have diverged significantly. Although less likely, the results

could also suggest that each of the three major clades represents a panmictic population

(see discussion).

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. I ______■ I ■ ui{ilL-u rl=0.044 iAuVQ v i . V f i , 1 / > , »lX , r t f f n y Pair-Wise GeographicPair-Wise Distance (km) ■ > > ■ > > 500 1000 1500 2000 u.i—4 urri‘2JAs.u.)LyJu.i—4 0 .0 0 0 0.052 0.104 0.078 0.130 Fig. I .5. .5. Pair-wiseFig.I genetic distance regressed against pair-wisesamples againstgeographic all other distanceeastern cladefor all eastern samples. clade != != 0.026 X (0 § 8 iO C o "S Q_ D 0 N> o

Reproduced with permission of the copyright owher. Further reproduction prohibited without permission r2=0.139 Pair-Wise Geographic Distance (km) 0 500 1000 1500 2000 o.ood 0.020 0.052r 0.104 0.130 Fig. t .6. .6. Pair-wiset Fig. genetic distance regressed against pair-wise geographic distance forall eastern clade samples against all central clade samples.

(0 ±

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. r RJ=0.0I8 ------0 0 1000 2000 3000 Pair-Wise Pair-Wise Geographic Distance (km) 0 .0 0 0 0.130 0.026 ro (O a> 0 1 0.052 «2 § 0.104 J % O Q Q 0.078 Fig. 1.7. Pair-wise 1.7. Fig. genetic distance regressed against pair-wise geographic distance for al! eastern clade samples against western clade samples.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 1500

______1000 Li

r2=0.085 iwmm& 500 Pair-Wise Geographic Distance (km)

- ^'Wwalii - 0 000 052 078 .

. 0 0.02C- 0 0 0.104 0.13a Fig. 1.8. 1.8. Fig. Pair-wise genetic distance regressed against pair-wise geographic distance for all central clade samples against all central clade samples. (0 c U»

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 500 1000 1500 2000 Pair-Wise Geographic Distance (km) 0 .0 0 0 0.052- 0.078- 0 .1 3Q 0 0 CO c 0 c CO o 0 o OL 3 % O o Fig. 1.9. Fig. 1.9. Pair-wise genetic distance regressed against samplespair-wise against geographic all western distance clade samples.forall central clade N> ■fc.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.

i l_l i ___ 1 ii 1 r2=0.622 l l j Pair-Wise Geographic Distance (km) ° - n 200 400 600 800 10001200 0.130 0.078 0.052 0.104 (/) (D 0) o 0> c Q. D e> Fig. 1.10. 1.10. Fig. Pair-wise genetic distance regressed against pair-wise geographic distance for all western clade samples against all western clade samples. N>

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 2500 3000 2000 samples. 1500 Pair-Wise Geographic Distance (km) 0 .000 0.026 0.052 0.130 0.104 0.078 E.bairdi CD CD a> (0 CD Q) 0) o c (0 o C Q. D O Fig. 1.11. Fig. 1.11. Pair-wise genetic distance regressed againstsamples pair-wise against geographic all distance for all eastern clade N) ON

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. I) I) I' l) r?=C».059 samples. Pair-Wise Geographic Distance (km) Distance Geographic Pair-Wise 00 1500 2000 2500 E.bairdi 0.026 0 . 0.078" 0.130r 0.104 11 11 (6 o

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 1500 1000 r2=(>.47() 500 samples. Pair-Wise Geographic Distance (km) Geographic Distance Pair-Wise 0

E.bairdi 0 .0 0 0 0.026 0.130 (D # 0.052 (/> c a> a> a> Q) O a 0.104 CL ~ 0.078 a Fig. 1.13. Fig. 1.13. Pair-wise genetic distance regressed against pair-wise geographic distance for all western clade samples against all

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. DISCUSSION

Evolution and Biogeography

Elaphe obsoleta is composed of three large mtDNA clades: 1) an eastern clade,

east of the Apalachicola River and Appalachian Mountains, 2) a central clade, west of

the Apalachicola River and the Appalachian Mountains and east of the Mississippi

River and 3) a western clade, west of the Mississippi River (Fig. 1.4). In addition, a

well-supported fourth clade comprised of E. bairdi appears as a sister group to the

clade west of the Mississippi River (Fig. 1.3). There is no relationship between intra-

clade pairwise genetic distance and intra-clade pair-wise geographic distance (Fig. 1.5-

1.13). In fact, some intra-clade sequences were identical despite being separated by

1,000 km. Although haplotypes within major clades are very similar, there is enough

intra-clade genetic variability to exclude the notion that each clade should be

considered a single panmictic unit. Additional arguments against panmixis are the

presence of genetically based color morphs (subspecies) (Bechtel and Bechtel, 1985)

confined to discrete geographic areas, as well as the low vagility of this species. A

better explanation of the low haplotype diversity in comparison with great geographic

distances is that it results from rapid range expansion from much smaller nuclei. Each

clade may have been composed of several populations with multiple haplotypes that

dispersed from southern refugia into northern areas as glaciers retreated 18,000 to 6,000

YBP (Huntley and Birks, 1983; Huntley and Webb, 1988, Huntley, 1990). (Fig. 4). This

scenario would account for the high haplotype diversity and the low genetic distance in

comparison with geographic distance.

The eastern and central clades are separated by the Apalachicola River. This

river is the site of morphological and molecular discontinuities within fish, amphibian,

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. and reptile species or clades (see reviews in Neill, 1957; Blainey, 1971; Swift et al.,

1985; Bermingham and A vise, 1986; Lawson, 1987; Avise, 1992; A vise 1996). At the

maximum rise in sea level during interglacial periods in the Pliocene and Pleistocene,

the Apalachicola River was embayed as a large saltwater channel well into Georgia and

Alabama, possibly up to the Fall Line in northern Georgia (Cooke, 1945; Neill, 1957;

Blainey, 1971). It was then that the Apalachicola River, in conjunction with the

Appalachian Mountains to the north, apparently served as a barrier separating the

eastern and central clades of E. obsoleta. However, the Wakulla Co., FL (26) sample is

anomalous, in that it is a member of the central clade found east of the Apalachicola

River. This may indicate that the Apalachicola River is no longer a barrier to east-west

dispersal of E. obsoleta or that limited amounts of dispersal have always occurred. It

has not yet been determined if the members of the eastern and central clade are

hybridizing.

The central clade is separated from the western clade by the Mississippi River, a

role the Mississippi has played in dividing the ranges of many species of fishes,

amphibians and reptiles in the central United States (Blair, 1958; Blair, 1965; Wiley

and Mayden, 1985; Mayden, 1988; Walker et al., 1998). The genetic distance between

the western and central/eastern clades is much larger than the genetic distance between

the eastern and central clades (Fig. 1.6, 1.7 and 1.9). Assuming the Mississippi River is

responsible for the initial divergence between the western and eastern/central clades, it

can be inferred from the phylogenetic estimate that it has served as a barrier to gene

flow for a longer period of time than has the Apalachicola River. Given the much

larger genetic distances, the Mississippi River may represent a more formidable barrier

to gene flow as well. During each interglacial period in the Pleistocene, the Mississippi

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. River experienced alluviation of valleys cut during the previous glacial stages. At that

time, the floodplain extended well beyond its current width (Thombury, 1965). This, in

combination with the cold waters of the melting glaciers, may have prevented dispersal

of E. obsoleta. However, two samples, East Baton Rouge Par., LA 2 (41) and

Tangipahoa Par., LA 1 (36) are members of the western clade found on the east side of

the Mississippi. In addition, one sample, Craighead Co. r Arkansas (59), belongs to the

central clade, but was found on the west side of the Mississippi River. This may

demonstrate that the Mississippi is no longer an adequate barrier to dispersal of E.

obsoleta.

The desert form, Elaphe bairdi, appears as a sister taxon to the western clade.

This species occupies xeric habitats and is not as dependent on mesic forest as is E.

obsoleta (Lawson and Lieb, 1990). Applying the Biological Species Concept, Olson

(1977) elevated this form to the level of species. Recognition of E. bairdi as a species

is consistent with the phylogenetic data presented here as it clearly represents an

independent lineage. However, it renders E. obsoleta paraphyletic given the

phylogenetic estimate presented here.

Taxonomy

The well-supported clades demonstrate that none of the currently accepted

subspecies represents a distinct evolutionary lineage (Fig. 1.3). As described in the

Introduction, many of these subspecies are diagnosed by color pattern. Apparently,

these color characters are labile and bear few clues to the phylogenetic history of

Elaphe obsoleta. In fact, if the primitive color pattern of E. obsoleta is blotched (this is

assumed because the outgroup, E. vulpina, is blotched as are all juvenile E. obsoleta)

then the northern black coloration has evolved independently several times. This dark

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. color pattern may have evolved multiple times in response to selection for enhanced

thermoregulatory capabilities in cooler northern environments (Braswell, 1977).

Although blotched rat snakes (E. o. lindheimeri and E. o. spiloides) show similar color

patterns on either side of the Mississippi or Apalachicola Rivers, the sharing of similar

color patterns does not appear to be an indication of close phylogenetic relationship.

This point is underscored by the fact that the oldest and most divergent split occurs in

the center of the range of E. o. lindheimeri. With respect to these mtDNA data, a

prohibitively large number of lineage sorting (deep coalescence) events would be

required to consider each subspecies to be an independent lineage. It appears that these

subspecies represent color pattern classes that are in disagreement with their

evolutionary history. In addition, preliminary factor analysis of morphological data

independent of color pattern confirms the notion that these subspecies are not well

differentiated when compared to the greater morphological differences found between

the clades identified here. Therefore, since the subspecies studied here do not conform

to the molecular-based phylogeny of this species, it is recommended that they be

eliminated from the taxonomy o f this group.

The taxonomy of a group should be consistent with its evolutionary history

(Wiley, 1981; Frost and Hillis, 1990). In light o f the mtDNA data presented here, it is

possible that the three geographically distinct clades of Elaphe obsoleta and the single

clade of E. bairdi represent four independent evolutionary lineages and thus constitute

distinct evolutionary species. Taxonomic recommendations will be deferred to Chapter

2 following evaluations of morphometric data brought to bear on this issue.

This study has demonstrated that the subspecies of E. obsoleta do not represent

distinct evolutionary lineages and underscores the danger of recognizing subspecies

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. based on few characters. It has been demonstrated that these poorly defined subspecies

actually mask the evolutionary history of the group. Therefore, describing or

recognizing subspecies from a few characters may not simply be a harmless handle of

convenience for museum curators, but may be detrimental to understanding

evolutionary history.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Chapter 2

Multivariate Statistical Analysis of Morphological Variation in theElaphe obsoleta Complex

INTRODUCTION

Using complete sequences of two mitochondrial genes, I demonstrated in Chapter 1 that

the subspecies of Elaphe obsoleta do not represent independently evolving lineages. It

was also shown that this species consists of three geographically distinct clades: 1) the

eastern clade, located east of the Apalachicola River and Appalachian Mountains, 2) the

central clade, located west of the Appalachian Mountains and the Apalachicola River

and east of the Mississippi River and 3) the western clade, located west of the

Mississippi River (Fig. 1.4). Elaphe bairdi, represents a fourth clade that is more

closely related to the western clade. This structure suggests that the current taxonomy,

which recognizes seven highly variable subspecies, may be in error with respect to the

evolutionary history of this group. Therefore, it may be prudent to recognize the four

molecular clades as four independently evolving lineages. In this chapter, the North

American rat snake complex is examined to determine if the morphology of this group

supports a taxonomic system that recognizes the four molecular clades or one that

supports the recognition of the seven subspecies.

Only a few studies have examined the morphology of Elaphe obsoleta.

Unfortunately, only superficial color pattern characters were used to support various

subspecific taxonomic names. In addition, most of these studies only examined the

North American rat snake in limited portions of their range and used very few

individuals and characters. All of the current subspecies are defined by the presence

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. or absence of blotches or stripes, or whether the ground color is black, gray, brown,

yellow or orange. Over the past 176 years, the following seven subspecies of Elaphe

obsoieta have been described based on those color patterns: E. o. obsoleta, E. o.

quadrivittata, E. o. lindheimeri, E o. spiloides,, E. o. deckerti, E. o. williamsi, and E. o.

rossalleni. Elaphe bairdi was considered a subspecies of E. obsoleta, but was elevated

to species status by Olson (1977). Another distinct form, E. o. parallela, was described

by Barbour and Engels (1942), but has since been considered an intergrade between E.

o. quadrivittata and E. o. obsoleta (Neill, 1949).

To understand what is currently known about the morphology and taxonomy of

E. obsoleta, brief descriptions of the key characters that are used to define these

subspecies are listed below:

Elaphe obsoleta obsoleta (Say, 1823) has a dark brown or black dorsum with

little evidence of any pattern (Wright and Wright, 1957; Conant and Collins, 1991). It

should be noted that many adult individuals within the range of this subspecies display

distinct dorsal blotches (Schultz, 1996). This poses a problem for the identification of

this taxa from other blotched subspecies (i.e. E. o. lindhiemeri and E. o. spiloides).

Elaphe obsoleta quadrivittata (Holbrook, 1836) has four dark dorsal stripes on a

ground color of yellow, or gray (Conant and Collins, 1991; Schultz, 1996). Many

specimens display blotches between the two most dorsal stripes (Schultz, 1996).

Elaphe obsoleta lindheimeri (Baird and Girard, 1853) has 25-35 large dorsal

blotches on a brown, yellow, or orange ground color. This subspecies shows

considerable variation in ground color (Conant and Collins, 1991; Schultz, 1996).

Distinguishing this subspecies from E. o. spiloides and E. o. obsoleta may be quite

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. difficult in areas where the distribution of the subspecies, as currently construed,

approach one another.

Elaphe obsoleta spiloides (Dumeril, Bibron, and Dumeril, 1854) resembles E. o.

lindheimeri , in that they both retain the juvenile blotched color pattern. The dorsal

ground color in E. o. spiloides tends to be gray or grayish-white, whereas that of E. o.

lindheimeri is brown, yellow, orange, or red. This subspecies may be very difficult to

distinguish from E. o. lindheimeri.

Elaphe obsoleta deckerti (Brady, 1932) has four brown longitudinal stripes on

an orange, yellow, or tan ground color, but is distinguished from E. o. quadrivittata in

that it retains the blotches found in juveniles (Neil, 1949; Wright and Wright, 1957).

Elaphe obsoleta williamsi (Barbour and Carr, 1940) appears as an intermediate

between E. o. spiloides and E. o. quadrivittata. The dorsum exhibits both blotches and

stripes on a ground color of white or gray (Wright and Wright, 1957; Schultz 1996).

Elaphe obsoleta rossalleni (Neil, 1949) is very similar to E. o. quadrivattata and

E. o. deckerti. The ground color may be orange, orange yellow, or orange brown. It

has four poorly defined longitudinal stripes. The presence of a red tongue in E. o.

rossalleni distinguishes it from the black tongued E. o. quadrivittata and E. o. deckerti

(Wright and Wright, 1957).

Elaphe bairdi (Yarrow, 1880 [in Cope, 1888]) was considered a subpecies of E.

obsoleta but was elevated to species status by Olson (1977). The tail in the species is

slightly longer than the tail in any subspecies of Elaphe obsoleta. It often has four

poorly defined longitudinal stripes and numerous dorsal blotches. The dorsal coloration

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. is generally brown or gray with a wash of yellow or orange at the edge o f each scale

(Conant and Collins, 1991). Superficially, E. bairdi it resembles E. o. quadrivittata.

All subspecies of Elaphe obsoleta display dorsal blotches as juveniles. As E. o.

obsoleta, E. o quadrivittata, E. o. rossalleni, E. o. deckerti, and E. bairdi develop, the

blotched pattern tends to become obscured. Adult E. o. spiloides and E. o. lindheimeri

display the juvenile blotch pattern. Adult Elaphe obsoleta williamsi retain the juvenile

blotches while expressing the adult stripes (Wright and Wright 1957; Schultz, 1996).

Ranges o f these subspecies are illustrated in Fig. 1.1. It has been assumed that

intergrades occur where the ranges of subspecies abut (Neill 1949; Schultz, 1996).

However, there are no intergrades between E. o. lindheimeri and E. o. bairdi (Lawson

and Lieb, 1990). Please note that in Fig. 1.1, the range o f E. o. spiloides and E. o.

lindheimeri has been altered from the maps in Conant and Collins (1991). Blotched

specimens with a gray ground color (E. o. spiloides) are found only in eastern Alabama

and southern Florida.

Only two short studies have attempted to examine the range of color pattern

variation in Elaphe obsoleta. Neill (1949), with no mention of actual specimens

examined, stated that E. o. obsoleta, E. o. spiloides (referred to in his paper as E. o.

confmis), E. o. williamsi, E. o quadrivittata. E. o. rossalleni, E. o deckerti, and E. o.

lindheimeri were all valid subspecies. No mention was made of E. bairdi. Based on

color pattern observations on less than 150 specimens, Dowling (1951, 1952)

considered only four subspecies: E. o. obsoleta, E. o. spiloides, E. o. quadrivittata, and

E. o. bairdi. Elaphe o. rossalleni, E. o. deckerti, and E. o. williamsi were placed in

synonomy with E. o. quadrivittata. Elaphe bairdi was considered a subspecies of E.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. obsoleta because Dowling (1951) assumed that this taxon intergrades with E. o.

lindheimeri.

Subsequently, Wright and Wright (1957) considered all seven subspecies valid

and placed them in their Handbook of Snakes. Conant (1975) also recognized all seven

subspecies, but omitted E. o. deckerti and E. o. williamsi in later editions of his field

guide (Conant and Collins, 1991).

Other studies have examined geographic variation of color patterns only in

limited portions of the range of Elaphe obsoleta. In their study of the herpetofauna of

southern Florida, Duellman and Schwartz (1958) considered E. o. rossalleni as a

subspecies distinct from E. o. quadrivittata based on color pattern. They regarded the

differences between the Florida Key populations of E. o deckerti and the mainland E. o.

quadrivittata to be trivial and thus did not recognize E. o. deckerti. Christman (1980),

when examining patterns of geographic variation in Florida snakes, found no characters

to support E. o. rossalleni, but did find the color pattern of E. o. deckerti to be distinct

from E. o. quadrivittata. He also maintained that E. o. spiloides was a valid subspecies

and that E. o. williamsi should be considered an intergrade between E. o. spiloides and

E. o. quadrivittata. The recognition of E. o. deckerti as an endemic form in the Florida

Keys was also supported by Paulson (1968). Olson (1977), considering color patterns

and scale counts, determined that intermediate forms between E. o. bairdi and E. o.

lindheimeri were rare hybrids. Based on this evidence, he suggested that E. bairdi be

recognized as a full species. Moreover, Parmley (1986) described differences in trunk

vertebrae between E. obsoleta and E. bairdi. Lawson and Lieb (1990) applied

allozyme, scale count, and color pattern data to demonstrate that there is only a narrow

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. zone of hybridization due to secondary contact between E. bairdi and E. o. lindheim eri,

thus supporting Olson’s claim that E. bairdi should be recognized as a distinct species.

This chapter represents the first attempt to examine a much broader range of

morphological variation in Elaphe obsoleta and E. bairdi based on 67 measurements o f

head and body scales and color pattern characters. The morphological data will be

examined to in light of the molecular phylogenetic advances that have been discovered

for this group to determine if concordance with the molecular clades exists.

MATERIAL AND METHODS

A total o f 1006 live and preserved specimens o f Elaphe obsoleta and Elaphe

bairdi was examined during this study (Appendix C). Samples were obtained

throughout the ranges of both species (Fig. 2.1). Specimens were classified into the

four molecular groups by the following criteria: 1) eastern clade if they were collected

east of the Apalachicola River and the Appalachian Mountains, 2) central clade if they

were collected west of the Apalachicola River and east of the Mississippi River, 3)

western clade if they were collected west of the Mississippi River, and 4) E. bairdi if

they were collected from southwest Texas and display characters representative of that

species. The subspecific identity of each specimen was also determined using the

traditional characters for each taxa described in the Introduction.

Descriptions of all meristic and mensural characters measured on each

specimen, and their abbreviations, are listed in Table 2.1. Characters that are discussed

in a long list are referred to by their number in Table 2.1. Many o f these characters are

illustrated in Fig. 2.2-2.S. Characters 3,4, 7, 8,9, and 10 were discarded prior to the

statistical analyses, because of lack of variation between specimens, clades or

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. I23

Eastern Clade (n=398)

44 1 8 .

Elaphe bairdi Central Clade (n= 32) 24 ' Western Clade (n=312) v (n=254) 17 10

Fig. 2.1. Map of the US showing the general location of specimens examined in this study .Specimens are grouped by clade. Values next to dark areas indicate the number of specimens examined from that locality. Individual specimens are plotted with a single dot

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 2.1. List of morphological characters used in this study. Length always refers to measurements made longitudinally to the long axis o f the body and width always refers to measurements made transversely to the long axis o f the body.

Meristic Measurements

1. Ventrals (V). Total number of ventral scales beginning with the first scale row that

contacts the first dorsal scale on both sides of the venter and not including the anal

plate (Dowling. 1951).

2. Subcaudals (SC). Total number o f subcaudals on one side, beginning with the first

scale behind the vent that contacts a subcaudal from the other side. The terminal

spine is not included (Rossman et al., 1997).

3. Supralabials Left (SLL). Supralabials on the left side of the head are counted from

the first scale posterior to the rostral scale along the mouth to the last enlarged scale

bordering the gape.

4. Supralabials Right (SLR). Supralabials on the right side of the head are counted

from the first scale posterior to the rostral scale along the mouth to the last enlarged

scale bordering the gape.

5. Infralabials Left (ILL). Inftalabials on the left side of the head bordering the mouth

are counted from the first labial posterior to the anterior genial scale to the last

enlarged scale bordering the gape.

6. Infralabials Right (ILR). Inftalabials on the right side of the head bordering the

mouth are counted from the first labial posterior to the mental scale to the last

enlarged scale bordering the gape.

7. Preocular Left (PROL). Number of preocular scales on the left side of the head.

8. Preocular Right (PROR). Number of preocular scales on the right side of the head.

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9. Postocular Left (POL). Number of postocular scales on the left side of the head.

10. Postocular Right (POR). Number of postocular scales on the right side of the head.

11. Temporals Left (TML). Number of temporal scales on the left side of the head.

Omitting the postocular scales, they include all scales found between the parietal

scale and the supralabial scales.

12. Temporals Right (TMR). Number of temporal scales on the right side of the head.

Omitting the postocular scales, they include all scales found between the parietal

scale and the supralabial scales.

13. Dorsal Scale Row 10 (DSR.10). Number of dorsal scales around the body,

beginning at the tenth ventral scale.

14. Dorsal Scale Row at midbody (DSR 50). Number of dorsal scales around the body

starting at the midbody ventral scale.

15. Dorsal Scale Row Penultimate (DSRPEN). Number of dorsal scale rows around the

body counted at the level o f the penultimate ventral.

16. Keel (K). The first dorsal scale row at midbody that exhibits a keel.

17. Dorsal Blotch Number (DB). Number of dorsal body blotches counted from the

neck to the level of the vent.

18. Lateral Blotch Number (LB). Number of lateral body blotches counted from the

neck to the level of the vent.

19. Wide Dorsal Blotch Width (DBW1). The number of scales counted transversely in

widest part of the dorsal blotch at midbody.

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20. Narrow Dorsal Blotch Width (DBW2)? The number of scale rows counted linearly

in narrowest part of the dorsal body blotch at midbody.

21. Long Dorsal Blotch Length (DBL1). The number of scales rows counted

transversely in the longest part of the dorsal body blotch at midbody.

22. Short Dorsal Blotch Length (DBL2). The number of scale rows counted linearly in

the shortest part of the dorsal body blotch at midbody.

23. Wide Lateral Blotch Width (LBW1). The number of scale rows counted

transveresely in the widest part of the lateral body blotch at midbody.

24. Narrow Lateral Blotch Width (LBW2). The number of scale rows counted

transversely in the narrowest part of the lateral body blotch at midbody.

25. Long Lateral Blotch Length (LBL1). The number of scale rows counted linearly in

the longest part of the lateral body blotch at midbody.

26. Short Lateral Blotch Length (LBL2). The number of scale rows counted linearly in

the shortest part of the lateral body blotch at midbody.

27. Dorsal Stripe Width (DSW). Number of scale rows counted transversely across the

dorsal body stripe.

28. Lateral Stripe Width (LSW). Number of scale rows counted transversely across the

lateral body stripe.

29. Supralabial Bars (SLBN). Number of barred or marked supralabials on the left side

o f the head.

30. Infralabial Bars (ILBN). Number of barred or marked inftalabials on the left side of

the head, not including supralabials darkened by the postocular stripe

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31. Lateral Ventral Blotch Number (VBLAT). The anterior-most ventral that contains a

lateral ventral blotch.

32. Medial Ventral Blotch Number (VBMED). The anterior-most ventral that contains

a medial ventral blotch.

Mensural Characters

33. Snout-Vent Length (SV). Measured from the anterior rostral tip to the posterior

margin of the anal plate.

34. Tail Length (T). Measured from the posterior margin of the anal plate to the tip of

the tail spine.

35. Head Length (HL). Measured from the rostral tip to the posterior apex of the

retroarticular process of the compound bone.

36. Parietal Length (PL). Length of the parietal measured along the median suture

from the posterior most point of contact with the frontal scale to the point of contact

with nuchal scales.

37. Anterior Parietal Width (PW). Measured from the contact point of the median

suture with the posterior-most extension of the frontal scale to the point where the

parietal contacts the ventral most postocular scale and the most ventral temporal

scale in the first temporal scale row.

38. Posterior Parietal Width (PWP). Measured at the narrowest posterior point in

contact with nuchal scales.

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39. Frontal Length (FL). Measured medially from the posterior suture point between

prefrontals to the posterior apex of the prefrontals.

40. Medial Frontal Width (FW). Measured from right supraocular-frontal contact to the

left supraocular-frontal contact located midway between the anterior-most point and

posterior-most point of the frontal scale.

41. Posterior Frontal Width (FWP). Measured across the posterior margin of frontal

scale from the contact point with the right surpaocular-parietal suture to the contact

point with the left supraocular-parietal suture.

42. Anterior Frontal Width (FWA) Measured across the anterior margin of the frontal

from the contact point with the right supraocular-prefrontal suture to the contact

point with the left supraocular-prefrontal suture.

43. Prefrontal Length (PRFL). Measured along the median suture from the posterior-

most contact point with the frontal to the anterior-most contact point with

intemasal.

44. Anterior Prefrontal Width (PRFWA). Measured on the left prefrontal from the

anterior prefrontal median suture connection with the intemasal to the dorsal-and

the anterior-most connection with the loreal.

45. Posterior Prefrontal Width (PRFWP). Measured on the left prefrontal from the

posterior prefrontal median suture connection with the frontal to the point of

contact with the loreal -preocular suture.

46. Intemasal Length (INL). Measured along the median suture from the posterior

connectio with the prefrontal to the anterior connection with the rostral.

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47. Anterior Intemasal Width (INWA). Measured on the left intemasal from the

median suture connection with the rostral to the point of contact with the anterior

nasal-posterior nasal suture.

48. Posterior Intemasal Width (INWP). Measured on the left intemasal from the

median suture contact with the prefrontal scale to the dorsal-and posterior-most

contact with the posterior nasal.

49. Eye Diameter (EYE). Measured at the widest horizontal point between preocular

and postocular.

50. Anterior Genial Length (AG). Measured from the anterior most point of contact

with the supralabials to the posterior most contact with the gular scales.

51. Posterior Genial Length (PG). Measured from the anterior most point of contact

with the anterior genial to the posterior most point of contact with the posterior

genial scale.

52. Intemasal Rostral Contact (INR). Measured across the width of contact between

both intemasals and the rostral scale.

53. Nasal Rostral Contact (NR). Measured along the anterior nasal contact with the

rostral scale from the contact point with the dorsal intemasal-rostral suture to the

supraiabial-rostral suture.

54. Rostral Height (RH). Measured from the rostral contact with the intemasal median

suture to the left ventral-most extension of the rostral scale.

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55. Rostral Width (RW). Measured from the left rostral contact point with the anterior

nasal-supralabial suture to the contact point with the right anterior nasal-supialabial

suture.

56. Posterior Nasal Length (PNL). Measured along the supralabial-posterior nasal

suture from the posterior nasal contact point with the supralabial-loreal suture to

the contact point with the anterior nasal-supralabial suture.

57. Anterior Nasal Length (ANL). Measured along the supralabial-anterior nasal

suture from the contact point with the supralabial-posterior nasal suture to the

contact point with the supraiabial-rostral suture.

58. Preocular Width (PROW). Height of the preocular measured from the dorsal-most

contact with supraocular-frontal-prefrontal sutures to the ventral-most contact with

the supralabials.

59. Dorsal Preocular Length (PROLD). Measured from the contact with the loreal-

prefrontal sutures straight across the preocular to the contact with the eye.

60. Ventral Preocular Length (PROLV). Measured from the preocular contact point

with the ioreal-supralabial suture to the contact point with eye and supralabial

suture.

61. Dorsal Loreal Length (LD). Measured from the loreal contact point with the

posterior nasal-prefrontal suture to the contact point with prefrontal-preocular

suture.

62. Ventral Loreal Length (LV). Measured from the loreal contact point with the

posterior nasal-supralabial suture to the preocular-supralabial suture.

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63. Loreal Height (LHT). Width o f the loreal measured from the loreal contact point

with the posterior nasal-supralabial suture to the contact point with the anterior

nasal-prefrontal suture.

64. Supralabial Length (LL). Measured from the posterior-most contact with the dorsal

body scales to the anterior most contact with the rostal scale.

65. Supralabial Height (LH). Width of the largest supralabial scale (usually the seventh

or eight supralabial scale) measured from the ventral margin to the dorsal-most

contact point with the temporal scale sutures.

66. First Dorsal Scale Length (DSR1). Measured on one dorsal scale in the first dorsal

scale row at midbody.

67. Vertebral Row Dorsal Scale Length (VR). Measured on one vertebral scale taken

at midbody in the vertebral scale row.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. without prohibited reproduction Further owner. copyright the of permission with Reproduced

PROLD 0 5 £ eC £

Fig. 2.3. Illustration of selected characters measured on the lateral and ventral head scales of Elaphe obsoleta and LBW 1

L B W 2

D S R 1

D B W 1

Fig. 2.4. Illustration of selected color pattern and body scale measurements made on Elaphe obsoleta and Elaphe bairdi.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. LSW DSW i— i n

Fig. 2.5. Illustration o f dorsal body stripe measurements made on Elaphe obsoleta and Elaphe bairdi.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. subspecies. All statistical analyses were performed using the program Systat 8.0 (SPSS,

1998).

Elaphe obsoleta and E. bairdi exhibit continuous growth, and a bias in size of

mensural characters between juveniles and adults occurs. Therefore, an attempt must be

made to minimize allometric influencesdue to differences in patterns of growth between

juveniles and adults (Thorpe and Leamy, 1983). Fig. 2.6 demonstrates that the change

in the HL/SV ratio diminishes near SV lengths o f500 mm. That is, the steep cline due

to changes in SV/HL begins to flatten at SV values of 500 mm. Therefore, only

specimens of with a SV length greater than 600 mm were included in the study, insuring

the exclusion of specimens exhibiting juvenile ontogenetic growth patterns. The change

in ratios of other mensural characters also decreases at SV lengths of 500 mm.

Adult specimens can vary in size from 600 mm to almost 2000 mm in this study.

Since size is assumed to be less heritable than shape, an attempt must be made to insure

that the differences among mensural characters in clades refer only to shape (Gould,

1966; Corruccini, 1975; Reist 1985). To produce a linear relationship between all

variables and reduce the effect of individual size variation, mensural characters were

transformed logarithmically (Hills, 1978; Thorpe and Leamy, 1983; and Sokal and

Rohlf, 1995). As demonstrated by Reist (1985, 1986), a univariate computation of

residuals provide the best estimate of shape of characters in ectothermic vertebrates

with continuous growth. Residuals of mensural characters were obtained by regressing

each variable against a character that is a good indicator of size. Therefore, the standard

residuals of the logio-transformed head (35) and body measurements (34, 66, and 67)

were obtained by regressing each against the

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. o o CM Arrow Arrow indicates O rV*” “» ‘ * O C'J"-'^7Sr i n -W&?- Elaphe Elaphe obsoleta. * * o E O c ■ M & . o 3 " " > CO o * o m r„-

1 1 i 1 h- CO m CO o o o o o o o o o o o o Figure Figure 2.6. Graph of HL/SV against SV for all samples of where where the change in HL/SV begins to diminish. AS/HH

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. log-transformed SV (33). Furthermore, the residuals of the log-transformed head

measurements (36-65) were produced by regressing each against the log-transformed

HL (35). Sexual dimorphism for all characters was tested using a Student’s /-test.

Also, male and female data were analyzed separately to minimize the possible effects of

sexual dimorphism.

All raw characters were first examined for statistical significance between the

four molecular groups (eastern clade, central clade, western clade and E. bairdi) using

ANOVA with a Bonferroni adjustment. Student's /-test was used to determine if

characters were significantly different between sex within each clade.

Canonical discriminant function analysis (DFA) was performed on meristic

variables and the residuals of the logio-transformed mensural variables. This technique

was used to determine if it is possible to differentiate among a priori groups by using

the available measurements (Manly 1994). This technique maximizes the separation

between groups and accounts for within-group variance and correlation. DFA has been

successful in differentiating closely related lineages of snakes using morphological data

(referred to as CVA in Thorpe 1976, 1980, 1983, 1987; Wuster and Thorpe 1992;

Wuster et al. 1995). DFA may help determine which morphological characters best

influence the inclusion of an individual into a specific molecular clade.

DFA was implemented on female and male data separately, as well as female

and male data combined. Four non-mutually exclusive sets of characters were

examined using DFA to determine which sets of characters best distinguish clades

morphometrically. The first set, Case 1, includes all mensural characters (34-67). The

second set, Case 2, includes non-color meristic characters and all mensural characters

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. (1,2,5, 11,13-16, and 34-67). The third set, Case 3, includes all meristic characters

and some color and mensural characters (I, 2, 5, 11, 13-17, 19, 21,23, 25,31,32, and

34-67). The fourth set, Case 4, excludes characters that do not appear strongly

significant as determined by ANOVA on raw data. All variables with F-ratio values

higher than 5.0 (significant at P < 0.001) will be included in Case 4.

Classification matrices based of DFA scores for each Case were produced to

determine how well individuals can be classified into their correct molecular groups.

Using all Case 2 variables, a classification matrix was produced from DFA scores that

attempted to maximize differences between subspecies. Specimens that could not be

identified correctly as one of the seven subspecies or appeared intermediate between

subspecies were not used. This eliminated bias in identification of subspecies and

allowed a fair comparison of the different classification matrices that attempt to place

individuals in their correct molecular clades or their correct subspecies based on all

morphological traits that exclude color pattern bias.

Principal components analysis (PCA) also was used on all Case 3 variables to

determine if groups can be separated morphometrically without an a priori hypothesis

of group membership. Significance between groups on each axis were tested using

ANOVA with a Bonferroni adjustment

It should be noted that certain characters were not used in the multivariate

analyses. No residual variable exists for SV (33) because it was used as the

independent variable to produce the residuals for characters regressed against it.

Certain variables were removed because they obviously replicate the same

measurement. That is, characters ILL (5) and ILR (6) refer to the number of infralabials

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. on the left and right side of the head, and characters TML (11) and TMR (12) refer to

the number of temporals on the left and right side of the head. Only the characters

taken on the left side of the head were used (ILL and TML). Also, the following color

measurements were eliminated from the DFA because they may be replicating other

characters or appear in so few snakes that they would severely limit the number

ofspecimens used in Case 3: 18, 20, 22,24, 26, 27, 28,29, and 30.

Standardized measurements were used to discuss differences between clades in a

practical sense. Therefore, HL and TL were divided by SV, and all head measurements

were divided by HL. These ratios were not used in any statistical analyses.

RESULTS

Significant sexual dimorphism (P < 0.05) was observed in the following characters:

eastern clade = 1, 19,33, 34,36, 37,39,40-43,45-47,49,50-52, 56,-58,60- 64,66, and

67; central clade = 2, 5, 6, 13-15, 17, 31, 33^11,43-47,49-52, 54-58, and 61-67;

western clade = 1, 2, 15, 17, 18, 33, 34, 36-38,41,44,45,49, 54-57, 61,62, and 65-67;

E. bairdi = 2 and 21. Characters in males tended to be larger than characters in females

except for the following: eastern clade = 1 and 19; central clade = 5, 6, and 14; western

clade = 1 and 15. The results of the ANOVAs between clades for each character

considering the sexes separately are displayed in Tables 2.2 and 2.3. In males, 83.6% of

the raw characters examined univariately differed significantly between clades. In

females, 80.3% of the raw characters examined univariately differed significantly

between clades. For males, the following characters had F-ratio values higher than 5.0

and were included in DFA for Case 4: 1,2, 5, 13-17, 19, 21, 23, 25,29-32, 35-38,40-

45, 47-49, 51, 52, 54, 55, 57, 59, 61-67. For females, the following characters had F-

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. The degree n o 14 175 131 136 145 224.933 380 Is* 285 32.979 152 2.16 643 2.16 ns. 345 90 45.090 228.748 41.340 W* n s. 355 242 105 42.483 n . 3.530 K. K. bairdi. 18 1360 194 569 194 144 1078.951 1649 1004.111 1620 164 10*17.439 n.s. 1.015 215.623 675 n.s. 245.425 610 238 ns. SV 602 16 144 10.354 13 |.;**« (_•••• |.;**« 3 766 !■:•*,c*** 0 W***,ll*** n.s. 0 610 W***||** 8.799 0 8.773 1092.16 18 1812.22 18 10.778 171.947 2.411 19 211.11 10.389 144 15 1.714 1.957 2 2 315 164 164 |;*«* (j*** yy*** 1.581 1.869 9 W***,U»** 8.970 0 w ..* l)» * . 238 238 0 0 TMI, I'MK 15 12 13.667 14 12.861 144 14 10 i:***,c***,w*** |;*** (j**« ||*»* |.;*.. c *** ||... 161 13 15 14 1750 w ***ll*** 0.890 0.767 II .K II W***,H*** 231 II II15 II 6 144 10 13 li***,C***,ll*** 163 |;*** (J.»« yy*** W***.ll*** II 436 11.385 8.916 10 10 130.091 178.862 40.547 22.562 11.1. W***,l)*** 236 104 14 14 18 18 3.583 09895 I'***, II* " 91 |.;*»* yy*** 96 286 13.278 92 126 150 5055| : . . . | | . . . 0 651 0.707 91 65 70.442 Subcaudals 18 246944 256 7 312 234 |;**« yy*»* |:**« (•••• ||*«* 4852 4672 0.908 5.877 W**M1*** 231.785 82 175 11.509 11.540 Ventral* w . . . |i««* 71.6)7 F Mean Max SI) Min /> MaxSI) /' 242 n Mean 229.403 82.769 12.736 Min 213 63 n 144 SI) /■ Min 220 Max 251 % nMean 163 Mean 232.941MaxSI) 243 3.917 89.147 99 5.694 0.646 13 0.570 Min 221/’ 72 n 2)7 ♦♦♦(/'cO.OOl). ♦♦♦(/'cO.OOl). number F-Ratio (/*), of specimens used for each character(n), mean ofeach character (mean), minimum character follow the format in Table 2.1. Clades are abbreviated as follows: E=Eastern, C=Central, W=Western, and B character was not applicable for that clade and n.s. indicates no significant difference exists between selected clades. Table 2.2. Results ofANOVAs performed on raw variables for males for the four clades ofrat snakes. Characters abbreviations ofsignificant difference between clades and the selected clade row is indicated **(0.00l<0.0l),(O.OK/'cO.OS), by * and value (Min), maximum character value (Max), and standard deviation (SD) are reported foreach clade. N.A. indicates that the

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Table 2.2, Continued N f C * C fN X fN * s P«* 5s Z r~. •* © © r~. 2 * 2 59 C iri fS f». — ^ S' ^ «£ ri X © • *C P- -C i-S ♦ n ^ c ^ # ^ c ^ ) £ — w ^ ^ »r> r* r, J C TI * c . S ^ C ^ C # *. ft rt • — /i f*i K c 2 -

^ c c • CJ — 55*r P- * • ^ fs *r ^ J rs»™ S - 2 V i * •^ K =. n i; r- « £ #■*. p * I N f'. N C C “ - *r - c c > - V

-J S pi Ok S •" r^ r* r» ^ r* f- c ^ »* g -r 06 ~ * • M ftC * I o *"> — 'C • N ^ c ? ^ ri C C - — f~. c* w> e iii

- . e ^ -• . — r» * * ^ r-- «/"v vn r - _ n V — V C K 5% ^ w 'S * J ^ O f1- f- w ^ TJ c w ^ * n w ^ o’ e — «n — iri d e — rs — r^oc - m - «»>,n < c

c ^ ® r- «N - IA C • _ O O w-t — J fi . _ w P rs v-i r- • J 5s! c w x J 5 X * C * • « -O -r - C ; m v n c d U — «*■

< U £ CN v© • ac v> * * _ ^ « f»l * * c w *r> I r c m «r o * K ? F r = : * C C A © ; n »ri f*i X - > — v» #•-. ac — S£ — v ri © — —

— * » - rn «o . - *n -r r j — P — f- p- f (s. — «^ — «r> rs 36 ~ ^ - C ^ O' - « — vi c c —

•o o> CJ *+-. wi r, • ff* c* , — r^. ^ «i - T " « C*4 W» — ^ £ r+. © '-t ->. p —; jj r, C « O' *' ~~— C X C - o U 5 5 5 ^~~

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Table 2.2, Continued z as» « Z as C S £ < z » e —— c r»4 es • «-i ^ ^ r-. rM © * -» — — ££ — — -» * © rM s ^ ^ — “ X J ^ n f*MJs4 ^ ***. • *0 'C ^ N C S f^‘ s£ « s£ f^‘ v> ^5 r>i— s; •»© r* r-* 96 _• © li.s a ^ a li.s s A C »A 5s a g x x g a c ex.* . ^ x ^ ^ e — i ^ m ©>7! • r»-‘©^ *r\ © ^ **•St , f

0 V»« — C V»» 0“ «— — © _ ! r'C N -^ *2! 2 !* £ 2 C S 5 2 = 2 t ™ ft S »^. N — IS — r r j r - <*. Os ■ <"**. - f r r t 3 -£ *■ T * 'T O*96 0 *. 61 * acr-: «^ — ^ w — Os ©— — ^ ©—— — e ^ * ? ^ * • — C T X 5> X * w-\ r* i .£ g ~ g .£ i r 5 ^ r* ^ o #m —— X C — ^ — 96 • ^ *+\ r+ r* 36 cha I —j •n ^ . 3 * ^ ^ * 5 o S' _ — v

~~^ c O' N ^ . • — C X fM x fn * * ~~c x O' d . :> ^ ©©©*-. •. ac < r s «n — iri > ■ > — w i n «r v — — <~~~~

~~C' © ^ - •= 2 £ * ^3i «r> cm T •> 5 S — ! cn — 5 r, c ? — r» — i~~

*/“i ^ * * 2 #

~~, ~M t£ IT to z • © © cn • • ©S *0 ^ sC •• ■ rs — **i ©’ ^ I © «S © ~~~

~~x is r» o> £ “ 2SR ©. V— N X ^C - -r — © «^v -r IN IN — IO C — in — to o e — — ro ©~~

~~n x «r r^ - »r in c ^ S'r-^to ^ * cn — IO — «T © C — IN — *r C S — IN~~

T3 0> 3 _» r^. ac «*• rN • _C sc ac »S £ ** “ • O' C*■*» r*» O X«r“. — r4 4 in K o c ^ io K 6 ! C 3 «N S .s£. 8 = 5^ 8.S 5^ 2 2 2 3 : 2 22 3: 2 2 2 3 : Min Max 3.22 SI) 4 85 /’ 0.464 (.’•*, W** 2.29 4.45 0.566 n. s 1.53 3 17 0.372 0 445 n. s. 1.38 0 473 2 82 n s. 2 32 0.267 3 88 n s. 1.15 2.2 WM* <_'•*, W** 1881 4.539 37% 0.714 2 36 5 27 CN JU JO 4$g £

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Table 2.2, Continued _2 ■S 2£ U «C x , > ■ 3£ . © p p © r*~.^.. IN IN IN IN — J — IN IN IN IN J — IN IN IN IN * I O = § I ^ vSJs; * w-i C? )P c c )P N ri fi I - ffi i r C rN : N O C ^ i/-i **» ini £ tr> * * ^ 2 IN N K o i o K N ac © p wv IN p; w-i fr O' C J C w-iO' fr <■* ? •i c j *i . f ^ *»i p ^ J Z 2 5 S r- ♦* r- S S.sS~ * c c r*» m - O p «r , x s f f — ff>‘ • y —i ; X • 'X 'i"‘ pn • I M I — li l — IN M IN — * ' ' n — in ^ V* — O' p — — — r-4 © ; — r-4 © — — «r x 2 Co © , OC © o _ < ^ r^. ~ — r< i n ^ f • r< * r** fr S ! • • X y _ *! S#‘ e - * : 3 2 2 2 = . * : 3 2 2 2 = . = : 3 2 2 2 § ©© ©®: : — — <-N — p o £ 5 S .£ 5 . ~ t »ii © e»ii«i ac © o © O r-*O© . «* n C © i © 'C pn Z2 ~ ^ ^ , • * s r C' © © - «r IN IN IN - Ui — • cs cn

c x M r-*" vi N * IN # * r- © © p P * © * © Vi w"v * © - r cn • i r A C < * C : r- _: o it i/% — p ^ “ x ^ — • r r — — 2?•* _ ^ u Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. without prohibited reproduction Further owner. copyright the of permission with Reproduced

Table 2.2, Continued c * > r © «**> O . ~ r -1 r*j © © . « 9. 64 ^ d ^ *ri e * e s s ac ac 3 n c<^ Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. without prohibited reproduction Further owner. copyright the of permission with Reproduced

Table 2.2, Continued £ J3f 1 >|: C/5 < : g; > : ©W% g © N - O - CM c% »r — — i» S — ^ — S* © ir» — «v n © «n «rv © ac i J 2 5 J o i*i — i c s */** vi «*© s w-i _ '—; £ § * s v-t • ^ c s>j ac nook ~ ^ v~ S K O J • * z z z z z z Z Z Z Z Z Z <<<<<< <<<<<< © © © r * >* © _ “ • — w*i d * — U ^ ^ • • ^ ^ __ * © — © * — * r • r- o C r* p ! * * « ^ 5 S Z r-» © f«* cm „ 5 = S • © cm © ♦, © cm 65 ^

z z z z z z z z z z<<<<<< z z <<<<<< CM— i !•> -o “ • «r» w. c*. “ - CM 1o —— CM • ! 2 2 2 3-=. 2 2 2 C» «c> j r 3C N O w CM w %©©©©♦" C p 1 S ’ *•’ ® . CM — *CV © CM U. ~ -

~~r — O — r* — „ — ri ri fC —ri O' ri n n — ac © ©— ^ Cl -T © VI — l f 2 2 2 5c* 3 -— a a -— 3 cm «■) VI O 5, — C ©. ©.~~

ns n* ns !;•••, CM, The degree 153 0 153 125.0 102 1300 30.658 2500 208.25 261.0 32.9 8 70.0 29.755 n. n. 327.0 84 213.310 JL_ hairdi. 1006.889 195.941 1473.0 605 163 280.0 1002.365 386 184 517 196 126 1020.0 185.815 40.160 1520.0 n.x85 i:»* 60 n.s. 6420 926.889 604.0 n. n. s. C»*. W» 610.0 _sy_ c*** 15.0 11300 1189 10.702 18.00 1560 14.0 15.0 148 148 5.0 W »«, !)••• W'", TMK C*M, W** n. s. 11.0 9.0 13.222 16.0 10.788 146 1679 1.991 1.739 1.775 2.199 2.194 9 9 9 W***, II ••• 7.0 6.0 610 0 w « »,» • • • 85 84 9.137 9.194 6 0 TMI. 6.0 4.0 13 667 13 10.0 14.0 15.0 11.797 143 0.494 856 1 2.088 626 153 W»»*. » ••• w . . . ()♦*. 0.849 0.749 JUL 15.0 15.0 16.0 15 015 150 13.0 120 13 889 13 14.0 100 |;**« \y*«* 11.776 146 |,;... c... 1)... l<;... c... (J... |,;... c... „»* 9 9 ill. 0.761 |]**** (_•••» VV*.» m |.; 94.0 3091 0.928 85.0 | : . . . |)*** 8 77.61770.0 12.76 10.0 831 12 100 94.0 89.0 60 85 83 700 8.060.0 8 0 84 c . . . w « » , 82 071 11.521 11.517 8.904 9.048 % .o 13.0 13.0 14.0 JiyfcsfitistelL. <_■•••, n*** <_■•••, 259.0 126 101242.0 125 123 124 126 9 220.0 223.0 5.817w . . . ))•*. 5.028 85 235 849 76.228 W*M |)M * 249.0 236.1 IS 236.1 226.0 /’ Max SI) 5.723 Min 238.0 Mean 24633 88.125 Max SI)/' 5.178 4.574 0.924 0.881 n MeanMin 231.388 n Min MaxSI) l> 226.0 Mean Max SI) 4.324n 5.542 0735 Mean Min /* n 148 w follow the format Tablein 2.1. Clades are abbreviated as follows: E=Eastem, OCentral, W=Westem, and B A. """(/^O.OOI). number F-Ratio (/*), of specimens used eachfor character (n), mean ofeach character (mean), minimum character

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Table 2.3, Continued I 5 © 9 5 c s s m r m © — — m r- > rsT " “ — •f « « ^ is n > in#n ^ ci « —« © - © —«n © £ a f Is f a £ ** C NX C ^ “ y i ^ ri c o > o c ri ^ :. t - - * r- r- — c X ^ ^ —^ — © * * * 1 f - ^© ^ s 2 *: :* - - £ © ; -r * * * © «n© J £ a ac i t

n f ^ > ^ fs *n — n — ^ rs J?S —«"» JL P r5*0 r: d > 9 E O ^ - n - p* —© — ^ p«*— < ^ — «n — © r-*— ^ »f »f ^ —*n? r* — c SScSS* £5 c 5 2 x v • *.r © ! © f*-. r» v - - n r- o 2 o r- n - ^ J 2 5 5 = T o T * p. • . p m r* • — i «—■ *• - rt © n i.ss» ^ © n C? .' . ? C 5 in v= • >n • 67 x x * © © © «n~s w — • •**.©*»© ^ n ci c *n c ^ — i r ^ >c© © © S Z Z c ^ * s’ f*i ^ 5^ /i - © r-- */-i ^ ^ n «r> n x . r X X n »**.*f n c i *n Ci 5 S «n (S If * C * < n o 7 9 > !•:• W"» n s !*•••, C**\ W*** li***, (.’•*, W** n s. c*. w*»* c*»,w*** li*,C*,l»*** Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. without prohibited reproduction Further owner. copyright the of permission with Reproduced

~~Table 2.3, Continued i l 2 =* Ul © s >£ © r >~~ o ® yS CCCff'l f IN J IT O *f ^ ^ ^ c r>»— / © © ^ © © «/S O— © a r * * *n rN ae © .» —• »r r. in x • c x in > ^ c ; S oc * ac © £J ^ ^ ^ pj s •s s s ■ 12 • ^ — o — y% « « 6 ! S ^ x d > d x i^i vS — 2 6 c i r - I — C — IN — a = * *« s 1 c C C C c T1 N - fr ^ r f — vC «r» — ac ' — r ^ — _ c222x e 2 2: 2 ^ c 2 2 2 x : 2 2 2 x n — — cn * #n * r^ w S .£ * /-' 3 .£ 5 5 .£ 3 /-' * S.£ wa oc oc : *r\

68 - C K ^ i r < X ; '« C O » N ae N * O C 1*1 O - MN __ — © rN © 5 x r - r> c r> - r x UI — ac ^ 5 © © ac rN — © __ r c —c * — c c «rt ^ r - o c «r i / * % — v% ac **• — — S r* r^. -r o -r r* r^. — S **• — ac * « m © n in © c «ri O' O' ^ vS © ^ © vS ^ O' *r#n*^So S ^ * n # r '* i ^ in i»nn » © V» ^ I. * N I*. N O 1 O; — ^ r» © © *r i *r © © r» ' «/"> O' O' 5 — 5 — m —: ***. © ^ CO' ' - O' 3C ' O © i * ' n j n »rC n *“> « in t O rs rs

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Table 2.3, Continued 2 : 2 < s a z as s ■ as * •3 s£ — u s ^ as g r» e I - C C C C C - IN — 5 •) ' t O' C “ ^ (•») -^ - o S^ ; I lC ? INTl C — iriinx oU o x n i i r i — ^ : — r* ^ . C IN «"N -T C* ^ 7 Z 5 5 ^C C " • • "T C C P^ A^ C # £ C *■» — ac — ^ •A ' P • o -r o cn I ; > r n < c r < ' «A i»J * S' ri ? C C i r ^ ^ * o o r^ r *n *ri . I « O > IN «A O — — 3 > T<> i - d « N S r ri

*25? w3 * 69 a. . — d n — i r x _ C •> . ~ r •*> SC . J 2 2 2 c c a *. * ac i*1. c c , I c c - c c IN ^ a *«*;c ac •«*■ _.-d c ac - p r*^ o' •r vd P * N C C N

~~i ^ ^ I t O tA IN ^ ^ lil u~~

~~_ C N - ^ = 2223; A M X X B r *• **• P *r>OB C i»i in i r r r -- r«* © — o ' n N — — m IA * T £ © O -T rN V - ^ Jv ^ 5\ p- rs rs p- 5\ £ 5 « 5 .£ S cn s *s — © © — *■>~~

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~~Table 2.3, Continued Ul ■ 2 > L Z r. ©~~

— © ^ — — rr — ^ ^ — r r — © ©r-: w*» ^ • " ( h C - C "T (N h* — — N N N - > - N N N — ^ sC — SS . ;» . SS — sC s i - © —i f-i rs rv* — «r © w> : i-“ c / — «/nac ? « —! x *r * c '■"* . n £ :.s 3C • ^ © o o © c*“:^ I - 9 — P3— « « T ^ ri— ^ rsi ^ — — — » — ? »N — — — — P S * = > P « • _ - N (N (N - 5 - (N (N N - eZZZSi. * C . > r* « ®. - C m n ©r*© © O* © © ff* irti ?» O* * © £ r * — ! i.s n C — fn — r* r* — - ' • e'. J P- p 1: m 70 * . *». f O c O fn — i r X . ' « f f « ' 222*^. S wi © S — CS £ m >£ r * x rs *r «r> •; C* r- 3 N 4 N w N --- J C rs *i v © © f**i cvi rs *rs g cH *“» r» © »♦ 3^ £ © " O ’ © • « ' f ' S <^i ^ * n • #nv* i.s CC ^ C in :*~i **: m 2 C C ^ fs « fs ^ . v « iri O r*i pi i r ^ ' — C N — — O' ?» N N n «N N — ^ 222Ec. c » c ^ i r i r 5» n « is c *r c is « wn ©v a* «r n i f d *f ri in r s • m ?• cs •r r4 r < r-4 *r © “ »£!;r“. «r — « « S s . S © - r o © I o O N O o /I M S C fS M N c s< ac ri r —

— ••*)••! *M ••"'•: «A .M u ( « u ill u * '« '( u s ..M «.A\ '•*•.")'•••:! **«M '•••.*")*•••}! * « Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. without prohibited reproduction Further owner. copyright the of permission with Reproduced

~~Table 2.3, Continued 5 -2 4£ 5: 2 ac y:~~

. i c © P ©• © i P > / fS-• IS fS © P — •r ac r K ci >, > C4 cvi C K - - — -r ^ *n f z wi » a — w-i ac «»i — g^SS — S ri ^ g * $ r - © O ac •" •" ac O © - r — $ cm SSSi. S S IS r*i t*i © a . ac © — © J i r* • ^ r-* © ri n © ; © © © - © -r — f j «f « n in S S d sj © r^ > ^ » ^ v» ^

u * > V > — 5* N N - IN O - I M © p p © 2 2 5 2 5 ^ = ; ♦ 5 ^5 ^ ^ ^ v n r- c - r n v» . * in ^ r* » - — f«K— ; •/■> © »/■> *n S . S s © - r rs ! ^ O J J ^ o * - o o £ ~ £ 3 < 71 — “"I ae n

~~# :~~

v • ^ Jv n n © «’ "t ’ « m C e , • - SSINISN i r © U* W - N M « wn © - - O --M * = 2 2 2 2 ? » . • r r» r •? * , _ ^ a ' ♦ £* © © r~ « Ck N h« N ©©©>—*: © © © ^ * _ ^ 2£ i * t i r * i r C !

s s r — li. — rs — ©s cs « « m co? o c ©«• m «r •! S' * - - U - - * S' •! :2222s. u £ ^ ^ ^ * > ^ r f Lil fN — *r o — r* *" sC vC ^ ^ vC sC *" o «/-j — © vo C p i C 1 5 • . £^^ £ S.S * 5 q q q O ^ * ^ «Nm ♦ **i

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Table 2.3, Conlinued -3 c © § I-N — 5 ©• o m 6 d m - J - X m id 06 --U N — 'T f o ^ c • • c ^ o fr *0 4 n - - n 4 — . . c © rN > ac © © © *n «f M U X - - ©•> r- m ' C N • N C5 O C' ! .£ 3 — « S 5 © ® nc — y in c* J « * ^ wn © © r - • ’ • - r © © wn ^ r-* O' © **J© * X o n n ^ * T C — © OC «T © ©* on - *i # # . © *-i w-v © ^ N «/■» fN rs* - * © ^ n c ^ — ac — «n © 5?% — — rs S C C H s © , © © © ^ -s 2 Z Z Z Z 2 Z Z Z Z 2 < < < < < < r m r © i* © r* m iri 7 2 2 Z ^ rs “ rs > n ^ m n — «n r ~ S■£ £~ ^ n r~. © ; 72

n r* n - o - o - n r* n x ■; P o ®! ■; o P •, x ©s © k © — ^ * r:® * ff!5 © » *. - © ***. f-* v» s© w —— in c 2 2 =2 zzzzzz rsi -? P O 3? * * 3? O P -? rsi < < < < < < c *r : c c « * es * °- St * — - © C © t-- —— 8.£ ■ , »N^ ^ 222xe. IN © ® © ® r r — •ri »* ® © s S~ 8 .s N 3 ; r IN © © in *r\

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~~Table 2.3, Continued -8 > * = ^ - * ' ' * - ^ = >* is~~

— — O — — ® » 1 r»: ® — v ^ n « •, , • «a n ^ v . :i. 3 5 2 5 r * C S' C — — r* o — « ps ps C ^ C *o *o C ^ C C a I .EI £~ * , c !»•. !»•. , c — — 73 zzzzzz < < < < < < 3?;t. 2 2 2 l 1 N - r O rs N f 9 - r - —* . O© rs< * «/■% i.E * ri C i r »*i 22233 »/■> © - r o - K i r r * (N c c c * — — «*• — c - r sc * o S>> W — VI 5 s p ^ 2 ^ p s 3.3er> * © © ratio values higher than 5.0 and were included in the DFA for Case 4: 1,2,5,11, 13-17,

~~19, 21,23,25, 30-33,36-38,40-42,43-45,47-49,51,52,54,57, 58,61,63-65. Only~~

~~characters with F-ratio values higher than 5.0 for both males and females were used in~~

~~the combined male and female data set in the DFA for Case 4.~~

~~The between groups F- matrix from the DFA showed significant differences~~

~~between all four molecular clades for all Cases using male and female data separately or~~

~~combined (P< 0.001). Wilks Lambda demonstrated that there was significant dispersion~~

~~among all of the molecular clades for all Cases using male and female data separately or~~

~~combined (P < 0.001). Three-dimensional DFA scatterplots for all Cases using male~~

~~and female data separately or combined are shown in Fig. 2.7-2.18. Although the three~~

~~clades of E. obsoleta appear to be grouped closely on these scatterplots, each clade still~~

~~occupies its own morphospace. The resolution between clades is clearest for all Case 3~~

~~and Case 4 variables and for male and female data analyzed separately. This may~~

~~indicate that color measurements along with other morphological characters are~~

~~important for distinguishing clade differences. The notched box plots are shown to~~

~~demonstrate that the clades are significantly different at either the first or second DFA~~

~~axis (Fig. 2.19-2.30). The notched box plots depict the morphological distances~~

~~between each clade for each axis and show where significant differences occur between~~

~~clades due to lack of overlap in the 95% confidence notches.~~

~~Three-dimensional plots and notched box plots for the PCA for Case 2 variables~~

~~are shown in Fig. 2.31-2.36. Significant separation between all clades is also~~

~~demonstrated with PCA. Using ANOVA with a Bonferroni adjustment, all clades were~~

~~significantly different at P < 0.001 on either the first or second principal component~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. D F2 • Eastern Clade o Central Clade ▼ Western Clade V Elaphe bairdi~~

~~Fig. 2.7. Three dimensional plot of the first three discriminant function scores based Case 1 variables for males.~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. CO U_~~

* • •

~~• Eastern Clade o Central Clade ▼ Western Clade v Elaphe bairdi~~

~~Fig. 2.8. Three dimensional plot o f the first three disciminant function scores based on all Case 2 variables for males.~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. • Eastern Clade o Central Clade ▼ Western Clade v Elaphe bairdi~~

~~Fig 2.9. Three dimensional plot of the first three discriminant function scores based on all Case 3 variables for males.~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. eo u.~~

~~-10~~

~~• Eastern Clade o Central Clade ▼ Western Clade v Elaphe bairdi~~

~~Fig. 2.10. Three dimensional plot of the first three discriminant function scores based on all Case 4 variables for males.~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. U_ Q~~

~~• Eastern Clade o Central Clade ▼ Western Clade Elaphe bairdi~~

~~Fig. 2.11. Three dimensional plot of the first three discriminant function scores based on all Case 1 variables for females.~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. CO U_ O~~

~~10 OF 2~~

~~• Eastern Clade o Central Clade ▼ Western Clade V Elaphe bairdi~~

~~Fig 2.12. Three dimensional plot of the first three discriminant function scores based on all Case 2 variables for females.~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. CO u_ Q~~

~~-10 -15 -20 -25 -30 Of 2~~

~~• Eastern Clade o Central Clade V Western Clade V Elaphe bairdi~~

~~Fig. 2.13. Three dimensional plot of the first three discriminant function scores based on all Case 3 variables for females.~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. o o~~

~~• • •~~

~~CO u_ O~~

~~14 16~~

~~• Eastern Clade o Central Clade ▼ Western Clade v Elaphe bairdi~~

~~Fig. 2.14. Three dimensional plot of the first three discriminant function scores based on all Case 4 variables for females.~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. w ▼~~

~~• Eastern Clade o Central Clade ▼ Western Clade V Elaphe bairdi~~

~~Fig. 2.15. Three dimensional plot of the first three discriminant function scores based on all Case 1 variables for combined male and female data.~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. CO u- 10~~

~~• Eastern Clade o Central Clade ▼ Western Clade Elaphe bairdi~~

~~Fig. 2.16. Three dimensional plot of the first three discriminant function scores based on all Case 2 variables for combined male and female data.~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. • Eastern Clade o Central Clade ▼ Western Clade v Elaphe bairdi~~

~~Fig. 2.17. Three dimensional plot of the first three discriminant function scores based on all Case 3 variables for combined male and female data.~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 10~~

~~• Eastern Clade o Central Clade ▼ Western Clade v Elaphe bairdi~~

~~Fig. 2.18. Three dimensional plot of the first three discriminant function scores based on all Case 4 variables for combined male and female data.~~

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. CLADE E bairdi. I 5 0 0 /vi A CM o £ CLADE E E C W B T Letters designating rat snakeclades are E = Eastern, C = Central, W = Western, and B = Fig. 2.19. Notchedbox plots ofthe first two discriminant functions based on all characters included in Case I for males. oo >4

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Fig. 2.20. Notched box plots of the first two discriminant functions based on all characters included in Case 2 for males. for 2 Case in included characters all on based functions discriminant two first the of plots box Notched 2.20. Fig. Letters designating rat snake clades are E = Eastern, C = Central, W = Western, and B = = B and Western, = W Central, = C Eastern, = E are clades snake rat designating Letters DF 1 (63.8%) C B W C E CLADE C B W C E . bairdi E. □ CLADE li. bairdi. O CLADE Letters designating rat snake clades are E = Eastern, C = Central, W = Western, B Fig. 2.21. Notched box plots ofthe first two discriminant functions based on all characters included in Case 3 for males.

Df 1 (47.0% ) 0 C B W C E CLADE (V* D i. L CM o c CO . bairdi. K. C B W C E CLADE DF 1 (55.2%) 10 Letters designating rat snake clades are E = Eastern, C = Central, W = Western, and B = = B females. and for IWestern, = W Case in Central, = C included Eastern, = E characters all are on clades based snake rat functions designating discriminant Letters two first the of plots box Notched 2.23. Fig. C B W C E J ______CLADE I ______I ______L Q UL CM 1 CM -4 5

------C B W C E 1 ------CLADE 1 ------bairdi. 1 ------L_ Fig. 2.24. Notched box plots of the first two discriminant functions based on all characters included in Case 2 Case in included characters all bairdi on based functions discriminant two first the of plots box Notched 2.24. Fig. for females only. Letters designating rat snakes clades are E = Eastern, C = Central, W = Western, and and Western, = W Central, = C Eastern, = E are clades snakes rat designating Letters only. females for DF 1(51.8%) . 10 CLADE o LL c\T CO CO o> ©"* P s c\i 1 CLADE H. ------1 « ------^ X £ K, K, bairdi. CLADE . . — r— — i f - 5 0 ‘10 E C W B x CM Q ° n. CO * ^ 10 ...... W T —1 J — ■“ 1 ------CLADE ------E E C W B "1 ...... - - 555 2^5 4s 0 10 -20 -10 -30 Fig 2.2S. Notched box plots ofthe firstLetters two designating discriminant rat functionssnake clades based are on E all= charactersEastern, C = Central,included W in Case= Western, 3 for and females. B = in D i b UL O'* vo u>

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Letters designating rat snake clades are E = Eastern, C = Central, W = Western, and B = = B and females. for 4 Western, = Case in W included Central, = C Eastern, characters = all E on are based clades snake functions rat discriminant designating two Letters first the of Plots Box Notched 2.26. Fig DF 1 (56.8 %) C B W C E CLADE s 0 CM o> O P s CM . bairdi. E. C B W C E CLADE a E. E. bairdi. CLADE ° 2 in B W CLADE CE 5 0 5 10 female data. Letters designating rat snake clades are E = Eastern, C = Central, W = Western, and B = Fig. 2.27. Notched box plots ofthe first twodiscriminant functions based on all characters included in Case I forcombined maleand vo LA

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. -10~~

~~CLADE CLADE~~

Fig. 2 .28. Notched box plots of the first two discriminant functions based on all characters included in Case 2 for combined male and female data. Letters designating rat snake clades are E = Eastern, C = Central, W - Western, and B = E. bairdi. B W E. E. bairdi. CLADE C E 5 0 5 -10 CO WB CLADE EC 0 5 15 female data. Letters designating rat snake clades are E = Eastern, C = Central, W = Western, and B = Fig. 2.29. Notched box plots ofthe first two discriminant functions based on all characters included in Case 3 for combined male and

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. K. K. bairdi. CLADE CLADE female data. Letters designating rat snake clades are E = Eastern, C = Central, W = Western, and B = Fig. 2.30. Notched box plots ofthe first twodiscriminant functions based on all characters included in Case 4 forcombined maleand VO 00

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. a-~~

~~• Eastern Clade o Central Clade ▼ Western Clade v EJaphe bairdi~~

~~Fig. 2.31. Three dimensional plot of the first three principal component scores based on all Case 2 variables for males only.~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. -4~~

~~• Eastern Clade o Central Clade V Western Clade V Elaphe bairdi~~

~~Fig. 2.32. Three dimensional plot of the first three principal component scores based on all Case 2 variables for females only.~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. • Eastern Clade o Central Clade ▼ Western Clade Elaphe bairdi~~

~~Fig. 2.33. Three dimensional plot of the first three component scores based on all Case 2 variables for combined male and female data.~~

~~101~~

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. B w CLADE E. E. bairdi. EC 1 3 4 2 0 1 2 3 B O n ■ 0s VO w CLADE EC 1 1 3 2 3 4 2 B in co o . o^* o^* O vP W CLADE C

3 2 4 -4 2 0 - -3 00 00 1 £ o O Lettersdesignating rat snake clades are E = Eastern, C = Central, W = Western, and B = Fig. 2.34. Notched box plots ofthe first three principal component scores based on all characters included in Case 2 for males. s

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. E. E. bairdi. CLADE B CM W CLADE C E 3 2 -1 ^ 1 ^ 0 CLADE 2~~

00 CM 1 Fig. 2.35. Notched box plots ofthe Lettersfirst threedesignating principal componentrat snake clades scores are basedE = Eastern, on all Ccharacters = Central, Wincluded = Western, in Case and 2 B for= females. o ut

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. CLADE K. K. bairdi. CLADE CO O -1 CLADE 1 3 2 0 4 -2 -3 •1 -4 Fig. 2.36. Notched box plots ofthe firstLetters three designatingprincipal component rat snake clades scores are based E = onEastern, all characters C = Central, includedW = Western,in Case and 2 B tor= females. h* o>

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. axis. Male, female, and combined data revealed that all clades were significantly~~

~~different except the eastern and western clade on the first principal component axis.~~

~~However, for males alone and the combined-sex data, ANOVA performed on the~~

~~second principal component axis revealed that all clades were significantly different.~~

~~All clades except the eastern and central groups were significantly different for female~~

~~data alone on the second principal component axis. Significant differences exist~~

~~between all clades on the third principal component axis for females only and~~

~~combined-sex data. The eastern and central clades were not significantly different on~~

~~the third principal component for males only.~~

~~Most characters had low (±0.01-0.9) or mid-ranged (±1.0-4.0) discriminant~~

~~functions (DF) (Tables. 2.4-2.15). The character HL (35) had high DF for both males~~

~~and females on the first axis for all cases. In addition, RH (54) was consistently large~~

~~for both males and females on the first axis for Cases 1-3. Males had high values for~~

~~AG (5), NR (53), and LL (64) on the first axis for Cases 1-3. Several other characters~~

~~had high DF functions for both sexes, but were specific to only one Case.~~

~~PCA loadings for the first three axes appears in Tables 2.16-2.18. The~~

~~characters PW (37), PRFWA (44), INWA (47), INWP (48), RH (54) and RW (55)~~

~~tended to have high loadings for both males and females on the first axis. All meristic~~

~~characters have negative loadings on the first axis for both sexes. Ventral s (1) and~~

~~subcaudals (2) have high positive loadings on the second axis for both sexes.~~

~~Molecular clade classification matrices based on DFA scores for males, females,~~

~~and combined sex data for all cases are shown in Tables 2.19-2.21. Total classification~~

~~scores for males, females, and combined sexes were very high (78-100%) and were~~

~~105~~

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 2.4. Canonical discriminant functions based on all Case 1 variables for males. Values in parentheses represent the proportion o f the sum o f the eigenvalues.

A xis C h aracter 1 {63 8% ) 2 {23.0% ) 3 {13 .2% ) TL 0.361 0.170 0.185 HL -0.880 0.207 0.197 PL -0.157 0.318 -0.302 PW -0.321 0.010 0.456 PWP -0.135 -0.118 0.413 FL 0.115 0.157 -0.340 FW -0.056 0.025 -0.382 FW P4 -0.004 -0.081 -0.067 FW -0.076 0.410 0.482 PR FL4 -0.143 0.221 0.112 PRFWA -0.049 0.213 -0.094 PRFWP -0.219 -0.020 -0.238 INL 0.273 0.087 0.055 INW A -0.154 -0.251 0.053 INWP 0.009 0.056 0.220 EYE -0.026 -0.113 -0.120 AG 0.380 0.314 -0.105 PG -0.097 -0.285 0.162 INR 0.127 -0.261 0.130 NR 0.333 -0.006 -0.227 RH -0.384 0.148 -0.502 RW 0.043 -0.219 0.076 PNL 0.259 -0.069 -0.064 A NL -0.274 0.140 -0.211 PROW 0.012 -0.126 -0.198 PROL 0.324 -0.024 0.109 PR O LV 4 0.145 0.106 0.055 LD 0.147 0.159 0.126 LV -0.039 0.066 -0.135 LHT -0.111 -0.087 0.166 LL -0.319 0.017 0.185 LH -0.001 -0.441 0.249 DSR1 -0.071 -0 .0 2 4 -0.064 VR -0.015 -0.208 -0.131

~~106~~

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 2.5. Canonical discriminant functions based on all Case 2 variables for males. Values in parentheses represent the proportion o f the sum o f the eigenvalues.

A xis C haracter______1(54 9% )______2 (32.0% )______3 (1 3 .1 % ) VENTRALS 0 .3 9 9 0.143 0.3 3 9 SUBCAUDALS 0.131 0.090 -0.588 ILLEFT -0.199 0.518 0.131 DSR10 0.096 0.180 -0 .180 D SR 50 -0.303 -0.027 -0 .1 7 9 DSRPEN -0.022 0.089 0.019 K 0 3 0 5 0 J 2 1 0.1 7 4 T L 0.191 -0.118 0 .0 5 0 HL -0.629 0.140 -0.227 PL -0 .152 0.235 0 .0 3 4 PW -0.279 -0.132 -0.377 PWP -0.212 -0.025 -0.253 FL 0.1 1 7 0.202 0 .1 4 2 FW -0.007 0.153 0.272 FWP 0.068 -0.088 0 .1 1 5 FW A -0 .144 0.138 -0.520 PRFL -0 .093 0.109 -0.169 PRFWA 0.100 0.061 0.185 PRFWP -0.200 0.132 0.1 6 5 INL 0.239 0.084 -0.132 INWA -0.191 -0.169 0.129 INWP -0 .010 0.018 -0 .210 EYE -0 .0 1 0 -0.094 0.153 AG 0.320 0.303 -0.079 PG -0.118 -0.309 -0.041 INR 0.050 -0.185 -0.053 NR 0 .3 0 5 -0.001 0.0 8 0 RH -0.411 0.235 0.248 RW 0.1 3 2 -0.120 0.082 PNL 0.221 -0.082 0.021 ANL -0 .1 9 4 0.155 0 .1 6 9 PROW 0.023 -0.088 0.093 PROLD 0.260 -0.078 -0 .104 PROLV 0.156 0.004 -0.083 LD 0 .1 5 6 0.095 -0.176 LV 0.004 0.051 0.128 LHT -0.093 -0.044 -0 .025 LL -0 .392 -0.115 -0.052 LH 0.043 -0.288 0.021 DSR1 0.014 0.084 0.069 VR 0.017 -0.114 0 .1 5 5 TML 0.243 0.233 0.168

~~107~~

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 2.6. Canonical discriminant functions based on all Case 3 variables for males. Values in parentheses represent the the proportion of the sum of the eigenvalues.

Axis Character 1154.6%) 2132.0%) 3 (13 4%) VENTRALS 0.028 0 3 8 0 0.481 SUBCAUDALS -0.011 -0346 -0309 ILLEFT -0 J0 5 -0.178 -0340 DSR10 -0.047 0.026 0347 DSR50 0.078 -0.436 -0.372 DSRPEN -0.288 0 3 1 0 -0.025 K 0 3 9 0 0.043 0 3 4 6 DB 0.048 -0.966 0.137 LB 0.624 -0334 0.762 DBW -0.093 -0.014 -0.181 DBL -0.425 0.021 0 3 0 8 LBW -0313 -0.054 0.000 LBL 0378 -0.481 0 3 4 6 VBLAT 0.286 0331 -0.410 VBMED -0.249 -0.149 0348 TL 0 3 7 6 0359 -0.345 HL -0.693 -0.848 0.465 PL -0.062 -0.777 0.064 PW 0 J6 0 -0.069 -0359 PWP -0.404 -0374 -0.416 FL 0.194 0.122 -0.126 FW -0.129 -0373 -0.123 FWP 0.138 0337 0301 FWA -0.496 -0.158 -0394 PRFL -0.032 -0.391 0.086 PRFWA -0.232 0.177 0 3 5 2 PRFWP -0.093 -0.405 -0.061 INL 0.130 -0387 0.159 INWA -0.287 0362 0.115 INWP -0.023 -0.046 -0.156 EYE -0.067 0314 0 315 AG 0.805 -0.077 -0.469 PG 0.050 -0.179 -0.016 INR -0.073 -0.095 -0301 NR 0399 0.031 -0385 RH -0.432 -0.784 0 3 7 8 RW -0.196 0335 0 3 2 0 PNL 0.308 0348 -0.066 ANL -0.217 0.295 -0.686 PROW -0320 0.347 -0371 PROLD 0.645 -0.176 0.128 PROLV 0.176 -0392 0.018 LD 0383 -0.199 -0.448 LV -0.318 -0392 0.809 LHT 0.171 0310 -0.106 LL -0.645 0.191 0 3 2 0 LH 0.064 0385 0.637 DSR1 0.225 -0.138 0 3 6 4 VR -0325 -0.192 0.055 TLT 0.663 0.044 0.340

~~108~~

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 2.7. Canonical discriminant functions based on all Case 4 variables for males. Values in parentheses represent the proportion o f the sum o f the eigenvalues.

Axis Character______I (47.0%) ______2(36.1%) 3 f 16.9%) VENTRALS 0.173 0.015 0.755 SUBCAUDALS -0204 0249 -0.095 ILLEFT -0290 -0.345 -0.122 DSR10 -0.195 0266 0 2 0 6 DSR50 -0.168 -0.072 -0277 DSRPEN 0.102 -0.041 -0.027 K 0.111 0.199 0.141 DB -0.456 0.580 0.519 DBW -0.151 -0.418 -0.126 DBL 0.048 -0.203 0 2 6 2 LBW -0.348 0.007 0.493 LBL -0265 0.175 -0.114 VBLAT 0-347 -0.032 -0208 VBMED -0.184 -0.063 -0.044 SLBN 0.024 0.024 0.178 ILBN 0.073 -0.092 -0310 HL -0.691 0.051 0.172 PL -0.419 0241 0 2 1 3 PW -0.428 0.178 -0.522 PWP -0279 0.171 -0213 FW -0.183 -0.138 0.108 FWP 0.119 0 2 1 0 0.074 FWA -0.142 -0.057 -0281 PRFL -0.408 0305 0.124 PRFWA 0272 -0273 0.407 PRFWP -0311 -0.108 0 2 3 2 1NWA 0254 -0.079 0.016 INWP 0.105 0.118 -0.075 EYE 0.135 -0.072 0249 PG -0.057 0.121 -0.174 INR 0.005 -0.134 -0252 RH -0.351 -0.121 0.172 RW 0.164 0.170 0 3 2 0 ANL 0.105 -0.087 -0.185 PROW 0.080 -0.029 -0.428 LD -0.147 0201 -0.191 LV -0.266 -0.135 0289 LHT 0.190 -0.304 -0.114 LL -0.000 -0.159 -0.114 LH 0.184 -0.098 0303 DSR1 0.131 0.094 0351 TML -0.044 0.184 0211

~~109~~

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 2.8. Canonical discriminant functions based on all Case 1 variables for females. Values in parentheses represent the proportion o f the sum o f the eigenvalues.

Axis Character______1 (55.2%)______2 (24.4%)______3 (20.4%) TL 0.150 0.157 0 3 4 5 HL -0.799 0 3 9 5 -0.098 PL 0.103 0383 -0.148 PW -OJOO -0319 0.025 PWP -0.156 -0.072 0 3 6 3 FL 0.085 0.124 -0 3 5 3 FW 0.015 -0.041 -0.175 FWP 0.004 0.068 0.160 FWA -0.117 0373 0.160 PRFL -0.096 0.175 -0.020 PRFWA 0.079 -0.141 -0.002 PRFWP -0.139 0.064 -0 3 0 7 INL 0.199 0.003 0.086 INWA -0.242 -0335 0 3 3 1 INWP -0317 -0.138 0 3 6 6 EYE 0.238 -0.179 -0.151 AG 0.092 0312 -0.228 PG -0.085 -0381 0.150 INR -0.233 -0.097 -0.026 NR 0.169 0.152 -0.164 RH -0.420 -0.074 -0.286 RW 0340 -0.063 -0.125 PNL 0.056 0.101 0.003 ANL -0.059 -0.157 -0.466 PROW -0336 -0.013 0.056 PROLD 0383 -0.189 -0.192 PROLV 0.00 0.00 0.00 LD 0.054 -0.064 -0.048 LV 0389 0.199 0.017 LHT -0338 0.246 -0361 LL -0387 0313 -0.157 LH -0.040 -0.339 0 3 1 1 DSR1 0.004 -0.169 -0.074 VR -0.003 -0.107 -0.172

~~110~~

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 2.9. Canonical discriminant functions based on all Case 2 variables for females. Values in parentheses represent the proportion o f die sum o f the eigenvalues.

Axis Character______1(51.8%) ______2 (32.6%)______3(15.6% ) VENTRALS 0244 0.085 0205 SUBCAUDALS 0221 0.033 -0.403 JLLEFT 0.065 -0297 0.126 DSR10 0.083 -0223 0.030 DSR50 -0.274 0.040 -0211 DSRPEN 0.019 -0.279 0.036 K 0.597 -0.018 0.021 TL 0.094 0.037 -0.129 HL -0.441 -0241 -0.121 PL 0.199 -0.197 0.086 PW -0292 02 1 8 0.111 PWP5 -0231 0.011 -0220 FL 0.118 -0.038 0.129 FW 0.022 0.075 0.116 FWP 0.024 -0.078 -0.017 FWA -0.142 -0251 -0.415 PRFL -0.085 -0.173 -0.080 PRFWA 0.070 0.040 -0.031 PRFWP -0.062 -0.079 0.104 INL 0.146 0.036 -0.092 INWA -0.233 0.073 -0.060 INWP -0.004 0.043 -0.094 EYE 0.167 0.029 0.152 AG 0.108 -0212 0.144 PG -0.152 0.099 -0.006 (NR -0.150 -0.036 0.143 NR 0274 -0.075 0.178 RH -0.478 0.023 0246 RW5 0218 0.078 0.095 PNL 0.088 0.103 -0.088 ANL -0.058 0.112 0.434 PROW -0211 -0.097 0.032 PROLD 0.140 0.041 0.178 PROLV 0.00 0.00 0.00 LD -0.119 0.143 0.043 LV 0229 0.028 -0.111 LHT -0.179 -0267 0.149 LL -0.156 -0.159 0.065 LH -0.079 0206 -0.326 DSR1 0.131 0.066 0.059 VR 0.039 02 3 6 0.180 TML 0.147 -0.139 -0.020

~~111~~

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 2.10. Canonical discriminant functions based on all Case 3 variables for females. Variables in parentheses represent the proportion of the sum o f the eigenvalues.

Axis Character 1 (55.4%) 2f26.2% ) 3 (18.4%) VENTRALS 0.513 0.827 0.750 SUBCAUDALS 0.300 -0.786 -0.126 ILLEFT -0.058 -0.376 -0.015 DSR10 0.803 0.342 0.217 DSR50 0.140 -2.793 0.694 DSRPEN 1.204 -1.789 0.598 K -0.168 0.697 -0.865 DB -2.569 1.414 •0.688 LB -1.510 1.923 -0.884 DBW 0.436 -2.214 1.069 DBL -1.876 3.113 -1.723 LBW -0.653 0.200 -0.055 LBL 0.249 1.501 -0.082 VBLAT 0.200 1.026 -0.358 VBMED 1.711 -1.165 0.739 SLBN 0.955 0.419 0.079 ILBN -0.758 -1.066 -0.273 TL 0.072 -0.688 1.348 HL 0.067 0.842 -1.520 PL -0.213 0.155 0.623 PW 0.518 1.380 -0.687 PWP -0.638 0.724 -0.429 FL 0.050 -2.519 1.170 FW 0.069 0.993 -1.368 FWP 0.317 0.974 -0.904 FW -1.281 -1.018 0.449 PRFL -0.163 0.591 -0.233 PRFWA -0.415 -0.054 -0.338 PRFWP -0.025 -0.458 0.840 INL -0.594 -2.318 0.879 INWA 0.511 0.741 -0.908 INWP 0.914 0.910 0.282 EYE 0.695 0.208 0.953 AG 0.542 -2.042 0.514 PG -0.019 0.316 -0.266 INR 0.384 1.381 0.649 NR •0.489 0.698 0.385 RH 1.732 0.960 -0.721 RW -0.160 0.399 0.578 PNL -1.084 1.485 -0.494 ANL 0.468 -2.424 1.902 PROW -0.334 -0.005 -0.289 PROLD -1.066 0.097 0.223 PROLV 0.00 0.00 0.00 LD -1.437 1.324 -0.587 LV -1.245 1.358 -0.020 LHT 0.688 -1.826 -0.947 LL 0.173 1.149 -0.898 LH 0.952 -0.902 0.029 DSR1 -2.265 2.053 -1.275 VR 1.925 -1.681 1.543 TML -0.087 1.298 -0.786

~~112~~

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 2.11. Canonical discriminant functions based on all case 4 variables for females. Values in parenthesis represent the proportion o f the sum o f the eigenvalues.

Axis Character I [56.8%) 2 [29.0% ) 3 [14 2%) VENTRALS 0.294 0.116 0 3 3 4 SUBCAUDALS 0203 02 6 7 -0.731 ILLEFT 0.120 -0.079 0205 DSR10 0351 0.056 0.101 DSR50 -0.252 -0.009 -0.634 DSRPEN -0.103 -0295 -0.193 K 0361 -0269 0323 DB 0.465 -0.191 0 2 5 6 DBW -0.371 0.055 0.059 DBL 0.041 -0226 0.756 LBW 0.035 -0.146 0.145 LBL -0212 0.178 0.155 VBLAT 0.058 -0.146 0.133 VBMED -0398 0.178 -0.188 [LBN 0256 -0.182 -0.044 HL4 -0.499 -0.621 -0.003 PL4 -0.012 0.014 -0.188 PW4 -0291 -0.100 0327 PWP4 -0.683 -0.095 0.274 FW4 0.173 -0.045 0.087 FWP4 -0.212 02 3 6 -0.645 FWA4 -0292 -0.632 0.133 PRFL4 -0.076 0.030 -0396 PRFWA4 0267 0.147 0364 PRFWP4 -0.104 -0.097 -0.023 INWA4 -0.044 02 4 2 -0.405 INWP4 -0228 0230 0.029 EYE4 -0.018 0397 -0.154 PG4 0.066 -0.082 0.217 INR4 -0393 0301 0.073 RH4 0.033 0365 -0.188 ANL4 0263 0.139 0.132 PROW4 0.168 -0.008 0294 LD4 0277 0.123 0.240 LHT4 -0.192 -1.064 0304 LL4 -0.003 -0.100 0.125 LH4 0.117 0.138 -0.002 TTML -0.050 -0.374 02 8 2

~~113~~

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 2.12. Canonical discriminant functions based on all Case 1 variables for combined male and female data. Values in parentheses represent theproportion o f the sum of the eigenvalues.

Axis Character ! (60.6%) 2 (25.0%) 3(14.4% ) TL 0 3 4 6 0.151 0 3 5 8 HL -0.783 0341 0.119 PL -0.048 0 3 6 7 -0.169 PW -0318 -0.125 0332 PWP -0.147 -0.120 0355 FL 0.125 0.161 -0.255 FW -0.096 0.033 -0331 FWP 0.026 -0.015 -0.020 FWA -0.057 0 3 3 9 0371 PRFL -0.099 0 3 1 4 0.027 PRFWA 0.015 -0.176 -0.147 PRFWP -0.173 0.063 -0314 INL 0346 0.022 0.069 INWA -0.201 -0328 0.144 INWP -0.099 -0.072 0.190 EYE 0.009 -0.085 -0.163 AG 0.298 0 3 2 4 -0.138 PG -0.124 -0348 0.160 INR -0.042 -0327 0.054 NR 0.286 0.040 -0.226 RH -0.356 0.130 -0.443 RW 0.103 -0.155 0.014 PNL 0.169 -0.013 -0.007 ANL -0.183 0.021 -0310 PROW -0.098 -0.056 -0.071 PROLD4 0 3 0 4 -0.153 -0.077 PROLV 0.188 0.159 -0.011 LD 0.071 0.033 0.034 LV 0.134 0.192 -0.067 LHT -0.211 0.002 -0.013 LL -0.339 0.137 0.128 LH 0.042 -0.396 0325 DSR1 0.002 -0.092 -0.106 VR -0.089 -0.129 -0.107

~~114~~

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 2.13. Canonical discriminant functions based on all Case 2 variables for combined male and female data. Values in parentheses represent the proportion o f the sum of the eigenvalues.

Axis Character 1C5I.1%1 2134.7%) 3 ( 14.2%) VENTRALS 0.118 0.078 0.112 SUBCAUDALS 0.240 -0.159 -0.460 il l e f t -0.043 -0.476 0 3 0 4 DSRIO 0.050 -0326 -0.109 DSR50 -0236 -0.019 -0.197 DSRPEN 0.034 -0.141 0.029 K 0317 -0.112 0.077 TL 0.113 0.032 -0.003 HL -0.663 -0.163 -0344 PL -0.023 -0362 0.069 PW -0307 0.172 -0.157 PWP -0.134 0.068 -0333 FL 0.125 -0.150 0.185 FW •0.081 -0.081 0.168 FWP 0.030 0.008 0.115 FWA -0.022 -0.165 -0.499 PRFL -0.083 -0.109 -0.135 PRFWA 0.057 0.024 0.082 PRFWP -0.184 -0.127 0.125 LNL 0225 -0.035 -0.082 INWA -0324 0.102 -0.043 INWP -0.056 0.073 -0.143 EYE 0.008 0.068 0.176 AG 0365 -0361 0.136 PG -0.137 0346 -0.051 INR -0.071 0.097 0.023 NR 0 3 7 8 -0.015 0.174 RH -0344 -0.167 0 3 0 6 RW 0.105 0.120 0.008 PNL 0.158 0.039 -0.008 ANL -0.158 -0.058 0.246 PRO -0.111 -0.015 0.054 PROLD 0385 0.144 0.095 PROLV 0.192 -0.047 -0.013 LD 0.013 0.003 -0.051 LV 0.186 -0.078 0.051 LHT -0318 -0.030 0.003 LL -0.342 -0.005 -0.140 LH 0.043 0369 -0.158 DSR1 0.053 0.002 0.061 VR -0.099 0.144 0.112 TML 0.133 -0332 0.141

~~115~~

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 2.14. Canonical discriminant functions based on all Case 3 variables for combined male and female data. Values in parentheses represent the proportion of the sum of the eigenvalues.

Axis Character I 152.4%) 2131.6%) 3 116 0%) VENTRALS 0.288 0.241 0.504 SUBCAUDALS 0.039 -0.107 -0.143 H i EFT -0202 -0.123 0.108 ILR1GHT -0.159 -0.246 0.051 DSR10 0.144 -0.130 0.194 DSR50 -0.127 -0.048 -0.443 DSRPEN -0.087 -0.195 -0.036 DSR1 0.144 0.112 -2.689 VR -0.119 0.079 2.733 K 0.243 -0.131 0 2 6 9 DBW -0.169 0.041 0.070 DB 0.498 -0.499 0 3 1 0 DBL -0.070 -0.034 0 3 4 6 LBW -0.017 -0.190 0240 LBL -0.075 0.011 0.022 VBLAT 0.166 0.162 -0 2 6 6 VBMED -0.221 -0.008 -0.077 TL 0.141 -0.111 -0.073 HL -0.399 -0.299 -0.192 PL 0.005 -0218 -0.066 PW -0270 -0.159 -0216 PWP -0239 0.011 -0.121 FL 0.125 0.107 -0.046 FW 0.042 -0.042 0.117 FWP -0.064 0.068 -0.118 FWA -0.171 -0.185 -0.125 PRFL -0.008 -0.167 -0.023 PRFWA -0.089 0.127 0236 PRFWP -0.162 -0.066 0.130 INL 0.187 -0.109 0.000 INWA -0.160 0.084 0.061 INWP 0.188 0.096 -0.056 EYE -0.023 0 2 0 4 0319 AG 0.137 -0242 -0.143 PG 0.044 0.027 -0.117 INR -0.050 0.127 -0.236 NR 0.325 0.004 -0.080 RH -0.214 0.060 0.051 RW 0.119 0.147 0.267 PNL 0.190 0.010 -0.065 ANL 0.098 0.158 0.004 PROW -0.048 -0.051 -0.101 PROLD 0.300 -0.052 -0.156 PROLV 0.124 0.063 0.013 LD 0.197 -0.012 -0.065 LV -0.036 -0.222 0.142 LHT -0316 0.014 0.083 LL -0.299 -0.069 -0.184 LH -0.072 0.054 0.179 DSRI -0.027 -0230 1.633 VR 0.068 0357 -1364 TML 0.073 -0260 0230

~~116~~

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 2.15. Canonical discriminant functions based on all Case 4 variables for combined male and female data. Values in parentheses represent the proportion o f the sum of the eigenvalues.

Axis Character I f50.9% l 2(32.1% ) 3 (17%) VENTRALS 0.270 0237 0.580 SUBCAUDALS 0.225 0.033 -0.118 ILLEFT -0.207 -0.328 0.139 DSRIO 0.117 -0.163 0.168 DSR50 -0.217 -0.178 -0.475 DSRPEN 0.027 -0.137 -0.023 K 0.322 0.007 0.255 DB 0.575 -0.325 0.356 DBW -0.384 -0.148 0.040 DBL -0.070 -0.089 0.390 LBW 0.000 -0.211 0273 LBL -0.007 0.031 -0.013 VBLAT 0.044 0.120 -0.093 VBMED -0.187 -0.034 -0.200 ILBN 0.046 -0.010 -0.131 PL 0.115 -0.205 0.014 PW -0.143 -0.125 -0.182 PWP -0.164 -0.161 -0.215 FW 0.056 -0.132 0.133 FWP 0.013 0.162 -0.143 FWA -0.107 -0.346 -0.191 PRFL 0.091 -0.130 -0.059 PRFWA 0.021 0.369 0.467 PRFWP -0.179 -0.193 0.076 INWA 0.050 0.134 0.136 INWP 0.117 0.112 -0.082 EYE -0.040 0.236 0.251 PG 0.054 0.097 -0.029 INR -0.240 -0.020 -0.102 RH -0.125 0.062 0.081 ANL 0.036 0.101 0.040 PROW 0.031 -0.011 -0.090 LD 0.082 -0.085 -0.032 LHT -0.253 -0.047 0.165 LL 0.037 0.046 -0.127 LH 0.060 0.247 0.213 TML 0.123 -0.215 0.171

~~117~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 2.16. Principal component loadings for all Case 2 variables for males.~~

Axis Character One Two Three VENTRALS -0.123 0.613 0.043 SUBCAUDALS -0.325 0.643 -0.094 ILLEFT -0.295 -0.067 0.672 DSR10 -0.321 0.023 0.628 DSR50 -0.271 0.168 0.378 DSRPEN -0.208 0.121 0.486 K -0.285 0.375 0.154 TL -0.341 0.415 -0.038 HL -0.582 -0.539 0.162 PL 0.304 0.138 0.396 PW 0.704 0.021 0.180 PWP 0.377 0.076 -0.180 FL 0.321 0.297 0.174 FW 0.586 -0.138 0.207 FWP 0.402 -0.130 0.240 FWA 0.525 -0.033 0.357 PRFL 0.265 -0.051 0.178 PRFWA 0.640 -0.377 -0.042 PRFWP 0.474 -0.349 0.178 INL 0.200 0.430 -0.137 INWA 0.636 -0.167 -0.114 INWP 0.638 -0.378 0.062 EYE 0.499 0.046 0.285 AG 0.305 0.347 -0.034 PG 0.146 0.151 -0.046 INR 0.440 -0.315 -0.211 NR 0.493 0.040 0.067 RH 0.617 -0.293 0.051 RW 0.717 -0.128 -0.069 PNL 0.362 0.174 -0.177 ANL 0.478 0.105 0.296 PROW 0.539 0.182 0.125 PROLD 0.208 0.529 -0.159 PROLV 0.227 0.456 0.039 LD 0.081 0.207 0.308 LV 0.268 0.293 0.226 LHT 0.477 0.076 0.101 LL 0.596 0.282 0.295 LH 0.590 0.138 -0.168 DSRI 0.592 0.270 -0.249 VR 0.579 0.354 -0.180 TML -0.264 0.003 0.415

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 2.17 Principal component loadings for all Case 2 variables for females.~~

Axis Character One Two Three VENTRALS •0.185 0.582 -0.073 SUBCAUDALS -0.350 0.558 -0.077 ILLEFT •0.181 0.123 0.683 DSR10 -0.330 0.033 0.586 DSR50 -0.236 0.119 0.285 DSRPEN -0.245 0.038 0.447 K -0.189 0.474 0.128 TL -0.332 0.362 -0.116 HL -0.545 -0.537 0.173 PL 0.254 0.177 0.284 PW 0.601 -0.025 0.098 PWP 0.374 0.137 -0.180 FL 0.265 0.032 0.245 FW 0.540 -0.216 0.237 FWP 0.520 -0.232 0.227 FWA 0.556 -0.157 0.212 PRFL 0.567 -0.024 0.234 PRFWA 0.695 -0.187 -0.071 PRFWP 0.497 -0.259 0.190 DML 0.234 0.384 -0.026 1NWA 0.544 -0.364 -0.003 INWP 0.644 -0.256 0.069 EYE 0.524 0.130 0.084 AG 0.420 0.241 -0.055 PG 0.257 0.007 -0.282 INR 0.475 -0.274 -0.133 NR 0.439 0.192 0.117 RH 0.702 -0.162 0.059 RW 0.724 -0.056 -0.122 PNL 0.436 0.156 -0.053 ANL 0.460 0.024 0.128 PROW 0.560 0.103 0.031 PROLD 0.419 0.675 -0.058 PROLV 0.419 0.675 -0.058 LD -0.075 0.136 0.470 LV 0.109 0.298 0.443 LHT 0.516 0.108 0.347 LL 0.584 0.179 0.224 LH 0.584 0.083 -0.325 DSR1 0.591 0.108 -0.066 VR 0.456 0.215 -0.074 TML -0.218 -0.028 0.591

~~119~~

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 2.18. Principal component loadings for all Case 2 variables for combined male and female data.

Axis Character One Two Three VENTRALS -0.031 0.094 0.030 SUBCAUDALS -0.262 0.666 -0.062 ILLEFT -0.245 -0.075 0.681 DSR10 -0.322 -0.032 0.618 DSR50 -0.246 0.112 0.360 DSRPEN -0.218 -0.021 0.482 K -0.149 0.264 0.155 TL -0.302 0.493 -0.063 HL -0.580 -0.526 0.121 PL 0.288 0.204 0.326 PW 0.657 -0.004 0.138 PWP 0.379 0.086 -0.171 FL 0.306 0.150 0.225 FW 0.568 -0.181 0.200 FWP 0.454 -0.167 0.200 FWA 0.541 -0.156 0.296 PRFL 0.357 -0.077 0.182 PRFWA 0.648 -0.354 -0.080 PRFWP 0.475 -0.371 0.152 INL 0.225 0.438 -0.050 INWA 0.591 -0.233 -0.097 INWP 0.632 -0.384 0.028 EYE 0.511 0.076 0.182 AG 0.359 0.281 -0.043 PG 0.206 0.147 -0.147 INR 0.448 -0.260 -0.223 NR 0.473 0.009 0.116 RH 0.643 -0.309 0.035 RW 0.716 -0.117 -0.113 PNL 0.387 0.220 -0.123 ANL 0.473 0.054 0.230 PROW 0.552 0.136 0.105 PROLD 0.294 0.475 -0.040 PROLV 0.316 0.386 0.143 LD 0.029 0.246 0.393 LV 0.208 0.266 0.342 LHT 0.492 0.108 0.209 LL 0.601 0.226 0.281 LH 0.586 0.094 -0.220 DSR1 0.594 0.266 -0.167 VR 0.525 0.382 -0.129 TML -0.248 -0.043 0.496

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Correct HXI % 97 Vo Vo 1 1 ii 0 0 98 1 0 94 w'~ 40 0 1 45 41 0 0 0 C 1 _.... 33 0 4 CASE Cancel % 1 1 99 36 1 1 100 1) 1) 0 100 0 0 1(H) 97 2 44 ______1 40 39 W 0 0 33 33 0 0 26 1! C 0 0 0 0 3 CASE 100 84 93 81 %Cancel 12 » 0 80 26 0 ...... "" ... 76 0 *'w' 24 1 ...... bainii. 113 118 84 12 97 i; 0 0 0 2 CASE C oitccI 92 84 75 14 •X) 7 % II 83 II 13 0 0 ...... 91 76 0 90 2 4 3 “ w ~ CASE J CASE no 18 5 0 0 C 1 107 3 i; Total 128 w I) into their assumed clades based on geographical location using the characters included in each Case. Letters representing clades are G i; Table 2.19. Discriminant function classification matrix for males only. The table shows the number ofspecimens correctly= classifiedEastern, C = Central, = W Western, = B /:’. c 17 87 12 Cladc

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 100 KM) 97 •X) %Correct 2 94 2 0 W II 34 0 0 0 1 32 36 C 0 10 0 12 2 35 2 0 It 0 0 £ASE4 100 2 2 100 13 %Correct 0 100 0 100 ______W 26 26 27 27 0 0 100 C 0 i; 8 0 0 CASE3 100 0 0 0 K. bairdi. K. 85 86 8 %Correct n 8 0 91 0 0 ______1 W 50 6 0 91 0 73 83 9 CASE2 3 1 43 58 It C 9 0 0 0 8 ______100 92 83 82 70 8 1) "/•Correct 8 0 0 ______54 44 W 9 0 74 11 1 3 C CASE I CASE 1 59 !•: 0 0 0 74 HI W I- 1) Tout Table 2.20. Discriminant function classification matrix for females only. The table shows the number ofspecimens correctlyclades are E = Eastern, C = Central, W = Western, = B C 14 67 classified into their assumed clades based on geographical location using the characters included in each Case. Letters representing Claile

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Correct 100 84 89 87 89 % 3 11 3 0 0 1 W 0 72 12 0 44 82 79 1- C 40 4 Correct 8795 2 2 6 66 91 94 % 3 II 3 100 0 0 84 8 0 0 79 CW 0 K. bairili. K. 1 3 76 0 50 85 K 4 45 3 0 0 0 100 81 % Correct % i'i 20 0 71 0 89 0 116 139 20 80 W 20 0 183 142 33 3 8 0 1: C 6 0 _ CASE 2 CASE 3 _ _ CASE 4 100 79 155 78 198 66 37 ...... 20 20 13CofTCCt % 0 0 89 118 0 10 136 29 183 151 37 4 0 0 __CASEJ_ __ 158 41 1- C "w*“ 0 w 4 specimens correctly classified into their assumed clades based on geographical Letters locationrepresenting using clades the charactersare E = Eastern,included C = Central, in each Case. W = Western, = B II Table 2.21. Discriminant function classification matrix forcombined male and female data. The table shows the numberof i; Clade C Total 203

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. consistently higher for Case 3 and Case 4. The eastern and central clades had the~~

~~lowest classification values. This is consistent with the high overlap in eastern and~~

~~central clade scores revealed in the scatterplots (Fig. 2.7-2.19) and the notched box plots~~

~~(Fig. 2.20-2.33), and the lower sequence divergences Fig. (1.5-1.13). Tables 2.22-2.24~~

~~reveal that classification matrices based on DFA for Case 2 variables that maximize~~

~~separation between subspecies produced low total scores for males, females, and~~

~~combined sexes (59-67%). This is much lower than the total values for the DFA~~

~~classification matrix on Case 2 variables for males, females, and combined sexes when~~

~~variation between clades is maximized (80-86%). The widest ranging subspecies, E. o.~~

~~obsoleta and E. o. lindheimeri, had the lowest classification scores. The insular form,~~

~~Elaphe o. deckerti produced the highest classification score (100%).~~

~~Ratios of mensural measurements are presented for males in Table 2.25 and~~

~~females in Table 2.26. The ranges of most characters overlap even when corrected for~~

~~size using proportional data.~~

~~DISCUSSION~~

~~The three molecular clades of Elaphe obsoleta and the fourth clade of E. bairdi~~

~~can be distinguished morphologically by using univariate and multivariate statistics.~~

~~The morphological groupings are concordant with the evolutionary lineages as revealed~~

~~in the molecular analyses (Fig 2.7-2.33). The morphological classification matrices also~~

~~support the separation of these molecular clades (Table 2.19-2.21). The eastern and~~

~~central clades are more closely related and are grouped more closely in morphological~~

~~space. The western and E. bairdi clades are sister taxa and are most~~

~~124~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 58 59 Correct %~~

~~69 51 100 0 12 6 0~~

~~0264 0 4 2 I 0 62 1 | 3 11 8~~

~~0 46 28 23 II ______~~

~~9 0 ______6 6 ______40003 16 40003 16 l i bairdi E. o. limi. E. o. ohs. E. o. quad. E. a ross. E. o. spU. E. o. will. E. o. deck. 2 2 0 4 3 16 I 00 0 0 0 0 0 0 5 43 12 9 5 2 00 0 0 0 0 4 0 100~~

3 14 72 20 10 3 I | 0 0 0 0 67 ______into their assumed subspecies using the data included in Case 2. Subspecies Table 2.22. Discriminant function classification matrix for males only. The table shows the numberof specimens correctly classified E. E. bairdi E . o. ross. 0 2 2 E. E. o. quad. E o .o b s . E. o. will. E o .d eck . Total E o .lin d . E a s p il. C/»

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 100 82 80 50 86 74 58 75 % Correct % 69 6 7 0 () 0 0 E. E. o. deck. 3 () 2 () 0 0 E. E. o. will. 0 0 1 1 1 1 15 9 0 0 0 4 0 0 0 E. E. o. spil. 13 8 0 0 0 1 1 1 27 0 0 2 0 55 0 ()0 14 1 0 15 42 8 4 0 0 0 E. E. o. lind. E. o. obs. E. o. quad. E. o. ross. 00 1 0 0 1 2 45 II 2 0 0 E. E. Ixtirdi 0 0 6 0 Total 8 62 E , o. ross. E. o. spil. E. o. deck. E. E. o. obs. E. o. quad. E. o. will. Subspecies E. E. bairdi E. o. lind. Table 2.23. Discriminant function classification matrix for females. theirassumed The tablesubspecies shows the using number the dataof specimensincluded correctly in Case classified2. into

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 100 100 % Correct % 57 7K K2 67 69 67 1 E. E. o. deck. 3 04 100 0 0 0 16 K 12 3 2 0 0 0 13 10 2 1 IK E. E. o. spil. E. o. will. 2 2 44 0 0 0 0 1 E. E. o. ross. 3 3 0 10 6 E. E. o. quad. 0 0 3 35 34 IK 0 155 55 31 113 E. E. o. obs. 0 41 0 134 E. E. o. lind. 34 0 0 1 1 100 00 0 0 0 0 0 0 E. E. Imirdi 20 0 3 0 0 0 0 0 Subspecies E. E. o. spil. E. E. o. will. E. o. deck. E. E. bairdi E. E. o. lind. Total 24 E. E. o. obs. E. o. quad. E . o. ross. Table 2.24. Discriminant function classification matrix for combined male andcorrectly female data. classified into their assumed The table subspecies shows the numberusing theof dataspecimens included in Case 2. tv)

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FW/HL 0.137 (0.013) 0.142 0.139 0.136 (0.008) 05 39 ) 021 . 0 FL/HL 0.225 (0.014) 0.227 (0.016) 0.220 0.243 ( 39 PWP/HL 0.088 (0.013) 0.087 (0.013) 0.082 0.079 (0.028) (0.013) PW/HL 0.209 (0.012) 0.214 (0.013) 0.211 0.196 (0.014) (0.009) ) 020 . PL/HL 0 0.216 ( 0.220 (0.023) 0.231 0.227 (0.019) 39 ) 002 . 0 HL/SV 0.034 (0.003) 0.034 (0.003) 0.034 0.037 (0.003) ( ) 022 . 0.218 0 (0.019) 0.231 0.214 (0.016) 0.250 ( TL/SV (0.015)

Species/Clade Table 2.25. Mean and standard deviation (in parentheses) ofmensural ratios measurements for males. E. E. alleghaniensis (Eastern Clade) E. spiloides E. obsoleta E. bairdi E. (Central Clade) (Western Clade)

~~> — — n 00~~

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. (0.011) 0.121 (0.010) 0.110 (0.011) 0.071(0.011) 0.120 (0.011) 0.0680.062 0.126 (0.015) 0.079 (0.013) 29 ) ) 010 010 . . 0 0 ( 0.106 ( PRFWA/HL PRFWA/HL PRFWP/HL INL/HL INWA/HL 0.151 0.151 0.114 (0.012) (0.012) 0.160 (0.013) 0.122 0.154 (0.014) 0.121 0.136 75 78 (0.010) (0.013) (0.017) 0.192(0.014) 0.162 (0.013) (0.014) 0.123 0.190 0.158 0.125 0.123(0.008) 0.188 (0.013) (0.009) 0.155 0.128 0.189 0.160 (0.011) Species/Clade FWP/HL FWA/HL PRFL/HL E. E. obsoleta bairdi E. (Western Clade) (0.014) E. E. alleghaniensis Table 2.25, Continued E. E. spiloides (Central Clade) (0.014) (Eastern Clade)

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. RH/HL (0.016) 0.152 (0.013) 0.162 0.013) 0.139 (0.018) NR/HL 0.082(0.009) 0.166 0.080 (0.008) 0.079 (0.008) 0,085 31 INR/HL 0.115 0.121 0.116 (0.014) 0.092 (0.012) (0.013) PG/HL (0.027) (0.013) 0.244 0.238 0.248 (0.025) (0.028) 0.222 (0.035) (0.021) 0.211 (0.021) (0.021) (0.015) 0.143(0.011) 0.149 0.248 0.273 (0.014) (0.021) 0.105 0.106 0.093 0.1090.109 0.149 0.256 (0.011) (0.010) (0.012) (0.011) spiloides (Eastern Clade) Species/Clade INWP/HL EYE/HL AG/HL E. alleghaniensisE. bairdi E. E. E. obsoleta (Western Clade) Table 2.25, Continued (Central Clade) (0.014)

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. without prohibited reproduction Further owner. copyright the of permission with Reproduced

~~Table 2.25, Continued *3 .Si U T3 cn ja 0 5: 2 Q- 2 a£ £ X z x j . < z a. o Q_ O a: x 0 o > o_ x C/3 4> 8. j j c c : to 5: | f I t s II o ® St o o . . o' .2 T3 © © U >.1 39 __ 1>~~

~~1/J ^ s 2 o o O •n SO o oo «« St 11 O in in S' : ^ ' o : : o <-> 2 u ? z W ■a ooo oo °° oo o o 999 69 E eg o -C oo o St o w 2 n - t s *2T««» o -a ( ® O g II S' : 1 !m ' ; ' © 3 131 o ^~~

(0.000) 0.005 0.005 (0.000) 0.005 (0.000) (0.00) 0.005 DSR1/SV VR/SV 0.006 0.006 (0.001) (0.000) 0.005 LH/HL 0.129 0.132 (0.013) (0.000) 0.121(0.013) 0.006 (0.014) (0.001) (0.006) 0.109 0.771 0.775 0.711 (0.019) (0.032) LHT/HL LL/HL 0.051 (0.007) (0.030) (0.006) (0.010) LV/HL 0.085 (0.010) 0.086 0.054 (0.009) 0.086 0.052 0.091 0.048 0.770 (Eastern Clade) Species/Clade Table 2.25, Continued E. alleghaniensisE. E. spiloidesE. E. E. obsoleta E. bairdi E. (Central Clade) (0.010) (Western Clade) (0.010) (0.007) (0.027)

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FW/HL 0.135 (0.012) 0.142 (0.013) 0.136 (0.010) FL/HL 0.223 (0.016) 0.222 0.140 0.231 (0.019) (0.013) (0.015 (0.017) 0.089 0.075 0.246 0.080 (0.014) 0.089 (0.012) PW/HL PWP/HL (0.012) (0.013) 0.194 0.209 (0.014) (0.014) (0.013) (0.009) PL/HL 0.209 0.208 0.216 (0.020 0.233 0.217 0.218 (0.002) (0.022) (0.003) (0.020) (0.003) (0.002) (0.020) 0.217 0.036 0.1970.204 0.035 (0.015) 0.038 (0.020) 0.229 0.034 (0.010) Species/Clade TL/SV HL/SV E. alleghaniensisE. E.bairdi E. E. obsoleta (Eastern Clade) spiloidesE. (0.025) (Western Clade) Table 2.6. Mean and standard deviation (in parentheses) ofmensural ratios measurements for females. (Central Clade)

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. without prohibited reproduction Further owner. copyright the of permission with Reproduced

Table 2.26, Continued ja -o M CJ £ < £ < 5 £ z £ < *J H 3 o V! •ft o *3 © Os ' o ' o tN OS ' o V~i ' o V> ST o ' .22 It £ Os s Os 2 e ® ® ~ ft > 5*5 . © .3 o o o «N © 4 o o ' o o © , o o U - C ; ; C - o' o ' o ' o Os P - 0 ' o sO r'i ' o o ' o t I c U 2 « 5 - - j j 2 J2 •o © o o © o ? 0 *“ ©_ © o o o fM Vi a> 134 oo IC IC oo oo ' o o ~ oo ' o «N ' o o sT ' o It o 2 a ' o o «r © o o - r ' o rT ' o o o . o 0.152 (0.014) 0.169 (0.015) 0.161 (0.011) 0.141 (0.018) NR/HL RH/HL 0.083 (0.010) 0.085 (0.011) 0.081 (0.011) 0.090 (0.008) 0.087 ^ 78 PG/HL PG/HL INR/HL 0.244 (0.026) 0.111 (0.013) (0.028) 0.209 0.247 0.120 0.234 (0.029) 0.114 (0.012) (0.059) AG/HL 0.253 (0.018) 0.257 (0.023) (0.018) 0.269 (0.022) EYE/HL 0.141 (0.013) (0.016) 0.140(0.011) 0.152 0.246 INWP/HL 0.106 (0.011) 0.113 0.150 (0.011) 0.092 0.109 (0.006) (0.012) ullegliuniensis spiloides (Eastern Clade) Species/Clade /.'. /.'. E. obsoletaE. Table 2.26, Continued E.buirdi (Western Clade) (Central Clade) (0.015)

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 0.053 0.053 (0.065) (0.009) 0.057 0.065 0.058 (0.007) (0.008) (0.008) 0.059 (0.008) 0.056 (0.007)0.064 (0.009) 0.084 (0.008) 0.087 (0.010) (0.011) 0.077 0.093 PROW/HL PROLD/HL PRQLV/HL LP/HL 0.126 (0.010) 0.132 (0.022) (0.013) 0.126 (0.012)0.124 (0.010) 0.069 (0.009) 0.065 0.064 (0.008) PNL/HL ANL/HL 0.057 0.063 (0.007)0.060 (0.008) (0.007) 0.054 (0.007)0.059 (0.008) RW/HL 0.200 (0.016) 0.209 (0.016) (0.009) 0.191 0.196 (0.013) (Eastern Clade) (0.016) Species/Clade E. ulleghaniensisE. Table 2.26, Continued E.buirdi E. E. obsolelu E. E. spiloides (Central Clade) (Western Clade)

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. VR/SV 0.005 0.005 (0.000) 0.005 (0.000) (0.000) 0.005 (0.000) DSR1/SV 0.005 (0.000) 0.006 (0.001) 0.006 (0.000) 0.005 (0.000) LH/HL 0.130 (0.016) 0.132 0.015) 0.119 (0.011) (0.009) 0.106 0.763 (0.028) (0.038) 0.048 (0.007) 0.053 0.782 (0.007) 0.052(0.005) 0.770 (0.026) 0.051 0.771 LV/HL LHT/HL LL/HL 0.084 0.086 0.085 (0.008) 0.100 (0.007) (0.005) (0.021) (Eastern Clade) (0.009) Species/Clade Table 2.26, Continued E. E. alleghaniensis E. spiloidesE. (Central Clade) E. obsolela (0.012) (Western Clade) E.buirdi

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. distantly related to the eastern and central clade (Fig. 1.3). However, E. bairdi is~~

~~morphologically much more divergent from the eastern and central clades than the~~

~~western clade (Fig. 2.7-2.33). The western clade is more similar morphologically to the~~

~~eastern and central clades than to E. bairdi. Elaphe bairdi may be morphologically~~

~~distinctive because it has evolved in an environment very different from that inhabited~~

~~by the other clades. All members of the eastern, central and western clades are found in~~

~~eastern US wooded habitats, whereas E bairdi is found in riparian habitats on the~~

~~Edward's Plateau (Schultz, 1996). It is possible that E. bairdi evolved its distinct~~

~~morphology in response to the xeric habitats of southwest Texas and northeastern~~

~~Mexico. Although these recently diverged groups can be distinguished~~

~~morphologically, the morphological distance between groups may betray the actual~~

~~evolutionary relationships of all four clades as revealed by using mtDNA (Fig 1.3).~~

~~DFA placed individuals into their molecular clades much better than into their~~

~~correct subspecies (Table 2.19-2.24). Difficulty arises in correctly identifying most~~

~~subspecies. Most of these subspecies could not be separated into groups by using the~~

~~morphological data presented in Table 2.1. All subspecies have been defined on one or~~

~~two color pattern characters that are inconsistent within their putative ranges. The two~~

~~widest ranging subspecies, E. o. obsoleta and E. o. lindheimeri, are the most difficult to~~

~~classify morphologically and genetically. The subspecies E. o. obsoleta cannot be~~

~~defined when using the subjective character black with little pattern. Many authors~~

~~have commented that blotched rat snakes appear well within the geographic range of~~

~~supposedly unpattemed E. o. obsoleta (Hurter, 1911; Hudson, 1942; McCauley, 1945;~~

~~Conant, 1951; Smith, 1961; Barbour, 1971; Minton, 1972; Mount, 1975; Vogt 1981;~~

~~138~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Dundee and Rossman, 1989; Oldfield and Moriarty, 1994). Numerous specimens~~

~~examined during this study that were collected in the range of E. o. obsoleta were~~

~~clearly patterned like E. o. lindheimeri or E. o. spiloides: KS-Douglas Co.: KU 558,~~

~~2450; WI- Iowa Co.: MPM 8805; NC-Macon Co.: NCSM 14345; IL-Fayette Co.: INHS~~

~~12494; OH-Montgomery Co.: LSUMZ 5825, 59265; NE-Cass Co. UNSM 549, 2236).~~

~~Very dark specimens of E. o. obsoleta in Virginia and North Carolina exhibit the same~~

~~stripes that are a key character for identifying E. o. quadrivittata (Mitchell, 1994;~~

~~Braswell, 1977) (multple examples of this pattern class can be found in the NCSM~~

~~collection: e.g., NC-Brunswick Co.: NCSM 2178, 3851, 12467, 15419, 16773). The~~

~~striped dorsal pattern and yellow ground color of E. o. quadrivittata grade into the black~~

~~color pattern of E. o. obsoleta in a large area of North Carolina, South Carolina and~~

~~Georgia (Braswell 1977). The brown, orange or yellow ground color described for E. o.~~

~~lindheimeri in Louisiana appears to lighten gradually into the gray ground color~~

~~described for E. o. spiloides in eastern Alabama. This gradation in color pattern makes~~

~~it very difficult to identify individual specimens to subspecies when obtained in central~~

~~or western Alabama (numerous examples of these intermediate pattern classes can be~~

~~found in the AUM collection: AL-Baldwin Co.: 414, 1030; Lee Co.: AUM 430, 1723,~~

~~2047, and 32509). Elaphe obsoleta spiloides west of the Apalachicola River and east of~~

~~Mobile Bay look like gray E. o. lindheimeri.~~

~~Elaphe obsoleta deckerti from the Florida Keys and E. o. williamsi from Levy~~

~~Co., Florida, can be classified by using DFA (Table 2.22-2.24). However, both of~~

~~these subspecies occupy small ranges and it may be possible to classify them~~

~~morphologically because of local adaptations. Dowling (1951) and Schultz (1996)~~

~~139~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. considered E. o. williamsi to be an intergrade between E. o. spiloides and E. o.~~

~~quadrivittata (Dowling, 1951, Schultz, 1996/ The pattern and range o f this form is~~

~~intermediate between E. o spiloides and E. o. quadrivittata (Dowling 1951; Christman~~

~~1980). Many E. o. quadrivittata from Alachua Co., FL examined in this study were~~

~~also heavily blotched and had stripes like E. o. williamsi. In preservative, these~~

~~specimens appear identical to the gray E. o. williamsi (see specimens UF 2734,2762,~~

~~14351, 19333,49264,49443, and 64983 from Alachua Co., FL). Genetically, E. o.~~

~~williamsi appears to be related to individuals as categorized E. o. spiloides from Taylor~~

~~Co., FL and Liberty Co., FL and clearly is a member o f the distinct eastern clade (Fig~~

~~1.3). E. o. william si also appears to be morphologically grouped with the eastern clade.~~

~~The southern Florida and Florida Keys form, E. o. deckerti, also appears~~

~~morphologically by distinct according to DFA (Table 2.22-2.24). However, it is very~~

~~difficult to assign specimens to that subspecies from the color patterns used to define it~~

~~Numerous specimens in the Florida Keys and southern Florida appear identical to E. o.~~

~~quadrivittata or E. o. rossalleni. The following specimens on the Florida Keys fail to~~

~~have the prominent blotches thought to distinguish E. o. deckerti from the faintly~~

~~patterned E. o rossalleni: FL: Monroe Co.: LSUMZ 34312, 28870; MCZ 12519. In~~

~~addition the red tongue that distinguished E. o. rossalleni from the black-tongued E. o.~~

~~deckerti also fails to be a reliable character. The following specimens within the range~~

~~of E o. rossalleni near the Everglades in Dade Co., FL have either black or mottled red~~

~~and black tongues: UF 113924, 113927; CM 51024. The traditional characters that~~

~~distinguish E. o. quadrivittata, E. o. rossalleni, and E. o. deckerti from one another~~

~~often fail. Genetically, E. o. deckerti forms a clade with the central Florida E. o.~~

~~140~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. quadrivittata. These striped forms are genetically part of the eastern clade and can be~~

~~classified morphologically within that clade (Fig 1.3).~~

~~From the univariate and multivariate analyses, it is apparent that head scale~~

~~shapes and scale counts vary greatly between clades (the specific differences are~~

~~described below). When examining scatterplots of Case 3 and 4 variables, it is evident~~

~~that detailed color pattern measurements aid in defining the bo undries between clades~~

~~more clearly (Fig 2.9, 2.10, 2.13,2.14,2.17, and 2.18). These color pattern~~

~~measurements should not be confused with the original vague color patterns used to~~

~~describe the subspecies. When examined in detail, it is clear that color patterns like DB,~~

~~LB, DBW1, DBL1, LBW1, and LBL1 vary substantially across the geographical~~

~~boundaries that were once thought to delimit subspecies with homogeneous color~~

~~patterns (Table 2.2 and 2.3). Superficially, the color pattern of E. o. lindheimeri was~~

~~considered to be the same on both sides of the Mississippi River. When examined in~~

~~detail, color patterns are clearly different in dorsal blotch number and shape. A similar~~

~~situation arises for E. o. spliloides at the Apalachicola River. By using the traditional~~

~~color characters, gray ground color and dark blotches, it might be assumed that this~~

~~subspecies has the same color pattern on both sides of the Apalachicola River.~~

~~However, the respective patterns present on specimens from east and west of the river~~

~~are very different in shape and number (Table 2.2 and 2.3).~~

~~Based on the findings presented here, it seems likely that subspecies described~~

~~from only a few color characters are unlikely to reflect the evolutionary history of the~~

~~taxa in question, especially in cases where individual subspecies are thought to cross~~

~~known geographic barriers. Examples from North American snakes may include~~

~~141~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Crotalus horridus, Agkistrodon contortrix, Lampropeltis getula, L. triangulum, and~~

~~Coluber constrictor. This study suggests that the taxonomy of a number of~~

~~polymorphic snakes in US may require revaluation.~~

~~Molecular data presented in Chapter 1 strongly suggest that none of the~~

~~currently recognized subspecies represent distinct evolutionary lineages. In agreement~~

~~with these data, it has been shown that most subspecies are also difficult to classify~~

~~morphologically using this data set presented here. Moreover, the traditional characters~~

~~used to describe these subspecies generally fail to adequately diagnose them.~~

~~Therefore, I suggest that the subspecies of E. obsoleta no longer be recognized. The~~

~~four molecular clades appear to represent four independent lineages, and thus form~~

~~distinct evolutionary species, each separated by geographical features known to~~

~~represent barriers to gene flow in other taxa as well (Blair, 1958; Blair, 1965; Wiley and~~

~~Mayden, 1985; Mayden, 1988; Walker etal., 1998). Moreover, the morphological~~

~~evidence reflects the separate evolutionary trends discovered in the sequences of the~~

~~two mtDNA genes. Because the taxonomy of this complex should reflect its~~

~~evolutionary history (Frost and Hillis 1992). I propose the following species names for~~

~~the four groups: eastern clade = Elaphe alleghaniensis-, central clade = E. spiloides;~~

~~western clade = E. obsoleta; E. bairdi = E. bairdi.~~

~~Although the range of values for many characters overlap among E.~~

~~alleghaniensis, E. spiloides, and E. obsoleta, descriptions of morphological trends will~~

~~be discussed below in the accounts for each species. Since the differences between E.~~

~~alleghaniensis, E. spiloides, and E. obsoleta are subtle, particular attention will be~~

~~placed on their various morphological differences. Elaphe bairdi is very distinct~~

~~142~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. morphologically relative to the other three species, and will be discussed only in terms~~

~~of its degree of differentiation from the other three species as a group.~~

~~Elaphe alleghaniensis (Holbrook)~~

~~Coluber alleghaniensis Holbrook, 1836: 111, pi. 20. Type locality: “summit of the Blue~~

~~Ridge in Virginia and Highlands of the Hudson.” (Holotype: ANSP 16792).~~

~~Coluber quadrivittatus Holbrook, 1836: 113-14, pi. 21.~~

~~Scotophis confmis Baird and Girard, 1853: 77~~

~~Coluber obsoletus lemniscatus Cope, 1888: 386 (Part).~~

~~Elaphe quadrivittata deckerti Brady, 1932: 5.~~

~~Elapie williamsi Barbour and Carr, 1940: 340.~~

~~Elaphe quadrivittata parallela Barbour and Engels, 1942: 103.~~

~~Elaphe obsoleta rossalleni Neill 1949, 1.~~

~~Nomenclature: The two oldest names for the type specimens found within the eastern~~

~~clade are Coluber alleghaniensis and C. quadrivittatus (Holbrook 1936). Elaphe~~

~~ailegheniensis is applied to the eastern clade because C. alleghaniensis appears in the~~

~~original edition of North American Herpetology (Holbrook 1836) two pages earlier than~~

~~does C. quadrivittatus. This decision has an added benefit in that E. quadrivittata is~~

~~typically associated with the yellow and black-striped form and would likely be~~

~~incorrectly applied only to those specimens by the layperson. The name E.~~

~~alleghaniensis has not been previously applied to nonmonophyletic subsets of the E.~~

~~obsoleta complex and is therefore less likely to create taxonomic confusion.~~

~~143~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Distribution: This species occurs east of the Apalachicola River in Florida, east of the~~

~~Chattahoochee River in Georgia, east of Appalachian Mountains, north to southeastern~~

~~New York and western Vermont and south to the Florida Keys (Fig 2.37).~~

~~Diagnosis: Elaphe alleghaniensis is a large colubrid with a snout-vent length averaging~~

~~1092 mm for males and 1020 mm for females (Table 2.2 and Table 2.3). The tail length~~

~~averages 242 mm for males and 213 mm for females. The large elongated head of this~~

~~species is well separated from the neck. This semi-arboreal snake has a laterally~~

~~compressed body slightly higher than wide. Both sexes of this species usually have~~

~~eight supralabials, 11 or 12 infralabials, one preocular, two postoculars, and eight or~~

~~nine total temporal scales (Tables 2.2 and 2.3). Keels begin at midbody on scale rows~~

~~three or four for both sexes. Males and females usually have 25 or 27 scale rows at the~~

~~level of the tenth ventral scale, 25 or 27 scale rows at midbody, and 19 or 21 scale rows~~

~~at the level of the penultimate ventral scale. Sexual dimorphism exists for many~~

~~characters, with males having the higher value for most characters except ventral~~

~~number and DBW1 (see Results).~~

~~Elaphe alleghaniensis tends to have a higher number of subcaudals~~

~~(males,mean=89.147; females, mean=82.071) than E.spiloides (males, mean=82.175;~~

~~females, mean= 76.228) and E. obsoleta (males, mean=82.769; females, mean=77.617).~~

~~Along with this character, the ratio of TL/SV is higher inE. alleghaniensis than E.~~

~~spiloides or E. obsoleta (Table 2.25 and 2.26/ When dorsal blotches are present, E.~~

~~alleghaniensis has a higher number (males, X=34.993; females, X=35.035) thanE.~~

~~spiloides (males, X=30.523; females, X=29.763) and E. obsoleta (males, X=31.353;~~

~~females, X=30.439). Rat snakes with smaller dorsal blotches have high numbers of~~

~~144~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. K. alleghaniemi.K. Fig Fig 2.37. Geographic distribution of~~

~~145~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission blotches. The DBW1 and DBL1 in E. alleghaniensis is smaller than E. spiloides and E.~~

~~obsoleta. Although not as significant, E. alleghaniensis has a higher number of shorter~~

~~lateral blotches than E. spiloides and E. obsoleta~~

~~The variation in head scale characters is noticeable between these three species~~

~~and is better viewed as a ratio with head length as the common denominator (Table 2.25~~

~~and 2.26.). Elaphe alleghaniensis differs from E. spiloides and E. obsoleta in having a~~

~~much shorter rostral height (RH), a narrower head, and longer muzzle. The narrower~~

~~head is reflected in the smaller size of the following dorsal head scales in males:~~

~~PW/HL, FW/HL, FWP/HL, PRFWA/HL, PRFWP/HL, INWA/HL, INWP/HL, and~~

~~INR/HL. These characters tend to be smaller in females as well, but with the addition~~

~~of FWA/HL. The following length variables are marginally shorter in both sexes o f E.~~

~~alleghaniensis : PL/HL, PRFL/HL. RW (as determined by the ratio: RW/HL) is~~

~~narrower and the LL (as determined by the ratio: LL/HL) is shorter than in the other~~

~~Elaphe species in both sexes of E. alleghaniensis. Differences between E.~~

~~alleghaniensis and E. bairdi are discussed in the E. bairdi species account.~~

~~Color Variation: Numerous color patterns can be found in this species. This species~~

~~may be black with no pattern, yellow with black or brown stripes, orange with little~~

~~evidence of any pattern, gray with brown or black blotches or have combinations of all~~

~~of these color patterns. The darkest adult specimens are found in eastern New York,~~

~~western New Hampshire, Connecticut, New Jersey, Maryland, eastern Virginia, North~~

~~and South Carolina (excluding the eastern coasts), and northern and central Georgia.~~

~~These specimens usually have black or dark brown ground colors. The dorsal pattern~~

~~may be visible as blotches on the dark brown ground color, or as white flecks that once~~

~~146~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. composed the border of faded blotches on a black ground color. Other specimens in the~~

~~northern part of the range are completely black with no trace of color. The patterned~~

~~specimens often have a series of medial dorsal blotches and alternating lateral blotches~~

~~on the side of the body, with most visible blotches usually occurring at midbody. The~~

~~dark northern specimens usually have two rows of ventral blotches that occupy two or~~

~~three scales and are visible anteriorly, but connect medially and produce an almost~~

~~black venter before midbody. Lateral blotches that occupy two or three scales are also~~

~~present on the anterior half of the venter. By midbody, these lateral blotches are usually~~

~~not visible due to the dark venter. The ventral surface o f the tail is usually black, but~~

~~some specimens retain a light medial stripe that is often present in juveniles. These~~

~~dark individuals usually have a black or brown head with white supralabials and~~

~~infralabials. Prominent dark bars are often found on the first six supralabials and the~~

~~first eleven, twelve, or thirteen infralabials. The ventral surface of the head is usually~~

~~white.~~

~~Specimens in southeastern Virginia often show the presence of dorsal and lateral~~

~~black stripes on a dusky dark gray or brown dorsal surface (Mitchell 1994). This~~

~~pattern appears to be intermediate between the dark northern specimens and the dark-~~

~~striped, olive, tan or yellow E. alleghaniensis of coastal North Carolina, South Carolina,~~

~~Georgia and coastal and central Florida. The intermediate color patterns from~~

~~individuals from North Carolina are described in detail by Braswell (1977). They~~

~~appear to have a dusky gray or olive ground color with two dorsal dark brown or black~~

~~stripes and two lateral brown or black stripes on either side of the body. The dorsal~~

~~stripes run from the neck to tail tip, whereas the lateral stripes begin at the neck and end~~

~~147~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. at the level of the cloaca. In addition, dorsal blotches are present in many specimens,~~

~~but the lateral blotches are often obscured by the dark lateral stripe. They tend to> have~~

~~diffuse or peppery medial ventral blotches that connect and form two rows from~~

~~midbody to the tail. Anteriorly, these blotches occupy two or three scales. Lateral~~

~~ventral blotches also occupy two or three scales and are obscured by the dark dorsum by~~

~~midbody. A prominent light medial line may be found on the ventral surface of the tail.~~

~~The head of these specimens appears dark olive and has dark labial bars that are similar~~

~~to those of black specimens in the northern part of their range.~~

~~The dusky, striped color pattern occurring in specimens from central North~~

~~Carolina, South Carolina and Georgia eventually grades into a lighter tan or yellow~~

~~form known from southeastern North Carolina, coastal South Carolina, coastal Georgia,~~

~~and northwestern and central Florida. Dusky gray or brown dorsal blotches are often~~

~~visible in these specimens. The ventral surface is usually much lighter on the anterior~~

~~third of the body. Light peppering or dark cresents formed on the posterior margin of~~

~~each scale may be evident from midbody to the tail. Dark ventral blotches may also be~~

~~present in some specimens. The lateral ventral blotches usually occupy two to three~~

~~scales and begin before midbody. By midbody, these lateral blotches usually join to~~

~~form a stripe that runs to the tip of the tail, but may not be present in all specimens. The~~

~~head and labial barring is often very light and may be absent in some individuals.~~

~~In southern Florida, the dorsal ground color may be orange or rust Specimens~~

~~may have completely immaculate, peppered, or blotched venters. Dorsal and lateral~~

~~blotches may be present or absent In addition, striping may also be absent or show no~~

~~148~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. color pattern. Specimens from the Florida Keys may be strongly blotched and appear~~

~~similar to the northern blotched black rat snakes.~~

~~In Levy Co. and Northern Citrus Co., Florida, E. alleghaniensis have a gray~~

~~dorsal ground color with black or brown blotches and stripes. This pattern grades into~~

~~the lighter yellow and striped snakes of central and northwestern Florida. They usually~~

~~have a blotched or peppered venter. Lateral ventral blotches are often clearly visible~~

~~from the neck to the cloaca and the tail may retain the light medial line displayed in the~~

~~juveniles. The gray head may be heavily peppered with small black flecks. The~~

~~anterior labial scales are often marked with large black bars.~~

~~To the west, from Taylor Co., to the Apalachicola River and Chatahoochee~~

~~River through central Georgia, the stripes are absent, but the gray ground color, dark~~

~~blotches, peppered head, blotched or peppered venter, and barred labials remain. These~~

~~specimens appear similar to the striped specimens from Levy Co., Florida. This gray~~

~~and dark blotched form grades into the black form of central and northern Georgia. A~~

~~postocular stripe that runs through the postocular scales, the first row of temporal~~

~~scales, and the seventh or eighth supralabial scale is present in most specimens. In~~

~~addition, a dark stripe may be present at the margin of the prefrontal and frontal scales.~~

~~The numerous adult color patterns found in Elaphe alleghaniensis grade into~~

~~one another over large areas. Furthermore, color patterns thought to be characteristic of~~

~~particular regions often occur in regions thought to be inhabited exclusively by rat~~

~~snakes with completely different color patterns, indicating that the color patterns are not~~

~~representative of natural groups. Given the geographically mosaic nature of E.~~

~~149~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. alleghaniensis color patterns, the exact boundary of any color pattern could not be~~

~~determined with accuracy.~~

~~Elaphe spiloides (Dumeril, Bibron, and Dumlril)~~

~~Elaphis spiloides Dumeril, Bibron, and Dumeril, 1854: 269. Type locality: “La~~

~~Nouvelle-Orleans”=New Orleans, Louisiana. (Holotype: MNHN 827).~~

~~Coluber obsoletus lemniscatus Cope: 1888: 386. (Part).~~

~~Nomenclature: The oldest type specimen found within the central clade is Elaphis~~

~~spiloides. Bianey (1971) indicated that the holotype could not have originated from~~

~~New Orleans, Louisiana, because the subspecific color pattern exhibited by the former~~

~~E. o. spiloides does not exist in southeastern Louisiana. The holotype was measured for~~

~~all characters listed in Table 2.1 and statistically analyzed. DFA clearly places the~~

~~holotype within the central clade (E. spiloides). Therefore, the holotype must have~~

~~originated within the range of the central clade.~~

~~Distribution: This species occurs east of the Apalachicola River and the Appalachian~~

~~Mountains and west of the Mississippi River. It is found south from Louisiana along~~

~~the Gulf Coast of the US to Florida, and north in western New York, east through~~

~~southern Ontario, Canada, southern Michigan, northeastern Indiana, central Illinois and~~

~~southwestern Wisconsin (Fig 2.38).~~

~~Diagnosis: Elaphe spiloides is a large colubrid with a snout-vent length averaging 1097~~

~~mm for males and 1007 mm for females (Table 2.2 and Table 2.3). The tail length~~

~~averages 229 mm for males and 195 mm for females. Like E. alleghaniensis , this~~

~~species has a large elongated head that is well separated from the neck. This semi-~~

~~150~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. spiloides. r Fig. Fig. 2.38. Geographic distribution of/i~~

~~151~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. arboreal species also has a laterally compressed body that is slightly higher than wide.~~

~~Males have higher values than females for most measurements and scale counts (see~~

~~Results). However, females have higher infralabial and temporal counts. Both sexes~~

~~usually have eight supralabials, 11 to 14 infralabials, one preocular, two postoculars,~~

~~and eight to ten temporal scales (Table 2.2 and Table 2.3). At midbody, keels begin on~~

~~scale row three or four for both sexes. Males and females usually have 25 or 27 scale~~

~~rows at the level of the tenth ventral scale, 25 or 27 scale rows at midbody, and 19 or 21~~

~~scale rows at the level of the penultimate ventral scale.~~

~~Elaphe spiloides differs from E. alleghaniensis by having fewer subcaudals,~~

~~fewer dorsal blotches, and a larger DBW, DBL, LBW, and LBL (Table 2.2 and 2.3).~~

~~This species generally has a smaller DBW and DBL than E. obsoleta. Although there is~~

~~considerable overlap in the ranges of mensural characters, E. spiloides tends to have a~~

~~broader head than E. alleghaniensis and E. obsoleta as reflected in the head scale~~

~~measurements. Male and female E. spiloides have larger values than E. alleghaniensis~~

~~for the following ratios: PL/HL, PW/HL, FW/HL, FWP/HL, PRFWA/HL, PRFWP/HL,~~

~~INWA/HL, INWP/HL, EYE/HL, AG/HL, INR/HL, RH/HL, RW/HL, ANL/HL,~~

~~PROW/HL, LHT/LV, LL/HL (Table 2.25). Female E. spiloides also have larger values~~

~~than E. alleghaniensis for the following ratios: FL/HL, FWA/HL, PRFL/HL, NR/HL,~~

~~PG/HL, PNL/HL (Table 2.26). Male and female E. spiloides have larger ratio~~

~~measurements than E. obsoleta for the following characters: PW/HL, PWP/HL, FL/HL,~~

~~FW/HL, FWP/HL, PRFWA/HL, INL/HL, INWA/HL, EYE/HL, AG/HL, PG/HL,~~

~~INR/HL, NR/HL, RH/HL, RW/HL, PNL/HL, PROW/ HL, PROLD/HL, PROLV/ HL,~~

~~152~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. LL/HL, and LH/HL (Table 2.25). Female E. spiloides tend to have higher values than~~

~~E. obsoleta for the following ratios: PRFWP/HL, ENWP/HL, and ANL/HL.~~

~~Color Variation: Elaphe spiloides does not vary greatly in the degree to which~~

~~specimens are blotched or completely black. Unlike, E. alleghaniensis , this species~~

~~does not have populations characterized by a striped color pattern. Most dark~~

~~specimens are found in the northern part of the range: northeastern Alabama to western~~

~~Pennsylvania and western New York across central and Northern Ohio, Indiana,~~

~~Michigan, Illinois, and Wisconsin. It should be stressed that this is only a trend in color~~

~~pattern and that clearly blotched specimens are found well within those States. The~~

~~black specimens usually retain some blotching well into adulthood, and traces of red,~~

~~orange or yellow may be found on the skin between dorsal scales. The head is usually~~

~~dark olive, brown, or black with dark supralabial bars. Medial blotches, dark flecking,~~

~~or peppering often occur before midbody on the venter. These markings may be visible~~

~~to the tail or obscured by a heavy wash of black pigment that often occurs over the~~

~~posterior half of the body. Lateral blotches on the ventral surface occupy two or three~~

~~scales and are usually obscured by the dark ventral pigmentation beginning before~~

~~midbody. Many specimens have a light medial line on the tail. The amount of ventral~~

~~blotching and black pigmentation is variable in dark northern specimens.~~

~~Distinctly blotched specimens are found throughout the ranged of this species,~~

~~but predominate in Louisiana, Mississippi, Alabama, Florida, western Kentucky,~~

~~western Tennessee, southern Illinois and southern Indiana. Near the Mississippi River,~~

~~specimens often have a tan or brown ground color. This tends to become lighter tan or~~

~~yellow in eastern Louisiana, yellow in Mississippi and eastern Alabama, and gray in~~

~~153~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. eastern Alabama and western Florida. However, individuals with a dark brown ground~~

~~color do occur in eastern Louisiana and Mississippi. The ground color is usually darker~~

~~in the northern part of their range. The blotches are usually black, dark brown, or dark~~

~~olive. The skin between the dorsal scales may be bright orange, red, yellow, brown, or~~

~~gray- The head is usually gray, olive, brown or black. Specimens in eastern Alabama~~

~~and western Florida usually have a gray head that is very heavily peppered or flecked~~

~~with dark pigment Blotched specimens may also retain a postocular stripe that runs~~

~~through the postocular scales, the first row of temporal scales, and the seventh or eighth~~

~~supralabial scale. In addition, a dark stripe may be noticeable at the margin of the~~

~~preffontal and frontal scales. Supralabial barring is also common in these blotched~~

~~specimens. Light, diffuse, peppery or solid black blotches are very common on the~~

~~ventral surface. The blotches usually occur before the 30th ventral scale and are often~~

~~visible to the cloaca. Occasionally, the ventral surface is heavily mottled posterior to~~

~~midbody obscuring the blotches. Blotches in some individuals appear as dark crescents~~

~~centered at the posterior margin of each ventral scale. Most specimens have a light~~

~~medial line on the ventral surface o f the tail.~~

~~Patterned and unpatterned dark specimens occur randomely throughout the~~

~~northern range of this species. Blotched individuals occur as far North as Wisconsin,~~

~~and very black individuals occur as far south as southern Louisiana. Many populations~~

~~considered to be black have distinctly blotched patterns on a dark brown dorsum.~~

~~Elaphe obsolete (Say)~~

~~Coluber obsoletus Say (in James), 1823: 140. Type Locality: “On the Missouri River~~

~~from the Vicinity Isle au Vache to Council Bluff.” This locality extends from~~

~~154~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. near Leavenworth, Kansas to Council Bluffs, Iowa. (Holotype: Presence is~~

~~unknown.)~~

~~Scotophis laetus Baird and Girard, 1853: 1977.~~

~~Scotophis lindheimeri Baird and Girard 1853: 74.~~

~~Nomenclature: Although missing, the oldest type specimen from the western clade is~~

~~Coluber obsoletus, collected somewhere between Cow Island (near Leavenworth),~~

~~Kansas and Council Bluffs, Iowa. This area is clearly within the range of the western~~

~~clade and the name Elaphe obsoleta should be applied to all members of that clade.~~

~~Distribution: Occurs west of the Mississippi River from south Louisiana along the Gulf~~

~~Coast to south Texas, west to central Texas on the Edward’s Plateau, and North through~~

~~Oklahoma, central and eastern Kansas, southeastern Nebraska, southeastern Iowa, and~~

~~extreme southeastern Minnesota (Fig 2.39).~~

~~Diagnosis: Elaphe obsoleta is similar to E alleghaniensis and E. spiloides. The~~

~~average adult snout-vent length is 1078 mm for males and 1002 mm for females (Table~~

~~2.2 and Table 2.3). The tail length averages 225 mm for males and 196 mm for~~

~~females. The head is large and elongated and is well separated from the neck. This~~

~~semi-arboreal snake also has a laterally compressed body that is slightly higher than~~

~~wide. Males tend to have a higher value for most characters (see Results). However,~~

~~females have a higher ventral scale count and a higher number of scale rows at the level~~

~~of the penultimate ventral scale (Table 2.2 and Table 2.3). Both sexes usually have~~

~~eight supralabials, 11 to 14 infralabials, one preocular, two postoculars, and ten or 11~~

~~temporal scales (Table 2.2 and Table 2.3). Keels begin at midbody on scale row three~~

~~or four for both sexes. Males and females have 25 or 27 scale~~

~~155~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. K. obsoleta.K. Fig. Fig. 2.39. Geographic distribution of~~

~~156~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. rows at the level of the tenth ventral scale, 25 or 27 scale rows at midbody, and 19 or 21~~

~~scale rows at the level of the penultimate ventral scale.~~

~~Elaphe obsoleta tends to have fewer subcaudals, fewer dorsal blotches and~~

~~fewer lateral blotches thanE. alleghaniensis (Table 2.2 and 2.3). Elaphe obsoleta tends~~

~~to have a larger DBW1, DBL1, and LBW1 than E. alleghaniensis and E. spiloides.~~

~~Compared to E. alleghaniensis, male E. obsoleta have larger ratio measurements for the~~

~~following characters. HL/SV, PL/HL, PRFL/HL, EYE/HL, AG/HL, RH/HL, ANL/HL,~~

~~LD/HL (Table 2.25). Most of these ratios are larger in females except for EYE/HL and~~

~~AG/HL, which are considerably smaller (Table 2.26). FW/HL, FWA/HL, and~~

~~PRFWP/HL are larger in female E. obsoleta than in female E. alleghaniensis. For both~~

~~sexes, most other characters tend to be of similar size or smaller than in E.~~

~~alleghaniensis (see E. alleghaniensis species description). Most mensural ratio~~

~~measurements are smaller in male E. obsoleta than E. spiloides except HL/SV, PL/HL,~~

~~FWA/HL, and LD/HL. Only HL/SV and LD/SV are larger in female E. obsoleta than~~

~~in female E. spiloides. The characters that are larger inE. spiloides than in E. obsoleta~~

~~are discussed in the E. spiloides account.~~

~~Color Variation: The color patterns in E. obsoleta are very similar to those found in E.~~

~~spiloides. Black E. obsoleta, with little pattern, are common in the following northern~~

~~parts of their distribution: northwestern Louisiana, central and western Arkansas, central~~

~~and western Oklahoma, Kansas, Nebraska, Missouri, Iowa and Minnesota. However,~~

~~heavily blotched specimens with black blotches on a dark brown ground color may also~~

~~be found in the northern regions. Specimens throughout the range of E. spiloides have~~

~~157~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. red, yellow, or orange colored skin between the dorsal scales. Very light flecks that~~

~~border the blotches in juveniles can often be seen in the dark black adult specimens.~~

~~Lateral ventral blotches usually occupy two or three scales and are obscured by the dark~~

~~ventral pigment just anterior to the midbody. Medial ventral markings may be in the~~

~~form of two connected rows of blotches, random dark peppering, or dark crescents~~

~~along the margin of each ventral scale. The ventral surface often becomes completely~~

~~black by midbody. Occasionally, specimens within the range of these dark specimens~~

~~have very light unpattemed venters. A light medial line on the ventral surface of the tail~~

~~is present in many individuals. The head is usually very black without any postocular~~

~~striping. Bars may be present or absent on the labial scales.~~

~~Blotched individuals occur in central and southern Louisiana, Texas, near the~~

~~Mississippi River in Tennessee, Arkansas, Missouri, Iowa, and Minnesota, most of~~

~~Oklahoma and southern Kansas. These blotched individuals often have a tan, brown, or~~

~~dark brown ground color and black or olive blotches. The blotches are often~~

~~ornamented with minute white flecks. The interstitial skin color may be orange, red,~~

~~yellow, or gray. Many specimens have blotched medial ventral surfaces that continue~~

~~uninterrupted to the tail. Often, individuals in western Louisiana, Texas, and Oklahoma~~

~~have the medial crescent pattern on the venter. A light medial line is also common in~~

~~many the patterned E. obsoleta. A postocular stripe and a bar along the posterior~~

~~margin of the prefrontal are visible in some specimens. Prominent dark labial barring is~~

~~present in most specimens, but labial barring may be faint or absent in some individuals.~~

~~Patterned and unpattemed black specimens occur sporadically throughout the~~

~~northern range of this species. Like Elaphe spiloides, it would be very difficult to~~

~~158~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. classify all members of this highly variable species as either purely black or completely~~

~~patterned.~~

~~Elaphe bairdi ( Yarrow)~~

~~Coluber bairdi Yarrow (in Cope), 1880: 492. Type Locality: Fort Davis, Apache~~

~~Mountains, Jeff Davis Co.: Texas. (Holotype: USNM 10403).~~

~~Nomenclature: This taxon was elevated to species status by Olson (1977), and no~~

~~confusion exists with respect to the name associated with the holotype.~~

~~Distribution: Southwestern Texas from the Edward’s Plateau to the northeastern~~

~~buttresses of the Sierra Madre Oriental to southern Nuevo Leon, Mexico (Shultz 1996)~~

~~(Fig. 2.40).~~

~~Diagnosis: Some of the morphological distinctions between E. bairdi and the other~~

~~members of the E. obsoleta complex have been described in the literature (Olson 1977;~~

~~Lawson and Lieb 1991). However, with Tables 2.2,2.3,2.25,2.26 and the descriptions~~

~~in this section, I will expand what is known regarding the morphology of this species.~~

~~Elaphe bairdi is a generally smaller and less robust than E. alleghaniensis , E. spiloides,~~

~~and E. obsoleta. Adult males have an average snout-vent length of 1004 mm and~~

~~females have an average snout-vent length o f927 mm. The head is distinct and well~~

~~differentiated from the body. Like the other Elaphe examined here, it has a laterally-~~

~~compressed body that is slightly higher than wide. Very little sexual dimorphism can~~

~~be found in E. bairdi. Males tend to have more subcaudals and a larger DBL1. Elaphe~~

~~bairdi has eight or nine supralabials, 13 or 14 infralabials, one preocular, and two~~

~~postoculars. Unlike the other E laphe examined here, the first keel at midbody begins on~~

~~the sixth dorsal scale row in adults. This is a diagnostic character for E. bairdi thatE.~~

~~159~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. E. E. bairdi.~~

~~i z '~~

~~1/ Fig. 2.40. Geographic distribution of~~

~~160~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. alleghaniensis, E. spiloides and E. obsoleta do not share. Males and females usually~~

~~have 25,27, or 29 scale rows at the level of the tenth ventral scale; 25,27 or 29 scale~~

~~rows at midbody; and 19 or 21 scale rows at the level of the penultimate ventral scale.~~

~~Elaphe bairdi usually has a higher number of ventrals, subcaudals, temporal~~

~~scales, dorsal blotches, and lateral blotches than the other three species (Table 2.2 and~~

~~2.3). The size o f the DBW1, DBL1, LBW1, and LBL2 is much smaller in this species.~~

~~The ratio, TL/SV, is also much larger in E. bairdi. The following ratios are smaller in~~

~~male E. bairdi compared to the other three Elaphe: PW/HL, PWP/HL, FW/ HL, PRFL/~~

~~HL, PRFWA/HL, PRFWP/HL, INWA/HL, DMWP/HL, PG/HL, INR/HL, RH/HL,~~

~~RW/HL, PROW/HL, LHT/HL, and LH/HL. These values are similar for females~~

~~except LHT/HL is not significantly smaller in E. bairdi than in the other three species.~~

~~The following characters tend to be much larger in both sexes of E. bairdi compared the~~

~~other Elaphe examined in this paper PL/HL, FL/HL, INL/HL, AG/ HL, NR/ HL,~~

~~PROLD/ HL, LV/HL. The small values for INL/HL, INWA/HL, INWP/HL, INR/HL,~~

~~RH/HL, and RW/HL and large values for INL/HL demonstrate that the muzzle in E.~~

~~bairdi is longer and narrower than E. alleghaniensis. E. spiloides, and E. obsoleta.~~

~~Color Variation: Elaphe bairdi tend to be more uniformly patterned than the other~~

~~three species. However, the ground coloration exhibits substantial intraspecific~~

~~variation. In some individuals, the ground color varies from brown to orange anteriorly,~~

~~and fades to gray or silver before the tail (Schultz 1996). In other specimens, a silver or~~

~~bluish-purple ground color occurs over the entire length of the body. Two dark dorsal~~

~~stripes run from the neck to the tail tip and two lateral stripes run from the neck to the~~

~~level of the cloaca. In some specimens, the stripes and/or blotches are very poorly~~

~~161~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. defined, and when blotches are present, they tend to be veiy light and difficult to see.~~

~~Medial ventral blotches usually occur before the first ventral scale in E. bairdi , but do~~

~~not often occur before the first ventral scale in the other three species of Elaphe~~

~~examined here. The medial blotches often become diffuse and form two diffuse rows~~

~~that run from midbody to the tail tip. A light postocular mask may be present in some~~

~~specimens. If barring on the labial scales is present, it is usually light.~~

~~162~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Summary and Conclusion~~

~~As demonstrated by the maximum likelihood and maximum parsimony~~

~~phylogenetic analysis of two complete mitochondrial gene sequences, the North~~

~~American rat snake complex {Elaphe obsoleta) is composed o f four distinct~~

~~evolutionary lineages. Each of the four lineages occurs in a specific geographic area: 1)~~

~~the eastern clade is located east of the Apalachicola River and the Appalachian~~

~~Mountains, 2) the central clade is located west of the Apalachicola River and the~~

~~Appalachian Mountains and east of the Mississippi River, 3) the western clade is~~

~~located west of the Mississippi River, and 4) Elaphe bairdi is located in southwest~~

~~Texas and northeastern Mexico. With respect to this phylogeographic analysis, the~~

~~former subspecies of Elaphe obsoleta do not represent distinct evolutionary lineages.~~

~~Because of reduced genetic variability ineach of the three clades of E. obsoleta, it~~

~~appears that populations within each of these three lineages expanded northward from~~

~~southern glacial refugia quite recently.~~

~~Univariate and multivariate analyses of 67 morphological characters scored~~

~~from 1006 specimens provided additional statistical support for the recognition of the~~

~~same four evolutionary lineages identified in the phylogeography study. Specimens can~~

~~be classified morphologically by using canonical discriminant function analysis into the~~

~~four molecular clades more accurately than they can be grouped into subspecific~~

~~categories. Moreover, the identification of these subspecies proved difficult when using~~

~~the traditional characters ascribed to them.~~

~~163~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. In light of the corroborating molecular and morphological evidence, I suggested~~

~~that the recognition of the subspecies of Elaphe obsoleta be discontinued. Instead, the~~

~~four molecular clades should be recognized as four species: 1) eastern clade=£.~~

~~alleghaniensis , 2) central clade = E. spiloides, 3) western clade = E. obsoleta, and 4) E.~~

~~bairdi — E. bairdi.~~

~~164~~

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~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix A~~

~~Tissue Examined~~

~~All tissues are from the United States. The first number is equal to the locality number~~

~~in Fig. 2. The following abbreviations in parentheses refer to tissues obtained from the~~

~~following: H = Louisiana State University Museum of Natural Science Herpetological~~

~~Tissue Collection, Band RLB = Blood From Living Snake taken by R. L. Lawson, CAS~~

~~= California Academy of Sciences Tissue Collection, FB= Shed Skins.~~

~~( I )RI: Washington Co. (FB-9); (2)CT: Middlesex Co.:Split Rock State Park (H-15889);~~

~~(3) CT: Middlesex Co.: Haddam (H-15890); (4)NY: Orange Co.: Port Jervis (H-8455); 5~~

~~MD: Frederick Co.: Brunswick (H-9349); (6) VA; Loudon Co.: Middleburg (H-3191);~~

~~(7) VA: Rockingham Co. (FB-14); (8) NC: Perquimans Co.: Hartford (H-8825); (9) NC:~~

~~Wake Co.: Raleigh (H-8826); (10) SC: Williamsburg Co. (FB 9); (11) SC: Jasper Co..~~

~~HardeeviHe (H-3384); (12) GA. Chatham Co.. Basin Road (old Hwy 17) (H-9475); (13)~~

~~FL: Putnam Co.: Rodman Resevoir, State Rd 310 at Deep Creek (H-15884); (14) FL:~~

~~Alachua Co.: Gainesville (H-3377); (15) FL: Broward Co.: Fort Lauderdale (H-3189);~~

~~(16) FL: Dade Co.: SW of Miami, Krone Ave (H-2229); (17) FL: Monroe Co.: Key~~

~~Largo, Tavemier (B13p35a); (18) FL: Monroe Co.: Key Largo, Tavemier (B13p35b);~~

~~(19) FL: Sarasota Co.: Sarosota City (FB-20): (20) FL: Pinnelas Co.: St. Petersburg,~~

~~Tampa Bay, Placido Bayou (H-7783); (21) FL: Hernando Co.:~~

~~173~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Brooksville, Hancock Lake (H-15028); (22) FL: Levy Co.: (B13p35c); (23) FL: Levy~~

~~Co.: HWY 24 and 345 (H-8775); (24) FL: Taylor Co.: Peny (H-3212); (25) Florida:~~

~~Wakulla Co.: US 98 and 319 (CAS 203079); (26) FL: Liberty Co., FL HWY 13 on the~~

~~west side of Ochlokonee River (CAS-203083); (27) FL: Walton Co.: SE of Freeport (H-~~

~~3276); (28) FL: Santa Rosa Co.: Blackwater River State Forest (H-3309); (29) AL:~~

~~Baldwin Co.: Bay Minnette (H-3190); (30) MS: Forrest Co.: Brooklyn (H-3186); (31)~~

~~MS: Hinds Co. (FB-3); (32) LA: Orleans Par.: New Orleans (H-8838); (33) LA: S t~~

~~Tammany Par 1.; Southern part of Parish at Pearl River, (34) LA; St (H-3379)~~

~~Tammany Par 2.: Abita Creek Preserve (H-15888); (35) LA: Tanghipahoa Par. 1:~~

~~Natlbany River and Tickfaw River (H-3188); (36) LA: Tangipahoa: Par. 2: Hammond~~

~~(H-3246); (37) LA: Terrebonne Par. 1: Houma; (H-8473); (38) LA: Terrebonne Par.2:~~

~~Houma (H-9338); (39) LA: East Feleciana Par.: Clinton (H-3209); (40) LA: East Baton~~

~~Rouge Par. 1: Baton Rouge (H-3169); (41) LA: East Baton Rouge Par 2.: Baton Rouge~~

~~(H-3306); (42) LA: Iberville Par.: Plaquemine (H-8824); (43) LA.St. Landry Par.: Hwy~~

~~71 (H-3280); (44) LA: Evangeline: Hwy 106 at Bayou Chico (H-15891); (45) LA:~~

~~Cameron Par..Johnson’s Bayou (H-8925); (46) LA: Natchitoches Par.: Derry (H-~~

~~15892); (47) TX: Comal Co.: New Braunfels (H-8678); (48) TX: Medina Co.. South o f~~

~~Medina Lake; (49) TX. Kerr Co.: 16 mi. South of Kerrville (H-8911); (50) TX:~~

~~Jefferson Davis Co.: 1 mi. east of McDonald Observatory (H-3381); (51) TX: Jefferson~~

~~Davis Co.: 1 mi. east of McDonald Observatory (H-3382); (52) TX: Palo Pinto Co.:~~

~~Brazos River (H-7572); (53) OK: Cleveland Co.: Vicinity of Norman (H-9337); (54)~~

~~KS: Sumner Co.: T33S, R1W, Sec. 6 (H-3394); (55) KS: Geary Co: T13S, R5E, Sec. 29~~

~~(H-3388); (56) MO: Greene Co: Springfield (CAS-162004); (57) AR: Madison Co.: 0.5~~

~~174~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. mi. east of Beaver Lake (H-15896); (58) AR: Garland Co.: Lake Little Switzerland CH-~~

~~14782); (59) AR: Craighead Co.: Jonesboro (H-14781); (60) IL: Pike Co.: 39°42’53”N,~~

~~90°39’16”W (H-9251); (61) IL: St Claire Co.: 8.5 km southeast of Mascoutah~~

~~(H14724); (62) IL: Gallatin Co.: 7 km north of Old Shawneetown (H-15031); (63) IL:~~

~~Johnson Co.: 2.5 mi. northeast of Belknap (H-15030); (64) TN: Decatur Co.: 1.5 mi.~~

~~northwest of Decaturville (H-3206); (65) TN: Grundy Co.: Savage Rd west of Co. Rd~~

~~399 (H-2286); (66) AL: Madison Co: Owana Cross Roads (H-8827); (67) AL: Talladega~~

~~Co.. 1.1 mil. South of Rendalia (H-3385); (68) AL: Lee Co.: Hwy 280 at Hwy 147 (H-~~

~~3345); (69) TN: Knox Co.: Knoxville (H-3376); (70) OH: Delaware Co.: Delaware (FB-~~

~~6); (71) OH: Stark Co.: Canton (CAS 208631 RLB13P27F); (72) WV: Wood Co. CH-~~

~~7572).~~

~~175~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B~~

~~Oligonucleotide Primers Used for Amplification and Sequencing of Elaphe vulpina, £ bairdi, and £obsoleta Cytochrome b and Control Regions~~

~~Amp. = amplification, seq. = sequencing. CR = control region. Note that HI6064 and~~

~~LI6090 are complimentary. Primer numbering refers to the 3' end when aligned with~~

~~Dinodon semicannatus sequence (Kumazawa el al., 1998).~~

~~Primer Primer Sequence Use Reference~~

~~Cytochrome b~~

~~L14910 5’-GAC CTG TGA TMT Amp. only This study~~

~~GAA AAC CAY CGT TGT-3’~~

~~L14919 5’-AAC CAC CGT TGT Amp. seq. This study~~

~~TAT TCA ACT-3’~~

~~LI 5324 5’-CCA TGA GGA CAA Seq. only This study~~

~~ATA TCA TTC-3’~~

~~LI 5584 5’-TCC CAT TYC ACC Seq. only This study~~

~~CAT ACC A-3’~~

~~LI5399 5’-TTA ATT GAG AAT Seq. only This study~~

~~CCGCC-3’~~

~~HI 6064 5’-CTT TGG IT T ACA Amp. Seq.This study~~

~~AG A ACA ATG CTT TA-3’~~

~~176~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Control Region~~

~~H690 5’-GTT GAG GCT TGC Amp. Seq. Kumazawa et al.~~

~~ATG TAT A-3’ 1996~~

~~L16090 5’-TAA AGC ATT GTT~~

~~CTT GTA AAC CAA AG-3’ Amp. seq. This study~~

~~H16602 5’-TTC CGG GCC ATT Seq. only This study~~

~~AAG ATG-3’~~

~~L16577 5’-GTT CTT TCC AAG Seq. only This study~~

~~ACC GCT-3’~~

~~177~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix C~~

~~Morphological Specimens Examined.~~

~~Elaphe alleghaniensis~~

~~Connecticut- Fairfield Co.: AMNH 17447. Hartford Co.: AMNH 119308, 119659,~~

~~130608, 130610-1, 134274-5, 136729. Litchfield Co.: AMNH 97248, 125047, 142245.~~

~~Middlesex Co.: AMNH 125048, 128045. New Haven Co.: AMNH 119309-11, 125049,~~

~~125050, 128046, 125051, 134277, 142652.~~

~~Florida- Alachua Co.: CAS 18200; LSUMZ 7469,42789,46945, 58198,58511; MCZ~~

~~19140; UF 2106,2140, 2149, 2734,2762-3,2976,4791, 8492,9205, 9825,12098,~~

~~14063, 14206, 14351, 14460-1, 16433,16460, 19276, 19330-1, 19333,21677,34002,~~

~~49261,49264,49329,49335,49344,49348-9,4944-4,49447,64983, 64987,64991-2,~~

~~64994, 65122,65546,65548,65550,65554,65562,65563,67532, 70563, 78860,~~

~~79974, 113265. Broward Co.: LSUMZ 57697, 57722. Citrus Co.: CAS 169854;~~

~~LSUMZ 80180; UF 21684,65551, 113277. Collier Co.: CM 26939, KU 145879,~~

~~176726; SM 13441; UF 16442, 21657,62891,65559,73172, 80024, 84482-4, 113280,~~

~~113281, 113283,113284. Columbia Co.: LSUMZ 58509. Dade Co.: ANSP 31324; CAS~~

~~204789; CM 37248,46740, 51024,66525; FMNH 204110-1; KU 68915; LSUMZ~~

~~18973; MCZ 12519,31785,42999,69125,180288 R-140321, R-140322; MPM 26221,~~

~~26222; UF 19273,65565,65567, 113292-3, 113295-9, 113300, 113303, 113924.~~

~~178~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. DeSoto Co.: MCZ 42394. Dixie Co.: CAS 192090; UF 16441, 16445,54164, 113306,~~

~~113307, 113309, 113310, 113312, 113313, 54163A, 54163B. Duval Co.: LSUMZ~~

~~58512; MCZ 6865,96679. Gilchrest Co.: LSUMZ 28868; UF 12091,64608, 113349.~~

~~Glades Co.; MCZ 170334. Hernando Co.: CAS 169468; LSUMZ 59642. Hillsboro Co.:~~

~~MCZ 6685. Indian River Co.; MCZ 39871-3, 168512. Leon Co.: AMNH 102321. Levy~~

~~Co.: CAS 175026; FMNH 245680; LSUMZ 7144, 7260, 28986; MCZ 46147-9,46301,~~

~~46302; UF 2824, 11142, 15960, 36522, 64340, 65561,65565, 113379, 11380-5,~~

~~113387, 113396-9,113400; UGAMNH 6370. Manatee Co.: CAS 192084-5. Marion~~

~~Co.. MCZ 12972, 12973,42392. Martin Co.: LSUMZ 58191. Monroe Co.: AMNH~~

~~73930; LSUMZ 28870,34312; MCZ 29335, R-180277; UF 8609,64995, 73462,~~

~~113421-2. Orange Co.: LSUMZ 80469, 80470, 80471. Palm Beach Co.: LSUMZ 9152,~~

~~58192. Pinellas Co.: CAS 17312; MCZ 18982. Polk Co.: CAS 192086. Putnam Co.:~~

~~LSUMZ 80181-2. Seminole Co.: LSUMZ 2019. Sumter Co.: CAS 173173. Suwannee~~

~~Co.. UF 74344, 113465. Taylor Co.: LSUMZ 57749; UF 113466, 113468-70. Wakulla~~

~~Co.. CAS 203079; LSUMZ 57748, 58073.~~

~~Georgia-Baker Co: ANSP 22251. Bleckley Co.: UF 11349-1. Brantley Co.: 113495.~~

~~Burke Co.: ANSP 30919. Camden CO.: ANSP 31191; UF 73469. Clinch Co.: UF~~

~~113502 . Cobb Co.: UF 2480. Columbia Co.: UF 113497. Decatur Co.: UF 12388.~~

~~Dekalb Co.: 4839. Emanuel Co.. ANSP 30908,30920-1; UF 7947. Glynn Co.: ANSP~~

~~31051. Grady Co.: UF 73465-7. Harris Co.: UF 16448. Irwin Co.. UF 113496. Liberty~~

~~Co.: CAS 14117. Lowndes Co.: ANSP 30895; UF 11443. McIntosh Co.: UF 84006.~~

~~Newton Co.: UF 113499. Rabun Co.: UGAMNH 6259,6315,6328-9,6334.~~

~~Richmond Co.: UF 16450-1, 113500-1. Screven Co.: 30922. Ware Co.: UF 2107.~~

~~179~~

~~permission of the copyright owner. Further reproduction prohibited without permission. Maryland-Frederick Co.: LSUMZ 44662.~~

~~New York-Dutchess Co.: AMNH 120078, 137475,142938. Orange Co.: AMNH 13777,~~

~~38752, 120079; LSUMZ 80471. Putnam Co.. AMNH 130189; CM 58466. Ulster Co..~~

~~UF 113524. Westchester Co.: AMNH 130190, 130787, 133024, 136676-7. Syntype o f~~

~~E. alleghaniensis: ANSP 16792.~~

~~North Carolina-Aveiy Co.: CAS 192088. Brunswick Co.: CM 21540; NCSM 1541,~~

~~2178,3851,6367, 12467, 12468, 12527,12528, 14671, 15026, 15383, 15599, 16773,~~

~~17923, 19525, 26248. Carteret Co.: CAS 43683. Cherokee Co.: CM 57174. Macon Co.:~~

~~NCSM 12771, 13047, 14160, 14161, 14268, 14344, 14345, 15922. Sampson Co.:~~

~~NCSM 2631, 8100, 10134-5, 10145, 10202-3, 14108, 15418-9, 16599, 16600, 19695,~~

~~25881, 26206,31848,39571,41287,41289. Transylvania Co.: CR 266. Wayah Bald~~

~~Mt.: CM 39615.~~

~~Pennsylvania-Center Co.: LSUMZ 46888.~~

~~South Carolina-Aiken Co.: CR 2633, MCZ-181268; SREL 126, 170, 1690, 3123,3132,~~

~~3225,3457-8, 3466, 4218,4223,4243,4246,4284,4362, -4; UGAMNH 6313, 6339,~~

~~6342,6348-9. Bamberg Co.: CR 2640, 3481,4217. Charleston Co.: AMNH 113049;~~

~~ANSP 3773; FMNH 4282,4766; SREL 1101; UGAMNH 6258. Fairfield Co.: CR~~

~~180~~

~~permission of the copyright owner. Further reproduction prohibited without permission. 2635. Jasper Co.: LSUMZ 58526. Lexington Co.: CM 16787; CR 2625,2631; FMNH~~

~~60560. Richland CO.. CR 263-4, 2624, 2632,2634,2636.~~

~~Virginia-Bedford Co.: CM 130179, 146302, 146431,146353, 146356, 146358, 146387,~~

~~146392, 146446-7, 146463, 146507, 146523-4, 146533, KU 68914.~~

~~Elaphe spiloides~~

~~Alabama-Baldwin Co.: AUM 414, 10310,10314, 10383,21011; LSUMZ 57698,~~

~~75698; UF 16443. Barbour Co.: UF 19277. Calhoun Co.: AUM 5348,6525-6,30795.~~

~~Cherokee Co.: LSUMZ 33070. Clarke Co.: Aum 12578, 22929-30. Cleburne Co.: AUM~~

~~80,2991. Coffee Co.: UF 108484. Dale Co.: LSUMZ 6709. Elmore Co.: UF 87565. Lee~~

~~Co.: AUM 31-3,415,430,766, 1723,2047,2091,2863,4729, 10385, 13681,27312,~~

~~32509, 32556; LM 9884; LSUMZ 58165, 58446. Macon Co.: LSUMZ 58459. Madison~~

~~Co.: LSUMZ 41189. Mobile Co.: AMNH 123900; ANSP 10893-4, 7798. Monroe Co.:~~

~~AUM 7133. Pike Co.. UF 108488. Randolph Co.: AUM 8957. Shelby Co.: UF 8499.~~

~~Talladega Co.. AUM 17107, 21608, 21676, 23021; LSUMZ 58527. Washington Co.:~~

~~AUM 58196.~~

~~FIorida-Escambia Co.: LSUMZ 15915; MCZ 169. Holmes Co.: LSUMZ 6508.~~

~~Okaloosa Co.: AUM 29748,30454; UF 16444,68180,72663. Santa Rosa Co.. AUM~~

~~29746; KU 82074; UF 67921, 113461-2; LSUMZ 58354. Walton Co.: LSUMZ 58196.~~

~~181~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Illinois-Adams Co.: MCZ 52. Bond Co.: 1NHS 5166,8175. Calhoun Co.: INHS 3392,~~

~~10614, 11450; UIMNH 16646-7,50840. Clark Co.: INHS 2598. Clay Co.: INHS~~

~~14011. Cole Co.: INHS 10097. Effingham Co.: INHS 3129, 884; MCZ 71839. Fayette~~

~~Co.: INHS 5165,12494, 13069. Gallatin Co.: INHS 1384,14012, 14014. Green Co.:~~

~~UIMNH 50838. Hamilton Co.. INHS 2266,2637. Jackson Co.: AUM 2001; CM~~

~~114334; FMNH 18655, 18656,155048, 204022,204037; INHS 1079, 1081-2, 1807,~~

~~2055, 2550, 3858; LSUMZ 58408-9; MCZ 181237, 181238. Jasper Co.: INHS 1299,~~

~~1950,2095, 14039. Jefferson Co.: 9287. Jersey Co.: 1744,1779, 1780,9901,11012-4,~~

~~11019, 11023, 11342,11345, 11448, 11451. Johnson Co.: INHS 13972. Lawrence Co.:~~

~~INHS 4727. Madison Co.: INHS 11020, 11022; UIMNH 16326, 33943. Monroe Co.:~~

~~INHS 3689, 4286,4753. Montgomery Co.. INHS 1301. Perry Co.: INHS 3363,9899,~~

~~9900, 13908. Piatt Co.: INHS 4248. Pike Co.: INHS 3650. Pope Co.: INHS 1386-7,~~

~~1388,2312,4987. Randolph Co.: INHS 4257. Richland Co.: INHS 14016. St. Clair Co.:~~

~~INHS 9898, 13453, 13454. Union Co.. FMNH 19262, 195485; INHS 1303, 1304, 1382,~~

~~1383, 1647, 2255-7, 5981, 7029, 8089, 10583, 11272; KU 69657; MCZ 181470.~~

~~Vermillion Co.: INHS 1302, 7028,9018, 10040, 10288,10622, 14004,5221. Wabash~~

~~Co.: INHS 5221. Washington Co.. INHS 4387. Wayne Co.. INHS 12744. White Co..~~

~~INHS 14015. Williamson Co.: INHS 6802.~~

~~Kentucky-Ballard Co.. KU 214415. Calloway Co.. KU 144760, 214413-4. Carlisle Co..~~

~~137763, 214398,214399. Fulton Co.: KU 206538, 214405-6. Grave Co.: KU 214410-1.~~

~~Hickman Co.: KU 144772,214416. Marshall Co.: KU 14476,214417.~~

~~182~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Loui si ana-Ascensi on Par.: LSUMZ 46834. East Baton Rouge Par.: LSUMZ 2832-3,~~

~~5857,6163,9470, 11888, 11904,12049, 12257, 12723-5,12727,23844,24795, 33063,~~

~~39161,49585, 56148, 56152, 57027, 57686, 57703,57838, 58193, 58348, 58399,~~

~~58407,59631,73935. East Feleciana Par.: LSUMZ 57744,58608. Iberville Par.:~~

~~LSUMZ 6508, 17719, 56497, 57723, 57736-7, 58194-5, 58416, 58229, 58445. St.~~

~~Charles Par.. LSUMZ 57687. St. Tammany Par.: LSUMZ 58521, 80221. Tangipahoa~~

~~Par.: LSUMZ 57695, 58163.~~

~~Michigan-Eaton Co.: CAS 74407.~~

~~Mississippi-Forrest Co.: LSUMZ 57688. Hancock Co.: CM 66533-4; MMNS 3478,~~

~~3482,3533, 3540. Harrison Co.: UF 16439-40, 16458. Pearl River Co.. MMNS 3205,~~

~~3467, 3486, 3494,3497-8, 3503, 3548. Smith Co.: LSUMZ 57721; UF 89487.~~

~~Wilkinson Co.: LSUMZ 57691.~~

~~Ohio-Montgomery Co.: LSUMZ 58525, 59625. Scioto Co.: INHS 9013.~~

~~Pennsylvania: Allegheny Co.: CM 774, 1620, 1764, 1966, 1976,4004, 5124,9923,~~

~~11457,20097, 27075,31395, 35329,35709, 140186.~~

~~Tennessee-Cumberland Co.: LSUMZ 58529. Decatur Co.: LSUMZ 57735. Hardeman~~

~~Co.. CM 19897. Knox Co.. LSUMZ 58162,58414,58510,59639. Lawrence Co.: CM~~

~~19900. Montgomery Co.: LSUMZ 58563. Morgan Co.: INHS 13276.~~

~~183~~

~~permission of the copyright owner. Further reproduction prohibited without permission. Wisconsin-Crawford Co.: MPM 2035, 2061. Grant Co.: CM 70347. Iowa Co.: MPM~~

~~8805. LaCrosse Co.: MPM 23355. Richland Co.: CM 75864; MPM 2027. Sauk Co.:~~

~~MPM 2137.~~

~~Elaphe obsoleta~~

~~Arkansas-Baxter Co.:ASUMZ 22353. Benton Co.: LSUMZ 73966. Clark Co.: ASUMZ~~

~~22692. Craighead Co.: ASUMZ 22689. Garland Co.: ASUMZ 22354, 22694. Lawrence~~

~~Co.. ASUMZ 22695. Logan Co.. ASUMZ 22693. Madison Co.: LSUMZ 80246.~~

~~Mississippi Co.: ASUMZ 22270. Poinsett Co.: ASUMZ 22690-1. Sevier Co.: LSUMZ~~

~~74497. White Co.: FMNH 37543; MPM 14371, 15705, 18632, 18846, 18903, 19101,~~

~~20712,21501.~~

~~Kansas-Barber Co.. KU 193399,211379. Bourbon Co.. KU 2432. Cowley Co.: KU~~

~~179036, 206174,216190. Douglas Co.: Ku 2424,2429-31, 2433-4, 2436-8,2442-8,~~

~~2450-3, 2456-60, 2725,2728, 7557, 28765, 38678, 153039, 188712, 207295. Geary~~

~~Co.: CAS 38678; LSUMZ 58530. Harper Co.: FMNH 95158; KU 186004. Sumner~~

~~Co.: KU 18886, 206231-33, 206400, 206490; LSUMZ 58607.~~

~~Louisiana-Acadia Par.: LSUMZ 58164, 74128. Bossier Par.: LSUMZ 58443; LSUS~~

~~4285; UTA 28252. Caddo Par.: LSUS 1322,2944, 5562, 5823, 5825, 5951,5991,6222,~~

~~6606; SM 10413, 10415. Lafayette Par.: LSUMZ 73888, 73897, 73911, 73954, 73955,~~

~~184~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 73956, 73958,74112,74122, 74495. Livingston Par.: LSUMZ 58345. Natchitoches~~

~~Par.: LSUMZ 58995-7. Point Coupee Par.: LSUMZ 57768. Rapides Par.: LSUMZ~~

~~58471. St. Landry Par.: LSUMZ 58230,73965,73973,74098,74116,74137, 73895,~~

~~73948,73959,73975,73977,73978,73981,74101,74103. Terrebonne Par.: LSUMZ~~

~~59418. Vermillion Par.: LSUMZ 59643.~~

~~Missouri-Butler Co.: INHS 10363. Douglas Co.: INHS 10586. Green Co.: CAS 162004.~~

~~Nebraska-Cass Co.. UNSM 2236, 8336. Gage Co.: UNSM 549, 552, 1408, 1439, 1446,~~

~~1840,6817, 8337. Johnson Co.: UNSM 2241. Nemaha Co.: UNSM 2237,2238. Otoe~~

~~Co.. UNSM 2244, 8335. Pawnee Co.: UNSM 7266. Richardson Co.: UNSM 550, 553,~~

~~1839,3981. Saunders Co.: UNSM 15456. Thayer Co.: UNSM 2242.~~

~~Oklahoma-Cleveland Co.: OMNH 550,642,2894,3751,5584,5882, 8105,8816,~~

~~10136, 13361, 15064, 15081, 18978, 20207,20209,20211,23195, 23247,23362,~~

~~26970, 28605, 29944, 33967,67163. Tulsa Co.: CM 61916.~~

~~Texas-Bexar Co.: AMNH 73366; ANSP 12129; CAS 30972-3, 31099-100; CAS-SU~~

~~17750-1; CM 22849; FMNH 3510; TCWC 42310; TNHC 55330. Brazos Co.: SM~~

~~10428. Kerr Co.: TCWC 205, 7240, 31069. Medina Co.: CM 19918; TCWC 38779,~~

~~42872; TNHC 42246; UTA 32197. Orange Co.: LSUMZ 80184. Robertson Co.: TCWC~~

~~6105. Tarrant Co.: UTA 7734, 8733, 10941, 14710, 17103, 19329, 25710,26467,~~

~~26468,28696,28697-99. Travis Co.: TNHC 98,610,947, 1632, 1705,2020-1,4584,~~

~~185~~

~~permission of the copyright owner. Further reproduction prohibited without permission. 5245, 5251-2, 5949,7230, 7243, 8786,9147, 10265, 10267, 18535-6,21881,22421,~~

~~29091-2,36336,42247-9,42681,42788,43104,46274,47182,47222,49948. Uvalde~~

~~Co.: TNHC 47636. Washington Co.: CR 429.~~

~~Mexico-Nuevo Leon: SM 8180-—Collecting locality is probably in error (£. Liner,~~

~~pers. com.).~~

~~Elaphe bairdi~~

~~TX-Bandera Co.: SM 104536; TCWC 79951; TNHC 25467. Boswue Co.: TCWC~~

~~36939. Brewster Co.: FMNH 26619, 27703,27844; SM 746, 12107. Jefferson Davis~~

~~Co.: AMNH 115718; CAS 7509; FMNH 27704; LSUMZ 58523-4; TCWC 42858,~~

~~66184; TNHC 33616. Kerr Co.. TCWC 42857,67282. Medina Co.: TCWC 48595,~~

~~48596. Pecos Co.: SM 13445. Real Co.: TCWC 30762. Val Verde Co.: LSUMZ 34527,~~

~~44461; SM 13272; TCWC 60528, 71050, 77161-2, 79746; TNHC 49251.~~

~~186~~

~~Reproduced with permission of the copyrightowner. Further reproduction prohibited without permission. Vita~~

~~Frank T. Burbrink was bom in Peoria, Illinois, on November 12, 1970. In 1993~~

~~he received his bachelor of science degree in biology from the University of Illinois in~~

~~Champaign-Urbana. In 1995 he earned his master’s degree in biology from the~~

~~University of Illinois in Champaign-Urbana in 1995. In order to escape the cold~~

~~weather of northern Illinois he moved to Baton Rouge, Louisiana, in August of 1995.~~

~~On May 19,2000, he will receive the degree of Doctor of Philosophy degree in zoology~~

~~from Louisiana State University in Baton Rouge. Frank is a consummate collector of~~

~~hillbilly, country, western swing, jazz, blues, and early rock and roll records from the~~

~~1920’s, 1930’s, 1940’s, and 1950’s.~~

~~187~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. DOCTORAL EXAMINATION AND DISSERTATION REPORT~~

~~Candidate: Frank T. Burbrlnk~~

~~Major Field: Zoology~~

~~Title of Dissertation: Systematlcs of the Polymorphic North American Rat Snake (Elaphe obsoleta)~~

~~Approved:~~

~~or Profesi~~

~~[Graduate School~~

~~EXAMINING COMMITTEE:~~

~~Date of Examination:~~

~~March 15. ZQQfl_____~~

~~Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.~~