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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. THE EVOLUTION OF LAMPYRIDAE, WITH SPECIAL EMPHASIS ON THE ORIGIN OF PHOTIC BEHAVIOR AND SIGNAL SYSTEM EVOLUTION (COLEOPTERA: LAMPYRIDAE)

DISSERTATION

Presented in Partial Fulfillment of the Requirements for The Degree Doctor of Philosophy in the Graduate School of The Ohio State University

Marc A. Branham

The Ohio State University 2002

Dissertation Committee: Approved by Dr. John W. Wenzel, Advisor Dr. Norman F. Johnson w. Co. Sl Dr. Woodbndge A. Foster Advisor ^ Dr. Douglas A. Nelson Department o f Entomology

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Copyright by Marc A. Branham 2002

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

The distribution, life history, and behavior of both larval and adult Lampyridae

are summarized, as well as larval and adult morphological characters that define this

family of beetles. The last larval and pupal stages of the North American firefly, Lucidota

atra (G~A~ Olivier 1790), are described and illustrated. The larva of L atra was

misidentified in the literature as a species in the genus P hotinus. A discussion of the

homology of abdominal sclerites in larval, pupal, and adult fireflies is provided. This

work represents the first cladistic analysis of genera in the family Lampyridae and other

closely related beetles. A monophyledc concept of Lampyridae is established. The

phylogenetic positions of the luminous cantharoid families [Omalisidae,

Rhagophthalmidae and Phengodidae] in relation to Lampyridae are discussed, as well as

the implications of the evolution of bioluminescence and photic s ign alin g in this group of

beetles. The Rhagophthalmidae appears to include D ioptom a and Diplocladon (formerly

located in Phengodidae) and the Phengodidae apparently includes Stenocladius (formerly

of Lampyridae). Harmatelia, Drilaster and P terotus are transferred to Elateroidea

m certae seeds and not included in Lampyridae where they were sometimes placed.

Through a phylogenetic analysis using adult morphological characters, it is shown that

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. the origin ofbioluminescence in cantharoid beetles appears to predate the origin o f the

family Lampyridae. The ability to produce and emit photic signals was first gained by

larvae and appears to function as an aposematic warning display, while subsequently

being gained in adults and used as sexual signals. This analysis also suggests that while

pheromonal sexual signals are used basally in the fam ily, these are used in conjunction

with, and then subsequently replaced by, photic signals in some Iampyrid lineages. Both

photic signals and the photic organs used to produce them have become greatly

elaborated in the fireflies that no longer employ pheromonal sexual signals. In addition,

the ability to produce a flashed sexual signal appears to have arisen at least three times in

the family Lampyridae. Convergent evolution also is evident in adult male photic organ

morphology. Further, it is recommended that individual signal-system components be

compared, rather than overall signal system complexity. The use of this strategy may

allow one to recognize and better interpret adaptive correlations despite convergence or

loss.

' Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. To Frank A. McDermott,

a pioneer in the study ofLampyridae and a man I would have liked to have met

“Bat there is no ram tonight no clouds, in fact to blot out the showering of starlight The moon has not risen from its sluggish daybed, and the only competition offered the stars in this clean night air is from lightning bogs over the drtchwater. They are leisurely about sharing their miracle, never overdoing the effect - one here, then another off in the distance, a fluid interval and then three airborne grace notes of yellow rr

Blue Rise By Rebecca Hill

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

I would like to acknowledge the members of my doctoral committee. First and

foremost, I am indebted to my advisor Dr. John Wenzel for continually challenging me to

look a little deeper and think cladistically. He is a wonderful mentor and Mend. I am still

amazed that he was brave enough to take on a student who studied not wasps, but beetles.

And to John’s credit, he has learned many difficult Iampyrid and cantharoid names. He

now knows a lot about beetles, for a hymenopterist! Dr. Norman Johnson’s impressive

knowledge of nomenclature, languages, geography and entomology has been invaluable

to me. One day 1 was in his office some four times... I appreciate his patience and

willingness to be of help. Dr. Woodbridge Foster is one of my favorite instructors. His

attention to detail is superhuman and his knowledge so broad that conversations with him

always leave me thinking. Dr. Douglas Nelson always encouraged me to think about

communication systems in the broadest terms, rather than only thinking about insects. It

is, of course, always important to place one’s work within a larger context. Doug’s quiet

and careful ways provide an excellent example of professionalism in science.

Dr. James Lloyd is both a Mend and mentor. I treasure the time I spend looking

out over a field of flashing fireflies with Jim pointing out this and that, or just talking

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. over beers about firefly staff nr fishing-1 often feel like Jim is the only one I can talk to

about the nitty gritty details of fireflies. Jim and his wife Dorothy have generously put me

up in a spare bedroom in their home during my visits to Gainesville, FL. They have

always treated me like family and I enjoy their company very much.

I would like to thank Dr, Hans Klompen and Dr. Peter Kovarik for our

discussions concerning morphology and the morphological similarities (though subtle)

between mites, histerid beetles and fireflies.

The following friends and colleagues have supplied particularly helpful ideas,

criticism, and friendship: Keith Philips, Todd Blackledge, Hojun Song, Chi Feng Lee,

and Ming Luen Jeng. I’d like to give special thanks to Miguel Archangelsky for teaching

me about scientific illustration and how to keep beetle larvae alive and happy in the lab.

Kurt Pickett should also receive special mention, as we arrived in the Wenzel Lab

together and were partners-in-crime for my entire stay at OSU. Kurt has always been

willing to help, listen, argue, and proofread whenever the need arose.

I would also like to gratefully acknowledge Mr. Bruce Leech, Ms. Susan Ward

and Ms. Karen Ronga, librarians at the Biological and Pharmacy Library, Ohio State

University, for their willingness to help me track down obscure references that were

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. oftentimes incorrectly cited. I often wondered if I could come up with a Iibrary-research

related problem that they could not solve; I never could. They are experts and I appreciate

them skills.

I would like to thank my wife Lori Sandhoidt and my brother Aric Branham for

their patient tolerence of my entomological endeavors. I would also like to thank my

mother, who never minded my filling up her freezer with bug jars when I was a boy.

I would also like to thank the following sources for supporting my research: the

Ohio State University Presidential Fellowship Fund, the Ohio State Graduate Student

International Dissertation Research Travel Grant Fund, a Ohio State Professional

Development Fund Travel Grant, and a National Geographic Society Grant-in-aid of

research. These funds and awards allowed me the opportunity to conduct research that I

would otherwise not have been able to do.

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

December 5,1966 Bom - Illinois, U.SA.

December 19% B.S. Organismal Biology University of Kansas Lawrence, Kansas

August 1995, M A Entomology University of Kansas Lawrence, Kansas

PUBLICATIONS

Archangelsky, M. and MA. Branham 2001. Description of the last larval stage and pupa of Pvropyga nigricans (Coleoptera: Lampyridae), and comparison with larvae of other known Photmini genera. Canadian Entomologist 133:1-10.

Archangelsky, M and MA. Branham 1998. Description of the preimaginal stages of Pvractomena borealis (Randall. 1838) (Coleoptera, Lampyridae) and notes on its biology. Proceedmgs o fthe Washington Entomological Society I00(3):421-430.

Branham, M A 2002. Lampyridae. in R.G. Beutel & RAJ3. Leschen, (eds.) Handbuch der Zoologie, Band IV Arthropoda: Insecta, Teilband 39, Evolution and Systematics. Waltyer de Gruyter, Berlin. [In Press]

Branham, NLA. 2002. A new technique for collecting glowworm fireflies (Lampyridae). Coleopterists Bulletin [hi Press]

Branham, NLA. and J.W. Wenzel 2001. The evolution of bioluminescence in cantharoids (Coleoptera: Elateroidea). Florida Entom ologist 84(4): 478-499.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Branham, M A. and M. Archangelsky 200G. Description of the last larval faster and pupa of Lucidota atra, (G.A. Oliver, 1790) (Coleoptera: Lampyridae), with a discussion of abdominal segment homology across life stages. Proceedings o f the Washington Entomological Society I02(4):869-877.

Branham, M A. and J.W. Wenzel 2000. The evolution of sexual communication in fireflies. XI* International Symposium on Bioluminescence and Chemiluminescence. Luminescence . 15(4): 202. [Abstract]

Branham, M.A. and M i). Greenfield 1996. Flashing males win mate success. Nature 381:745-746.

Branham, MA. 1993. A new eastern record for Cypherotylus califomicus Lacordaire in the United States (Coleoptera: Erotylidae). Coleopterists Bulletin 47(l):8I-82.

FIELDS OF STUDY

Major Field: Entomology

Specialization: Systematics of Coleoptera, especially phylogenetics and behavior.

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

Abstract------ii

Dedication ------iv

Acknowledgments ------v

Vita------viii

List of Tables ______xv

List of Figures ------.xvi

I i

Introduction ______I

The Evolution of Bioluminescence in Cantfaaroids: The Origin and Function of Photic

Organs in Firefly Relatives ______6

Introduction ______6

Materials and Methods ______.______7

Results------8

Discussion ------9

Testing the Monophyly of Existing Families ______9

Lampyridae ______9

Other Cantharoid Families-______12

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Phylogenetic Relationship Between Lampyridae and Phengodidae ______13

The Evolution o f Bxohnninescence in Non-Iampyrid Cantharoids______13

Photic Organ Evohidon in Non-Iampyrid Cantharoids ______16

Adah Females______._____18

Adult M ales ------19

Luminescence and Life Stages ------21

Evolution of Photic Signaling in Non-Firefly Cantharoids ______22

Conclusion ______—....------23

r 'h a ntp r ^ 44

An Overview of the Family Lampyridae. ______44

Distribution ______44

Biology ______44

Adults. ______44

Larvae______48

Description ______52

Adults — — ______...... _...... ------o2

Larvae______54

Phytogeny ------56

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Description of the Larva and Pupa of Lucidota atra (G. A. OLIVIER 1790): Abdominal

Segment Homology and Location of Photic Organs Across Life Stages. ______71

Introduction— ..— ------71

Materials and Methods ______72

Results------73

Description of last larval instar ______73

Length------73

Head capsule...... ^1^1

Antenna ------74

Mandible ...... 5

Labium ....______75

Maxilla______76

Thorax______76

Legs ------77

11 il ...... ------77

Description of Pupa ______78

Head ______78

Abdomen ______79

Spu . ------... ------~— ...... — —------8 0

Discussion ------80

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Traditional Perspective, and Modem View ______81

Abdominal Sclerites in Lampyridae ______82

The Evolution of Firefly Signal Systems and Photic Organ Morphologies ______95

Introduction ______95

Materials and Methods _____ ™.______101

Results------102

Topology o f the Tree ______102

Levels of Homoplasy in the Analysis ______103

Discussion ______104

Taxonomy of the Lampyridae ______104

An Ancient Orign of Bioluminescence in the “Cantharoidea” ______106

Larval Luminescence as an Aposematic Display ------107

Carry-Over of Luminescence into Adults ______109

The Pattern of Flightlessness in Females Within Lampyridae ______113

Evolution of Chemical Signals vs. Visual Signals ______115

Evolution ofPhotic Signaling Systems ______117

Signal System Components ------119

Sedentary, Synchronously Flashing Aggregations of Fireflies ------120

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Evolution of Photic Organs ------122

Conclusion ------126

Bibiography ______144

Appendix A______163

AppUldlX U» 1 6 I

Appendix C------172

xnr

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

Table

1 Some notable papers showing the previous taxonomic placement of taxa that this study relegates to “mcertae sedis ” status......

2 A comparison of cantharoid taxa included in this analysis with special reference to photic organ morphology across life stages and the presence of neotenic characteristics in adult females .

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

Fignre Page

1. Strict consensus of280 most parsimonious trees (848 steps, CX 0.16, RX 0.57)...... 32

2. The non-lampyrid clades of the strict consensus tree with both Bremer Support values and the number of synapomorphic characters plotted at each node ...... 34

3. The evolution of bio luminescence in cantharoids. The condensed strict consensus tree with two origins of luminescence and one loss plotted 36

4. The evolution of larval bioiuminescence in cantharoids ...... 38

5. The evolution ofbioiuminescence m female cantharoids ...... 40

6. The evolution of bioiuminescence in male cantharoids ...... 42

7. Some of the variation found in Iampyrid antennal morphology: (A) Serrate, Pyrocoelia paretexta, left antenna, ventral view; (B) Bipectmate, Psilocladus sp.. right antenna, dorsal view; (C) Flabellate, Dodacles plumose, left antenna, ventral view; (D) BiflabefTate, Lam procera sp., left antenna, ventral view; (E) also

xvi

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Biflabellate, Lucio splendens, right antenna, ventral view; (F) Filiform, Photinus p yra lis, left antenna, ventral view; (G) Capitate, Petalacmis praeclarus , left antenna, anterior view...... 59

8. The falcate type adult mandibles found in Lampyridae, Lucidina biplagiata ...... 61

9. The somewhat shorter and more robust adult mandible type, Photinus pyralisJ83

10. The specialized or modified type adult mandible, Phaenolis ustulatus 65

11. The trilobate type aedeagus found in Lampyridae, Photinus pyralis 67

12. The aedeagus of Photuris species possess two long, thin, lateral filaments that are attached to the phallobase, Photuris divisi...... 69

13. Lucidota atra; fifth instar larva. (A) Right antenna, dorsal view. (B) Right mandible, dorsal view ...... 87

14. Lucidota atra; fifth instar larva. (A) Habitus. (B) Head capsule, dorsal view. (C) Head capsule, ventral view ...... 89

15. Lucidota atra; fifth instar larva. (A) Labium, dorsal view. (B) Left maxilla, dorsal view ...... 91

16. Lucidota atra; pupa. (A) Ventral view. (B) Dorsal view ...... 93

17. The lampyrid clade of the strict consensus tree with both Bremer Support

xvii

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Values and the number of synapomorphic characters plotted at each node...... 128

18. The evolution of pheromone nse in Lampyridae ...... 130

19. The evolution of photic signals, produced by either sex, in Lampyridae is represented by a single optimization of four origins and four losses 132

20. The evolution of signaling systems that use a combination of both pheromonal and photic signals ...... 134

21. The evolution of sexual signal systems, sedentary or active primary signalers, and the sex of the primary signaler ...... 136

22. Adult male photic organ evolution on abdominal ventrite seven (true abdominal segment eight) ...... 138

23. Adult male photic organ evolution on abdominal ventrite six (true abdominal segment seven) ...... 140

24. Adult male photic organ evolution on abdominal ventrite five (true abdominal segment six) ...... 142

xviii

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

INTRODUCTION

Fireflies are an exciting group through which to study the evolution of

communication, and, in particular, the evolution of bioluminescent signals. Most

bioluminescent organisms are marine and are not only difficult to study, but have been

the focus of biological investigations only relatively recently, e.g., William Beebe was

the first to use a bathysphere for scientific observations in the bathypelagic environment

in 1930 (Ellis 1996). In contrast, fireflies have enjoyed a much longer history of study,

dating back to the fifthteenth and sixthteenth centuries (Newton 1957) and today still

serve as the model system for the study of bioiuminescence. Firefly specimens are

available from many diverse geographic regions and when found alive in the field, they

can generally be observed quite easily for long periods of time. Adult fireflies also

possess an amazing diversity of sexual signals, ranging from the use of long-range

chemical signals (pheromones), to pheromones used in conjunction with photic glows, to

only photic glows, pulses and flashes. Like other aspects of firefly luminescence, i.e., the

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. chemilummescence involved in the production of tight and behavioral roles of these

photic emissions, studies of the evolution of firefly sexual signals will often be the

standard against which other bioluminescent systems win be compared.

Fireflies are well known to most people because of their charismatic

bioluminescent displays. Fireflies are the only bioluminescent organism s most people

will ever encounter. Therefore, when someone chances upon a luminescent beetle that is

not a firefly, it is commonly mistaken for one. bisect taxonomists are no exception and

have placed many non-fireflies into the same family (Lampyridae) solely on the insects’

ability to produce light. As an example, taxonomists have always considered another

luminescent beetle family, Phengodidae, to be the family most closest related to

Lampyridae because of the presence o f glowing paedomorphic "‘larva-like” females in

both families. In addition, several genera, e.g., Drilaster, Harmatelia, Pterotus,

Stenocladius, and Dioptoma, have proved to be difficult to place within any of the

existing cantharoid families and at various tunes have been placed within Lampyridae

(Oliver 1910a, 1910b; McDermott 1964,1966; Crowson 1972; Lawrence & Newton

1995). One of the goals of this study was to examine the composition of the family

Lampyridae to determine its limits, specifically to ascertain which taxa should be

included within Lampyridae and what are the limits if the family is to be monophyletic.

•>

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. There are many benefits to conducting a phylogenetic analysis when addressing

issues of monophyly and character-state transformations. When investigating whether a

group is monophyletic, one can more or less simultaneously compare ail available data in

order to reconstruct a pattern of relatedness, rather than relying on only a couple of easily

diagnosable characters to delimit groups. While a few characters might be easy to

diagnose, if they are convergent, they are likely to mislead systemadsts searching for the

real evolutionary pattern. The “presence of photic organs” is one such character. The

study of signal evolution requires a phylogenetic framework within which character-state

transformations are placed in a series. Without a phytogeny, one could only speculate as

to the correct order of character-state changes or whether various s ignal system

components are ancestral or derived, or convergent or homologous. In addition, through

the identification of convergent and homologous signal components, investigators can

more accurately compare and contrast components of various signal systems that are the

same (homologous) or similar (convergent).

hi order to define the limits of Lampyridae and to study signal evolution within

the family, a phylogenetic analysis was conducted using morphological characters coded

from male specimens. Morphological data from larvae and females, as well as behavioral

data, were not incorporated because data from these sources are lacking. Only 23% of the

current Iampyrid genera have larvae known for at least one species, female lampyrids are

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. not commonly collected and are therefore not well represented in collections, and

behavioral data, where present, are generally incomplete. The collection and

incorporation of molecular data into this data set is planned for the near future. Rather

than examining signal evolution within one or two genera of fireflies, this study focuses

on the broader evolutionary patterns of signal systems across higher-level taxa, i.e.,

genera within Lampyridae, as well as the general occurrence of luminescence in families

closely related to Lampyridae.

Morphological characters based on antennal and photic organ morphologies were

used in this analysis and clearly represent morphological elaborations associated with

pheromonal and photic signal systems, respectively. Some have argued that the inclusion

of characters under study in an analysis appears circular, and is, therefore inappropriate.

However, it is widely held that including such characters is not circular and that it is, in

fact, inappropriate to exclude these types of data from the analysis, as they are

phylogenetically informative.

My dissertation research into the evolution of Lampyridae is designed around

three areas o f inquiry: I) the phylogenetic placement of the Lampyridae among its close

relatives and the implications of this pattern to the evolution of bioluminescence in

fireflies, 2) defining the features, ecology and habits of Lampyridae and 3) investigating

both signal system evolution within Lampyridae and the photic organ morphology used

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. to produce various types of photic signals. The specific goals within each of these three

levels will be outlined and addressed within each o f the foUowing chapters

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

THE EVOLUTION OF BIOLUMINESCENCE IN CANTHAROEDS: THE ORIGIN

AND FUNCTION OF PHOTIC ORGANS IN FIREFLY RELATIVES.

INTRODUCTION

The common and conspicuous bioLuminescent displays of adult fireflies have been

marveled at by man throughout history and have long been recognized as displays of

courtship. In 1647, Thomas Bartholin related an observation of Carolus Vintimillia that

“nature had endowed them [female fireflies] with a vigorous light in order that they could

call the males at night with their shine” (Harvey 1957). Bishop Heber in hisTour

through Ceylon remarks: “Before beside us and above, the firefly lights his torch of love”

(Harvey 1940). However, the less conspicuous bioluminescent emissions of less well-

known beetles seem to have escaped notice by most. This phylogenetic analysis focuses

on the origin of luminous habit and the evolution of luminescence. Therefore, the taxa

chosen for this analysis most heavily represent the breadth of Lampyridae with an equal

number of luminous and non-Iuminous genera in the cantharoid lineage. This analysis

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. establishes the limits of a monophyletic Lampyridae, places other luminous taxa that are

thought to be closely related to fireflies, and investigates possible origins and losses of

luminescence in taxa related to Lampyridae.

The superfamily Cantharoidea was combined into the Elateroidea when Lawrence

(1988) redefined Elateriformia. The present analysis includes most of the families that

formerly composed the Cantharoidea of Crowson (1955,1972) (his included

Brachypsectridae, Omalisidae (= Omaiysidae, Homalisidae), Karumiidae, Drilidae,

Phengodidae, Telegeusidae, Lampyridae, Cantharidae, Lycidae, Cneoglossidae,

Plastoceridae and Omethidae). I refer loosely to the taxa used in this analysis as

“cantharoids,” as they have been treated historically as a monophyletic group within the

Elateroidea (Lawrence 1988).

MATERIALS AND METHODS

Eighty-five exemplar taxa were selected to represent a diversity of Lampyridae

and outgroup families. Selection of taxa included as many subfamilies and tribes as

possible within Lampyridae, based on the classification schemes of Crowson (1972) and

Lawrence & Newton (1995), and II subfamilies within 9 other families, based on

Lawrence & Newton (1995) (Appendix I). Specimens of Elateridae were not included in

this analysis, as they are too distantly related to the taxa considered here (Lawrence

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 1988). Seventy four male morphological characters with a total of 212 character states

were used in the analysis. Inapplicable characters were coded as while m issin g

characters were coded as “7”. All characters were analyzed under equal weights with 20

multistate characters coded as additive (see Appendix 2,3). Plastoceridae was designated

as the root of the tree based on Lawrence’s (1988) phylogenetic analysis of the

Elateriformia. The parsimony ratchet (Nixon 1999) (consisting of 100 iterations,

weighting 12% of the characters) was implemented in Nona (Goloboff 1993), run w ithin

Winclada (Nixon 2000). The most parsimonious trees discovered were used as the

starting place for a more exhaustive search using the “max*” command within NONA.

The “best” command was then used to eliminate sub-optimal trees. A strict consensus tree

was then calculated from these most parsimonious trees. Bremer support (Bremer 1988,

1994) was calculated using NONA, and the search was set to a Bremer support level of 5,

with four runs, each with a buffer o f5000 trees.

RESULTS

The parsimony ratchet returned trees of 818 steps. Starting from these 52 trees,

“max*” and “best” gave 280 most parsimonious trees of 818 steps. A strict consensus

(Figure 1) of these 280 trees collapsed 13 nodes and produced a consensus tree of 848

steps (ci = 0.16, ri = 0.57). Bremer values listed in Figure 2 indicate the number of steps

that are required, up to 5, to find the closest tree that does not contain that particular node.

Lampyridae is monophyletic, with the exception of a few taxa that have been of

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. controversial affinity ( Harmatelia, Drilaster, Pterotus, and Stenocladius^) Two genera

currently classified, as phengodids (Dioptoma and Diplocladori) were placed with

Rhagophthalmids in tins phylogenetic analysis. The families Drilidae, Omalisidae,

Lycidae, Omethidae, Teleguesidae and Phengodidae appear to be monophyletic. The

monopbyly of Cantharidae is not supported.

DISCUSSION

Testing the Monophviy of Existing Families

Lampyridae

In view of this phytogeny, Lampyridae as defined by Crowson (1972) and

Lawrence and Newton (1995) is not monophyletic. The three synapomorphies that define

the base of Lampyridae are these: covered head position, oblique attachment o f trochanter

to femora, and wing vein CuAl intersecting MP above fork (Kukalova-Peck & Lawrence

1993). The genera Harmatelia, Pterotus, Drilaster (O totreta ) and Stenocladius are

currently classified as Iampyrids (Lawrence & Newton 1995, after Crowson 1972) though

in this analysis they are clearly placed outside of the family Lampyridae. This is not

entirely unexpected, as several previous authors (LeConte 1859, McDermott 1964,

Crowson 1972) who have examined some of these taxa have viewed them as possessing

questionable affinities to existing families. LeConte (1859) placed Pterotus obscuripennis

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. in Drilidae and then later (1881) moved it to Phengodidae. Of the genus P terotus,

LeConte (1859) stated, “A singular genus, which 1 have described at length from my

inability to place it properly. It seems to have a mixture of characters belonging to the

Lampyrides, Telephorides and Drilids, but from the small size of the posterior coxae is

probably better placed with the latter ” McDermott (1964) also mentions the difficulty he

encountered in trying to place some of these taxa, “Both P terotus and H arm atelia share a

large degree of similarity between some characters. Also neither Sts strictly to the

accepted Iampyrid characteristics and both have some suggestion of phengodid affinities.

Combining these two genera in the sub-family Pterotinae is admittedly arbitrary but

nevertheless serves to bring them together as transitional forms.” Crowson (1972) wrote

that the generaP terotus and Ototretadrilus were the most phengodid-like and probably

the most primitive firefly genera he had studied. (Specimens of Ototretadrilus were not

available to me.)

The genus D rilaster was originally described in the family Drilidae by

Kiesenwetter (1 8 7 9 ),and O totreta was originally described in the family Lampyridae.

The synonymy between D rilaster and O totreta was noticed by Nakane (1 9 5 0 ) (see also

Sato 1 9 6 8 ).However, Asian workers continued to use the older name ( D rilaster ) but

moved it to Lampyridae. American and European workers continued using O totreta, as it

appeared to be the valid nam e in McDermott (1 9 6 4 and 1 9 66).Therefore, not only should

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. American and European workers discontinue the use of the name “ O totreta.” bat these

taxa should no longer be associated with the family Lampyridae. For a short history of

the taxonomic placement of D rilaster and O totreta, see Table I.

According to this phytogeny (Figure I), if Harmatelia, Pterotus, Drilaster and

Stenocladius were to continue to be considered fireflies, the families Lycidac,

Cantharidae, Phengodidae, Omethidae and Teleguesidae would need to be synonymized

with Lampyridae. This seems drastic, given the peripheral significance of the genera and

the traditional affection for the families. H arm atelia, Pterotus , and D rilaster should be

removed from Lampyridae and given the taxonomic label “Elateroidea incertae seeds"

awaiting further study to place them properly. D rilaster may not be monophyletic. This

analysis clearly places Stenocladius sp. in the family Phengodidae. Ohba et. aL (1996)

studied the external morphology of Stenocladius larvae and found that they did not

possess an epicranial suture on the dorsal surface o f the larval head. This suture is well

developed in larvae of Lampyridae. They hypothesized that the fused dorsal surface of

the larval head inStenocladius is more closely allied with Phengodidae than Lampyridae.

My analysis supports this association. Because the clade containing these taxa is

unresolved, there is no information for the appropriate placement o f Stenocladius among

the subfamilies ofPhengodidae (Figure 2).

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

The families Plastoceridae, Drilidae, Omalisidae, Rhagophthalmidae, Lycidae,

Omethidae, and Teleguesidae are supported as being monophyletic in this analysis, but

include very few representatives. Phengodidae and Cantharidae are not supported as

being monophyletic. The four other phengodid taxa used in this analysis, other than

Dioptoma adamsi and Diplocladort sp. are a monophyletic clade. The analysis placed

D ioptom a and Diplocladort at the base of the clade containing the family

Rhagophthalmidae. The Bremer support value for the base of this clade (Figure 2) is high

(>5), indicating strong support. The seven synapomorphies that define this clade are

these: twelve antennomeres in male antennae, thud antennomere long, basal antennal

flagellomeres not symmetrical with apical flagellomeres, mandible apices acute (inside

angle < 90 degrees), emargmate eyes, eyes posterior-ventrally approximated, and wing

vein MP3 not contacting MP1+2. Therefore, I propose moving the genera D ioptoma and

Diplocladort out of Phengodidae and into Rhagophthalmidae. On the other hand,

Cantharidae does not seem to be supported as monophyletic in this analysis, and none o f

the four cantharid taxa included in the analysis form a clade. Cantharidae needs to be

farther examined in relation to other taxa and sampled more thoroughly within a

phylogenetic context before a taxonomic change is made.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Phylogenetic Relationship B etw een T.ampyridae and Phengndiffae

The family Phengodidae is composed of bioluminescent species that commonly

resemble fireflies in their general appearance and are usually also found to be sympatric

with many firefly species. Even though phengodid beetles share aspects of their biology

with Iampyrids (larviform females, the use of pheromones and luminescence),

phengodids historically have been seen as a group taxonomically distinct from

Lampyridae. However, it is probably the similarities between Lampyridae and

Phengodidae, with bioluminescence being one of the most obvious, that have linked them

as closely related taxa in the eyes of many cantharoid workers. ‘‘Within Cantharoidea,

Phengodidae and Lampyridae appear to be directly related, so that the luminosity of both

groups can plausibly be attributed to inheritance from a common ancestor...” (Crowson

1972). The present phylogeny provides evidence that Phengodidae is not sister to, or

basal to, Lampyridae. In addition, it shows that Rhagophthalmidae and Omalisidae are

the bioluminescent families that are sister, or basal to Lampyridae, respectively (Figure

3)-

The Evolution of Biolnmwiescence in Non-lampyrid Cantharoids

Bioluminescence in the order Coleoptera is known to occur in Elateridae,

Staphilinidae (Costa et. a t 1986) and four cantharoid families: Omalisidae,

Ragophthalmidae, Phengodidae and Lampyridae (Lloyd 1978). Several recent studies

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. have provided hypotheses concerning the evolutionary relationships within or around

Cantharoidea (Crowson 1972, Potatskaja 1983, and Beutel 1995) using a variety of

techniques, characters and exemplar taxa. P lottin g luminescence (larval or adult) onto

these different trees, supports interpretations rang in g from three origin s, to one origin and

three losses. However, these studies treat families as single units. Therefore, if

luminescence arises only once in each of the three luminescent families, (Omalisidae,

Phengodidae, and Lampyridae, with Rhagophthalmus species treated as phengodids),

three separate origins would be the maximum number of steps. Conversely, a single

origin would be the minimum number of steps if all fam ilie s were treated as being

monophyletic. Crowson (1972) proposed a dendrogram for the relationships between the

cantharoid families (Omethidae, Cantharidae, Plastoceridae, Lycidae, Omalisidae,

Drilidae, Telegeusidae, Phengodidae, and Lampyridae). This scheme predicts two

character optimizations of three steps each. One optimization poses three origins for

bio luminescence, while the second poses two origins and one loss of bio luminescence.

Potatskaja (1983) proposed a dendrogram for the relationships between the cantharoid

fam ilie s (Brachyspectridae, Cantharidae, Phengodidae, Drilidae, Omalisidae, Lycidae and

Lampyridae) based on larval mouthpart characters. Potatskaja concluded that two

lineages, one termed “cantharid” (composed of Phengodidae, Drilidae, Omalisidae,

Brachyspectridae, and Cantharidae) and the other “tycid” (composed of Lampyridae and

Lycidae) originated from a phengodid ancestral form. No specific taxon was designated

as the root. In reference to the origin of bioluminescence, PotatakajaTs topology predicts

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. two optimizations of three steps each: three separate origins, or two origins and one loss.

In 1995, Beutel proposed a phylogenetic analysis of Elateriformia based on 27 larval

characters (33 states). Within his analysis Cantharoidea was represented by seven taxa,

one species per each of Brachyspectridae, Cantharidae, Drilidae, Omalisidae,

Phengodidae, Lampyridae, and Lycidae. All of the cantharoid taxa were placed in the

same clade except for Cantharidae, winch was placed close to Elateridae, rather than with

the rest of the cantharoids. The clade containing the bioluminescent cantharoid taxa was

poorly resolved in the consensus tree and predicts a single topology of one origin and

three losses.

My analysis suggests a single solution, considering a taxon to be luminescent if

any life stage is luminescent. There are two origins of bioluminescence with one loss:

luminescence arose once basaQy, early in the evolutionary history of the cantharoid clade,

and was subsequently lost and then later regained in the phengodids, see Figure 3. Taxa

in which luminescence was regained under this scenario are currently classified as

belonging to the family Phengodidae, (Cenophengus palltsus, Phrixothrix reducticomis,

Pseudophengodes pulchella, and Zarhipis mtegripermis), as well as Stenocladius sp.,

which I consider to be a phengodid and herein propose its inclusion in this family. The

seven synapomorphies that define Phengodidae are these: tibial spurs are absent,

bipectinate antennae, distal margin of antennal flagellomeres approximating proximal

m argin in width, antennal lobes produced from basal region of flagellomere, two

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. elongated antennal lobes per flagellomere, narrow juncture between flagellomere and

antennal lobe, and juncture between lateral and bind m argins of pronotum are truncate

(=90 degrees). Therefore, all luminescence in the cantharoid lineage is homologous

except for that of Phengodidae, which is a reversal to luminous habit Additionally, all

known luminescent cantharoid taxa have lum ino u s larvae, and in Omalisidae the larvae

are luminous, but either adult is not (Crowson 1972). The fact that Omalisidae is the most

basal of all the bioluminescent cantharoids indicates that luminescence arose first in the

larvae and then subsequently in the adults (Figure 4).

Photic Organ Evolution in Non-Iampyrid Cantharoids

Larvae

Of the larval photic organs, only those of Phengodidae and Lampyridae have been

studied in detail. The larvae of Omalisidae have never been studied, and the larvae of

Rhagophthalmidae have been studied and described only recently (Wittmer and Ohba

1994), though no morphological, physiological or histological work has been published

on this group. Therefore, from what is currently known from evidence scattered among

the taxa, the pattern of two luminous spots per segment on larvae is the most ancient and

common larval photic organ pattern in the cantharoid lineage (Figure 4). The number of

luminous segments varies, but all known luminous cantharoid larvae bear pairs of

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. luminous photic organs. While most Iampyrid larvae bear a single pair of photic organs

on only the eighth abdominal segment, larvae of the other taxa generally possess a pair of

photic organs on each abdominal segment with additional pahs sometimes present on the

larval thorax (Table 2). While the larvae of many genera of luminous cantharoids are not

yet known, all species known to be luminous as adults are also luminous as larvae.

Therefore, while only some larvae are known horn the families Omalisidae,

Rhagophthalmidae, Phengodidae and Lampyridae, the larvae of all species in these

families are hypothesized to be luminous (Figure 4).

Crowson (1972) hypothesized that Barber's (1908) luminous larva horn

Guatemala, described as Astraptor sp., could have been a large female larva or a

Iarviform female o f Tetegeusis . Sivrnski (1981) points out that in a later unpublished

manuscript Schwarz and Barber identified the single specimen as the phengodid

Microphenus gorhami. Therefore, as for as is known, the family Teleguesidae does not

contain any luminous taxa. Barber (1908) mentions that there was a single photic organ in

the head which produced a red light that was thrown directly forward and hence was not

easily seen from above. This specimen seemed to have no other photic organs, though it

was observed in the daytime and not for long. A red head-light is known to be present

only in other phengodid larvae (Vrviani & Bechara 1997).

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

Females of luminous cantharoid taxa, excluding Lampyridae, generally possess

the same photic organ morphology as their larvae, which is generally paired, luminous

spots on the post-lateral margins on some of the thoracic and each of the abdominal

segments (Table 2), Females, and males of the family Lampyridae vary in photic organ

morphology (Lloyd 1978), perhaps due to sexual selection, as the females of many firefly

species attract mates via a luminescent sexual signal system (McDermott 1917, Schwalb

I960, Lloyd 1978 and 1979, Branham & Greenfield 1996, Vencl & Carlson 1998). While

the luminescent sexual signals of fireflies have received considerable attention,

pheromones are the dominant sexual signals used in courtship in most cantharoids,

including Phengodidae, and also are used by many lampyrids (McDermott 1964, Lloyd

1971). Therefore, (with the exception of the family Omalisidae and the genus Drilaster),

the pattern of bioluminescent evolution in the larvae (Figure 4) and the pattern found in

the females of luminous taxa (Figure 5) are very similar. The fact that Omalisidae is the

basal-most luminous cantharoid taxon, and both adult males and females are not

bioluminescent, suggests that bioluminescence first arose in larvae and later in adults.

Rhagophxhalmus ohbai females, in addition to retaining the larval pattern of

photic organs, also possess a novel photic organ on the eighth ventrite, which is used in

courtship, see Table 2 (Ohba ef. aL 1996a). After using the ventral photic organ on the

eighth ventrite for courtship, females curl around their eggs and glow from ten sets of

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. paired photic organs located at the lateral m argins of the ten luminous body segments,

(see Table 2), which serve as an aposematic warning display (Ohba e t aL 1996a, Chen

1999). Rhagopthahnidae appears to be the sister of Lampyridae, which is the only other

cantharoid family known to contain females that employ photic sign a ls in courtship. In

addition, based on the presence of extremely well developed eyes in rhagopthalmid males

and the lack of greatly elaborate bipectmate antennae, such as those found in

Phengodidae, I believe that photic signals are the primary mode of sexual s ignaling in this

family. Crowson (1972) incorrectly cited Green (1912) as reporting that the female of

Harm atelia is apterous and Iarviform. Green (1912) states that, “I have not yet succeeded

in determining the female of this beetle, and it rem ains uncertain whether the other sex is

an apterous grub-like creature, or whether it is in the form of another beetle.”

Adult Males

All known bioluminescent adult cantharoid males are restricted to the families

Lampyridae, Rhagophthalmidae (as defined here) and Phengodidae (as defined here), as

well as the genera H arm atelia and Dioptoma. While the exact number and position of the

photic organs varies, they are generally found in pairs on one or more of the thoracic

segments, and on each o f the first eight abdominal segments. Male photic organs, like

those of larvae and females, are found near the lateral marg in s of these body segments

(Table 2). The more dorsal position of these lateral photic organs in the adult males

versus larval males is probably due to the lateral tergites, found as plates in the dorsal

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. region on the side of the larval abdomen, becoming fused to the larval tergites to form a

single large plate covering the entire dorsal surface of the adult male abdomen. One

exception is Pseudophengodes pulchetta, which on the ventral surface of the eighth

ventrite bears a large photic organ that seems to be used in courtship. Until recently, the

only phengodid genus that was known to contain adult luminescent males was

Pseudophengodes . However, VLviani and Bechara (1997) discovered through rearing

experiments that phengodid males in the tribe Mastmocerhn ( Brasilocerus , Euryopa,

Mastinocerus, Mastinomorphus, Phrixothrix, Stenophrixothrix, and Taxinomastmocerus )

are luminous throughout the adult stage and that the luminescent emissions seem to serve

a defensive rather than courtship function. No adult phengodid males in the North

American tribe Phengodini ( Phengodes and Zarhipis) are known to be continuously

luminescent through the entire adult stage. Even though there is little variation found in

m ale photic organs outside of Lampyridae, the scattered occurrence of photic organs in

males clearly seems to indicate multiple origins (Figure 6).

J.W. Green’s first published observations (1911) of live Harmatelia bilinea males

did not include any notice of luminescence, even though Green was specifically looking

for evidence that this insect was luminescent. The following year (1912), Green published

that he had observed two specimens that “exhibited a distinct light when examined in a

dark room.” The fact that Green had examined many Harmatelia specimens without

noticing any photic emission may be an indication that these males are not luminescent

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. throughout their entire adult life. It is well knows (McDermott 1965, Lloyd I978,Vivani

& Bechara 1997, Branham & Archangelsky 2000) that some Iampyrid species that are not

luminescent as adults retain the ability to glow via larval photic organs for a short tfme

alter they have eclosed and are still teneraL McDermott (1965) hypothesized that the

males of both P terotus and H arm atelia might have the ability to produce light only

briefly after eclosion, as does the firefly Lucidota atra. McDermott also mentioned in tins

same work that H.S. Barber (unpublished observation, confirmed by J.E. Lloyd) observed

thatPhengodes males also have the ability to produce light shortly after eclosion. This

photic carry-over into the adult, while only temporary in some taxa, is suggestive of a

larval origin of the photic organ and its carry-over into the adult.

Some Rhagophthahnus ohbai males are known to be luminous from paired spots

along the lateral edges of ten body segments, (see Table 2). These males are weakly

luminous (Chen 1999) and evidently are not always observed (Ohba et. al. 1996a). It

seems likely thatRhagophthahnus males have only a temporary ability to produce light

immediately following eclosion, and luminescence is not used in courtship (Ohha et. aL

1996a).

Luminescence and Life Stages

The phengodid genus Phrixothrix is the only luminous non-Iampyrid cantharoid

in which larvae, adult males, and adult females are known to be luminescent throughout

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. all life stages (Table 2). Luminescence throughout all life stages of P hrixothrix hirtus is

essentially the same. The photic organ morphology appears to be identical between all

life stages with the exception that the head lantern is lacking in the adult males. Therefore

the lateral lanterns of all stages appear identical and the head lanterns of the larvae and

females are identical. In addition, the photic emission spectrum is essentially the same for

each type o f photic organ, regardless of life stage (Costa et aL 1999). An additional

example of similarity in the emission spectra and a possible connection between larval

and adult luminescence was found by Viviani and Bechara (1997), who argued that,

“Continuance of the same bioluminescent color in the lateral lanterns of larval, pupal, and

adult stages of Mastmomorhus sp.l and P. heydeni suggests conservation o f the same

luciferase iso-form throughout its life cycle.” The pattern of photic organ morphology

appearing to be more or less identical across life stages, along with similar photic

emission spectra being emitted from these organs, supports the hypothesis by Crowson

(1972) that luminescence first evolved in larvae and was then “carried over into adults.”

Evolution of Photic S ig n a lin g in Non-Firefly Cantharoids

Sivinski (1981) provides a detailed synopsis of the various theories that have been

proposed for the function of larval luminescence and the evidence supporting each. While

it is now generally accepted that lampyrids are chemically defended and larval photic

em issio n s probably function as aposematic displays (Belt 1874, Lloyd 1973b, Sydow &

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Lloyd 1975, Eisner et. aL 1978, Underwood e t aL 1997, Knight et. aL 1999, De Cock &

Matthysen 2000), there exists much less information concerning whether other larval

cantharoids are distasteful as well, though it appears that at least some phengodids are

chemically defended (Burmeister 1873, Harvey 1952, Sivinski 1981). It is interesting that

almost all larval photic organs are paired and are located on the sides of the abdomen or

on the eighth abdominal ventrite where the photic emissions are readily seen from the

side or from above. This is most consistent with the aposematic warning display

hypothesis. The exception to this rule is the pair of medial photic organs on the head of

some phengodid larvae such as Phrixothrix (Table 2). The photic emissions from these

organs were measured by Viviani and Bechara (1997) and were found to be in the range

of574-636nm, well into the red range. Electroretinograms of Phrixothrix larvae showed

that these larvae have a spectral sensitivity shifted to the red (ViL Viviani, E.J.H.

Bechara, D. Ventura and A. Lail unpublished data, Viviani & Bechara 1997). Viviani and

Bechara (1997) hypothesize that these red-emitting head-mounted photic organs provide

an illumination function, which may help in locating prey that do not possess spectral

sensitivity shifted to the red, and that the lateral photic organs serve an aposematic

defensive function.

As the basal luminescent taxa possess only lateral photic organs, it is probable

that the first function o f larval luminescence was as an aposematic warning display.

Larval photic organs were then lost in Lycidae, Omethidae and Cantharidae, and then

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. reappear in Phengodidae. ha some phengodids, a photic organ arose on the head and

produced red light which was used for illuminating prey (Viviani & Bechara 1997).

The function of luminescence in the adult phengodids is not well understood.

Available data suggest that bioluminescence, produced by the lateral organs o f the adult

males and females in these two families, seems to serve in aposematic defense rather than

mate attraction (Rivers 1886, Tiemann 1967, Sivinski 1981). However, the continuous

glow produced by the ventral photic organ on the eighth abdominal ventrite in

Pseudophengodes is also consistent with use as either illumination of the surroundings

during flight or intersexual communication (Viviani & Bechara 1997).

The function of luminescence in Rhagophthahnus ohbai was studied by Ohba et.

aL (1996a), who provide evidence that the emissions of the lateral photic organs in

females serve an aposematic warning function, illumination while the female guards her

eggs, while the ventral photic organ on the eighth abdominal segment seems to function

exclusively in a courtship context. Hence, in this family the paired lateral and the ventral

photic organs are used independently in separate contexts: defense and courtship. Across

all luminescent cantharoid taxa, photic organs used to produce sexual signals seem to be

restricted to the ventral regions of the body. It is also interesting to note that the eighth

abdominal segment is consistently associated with the location of such photic organs. The

reason for this association remains unknown.

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

My phylogenetic analysis suggests that bioluminescence arose twice within the

cantharoid lineage and was lost once. The first origin of luminescence in the lineage was

ancient and luminescence first arose in larvae where it served as an aposematic warning

display. Luminescence was retained in the larvae of the Rhagophthalmidae and

Lampyridae and was likely carried through the pupae into the adult stage, where it

became functional in some taxa and not others. While photic signals are used in mate

attraction in the Rhagophthalmidae, adult photic signals reached their greatest

sophistication in the adults of the family Lampyridae, where photic signals are used in

intraspecific communication, and both photic organs and photic signals became greatly

elaborated under the context of sexual selection. The second origin of luminescence

occurred in the family Phengodidae, where its function in both larvae and adults is as an

aposematic warning display. In some phengodid taxa, luminescence has become

elaborated to serve possibly as an illumination device for locating prey. In addition, males

of the genus Pseudophengodes possess a Iampyrid-like photic organ on the eighth

abdominal ventrite, which glows continuously and likely serves either to illuminate

potential landing sites or to function in courtship. While some researchers have

previously hypothesized that the families Lampyridae and Phengodidae are close relatives

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. and share the charismatic ability to produce bioluminescent signals, these two fernilfps

are perhaps more interesting than previously thought, because they are not closely related

and then: bioluminescence is convergent

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. TABLE 1

A summary of the various classification schemes, from the earliest works to recent,

showing the various taxonomic placement of taxa that this study relegates to “ Incertae

S ed is” status. Authors have had difficulty in placing these taxa within existing families

because they share various characteristics with several differentfam ilie s, A phylogenetic

analysis including these problematic taxa (this analysis) has the advantage of using all

available characters to place taxa. However, while this analysis placed Stenocladius

within Phengodidae, and Dioptoma and Diplocladon within Rhagophthalmidae,

D rilaster , H arm atelia and Pterotus were not placed within any o f the existing cantharoid

families and therefore appear to represent lineages independent o f the existing cantharoid

lineages.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. )" is used in )". The varied Drilaster Drilaster* (this analysis) Pterotus Stenocladius Diplocladon Dioptoma M, M, Branham Harmatelia (=■ Elateroidea Inceriae Sedls Inceriae (**Drilaster Phengodidae Rhagophthalmidae

Ototreta Ototreta while "

Drilaster)* (= (1995) Ototreta, {Gorham 1883} Ototreta and to eliminate further confusion, Additionally, I Harmatelia Pterotus Stenocladius Dioptoma Diplocladon Lawrence& Newton (after Crowson 1972) Ototretinae Pterotinae Rhagophlhalminae Lampyridae Phengodidae Drilaster status. , , Sato (1968) confirmed this synonymy while pointing out that Ototreta (1964,1966) imertaeSedis is used in Asia without reference to TABLE 1 {Pascoe {Pascoe 1860} (1964)(1966) F, F, McDermott Ototreta Harmatelia Pterotus Dioptoma Luciolinae (1966) Pterotinae (1964,1966) Rhagophlhalminae Ototretinae (1964) and Lampyridae {-Drilaster)* Drilaster retaining the name to cede the priority ofthe name Ototreta, Drilaster and E. E. Oliver (1910) a, b Drilaster Ototreta Harmatelia Crowson ( 1972) as well as Lawrence and Newton ( 1995) retained McDermott's use of“ Luciolinae Megalophthalminae Drilaster Stenocladius Lampyridae (a) Drilidae (b) Drilidae (b) Ototreta, be removed from Lampyridae and be given Elateroidea

Drilaster has priority over Early Early Work {E, Olivier{E, 1900} {Fairmaire {Fairmaire 1878} {JCiesenwetter {LeConte {LeConte 1859} {Walker {Walker 1858} Ototreta Drilaster Stenocladius Pterotus Harmatelia 1879} Lampyridae Drilidae Elateridae ♦ McDermott ♦ ( 1964,1966) synonymized Europe and (lie United States, I refer to this genus as use ofthese generic names has led to some confusion: the genus Drilaster propose that

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. TABLE2

A comparison of cantharoid taxa included in this analysis with, special reference to photic

organ morphology across life stages and the presence of neotenic characteristics in adult

females.

2 9

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

Mates; Morphology Organ Photic None None None margin of prothorax. One pair One prothorax. of margin on lateral margins of all 8 abd. 8 all of of margins pair lateral one on and segments, 4 p, organs along doisal hind doisal along organs 4 p, 1912) (Green seg.5-7 None dorsal organs on each abd. each on organs dorsal paired p. organs on lateral on and seg. organs p. abd. additional each an of paired with margins mesonotum on pronotum organ central 1999) (Chen Weak light visible from sets of sets from visible light Weak None None (Viviani & Bechara 1997) Bechara & (Viviani on prothorax and paired p. paired and dorsolaterally prothorax on located organs A single dorsomedian p.organ dorsomedian single A segments abd. and thoraslc on

et. el. et. el. et. Rsmalei Photic Organ Morphology Organ Photic None None None Roundly quadrate p. organ, p. quadrate Roundly almost completely occupying completely almost 1996a) the venter of penultimate abd. penultimate of venter the segment (Green 1912) (Green segment p. organs on each body each on organs p. segment except head and last and head except segment body segment (Haneda 1950) (Haneda of segment margins body postlateral on organs (Wittmer 6 sag. abd. A medial dorsal and two lateral two and dorsal medial A None None single p. organ on ventrite of abdomen; and ventrite on mesothorax organ p. single Ten sets of paired p. paired of sets Ten None None None 1973, Harvey 1952) Harvey 1973, ??? posterolateral margins of of margins posterolateral segment (Halverson (Halverson segment Two medial organs on head and head on organs medial Two eleven pairs of p. organs in the in organs p. abd. of pairs 9M the eleven through thoracic

are

1966) Seleste unicolor Seleste TABLE 2 (Continued on following page,) et- at. et- Femiles; Alate or Larvlform abdomen (Crowson 1972) (Crowson abdomen abdomen (Lawrence 1991a). (Lawrence abdomen of Females "larviform’ (Barker 1969) (Barker 1972) "larviform’ (Crowson abdomen Wingless, elytra shorter than shorter elytra Wingless, Wingless, elytra shorter than shorter elytra Wingless, than shorter elytra Wingless, Neotenio: larviform with the with larviform Neotenio: exception of adult antennae and antennae Into adult of subdivided exception (tarsi legs Larviform: possessing larval possessing Larviform: larsomeres and daws) (Lawrence daws) and 1995) larsomeres Newton & antennae and legs (Lawrence & (Lawrence legs and antennae 1995) Newton Neotenic: larviform with the with tarsi and larviform antennae, 8-seg. of tarsomeres Neotenic: Into exception subdivided ClSWS (Costa Nearly always alate always Nearly (Lawrence 1991) (Lawrence Unknown Unknown None Larviform Larviform Alate, occasionally reduced wings reduced occasionally Alate,

1996a) et. 1973, Harvey 1952) Harvey 1973, et. el. et. Larvae: Photic Organ Morphology Organ Photic Unknown None Paired p. organs on sides of abdomen of sides on organs p. 1891b) Paired (Lawrence Unknown A medial dorsal and two lateral p. lateral two and dorsal medial A 1973) a/. et. (Halverson organs on each body segment except segment body each segment on body organs last and head mesothorax to S'* abdominal segment from segment: abdominal each on S'* to of. organs (Ohba mesothorax total). sets (10 NoneUnknown Alate None Unknown Unknown A median dorsal and two postlateral p. postlateral two and dorsal median A Halverson Halverson Two medial organs on head and eleven and head on 9 the organs 1997, medial through Two Bechara & thoracic 2* of (Viviani margins segment abd. pairs of p. organs in the posterolateral the in organs p. of pairs Cenophenyus Dloptoma Dlplocladon Rhagophthalmus Phrixothrix Plaetoceridae DrWdee Omalleidae Rhagophtalmidae Lycidae Phengodldae Cantharldae Omathidae Telegueeldae Q

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

1887) el al abd. segment abd. Or Meles; Morphology Organ Photic (Viviani & Bechara 1887, M. segment 1887, abd.

al.

(Tiemann 1907, (Tiemann 11 luminous bands: 1 each at each 1 bands: luminous 11 Fsmsls; Morphology Organ Photic abd. on spots luminous 1-3 tergites. Paired p. organs (1 organs p. Paired tergites. 1887) base of meso- and metathroax, abd. and last the meso- of but base all on and spot per side): on upper lateral upper on Also, side): 1-8. sags, per abd. spot of surfaces sterna 2-8. 2-8. sterna Rivera 1880) Rivera entire body - no specific photic specific - no body entire organs present. (Ohba e(, e(, (Ohba present. organs A diffuse glow emitted from emitted glow diffuse A Paired p. organ on 7” abd. 7” on organ p. Paired segment (Dean 1878) (Dean segment

1887) el al. Females; Alate or Larviform Larviform Neolenlc: larviform with the with larviform Neolenlc: exception of tarsi subdivided into subdivided tarsi of exception (Ohba tarsomeres and claws. and tarsomeres Larviform?; Dean (1878) states that states (1878) Dean Larviform?; Alate (some larviform?) (some However, Alate of presence larviform. the and fully are Indicates antennae females II adult None Plate eyes, his compound paired claws on each leg. each on claws paired

abd. segment abd. abd. segment abd. 7* T a/. 1886b) a/. el. Larvae: Morphology Organ Photic Unknown Unknown Unknown 11 luminous bands: 1 each at base at each 1 bands: luminous 11 lateral surfaces of abd. sags. 1*8. sags. abd. of surfaces lateral 1907) (1lemann of meso- and metathroax, and on all on and metathroax, and meso- of but the last abd. lergltes. Paired p. Paired lergltes. upper on abd. side): last the per but spot (1 organs (Ohba (Ohba Paired p. organ on on organ p. Paired Unknown Unknown Unknownof side each on organ p. One Paired p. organs on 8* abd. seg. abd. 8* on organs p. Paired Paired p. organ on on organ p. Paired (Ohba 1883) (Ohba 1878) (Dean Paaudophengodea Zarhlpla Haimatalla Stanooladlua Ptemtua Drilaaler TABLE 2 (Continued) "Inoartaa Sadia"

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FIGURE 1

Strict consensus o f280 most parsimonious trees (848 steps, C i. 0.16, RX 0.57). The

limits of the cantharoid out groups and the family Lampyridae are delimited by the

vertical bars on the far right of figure.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. a.CO 3 es s o3 2 Sa

s 'Ok O Lam pyrldae

F IG U R E 1

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

The strict consensus with the Iampyrid clade collapsed and represented by Lampyridae.

Numbers located under the nodes are Bremer Support values (set at a max. Bremer value

o f 5). Numbers located above the nodes present the number of synapomorphic characters

at that node.

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FIGURE 2

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FIGURE 3

The evolution, of bioluminescence in cantharoids. The condensed strict consensus tree

with two origins of luminescence and one loss plotted.

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FIGURE 3

3 7

I Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FIGURE 4

The evolution of larval bioluminescence in cantharoids. The strict consensus tree with the

lampyrid clade collapsed. The known presence of larval luminescence is indicated by

bold branches, with the hypothesized presence of larval luminescence indicated by dotted

branches.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 3 S 2 3

Q. CO N4 Ilil

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8C o e a <2 t» m o»b • o S 'S 3 -JS 5 5 ° o » 3 3 S ^ 3 3 « •! 5 ^ 2 Q

FIGURE 4

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FIGURE 5

The evolution, of bioluminescence in female cantharoids. The strict consensus tree with

the Iampyrid clade collapsed. The known presence o f luminescence in female*; is

indicated by bold branches, with the hypothesized presence of luminescence in females

indicated by dotted branches.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. £ Q.COM S 3-5

' 5 * a s s a | • 5 P 2 a ®S ® § p . MOi 3 C

-» J* §1Q» 5 II 5 o s l J s j t | N § l

E!5'S S S fe Q to O O t

FIGURE 5

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FIGURE 6

The evolution ofbioluminescence in male cantharoids. The strict consensus tree with the

lampyrid clade collapsed. The known presence of luminescence in males is indicated by

bold branches, with the hypothesized presence of luminescence in males indicated by

dotted branches.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. * a £ i g U

a a 5

s g « Q « O O t

FIGURE 6

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

AN OVERVIEW OF THE FAMILY LAMPRIDAE.

Distribtrtion

Eighty three genera and approximately 2000 species are described within the

family Lampyridae. These are distributed worldwide with the greatest diversity of

species occurring in the Oriental as well as the Neotropical regions. The two largest

genera are the Old World Luciola and the New World Photm us (McDermott

1964,1966; Lawrence 1982). While species diversity is higher in regions that are

predominately moist and relatively humid, a minority of species is known to occur in

arid habitats, i.e., Microphotus, some Pleotom us, etc.

Biology

Adults

Only a percentage of adult lampyrids are luminous, and genera that lack the

ability to glow as adults are generally, though not always, restricted to the basal

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. region of the family and rely solely on long-range pheromones for mate attraction

(Branham & Wenzel submitted). The luminous signals of adult males are produced

from photic organs of various shapes (one spot, paired spots, stripe, entire surface of

ventrite, etc.) located on abdominal ventrites 5-7 (true abdominal segments VI-Vni).

The photic organs o f adult females also appear on ventrites 5-7, though females

generally possess photic organs of reduced size and have fewer luminous ventrites

than males of the same species. Complex luminous sexual signals, consisting of

single flashes or trains of multiple flashes, appear to be associated with more

structurally complex photic organs, which have both increased innervation and

tracheal supply (Buck 1948). These complex organs are large and commonly occupy

the entire ventral surface of the abdominal segment on which they appear (B ranham

& Wenzel, submitted).

Luminescence in adults is typically used in sexual s ignaling to communicate

species identity and facilitate pair formation. These photic emissions vary in

complexity from glows produced by the female to attract non-luminescent males to

flash-answer dialogues between both sexes of the same species, employing critical

tim ing parameters (Lloyd 1966,1971). While glows are used by some species as

sexual signals, communication protocols that employ flash-answer signals are known

to have evolved at least three times in the family (Branham & Wenzel, submitted).

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. “Intermittent flash synchrony” is sometimes known to occur in populations, or

even in groups of males within a population, of some Photm us species which

normally produce asynchronous flashes (Mast 1912; Blah 1915; Otte & Smiley

1977). The intermittent synchrony found in Photm us fireflies is produced by flying

males. However, perhaps the most spectacular examples of synchronous flashing are

produced by sedentary aggregations of synchronously flash in g fireflies of several

species of the Old World Luciola, which occur in tropical Southeast Asia, from the

Philippines to eastern India and Sri Lanka (Buck 193S). These synchronous

aggregations of fireflies consist of both sexes, which are drawn to the aggregation

sites from some distance and remain there, signaling at night, for weeks if not months

on the same vegetation (Case 1980; Ohba 1984; Lloyd et al. 1989). Many models

have been proposed to explain the possible mechanisms/protocols by which

individuals in the population are able to maintain such a high degree of synchrony

(Lloyd 1973a; Buck & Buck 1978; Buck & Buck 1980; Otte 1980).

Within Lampyridae, morphological structures adapted to form copulation

clamps are known from only a few species of P teroptyx and Luciola. A copulation

clamp partially formed from the deflexed elytral apices, is restricted to males of

P teroptyx from New Guinea. To form the top of the clamp, the tips of the male’s

elytra hook around the anterior edge of the female’s sixth abdominal terghe. The

bottom of the clamp is formed from the distal margin of the male’s seventh stemite,

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. which pushes upwards against the ventral surface of the female’s abdomen (Lloyd &

Wing 1981, Wing et al. 1983, Ballantyne 1987a). Males o f non-New Guinea

P teroptyx and Luciola pupilla do not possess deflexed elytral apices but show clamp-

like modifications to the end of the male abdomen (Ballantyne 1987b). Interestingly,

the modifications of the abdomen in non-New Guinean Pteroptyx do not appear to be

homologous with those found in their New Guinea relatives (Ballantyne

I987a,1987b).

Aggressive mimicry is known to occur in the females of the Photuris

permsybanica-versicolor group. These females respond to the flash patterns of

conspecific males with their own species-specific signals. Following a short flash

dialogue, male Photuris land near the females and mating ensues (Barber 1951).

Either the mechanical act of mating and/or substances transmitted from male to

female during mating cause the females to replace normal sexual signaling behavior

with aggressive mimicry signaling behavior (Zom & Carlson 1978). After this

behavioral switch, females fly down from the trees or high vegetation where they had

been mating to land on the ground or short vegetation, where sympatric Photinus or

Pyractomena females are perched and respond to the flashes of their own conspecific

males (Lloyd 1965,1984). The P hoturis “femmes fatales” then produce flashes that

approximate the female response of the sympatric Photm us or Pyractomena, and,

when their heterospecific males land near them to mate, thePhoturis females quickly

4 7

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. seize and devour them (Lloyd 1965,1984). While it was first hypothesized that these

aggressive mimics gained valuable protein from feeding on other fireflies, Eisner et

al. (1997) discovered that Photuris versicolor females acquire defensive steroids

called lucibufagins- High levels of lucibufagins are common in P hotm us fireflies, but

both P hoturis males and females appear to lack lucibufagins upon emergence from

the pupa. In addition, Eisner et al. (1997) cites evidence that P hoturis aggressive

mimics endow their eggs with the newly acquired defensive steroids.

Spermatophores are found in some species of fireflies in which females mate

more than once. Nutrients derived from these spermatophores have been shown to be

incorporated into developing oocytes. Conversely, in species where females mate

only once, the eggs are fully developed immediately following female eclosion and

males do not form spermatophores (Wing 1985, van der Reijden et al. 1997, Rooney

& Lewis 1999,2002).

Larvae

Lampyrid larvae are common in mesic environments, where they are found

along the marg in s of streams and ponds, as well as in leaf litter or rotten logs and

under rocks. In arid regions, larvae are commonly active above ground immediately

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. following rains. All known. Iampyrid larvae are luminous. In fact, the origin of larval

luminescence predates the origin, of the family Lampyxidae (Branham & Wenzel

2001). Larvae produce light via paired photic organs located on the ventral surface of

the eighth, abdominal segment. The only larva that is known to vary from this photic

organ morphology is Lamprohiza delarouzei Jacq-DuV., which has two pairs of

photic organs (one pair each on abdominal segments 2 and 6) (Balduf 1935), and

Lamprohiza splendidula L., which have 3-12 luminous spots on abdominal 2-6

(Schwalb I960).

Though luminescence is employed in the eggs, larvae, and most pupae of

Iampyrid species, the adaptive significance only of larval luminescence has received

considerable attention (McDermott 1964, Crowson 1972, Siviniski 1981). The fact

that fireflies are known to sequester defensive steroids and are commonly rejected by

naive predators suggests that larval luminescence functions as an aposematic warning

signal (Kloff et aL 1975; Eisner et al. 1978; Underwood et aL 1997; Knight 1999).

Some genera, such as Photmus, are subterranean as larvae while others, i.e..

P hoturis, are active on the surface in leaf litter and still others, i.e., Pyractomena, are

semi-aquatic and are able to stay submerged for a short period of time while foraging

for food (Buschman 1984b). Some species in the genus Luciola are fully aquatic,

respiring through the use of tracheal gills and feeding exclusively on one or two

4 9

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. species of aquatic snail (Aimandale 1900, Blair 1927, Okada 1928). The larvae, pupae

and adults of both sexes of Pleotomodes needhami Green have been collected in the

nests of several species of ants. P. needhami is thought not to feed on the ants or their

brood, and the ants appear to ignore the fireflies (Sivinski et al. 1998).

Schwalb (1960) discovered that larvae locate prey via chemical cues and seem

to have the ability not only to follow old snail slime trails but also to determine the

polarity of these trails in order to follow them to the sn a il. Once it locates the prey,

but before feeding, a larvae injects digestive juices from the midgut through a channel

in each mandible into the prey. The digestive juices paralyze the prey and then liquefy

its tissues, which are then ingested through the larva’s oral cavities (Fabre 1913,

Vogel 1915). Some larvae are very specific as to which prey items they will take

(several species of Luciola appear to feed exclusively on only one species of aquatic

snail), whereas larvae of Photuris are known to feed on varied live prey, “scavenge”

on dead insects, worms, and apparently even feed on the fructose of berries

(McDermott 1964, Buschman 1984a).

The tenth abdominal segment of all Iampyrid larvae bears eversible

filamentous appendages (pygopods) used as holdfast organs. Each of these

filamentous, tubular organs is covered with minute hooks that adhere to all types of

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. substrates. The holdfast organ also aids in locomotion and in the grooming of the

larval head after feeding (Parva 1919, Bugnion 1933, Domagala & Ghiradella 1984,

Archangelsky & Branham 1998).

The number of larval instars appears to vary in Lampyridae. While

Archangelsky and Branham (1998) found five larval molts in Pyractomena borealis

(Randall), which were all reared under the same photoperiod, Buschman (1977) found

instar number to vary from 4 to 9 in Pyractomena lucifera (Melsheimer) under

different photoperiod regimes. Higher numbers of larval instars seem to be present in

larvae under shorter photoperiods. Naisse (1966) and Buschman (1977) found that the

female larvae of Lampyris noctiluca L. and Pyractomena lucifera have one more than

male larvae of the same species. Lampyrid larvae can take from several months to two

years to reach pupation (Williams 1917, Hess 1920). Lampyrids pupate underground

in excavated cells, on the surface in covered cells called igloos, or in natural cavities

in dead logs (Archangelsky & Branham 1998,2001; B uschm an 1984a; Branham &

Archangelsky 2000; Hess 1920; LaBeOa & Lloyd 1991). Those species that are

aquatic or semi-aquatic as larvae (some Luciola and Pyractomena, respectively) exit

the water prior to pupation and construct mud cells. Some Aspisom a species have

arboreal larvae that transform into cryptic pupae, which hang like “Lepidoptera

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. chrysalids” (LaBeila & Lloyd 1991). Most known pupae are luminescent (even if they

are not luminescent as adults) and increase the intensity of light they produce when

disturbed.

Description

Adults

7-40mm in length, elongate oval and generally flattened. Head is completely

or partially concealed from above by the pronotum. Compound eyes present, posterior

region generally not emarginated. Ocelli absent. Antennal insertions flush with head

capsule and approximate. Antennal morphology varies from filiform, to serrate,

bipectinate, flabellate, biflafaellate, and capitate (Figures 7A-7G). 8-11, or 13

antenomeres male antennae (with 11 being the most common). Basal antennal

flagellomeres symmetrical with apical flagellomeres. Frontoclypeal suture absent or

incomplete. Mandibles usually falcate (Figure 8), though sometimes shorter and

somewhat robust (Figure 9) and in a few genera are completely glabrous with the

exception of the distal tips (Figure 10), (this condition has been called the

“specialized" or “modified” condition by early workers). Mandible without mola.

Maxilla with a distinct galea and Iacinia, with the apex of the galea densely setose.

Lacinia without spines. Maxillary palp 4-segmented with apical maxillary palpomere

alm o s t always securiform Labial palp 3-segmented with apical labial palp almost

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. always securiform. Anterior-most margin of the pronotal shield appearing rounded or

otherwise pointed and round. Mesal m argins of metepistema are straigh t or nearly so.

Base of prothorax approximating elytra! bases. Elytra present, or reduced in males;

present, reduced or absent in female. Elytra not reticulate. Epipleura distinct and

usually wide at the base of the elytra. Hind wings present, reduced or absent. Radial

cell usually well-developed. Anal lobe of hind wing absent Procoxal cavities

contiguous. Metacoxal cavitites contiguous or nearly contiguous. Tibia! spurs are

usually indistinct or absent Tarsal formula 5-5-5. Claws are paired and simple or

toothed or bifid, without setae near base. Scutellum distinct and sclerotized. Most

adult male lampyrids possess a hologastrous type abdomen with 7 or 8 visible,

ventral, abdominal segments (ventrites), with S ventrites found in the majority of

genera family-wide. In taxa with 8 ventrites, the 8* is the term inal ventral sclerite (the

8* ventrite, when present, is sometimes concealed beneath the 7* ventrite).

Abdominal stemite 2 is generally the first ventral visible abdominal segment (=

ventrite 1), as stemite I has become internalized. Therefore, in males of most firefly

species, the true abdominal segments 0-EX correspond to the visible ventrites 1-8.

Males of the subfamily Luciolinae bear only 6 ventrites and the paedomorphic

brachypterous males of the European species Phosphaenus hemipterus Fourcroy more

or less retain a larval-type abdomen with stemhes 1-9 being visible. In most firefly

species, adult females generally possess one ventrite fewer than males of the same

species. Ventrite 1 is not completely divided by metacoxae. The first abdominal

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. segment is delineated by the terghe only, except in Phosphaemis henripterns and

Photinus gramilatus. Spiracles on the edges o f ventrites or in the emargmations of

abdominal segments. Photic organs present in the one or both sexes (adults) of some

species. When present, photic organs are generally located on ventrites 5,6 and 7

(abdominal segments VI,VII, and VTH). Aedeagus trilobate: phaQobase, parameres,

and penis/median lobe (Figure II). The tips of the parameres may sometimes form

sharp “hooks,” as seen in Tenaspis, Erythrofychnia and Macrolampis. The paramere

tips in Diaphanes are “hooked” but are blunt rather than sharp. In Tenaspis , the

parameres are basely robust while the apical halves become greatly narrowed and

curved, forming long, slender hook-like extensions. The aedeagus of Photuris

possesses two long, thin, lateral filaments that are attached to the phaQobase with a

slightly bulbous apical tip (Figure 12). Two similar lateral filaments are also present

in Vesta species, but do not arise from the phaQobase, but rather, one lateral process

arises from the medial region on the outside surface of each of the two parameres.

The female genitalia bear stylx on the distal apex.

Larvae

5-65mm in length. Body is onisciform, fusiform, or subparaQeL Tergal

margins of larvae are explanate in some taxa ( AtypheUa, Lamprigera, Photuris, etc.),

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. while not explanate in the larvae o f others {Photinus, Pyractomena, Luciola,

Lucidota, Pteroptyx, Colophotia, etc.). Terghes are more or less heavily sclerotized

and pigmented Head subcylindrical, narrower than the thorax and when fully

retracted, is generally concealed from above by the prothorax. The muscular tube that

surrounds the head, unless head is fully extended, commonly conceals occiput

Epicranial and frontal sutures present A single simple eye (stemmata) is located on

each side of the head just caudad of the antenna! base. Head capsule is not fused

ventraily, or very narrowly closed Antennae are 3-segmented and retractable.

Sensonum present on the distal end of the preapicai antenna! segment and is shorter

than or equal in length to the apical antenna! segment (Figure 13a). Clypeus and

Iabrum indistinct Mandibles falcate with an internal channel running from the

mandible base to it's opening on the distal margin just before the apex (Figures I3b).

When viewed in cross section, this internal channel is located within the chitinous

wall of the dorsal surface of the mandible. Incisor edge of mandible usually bearing 1

or 2 heavily sclerotized retinacula. The mesal region of the mandibular base bearing

setae or spines. Cardo sclerotized and distinct Galea 2-segmented. Maxillary palp 3

or 4 segmented Labial palp 2-segmented Stipes broad and elongate. Gula wider than

long. Pronotal tergite subelliptical and narrowed anteriorly. Thoracic terghes

generally divided by a sagittal line (Figure 14a). Meso and metathoracic tergites

subquadrate, slightly wider than long. Three pairs of articulated thoracic legs, each

being 5-segmented (including the simple pretarsus). Femora and tibiotarsi often

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. bearing I or 2 rows of strong setae on inner margin. Legs may be either long and

slender, or short and stout One pair ofbiforous spiracles are present on the

mesopleura. 10-abdominai segments are present, with segment 10 often being

concealed horn above. Abdominal terghes 1-8 generally divided by a sagittal ling,

which is lacking on terghe 9 (Figure 16). Segment 10 is reduced to a short sclerotized

ring that bears the tubular, eversible/retractable filamentous holdfast organs

(pygopods), which are covered with minute hooks. Biforous spiracles are present on

abdominal pleurites 1-8. In fully aquatic Iampyrid larvae, sac-like gills are present on

sides o f abdominal segments 1-8. A two-spotted photic organ is always present on

abdominal segment 8, with one luminous spot appearing on each of the two

abdominal pleurites. While these luminous spots are located on the pleurites and

directed ventrally, they also can be seen glowing dorsally via light transmitted

through the 8* abdominal terghe. The larvae of Lamprohiza bear additional paired

photic organs on ventrites anterior to the 8* segment.

Phytogeny

To date, the only family-wide phylogenetic analysis ofLampyridae has been

conducted by Branham and Wenzel (2001). This analysis was conducted using 58

Iampyrid taxa, representing 7 of the 8 Iampyrid subfamilies (specimens of

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Ototretadrilinae were unavailable for study) as well as 27 outgroup taxa representing

9 other cantharoid families (Lawrence & Newton 1995; Wittmer & Ohba 1994).

Historically, Phengodidae was the hypothesized sister group of Laxrtpyridae due to the

presence of bioluminescence in ail known larvae and m anyof the adults in both

families, which was presumably inherited from a common bioluminescent ancestor

(Crowson 1972). However, a phylogenetic analysis of the cantharoid fa m ilies

Plastoceridae, Drilidae, Omalisidae (=Homalisidae), Rhagnphthalmidae. Lycidae,

Cantharidae, Omethidae, Telegeusidae, Phengodidae, and Lampyridae places

Phengodidae as a distant relative to Lampyridae and hypothesizes that the use of

biohuninescence in both fam ilie s is convergent (Branham & Wenzel 2001).

This phytogeny also indicates Lampyridae is not monophyietic as defined by

Crowson (1972) and Lawrence and Newton (1995) and places the Iampyrid genera

Harmatelia, Pterotus, Drilaster (=Ototreta), and Stenocladhts outside of the family

Lampyridae. hi addition, while Stenocladhts is placed within Phengodidae, none of

the other three genera are placed within existing families. This is not surprising, as

these taxa are known to possess questionable affinities to existing cantharoid families,

and their placement within Lampyridae appears arbitrary (LeConte 1859; McDermott

1964; Crowson 1972). Therefore, to preserve the monophyly of Lampyridae,

Branham and Wenzel (2001) removed these genera from Lampyridae and placed

Drilaster (—Ototreta), Harmatelia, and Pterotus in “Haleroidea incertae sedisn and

Stenocladhts into “Phengodidae Incertae Sedis .” Under this new arrangement, the

5 7

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. synapomorphies defining the base ofLampyridae include: head covered by pronotum,

oblique attachment of the trochanter to femur and wing vein CuAl intersecting MP

above fork (Kukalova-Peck & Lawrence 1993). The monophyly of only 2 (Photurinae

and Ludolinae) of the 7 Iampyrid subfamilies is supported by a modem phylogenetic

analysis (Branham & Wenzel 2001). This finding supports the views of McDermott

(1964) and Crowson (1972) who considered the taxonomy ofLampyridae to be

helpful in the identification of taxa though not in delineating natural groups.

The phylogenetic pattern within Lampyridae and closely related taxa suggests

that luminescent signals first evolved in larvae as aposematic warning displays and

were then carried over into the adults, where they are used as sexual signals. It also is

likely that luminous signals in adult lampyrids function simultaneously as species-

specific sexual signals and warning displays (Branham & Wenzel, submitted).

Patterns of photic organ morphology in larvae and adult fireflies suggest a carryover

of photic organs from larvae into adults. All Iampyrid larvae possess paired photic

organs, a morphology which is also found in paedomorphic adult lampyrids. Adult

fireflies, which produce complex flashing signals, possess larger photic organs,

appearing on multiple abdominal segments, than fireflies not producing flashed

signals (Branham & Wenzel, submitted).

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FIGURE 7 Some o f the variation found in Iampyrid antenna! morphology: (A) Serrate, Pyrocoelia paretexta., left antenna, ventral view; (B) Bipectinate, Psilocladus sp.,

right antenna, dorsal view; (C) Flabellate, Dodacles plumose , left antenna, ventral view; (D) Biflabellate, Lam procera sp., left antenna, ventral view; (E) also Biflabellate, Lucio splendens, right antenna, ventral view; (F) Filiform, Photinus

pyralis, left antenna, ventral view; (G) Capitate, Petalacmis praeclarus, left antenna, anterior view. Scale bar = 1mm.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FIGURE 8 The falcate type adult mandibles found in Lampyridae, Lucidma biplagiata.

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

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FIGURE 9 The somewhat shorter and more robust adult mandible type, Photinus pyralis.

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

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FIGURE 10 The specialized or modified type adult mandible, Phaenolis ustulatus.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FIGURE 10 66

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FIGURE II The trilobate type aedeagus found in Lampyridae, Photinus pyralis.

6 7

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FIGURE 11 68

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FIGURE 12 The aedeagus o f Photuris species possess two long, thin] lateral filaments that are

attached to the phallobase, Photuris drvisL

6 9

DESCRIPTION OF THE LARVA AND PUPA OF LUCWOTA ATRA (G. A.

OLIVIER 1790): ABDOMINAL SEGMENT HOMOLOGY AND LOCATION OF

PHOTIC ORGANS ACROSS LIFE STAGES.

INTRODUCTION

The genus Lucidota, as defined by Laporte (1833) and fixed by Motschulsky

(1853), is restricted to the New World and contains some 64 described species. The

genus ranges from the United States to Argentina. Lucidota atra (G.A. Olivier) occurs

from the northeastern United States to Central America (McDermott 1966). The larva

of this species was first described by H. F. Wickham (1895), but because larvae in the

tribe Photinini are difficult to distinguish (LaBeila & Lloyd 1991), a more detailed

larval description is required. Peterson (1951) apparently incorrectly identified the

larva on which he based his drawing of "Photinus sp." in his book "Larvae of

Insects.”

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Upon examining "Photinus sp.n in Peterson's larval collection at The Ohio State

University and comparing it with larvae I had reared, I discovered that Peterson's

larva is actually L atra .

For this study, last instar larvae were collected in early spring and kept until

eclosion, thus allowing a positive identification from the adult. No larval descriptions

exist for other species of this genus, most likely due to difficulties in rearing firefly

larvae (Archangelsky & Branham 1998).

MATERIALS AND METHODS

Seven last instar larvae were collected in a rotting tog on April 6,1993 outside

of Lawrence, Kansas, and kept in a glass jar with damp wood from the log, along with

some terrestrial snails collected in the same wood. Empty snail shells were removed

from the jar every few days. The wood inside the jar was inspected for moisture

content periodically. When the wood appeared to be drying out, it was moistened with

distilled water. To further simulate the inside of the log, the jar was wrapped with

paper to reduce light entering the jar. No special requirements were necessary for

pupation.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Three larvae and one pupa were fixed In boiling water and transferred to 70%

EtOH. In order to study the larval morphology, a specimen's head, mouthparts and

antennae were dissected, cleared in lactic acid, and mounted on microscope slides

using Hoyer's as the mounting medium. The descriptions and drawings were done

using a Wild MS dissecting microscope and a Zeiss Axioscope 20 compound

microscope, both with a camera lucida.

RESULTS

Description of last larval instar

Length

13.0 to 15.0mm. Body elongate, fusiform, slightly flattened dorsoventrally

(Figure 14,A). Whitish ventrally with pink along sides of thorax and abdomen.

Sclerotized regions uniformly light to dark brown and granulose. AH tergites, accept

abdominal tergites 8 and 9, bearing 3 light colored stripes that are more or less

parallel to the longitudinal axis of body.

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

Prognathous, subquadrate, dorsoventraly flattened, and robust (Figure 143);

retractable within, thorax. Labrum and clypeus fused. Epicranial suture present as well

as frontal sutures that extend to bases of antennae. One pair of lateral stemmata,

posterior to base of antennae. Head capsule not fused ventrally (Figure 14, C).

Antenna

3-segmented, partially retractable w ithin membranous base (Figure 13, A);

originating on latero-apical edges of head capsule. Basal segment widest, attached to

membranous base, median portion of dorsal surface covered with medium length

setae pointing anteriorly, lateral pointing setae on anterior third of segment

approximately 2 to 3 times a long as setae in medial region. Second segment shorter

than third, narrower, evenly covered by long setae, carrying a large globular

sensorium slightly longer than third antennomere. Third segment very short, stout,

with several short setae, an apical spine, and a small globular sensorium on inner

surface just below the antennal apex.

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

Symmetrical, strongly falcate, with an inner channel opening subapicaDy on

outer edge (Figure 13, B). Retinaculum present, forming 2 inner teeth on the apical

third of mandible. Basal third of the retinaculum covered with a dense brush of setae.

Medial region of mandible covered by a single row o f long setae pointing inward

toward the retinaculum, perpendicular to the inner channel of the mandible; I long

seta parallel to apical point of the mandible, just anterior to row of setae located

medially. One 4-pronged seta or sensory appendage on outer margin of mandible, just

before channel opening; outermost prong of this seta longer than the other 3.

Labium

Closely attached to maxilla, formed by a short and strongly sclerotized

prementum, men turn (distaily membranous) and submentum (fused to mentum).

Prementum heart shaped, in both dorsal and ventral views with distal apical cleft

(Figure 15 A); in dorsal view, bearing 2 basal regions of very fine setae, with longer

setae present on the segments of the palp; 2 brushes of fine cuticular spines present on

each side of prementum. Palpus 2-segmented; basal segment short, bearing several

spines, second segm ent twice as long as first, pointed and somewhat forked, with a

single spine (Figure 15,A).

75 i j ! Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Maxilla

Apical region (Figure 153)- Basal region (Figure 14,C). Long and robust,

closely attached to labium. Cardo (Figure 14,C) irregularly shaped, bearing no setae.

Stipes (Figure 153) very broad, ventral surface covered with setae and bearing a

single long seta; dorsal surface bearing 2 long setae. Galea large, 2-segmented, basal

segment very long, 3 times as long as second segment and lacking setae; distal

segment short, conical and bearing several short setae with I seta on distal apex of

segment. Lacinia large, twice as long as first segment of the galea, inner surface

covered with a thick brush of cudcnlar spines. Palpus 3-segmented, basal segment

largest, subquadrate, longer than other 2 segments combined, distal two-thirds

covered with medium to long setae; second segment wider than long and bearing

medium length setae; distal segment subconical without setae, bearing a globular

sensorium-type structure.

Thorax

Prothorax subcircular, wider at base, containing retracted head when larva is

in repose. Meso- and metathorax subrectangular. Thoracic tergites subdivided by

sagittal line. Fach segment with pleural area formed by an upper Iaterotergite, below

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. it an epimeron and epistemum separated by pleural suture; mesothoracic lateroterghe

subdivided, anterior plate smaller, carrying mesothorasic spiracle. Prostenmm

medium sized; meso- and metastema smaller, narrow, subdivided into an anterior

basistemum and a posterior stemelium. I pair of bifbrous spiracles present on

mesopleuron.

Legs

5-segmented, coxae long and cylindrical, robust; trochanters small,

subtriangular in lateral view; femora long and cylindrical, widening slightly apically,

with a single long seta in medial inner portion; tibiotarsi as long as femora, tapering

towards distal end; pretarsi strong, simple, with a pair of stout setae at base. Double

row of strong setae on inner margin of tibiotarsi, lacking on inner margin of femora.

Abdomen

10-segmented, segments 1 to 8 similar in shape, tapering toward end; each

tergite subrectangular, tergites 1 through 8 divided by a sagittal line and 2 lighter

colored lines parallel to sagittal line; lateral portions of tergite 8 lightly colored;

lateral portions of tergite 9 tightly colored and without sagittal line; segment 10 a

narrow ring surrounding anal region, carrying holdfast organ. Pleural areas well

7 7

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. developed, segments 1 to 7 subdivided, upper plate large, suboval, carrying spiracles,

lower plate small, narrowly subtriangular; pleuron 8 with only I suboval plate

carrying a spiracle; pleural areas of segments 9 and 10 reduced Abdominal sterna

large, subquadrate, narrowing towards end of abdomen. Postero-lateral comers of

stemite 8 bearing a twin-spotted photic organ. Color pattern sim ilar to that of thorax.

Biforous spiracles present on pleurites I to 8,

Description of Pupa

Female, one day old Slightly curved, ventraily concave; young pupa white,

older pupa approaching charcoal in color. Length: 10.0 to 11.0 mm

Head

Completely covered by pronotum in dorsal view (Figs. I63X white. Eyes

small, on sides of head; antennae inserted in front of eyes, serrate with 11 obvious

segments, extending in length to metacoxae; antenna and mouthparts white.

78 i

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

Pronotum large, subtxiangular, slight emargination on either side o f anterior

apex, covering head; white or cream. Meso- and metanotum shorter than pronotum,

subrectangular, carrying wing pads on sides; posterior medial portion of mesonotum

coming to a point, point lacking on metanotum. First and second pair of legs fully

visible in ventral view; third pair of legs almost completely covered by wingpads,

only metatarsus visible.

Abdomen

Segments wider than long, white. Tergite I with postero-lateral comers

pointing perpendicular to sagittal axis of pupa; postero-lateral comers of tergites 2

through 8 coming to a point and directed posteriorly. Pleurites fused to the stemites

(except for pleurite 1 which bears spiracle 1) thus forming lateral margins of

abdominal "ventrites" (see Discussion.) First stemite lacking, first ventrite (stemite 2)

partially visible, rem aining ventrites fully visible, 7 total in female pupa, (male pupa

with 8 ventrites total.) Medial-lateral comers of ventrite 7 (stemite 8) bearing a twin-

spotted photic organ-

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

9 pairs; First on pleuron of mesothorax, remaining 8 on abdominal segments 1

to 8 .

DISCUSSION

B io lo g y

The activity period of L atra adults ranges from early June to late July, and

the larval life is suspected to be approximately two years long, since both large and

small larvae have been found together during m idsum m er (Balduf 1 9 35). However, as

this species has not been successfully reared from egg to adult, the two year life cycle

remains speculative. Because L atra larvae are typically found in rotten logs and

stumps from the fall through early spring, it is assumed that these are the larvae that

overwinter (Williams 1917, MacDermott 1964, MAB personal observation). It should

be noted, however, that stumps and logs are places where coleopterists typically look

for beetle larvae. Both larvae and pupae produced a glow from a two-spotted photic

organ when disturbed. The two-spotted photic organ is located on the eighth stemite

of the larvae and seventh ventrite (eighth stemite) o f the female pupa. Therefore, it is

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. expected that the male pupa would have a similar organ on its seventh v entrite, as the

seventh ventrite of adult males of this species bear such an organ, though the ability

to luminesce seems to diminish shortly after eclosion (MAB personal observation).

Because no L. atra larvae have been found fora g in g in the open, they may be

subterranean in habit Adults usually fly during the day, and males follow pheromone

plumes to the females (Lloyd 1972). The female is typically up to one-third larger

than the male.

Traditional Perspective, and Modem View

Peterson (1951) included a side view drawing of a lampyrid larva along with a

mandible and antennae, which was labeled "Photinus sp.n This same drawing was

included by LaBella and Lloyd (1991) and thereafter has been used as an example of

a P hotinus larva. Upon comparing my L atra larvae with the actual specimen upon

which Peterson based his chawing (in the Peterson Larval Collection, The Ohio State

University, Columbus, Obio), they are identical matches. Additionally, Peterson’s

determination label in the vial with this specimen reads "Photinus sp. ?.” This

question mark on the determination label, evidently put there by Peterson himself,

was most likely accidentally overlooked, and, thus for some 49 years, the specimen

has been misidentrfied as P hotinus sp., ratherthan Lucidota atra. This mistake is very

easy to make due to the great morphological similarity between genera in the tribe

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Photfrrim, to which bothP hotinus and Lucidota ace assigned. The only definitive

method to associate larvae and adults is to rear the larvae.

Abdominal Sclerites in Lampyridae

Considerable confusion has occurred in discussions concerning the number of

abdominal sclerites of adult fireflies. I believe that this confusion has been caused

largely by the fact that Iampyrids possess a varying number o f visible ventral

abdominal sclerites, and there is a lack of accuracy in defining the terms used in

descriptions and discussions of the abdomen. Without both an understanding of the

homology among abdominal sclerites and the use of accurate terminology,

morphological investigations of Iampyrids are bound to remain confused.

The description o f both larval and pupal states of L atra is instructive for

following the reduction and fusion of various abdominal segments and sclerites from

the larval stage, where all abdominal sclerites are present and obvious, to the pupal

stage, where the effect of internalization, reduction and fusion can be first detected. In

all firefly larvae currently known, there are ten abdominal segments, with the tenth

being quite small and, therefore, commonly overlooked. Each abdominal segment

bears a tergite, distinct pleurhes that bear the spiracles (segments one through eight),

and a stemite. Identification of abdominal segments and sclerites in the larvae is not

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. difficult. Reduction of abdominal segments and the internalization o f sclerites in the

adults, however, can make it difficult to find homology among abdominal segments.

In Coleoptera, the adult abdomen is usually composed of ten segments in the

male (with the tenth often being highly reduced or fused with the ninth), and nine in

the female (with the ninth being modified to form the genital segment) (Lawrence and

Britton 1991). As was pointed out by Green (1956), the first abdominal segment in

adult Lampyridae is indicated only by the first abdominal tergite, except females of

Photinus gramdatus (Green, p597). However, after investigation I concluded that the

pleuxite bearing the first abdominal spiracle also is present, though in reduced form,

in pupal L atra and the adults of some Iampyrids. In the adult, the first visible ventral

scierite actually is that of the second abdominal segment, as the ventral portion of the

first abdominal segment usually is so internalized and reduced that it is not visible

ventrally. This condition is termed a "hologastrous type abdomen” (Nichols 1989).

Additionally, in adult Iampyrids the abdominal pleurites are fused to the stemites,

thus form ing a continuous ventral plate. The median half of this ventral plate was the

larval stemite, and the lateral regions on each side were the pleurites. Green (1956),

therefore suggested "...it would be incorrect to refer to the ventral segments of the

abdomen as stemites." Lawrence and Britton (1991) use the term "ventrites" to denote

stemites that are externally visible.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. In light of these two situations, i.e., the absence of stemite (I) and the fusion of

stemites with pleurites, while also keeping with. Green (1956) and Lawrence and

Britton (1991), I adopt the term "ventrites" to denote the visible ventral sclerites in the

L atra papa, which like other firefly species, has the same abdominal morphology as

the adult In most firefly species, the female has one ventrite fewer than the male,

with species in the Luciolinae being the exception, the male having six and the female

having seven ventrites. McDermott (1964) stated that "The Lampyridae may be

defined as that family of the Cantharoidea having usually seven visible ventral

abdominal segments in the male." Apparently, McDermott did not count the ninth

abdominal segment, when visible, as a visible ventral segment. This may be due to

the small size if the ninth ventral sclerite in relation to the other ventral sclerites and

the fact that it is usually the terminal ventral sclerite. It is my present conclusion that

the sclerite of the ninth abdominal segment (ventrite eight) needs to be counted as a

"ventrite1’ when visible. Depending upon whether the eighth ventrite is concealed

under ventrite seven or exposed, adult males will have either seven or eight ventrites

(MAB, personal observation), with eight ventrites being found in the majority of

genera, family-wide, exam inedby MAB. Therefore, in males of most firefly species,

ventrites one through eight correspond to abdominal segments two through nine. The

only known exception is for members of the subfamily Luciolinae, which bear only

six ventrites (McDermott 1964, Ballentyne I987a,b) and the paedomorphic

brachypterous m ale of the European species Phosphaenus hemipterus (MAB personal

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. observation.) La the Luciolinae, the last segment exposed in the male is ventrite six

(abdominal segment VII), with segment VM apparently reduced, or altogether lost,

with segment nine forming part of the aedeagai sheath, which is retracted into the

abdomen (Ballentyne 1992). The male of Phosphaemis hemipterus retains a larviform

type abdomen. Therefore, it is no surprise that Torre-Bueno’s (Nichols 1989)

definition of “stemite = ventrite” is insufficient in conveying the homology of ventral

abdominal segments in adults of Lampyridae.

Even though "segmental fusion" does not seem to occur in the firefly

abdomen, the use of the term "ventrite" should be used with care to avoid confusion

of the homology of various abdominal segments. However, the use of the term

"ventrite" to denote only visible abdominal segments in the adult, while also keeping

in mind (and mentioning a point of reference) that "ventrite one" is actually the

ventral sclerite o f the second abdominal segment (in almost all cases), is simply good

nomenclature and serves to avoid confusion concerning reference to a particular adult

abdominal segment.

CONCLUSION

The firefly larva labeled as "Photinus sp." by Peterson (1951) is actually the

larva o f Lucidota atra, which is herein redescribed in greater detail than the original

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. description (Wickham 1895) in order to facilitate larval identification. Through

rearing tins species from larva to adult, it was possible to investigate the homology of

abdominal segments and track possible fusion or reduction events that led to a

decrease in number of visible ventral abdominal sclerites in the adult Fusion of both

abdominal segments and "ventrites" are not known to occur in currently studied

lampyrid taxa.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FIGURE 13

Lucidota atra, fifth instar larva. (A) Right antenna, dorsal view. (B) Right mandible,

dorsal view. Scale bars — 0.2mm. (A and B, drawn by M. Archangelsky)

Lucidota atra, fifth, instar larva. (A) Habitus. Scale bar = 5mm. (B) Head capsule,

dorsal view. (C) Head capsule, ventral view. Scale bar = lmm. (A, B, and C, drawn by M.

Branham)

Lucidota atra, fifth instar larva. (A) Labium, dorsal view. Scale bar= 0 1 5mm. (B)

Left maxilla, dorsal view. Scale bar = 0.23mm. (A, drawn by M. Arcbangelsky and B, drawn

by M. Branham)

Lucidota atra, pupa. (A) Ventral view. (B) Dorsal view. Scale bar= 4mm. (A. and B,

drawn by M. Archangelsky)

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

f i g u r e 16

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. CHAPTER 5

THE EVOLUTION OF FIREFLY SIGNAL SYSTEMS AND PHOTIC ORGAN

MORPHOLOGIES.

INTRODUCTION

The bioluminescent displays of fireflies have captured the attention of Man for

thousands of years. While fireflies have appeared often in poems, songs and stories of

folklore in many diverse cultures (Harvey 1957), only since the mid 1800s has it beat

known and reported in print that the luminous displays serve to communicate an

individual’s sex, species, and exact location (Mast 1912, Barber 1951, McDermott

1958, Papi 1969, Lloyd 1964,1971). Osten-Sacken (1861) appears to have written the

earliest known published description of firefly flash patterns as sexual signals. Other

early citations include McDermott (1911) and Mast (1912), each of which speculates

on the evolution of these signals. Recent research by Branham and Greenfield (1996)

indicates that these bioluminescent signal systems are under sexual selection like

acoustic and non-bioluminescent visual systems. Some characteristics of these

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. bioluminescent sexual signals vary within a species, while others are more or less

static, and females discriminate between males on the basis of such s ignal parameters

(Branham & Greenfield 1996).

The family Lampyridae has the greatest diversity o f photic organ

morphologies in the order Coleoptera (Buck 1948, Lloyd 1971), but luminescence is

not limited to the family, nor do all fireflies produce light as adults. Fam ilie s closely

related to Lampyridae that are luminescent in at least one life stage include

Rhagophthalmidae, Phengodide and Omalisidae. Wittmer and Ohba (1994) elevated

thelum ino u s genus Rhagophthalmus Motschulsky to family status, previously having

been classified either as Lampyridae (McDermott 1964,1966) or Phengodidae

(Crowson 1972). Branham and Wenzel (2001) moved D ioptoma Pascoe and

Diplocladon Gorham out of Phengodidae and into Rhagophthalmidae. All known

larvae of Lampyridae, Phengodidae, Rhagophthalmidae, as well as the genus

O m alisus Geoffroy (— Omalysus, Homalisus), in Omalisidae, are luminous.

Omalisidae is the only one of the four luminous families that lack bioluminescent

adults. The glowing click beetles, Pyrophorinae, Elateridae, also are bioluminescent

as both larvae and adults but are excluded from this analysis, because they are too

distant to be informative(Lawrence 1988, Beutel 1995). Plastocendae on the other

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. hand is more closely related to the families of interest, those comprising the old

superfamily Cantharoidea, and seemed the most logical choice for a distant outgroup

(Lawrence 1988).

Firefly larvae do not use luminous s ign als for sexual purposes, as they are

immature and cannot reproduce during this life stage, so there has been much

speculation as to the function of luminescence in larvae. One of the most widely held

hypotheses is that Iampyrid larvae use their photic emissions as aposematic displays

(Belt 1874; Cowles 1959; Crowson 1972; Sivinski 1981), and it has been shown that

fireflies possess distasteful steroids, termed lucibufagins, in their haemolymph (Blum

& Sannasi 1974, Eisner e t al. 1978,1997). Underwood e t al. (1997) tested the

hypothesis of aposematic display and concluded that a predator, such as a mouse,

could associate a bioluminescent glow with a distasteful substance.

In some individuals, usually females, as seen in Photuris species, the adults

bear “normal” adult photic organs on the fifth and six visible abdominal ventrites

(true abdominal segments six and seven) and in addition, also sometimes bear a

functional larval-type photic organ on the seventh ventrite (stemite VIII) (Hess 1922).

In the case of still other fireflies, those species that employ sophisticated photic

signals produce them from morphologically advanced photic organs that possess

increased amounts of tracheation, innervation, and the possession of a reflective layer.

9 7

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Such organs are found in Photinus, Photuris, Bicettonycha, etc. (Buck 1948), and are

generally confined to the fifth and sixth visible ventrites (true abdominal segments six

and seven) of the adult (Hess 1922, Branham & Archangelsky 2000). Adults in the

subfamily Luciolinae ( Luciola, Colophotia, Pteroptyx, etc.) also produce complex

photic signals, though the abdomen has undergone reduction of the eighth stemite in

tins clade of fireflies (Ballentyne 1992).

While other secondary functions for firefly luminescence have been posed,

such as illumination of substrate during landing, (Lloyd 1968), the primary function

of luminous behavior in adults is to facilitate pair formation (McDermott 1911,1912;

Mast 1912; Buck 1937; Lloyd 1966). Most behavioral research has focused on

Nearctic species {Photinus, Photuris, Pyractomena, etc.), most of which possess a

signal system in which males fly while broadcasting their species-specific flash

pattern, and females are sedentary and respond to the males' s ignal with a species-

specific signal after a species-specific delay. A short flash dialog commonly ensues

until either the male departs or lands near the female, where m ating may occur. This

type of photic communication system has been termed “Signal System II” by Lloyd

(1971), who has noted that this is one of the many significant discoveries made by

FA. McDermott (Lloyd 1990). These signals are commonly termed “critically timed

signals,” because the timing parameters of the males' flash patterns show little

variation, and the species-specific delay after the males' last flash to the beginning o f

9 8

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. the females7 response is critically timed. Courtship is compromised when this timing

is violated as signals fail to communicate species identity, hi other Nearctic firefly

species, such as those ofMicrophotus, only the sedentary female glows while the

non-Iuminous male flies in search of the glow. In still other species, such as

Pleotomus pattens, the sedentary female produces a pheromone that acts as a long-

range sexual signal and presumably glows to help males easily locate the source of

the pheromone at close range. This type of communication system has been termed

“Signal System I” by Lloyd (1971), who also hypothesized that Signal System II is

most likely derived from Signal System I. Still other firefly species do not produce

light as adults and may rely exclusively on pheromones for pair formation, i.e.,

Lucidota atra (Lloyd 1972a).

Firefly species in other parts of the world have mating systems that deviate

from those of Signal System I and II, studied in North America. The behavior of

many Japanese fireflies has been documented through the work of Ohba (1980,1986)

and Ohba and Goto (1992a, 1992b, 1993). Ohba (1983) proposed a classification

scheme for the various mating systems found in Japanese fireflies, designated

according to representative taxa. In decreasing order of complexity, they are these: the

HP system (for Hotaria parvula), in which both sexes produce photic pulses with

critical timing parameters (Signal System It); the LL system (for Luciola lateralis), in

which males use a single pulse that does not employ critical parameters, though there

9 9

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. is critical tuning in the female’s response to the male s ignal; the LC system (for

Luciola cruciata), in which males use a single long pulse, and also synchronous

flashing, but do not employ critical parameters; the PR system (for Pyrocoelia rufa\

which is composed of a continuous glow and pheromones (Signal System I); the CR

system (for Cyphonocerus ruficollis), in which pheromones are used in conjunction

with a weak glow; and the LB system (for Lucidina biplagiata), which is adiurnal

signal system composed entirely of chemical signals (pheromones.) A dendrogram of

eight species of Japanese fireflies representing three types of communication systems

(HP, LC and LL) was subsequently constructed based on electrophoretic analysis of

allozymes using Nei's genetic distance by UPGMA (Susuki e t al . 1996). This study

showed that the LC system was basal to the LL system in one of the two sister clades

and the other clade was composed of taxa using the HP signal system. This

dendrogram pattern of signal evolution is not very informative. Susuki (1997) later

expanded his phylogenetic analyses of Japanese firefly mating systems to combine

mitochondrial 16s ribosomal RNA data with allozymes. Phylogenetic trees were

constructed using the Neighbor-Joining, Maximum Parsimony, and Maximum

Likelihood methods, but only one tree, produced by the Neighbor-Joining method

was presented- While character optimization is ambiguous in several parts of this tree,

only one optimization of characters involved in signal system evolution was presented

and discussed. The optimization presented shows a trend from diurnal basal taxa that

employ only chemical sign als (LB), or taxa using chemical signals and photic glows

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. (CR and PR) in one of the two major clades, to taxa using only chemical signals (LB)

leading to taxa that use only critically timed photic Sashes (HP), which then gives rise

to taxa that produce discrete flashes while having lost critical timing parameters (LC

and LL), in the second ciade.

MATERIALS AND METHODS

For specifics of which taxa were used in the analysis, as well as the methods

used in phylogenetic reconstruction, see Chapter 2, Materials and Methods. The

superfamily Cantharoidea was combined into the Elateroidea of Lawrence (1988).

This analysis includes most of the fam ilie s that formerly composed the Cantharoidea

of Crowson (1955,1972) (viz: Brachypsectridae, Omalisidae (= Omalysidae,

Homalisidae), Kanuniidae, Drilidae, Phengodidae, Telegeusidae, Lampyridae,

Cantharidae, Lycidae, Cneoglossidae, Plastoceridae and Omethidae). Because of the

historical uncertainty of the composition of Lampyridae, 1 included many outgroup

taxa that might resolve questions regarding whether certain characters of Lampyridae

would be best considered symplesiomorphies of cantharoids or synapomorpMes of

Lampyridae. I loosely refer to the taxa used in this analysis as “cantharoids,” as they

have generally been thought of as a monophyIetic group within the Elateroidea

(Lawrence 1988).

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Selection of taxa for the outgroup was made by including taxa from each

subfamily within each of the outgroup families when possible, based on Lawrence &

Newton (1995). Plastoceridae was designated as the root of the tree based on

Lawrence’s phylogenetic analysis (1988) of the Elateriformia.

RESULTS

Topology of the Tree

For specifics on the output of the Parsimony Ratchet and the NONA tree

searches, see Chapter 2, Results section. The consensus tree is fully resolved, except

for the four phengodid taxa and Stenocladius sp. and a trichotomy of M aithinus

occipitalis (Cantharidae) and the adjacent clade o f Pseudotelegeusis and Telegeusis

with the adjacent clade of Phengodidae plus Stenocladius sp. in the outgroup. The

ingroup of the consensus tree is fully resolved except for a polytomy of nine lampyrid

taxa and two trichotomies: a) Bicellonycha amoena, Photuris divisa, and P.

brunnipennis, and b) Luciola lateralis, L. salomonis and the adjacent

Luciola/Colophotia/Pteroptyx clade, (Fig. I). Bremer support was evaluated using

NONA and the search was set to a Bremer support level of 5, with four runs (each

holding 100 trees) and a total hold o f 5 0 0 0 trees. Bremer values for the ingroup, listed

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. in Figure 17, indicate the number of steps that are required, up to 5, to derive a tree

that does not include the particular node of interest.

Lampyridae is monophyletic with the exception of a few taxa that historically

have been of questionable affinity {Harmatelia, Drilaster, Pterotus, and

Stenocladius .) With these four taxa excluded from Lampyridae, the family is

monophyletic. Rhagophthalmidae is shown in this analysis to be monophyletic and

forms a sister clade to Lampyridae in my sense. Brachylampis sanguinicollis and

Psilocladus sp. compose the basal stem of the lampyrid clades. The family is

composed of two major clades, one with EHychnia corrusca in a basal position and

the other composed of two sister clades with Macrolampis acicularis and Robopus sp.

defining the basal position of each.

Levels of Homoplasy in the Analysis

The Consistency Index of this analysis (consensus tree Ci.=0.16) may appear

low, but it is unremarkable when one considers the number of taxa used in the

analysis (85 total taxa) (Sanderson & Donoghue 1989). Antennal morphology has

been suspected of being very homoplastic within Lampyridae (McDermott 1964).

However, this analysis does not show that antenna! morphology is any more

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. homoplastic than any of the other characters used in the analysis. When the CX of

characters are compared (see Appendix: B), both antenna! charactersand non-antermal

characters show the same amounts of variation.

DISCUSSION

Taxonomy o f the Lampyridae

McDermott (1964) created what many in the field hold to be the first of the

modem classification schemes for the family Lampyridae. In this work, McDermott

clearly pointed out that he believed the classification scheme he was proposing was

not natural, though he regarded it as “perhaps being of some utility.” McDermott

revised his classification of Lampyridae a few years later (McDermott 1966).

Crowson (1972) revised McDermott's (1966) classification through a “possibly

unproved scheme of subfamilies” by removing some taxa from Lampyridae and

placing them in Omethidae and Phengodidae (thereby eliminating the Matheteinae

and Rhagophthalminae of McDermott's Lampyridae), and moved Ototretadrilns out

ofDrilidae and into Lampyridae (thereby creating Ototretadrflinae). Additionally,

Crowson added Cyphonocerinae and Ototretmae to Lampyridae by further

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. subdividing previously existing subfamilies. Lawrence and Newton (1995) have since

adopted Crowson’s classification scheme for Lampyridae in their work on the higher

level classification of Coleoptera.

When comparing my phylogenetic analysis to McDermott’s (1966) and

Crowson’s (1972) classifications, one finds that my analysis supports the monophyly

of only two Iampyrid subfamilies, Luciolinae and Photurinae. Furthermore, the

genera included in these two subfamilies are the same for both classifications.

Therefore, it seems obvious that the taxa included within these two subfamilies have

long been identified as distinct from the rest of Lampyridae. The fact that the existing

classification for Lampyridae does appear largely artificial does not come as a

surprise, as this has been alluded to by various authors (McDermott 1964, and

Crowson 1972). A recent comparison of the larval morphology of several genera of

lampyrids (Archangelsky & Branham 2001), also is consistent with the need to revise

the current classification.

My analysis shows that, as previously defined, Lampyridae is not

monophyletic: Drilaster, Harmatelia, Pterotus, and Stenocladhts do not belong

within Lampyridae. These four genera have a history of being moved between various

families that are phylogenetically related to Lampyridae, see Table 1 in this

dissertation (Branham & Wenzel 2001). This phyiogeny places Drilaster, Harmatelia

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. and P terotus outside o f Lampyridae but does not place them within any existing

family groups. Stenocladhts is placed outside ofLampyridae, but in an unresolved

polytomy that is composed of taxa currently in the family Phengodidae (Figure 3).

Given this pattern, my solution is to formally move Drilaster, Harmatelia and

P terotus to “Elateroidea m certae sedisT status and to move Stenocladhts into

“Phengodidae mcertae seeds”, see Table 1 in this dissertation (Branham & Wenzel

2001).

An Ancient Orign of Biotummescence in the “Cantharoidea”

My phylogenetic analysis predicts at least two origins and one loss of

luminescence in the cantharoid lineage (Figure 3). The first origin was very ancient

within this lineage and applies to the luminescence found in Omalisidae,

Rhagophthalmidae and Lampyridae, plus the “Elateroidea insertae seeds” genera

Harmatelia, Drilaster and Pterotus (Branham & Wenzel 2001). Also, all known

luminescent cantharoid taxa have luminous larvae, while Omalisidae is the only

family in which luminescence is present in the larvae and not in at least one adult

(Crowson 1972). The fact that Omalisidae is the most basal of all the bioluminescent

cantharoids indicates that luminescence arose first in the larvae and then subsequently

in the adults and that the origin of luminescence in cantharoid taxa predates the origin

of the first Iampyrid (Figure 3). This finding is supported by Crowson’s (1972)

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. comment, “The fact that there are no well established instances of cantharoid

beetles luminous as adults but not as larvae, suggests, though it is hardly sufficient to

prove, that luminosity first arose in the larval stage.”

Larval Luminescence as an Aposematic Display

Lampyrid larvae are thought to produce photic signals as aposematic displays

to convey to potential predators that they are chemically defended (Belt 1874,

Crowson 1972, Sivinksi 1981, Tyler 2001o,h). The chemical substances responsible

for this defense were identified by Eisner e t al. (1978,1997) as defensive steroids

called lucibufagins, which typically serve as cardiotonic agents. Lucibufagins are

structurally related to the bufodienolides of toads and the cardenolides of plants

(Fieser & Fieser, 1949; Budavari e t a l., 1996) and at low concentrations induce

nausea and vomiting (Kaiser & MichI, 1958; Kelly & Smith, 1996). Blum and

Sannasi (1974) and Kloft e t al. (1975) demonstrated that fireflies display reflex

bleeding. Reflex bleeding in adults seems to be the mechanism by which fireflies can

deter potential predators without sacrificing their lives, as lucibufagins seem to be

contained in the effusing haemolymph (Blum & Sannasi 1974). Knight e t aL (1999)

discussed how ingestion of a single P hotim is rapidly killed lizards in the Australian

genus Pogona (Agamidae). “The lizard immediately ate the firefly and within 60-90

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. min began showing oral gaping movements. These became more Sequent over the

next 3 0 mm, bat there was no vomiting. The animal also underwent a color change,

horn tan to black, in the ventral region of the neck and abdomen, and the back of the

tail. In the coarse of gaping, the lizard tended to protrude and bite its tongue. It then

became quiescent and died.” Knight e t al. (1999) also pointed out that firefly

toxicosis has been involved in the death of a lizard native to Caucasus ( Lacerta

derjugini), several African chameleons ( Chamaeleo pardalis ) and a hylid frog

(Litoria caerulea) native to Australia. Lizards that are naturally sympatric with

Photinus, such as those in the genera Anolis, Sceloporus, and Eumeces are known to

reject these fireflies prior to ingestion (Sexton, 1960,1964; Lloyd, 1973; Sydow &

Lloyd, 1975). Other predators such as mice can leam to associate luminescence with a

distasteful substance (Underwood e t aL 1997). There is evidence (Lloyd 1989) that

bats feed on fireflies, though there appears to be no evidence concerning the bats’

reaction, if any, to eating fireflies or isolated lucibufagins.

Both adults and larvae in the firefly genera Photinus and P hoturis are quite

distasteful and effective in preventing repeated tastings by humans (NLAJ3. personal

experience), and both the intensity and frequency of larval glowing, correlates with

increasing harassment by a potential predator. While more studies of larval photic

emission as aposematic displays are needed, current evidence is clearly suggestive of

an aposematic function.

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Carry-Over of Luminescence into Adults

McDermott (1964) and Crowson (1972) discussed the fact that many

cantharoid beetles are known to carry larval characters into the adult stage. The most

well known examples of this are the adult paedomorphic females found in the

families Drilidae, Rhagophthaimidae, Phengodidae and some Lampyridae. While

only a few firefly species possess fully Iarviform females (those which are

morphologically identical to larvae except for the possession of copulatory organs),

such occurrences of paedomorphosis are also found in the females of the drilid genus

Selasia, the rhagophthamid genus Diplocladon, and all known phengodid females

with the exception of Stenocladhts (Branham & Wenzel, 2001). Adult female

Lampyridae can possess a wide range of paedomorphic features, which range from

the possession of only a few to an entire suite of larval characters. However, some

authors have incorrectly identified females of some species as being ‘iarviform” due

to lack o f wings, when in reality the females were either brachypterous, brachypterous

and physogastric, or simply apterous.

A so called paedomorphic character commonly found in adult fireflies is the

two-spotted photic organ on the ventral surface of the eighth abdominal segment. All

known Iampyrid larvae possess a photic organ of two luminescent spots on the eighth

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. abdominal segment In addition to this condition, the larvae o f Lamprohiza possess

additional pairs of photic organs on abdominal segments 2-6, and sometimes these

spots are not symmetrically arranged on segments of the abdomen (Balduf 1935,

Schwalb 1960, and Buschman 1988). Buschman (1988) pointed oat that the fairly

uniform morphology of paired larval photic organs contrasts greatly with the diverse

morphologies found in adult fireflies and hypothesized that the lack of variation in

larval photic organ morphology might be due to the two luminescent spots

functioning as “'eye-spot or false-head displays.” The larval photic organ morphology

appears to be homologous with paired photic organs on the 8th abdominal segment in

adults, as they are in the same location in the abdomen, have the same shape, and

histology, and produce the same type of photic emissions—long continuous glows

(Hess 1922, Buck 1948, McDermott 1964, Branham & Archangelsky 2000). In

addition, the emission spectra of the light produced by both adults and larvae of

Photuris are identical, with a maxima at 552.4 nm (Coblentz 1912, McElroy &

Seliger 1966). Paired photic organs are not restricted to larviform females, as they

appear in some brachypterous/physogastric females, fully alate females, and some

fully alate males as well.

If biohunmescence in adult lampyrids first arose as a carry-over from the

larva, then presumably it would persist throughout the pupal stage, as it is known to

do in some Phengodidae and Lampyridae today (Crowson 1972, Viviarri & Bechara

1997,Costa e t al. 1999).This photic organ morphology appears throughout the pupal

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. stage and sometimes into the adult. In the case of fireflies that are noil-luminescent as

adults, such as Pyropyga (Branham and Archangelsky, unpublished), and Lucidota

atra (Branham & Archangelsky 2000), the paired “larval-type” photic organs are

present in the adults and are functional only for a short time after eclosion. Usually

after 24hrs. following eclosion, and after the cuticle has hardened, the paired photic

organs appear non-functional, and either disappear completely or remain as twin spots

on the abdominal cuticle of the adult (McDermott 1964, MAB personal observation).

The larval-type photic organ is much more obvious in the adults of some fireflies. In

the genus Robopus, larval photic organs are retained as the only functional photic

organs in adult males and females, are larger than those found in the larvae, and

produce bioluminescent sexual signals used in pair formation (Barber 1941,

McDermott 1964, Famworth 1973 and MAB personal observation). In addition, the

function of both larval and adult photic organs in Robopus species, Le., the type of

glow produced and its control, appears to be the same (Buck 1942; M A B in prep.).

The larval carry-over of photic organs is further supported by the fact that all

known Iampyrid larvae that have been studied histologically have the same type of

photic organs, which were termed “TypeH F by Buck (194S), with the exception of

Lamprorhiza (Phausis) delarouzei, which was studied by Bugnion in 1929 and

appears to have “Type II” photic organs. It should also be noted that Lamprorhiza

splendidula larvae possess “Type HP, not “Type IF photic organ histology. While it

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. may be possible for two different types of histological photic structures to be present

in the larvae of one genus, I propose that the larval photic organs of L. delarouzei

need to be re-examined. Oertel e t a l (1975) investigated the ultra structure/histology

of the larval light organ in Photuris pennsylvanica DeGeer and found many

similarities between larval and adult photic organs, and that the triggering of light

emission is slower in the larva where innervation is direct, than in the adult, where

innervation is indirect. Oertel e t a l (1975) point out that in P. pennsylvanica, “the

larval light organ is 100 times slower than the adult light organ in its response to

neural excitation and the larval glow is about 50 times as long as that of the adult.”

This makes perfect sense in an evolutionary context when one considers the types of

selection pressures that would shape both types of photic emissions. Larval photic

emissions are used as aposematic warning displays and adult photic emissions are

used as sexual signals that should be expected to evolve rapidly under sexual

selection. It should be pointed out that the adult photic organs that were being

compared to the larval organs in the Oertel e t a l study, were not those in the 8th

abdominal segment They were the derived adult photic organs found anterior to the

8 th abdominal segment

I contend that there has been a carry over of the larval photic organ condition

into the adults of some lampyrid taxa. Therefore, it is my hypothesis, and also that of

McDermott (1964) and Sivinski (1981), that luminescence first arose in larval

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. cantharoids, was probably maintained through an aposematic function in larvae and

later was co-opted as a sexual signal (perhaps as both a warning and sexual signal) in

the adults of several cantharoid families while reaching it greatest elaboration and

refinement in fireflies.

The Pattern of Flightlessness in Females Within Lampyridae

Crowson (1972) speculated that luminescence in adult cantharoids may be in

some way related to the ability of cantharoid taxa to possess paedomorphic

characters. One suite of likely paedomorphic characters has to do with wing reduction

or loss in females. Extreme cases of wing reduction or loss also seem to be associated

with the possession of the two-spotted larval-type photic organs on the ventral surface

of the females eighth abdominal segment.

Because the basal region of the lampyrid clade is composed of firefly species

with alate females and the various forms of paedomorphic females (larviform,

brachypterous, and/or physo gastric forms) are scattered throughout the lampyrid

clade, it appears that these paedomorphic forms have arisen multiple times within the

family While losing wings and then regaining them has been argued as extremely

improbable, wings can be lost and then regained in water striders, Gerridae, by

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. polymorphism (Andersen 1993). Andersen has shown that definite associations exist

between loosing the ability to fly (doe to brachyptery) and durational stability of

habitats, and was able to infer that the ancestor of this group of water striders was

flightless or permanently dimorphic. While I have no direct evidence that the same is

true within Lampyridae, all of the cantharoid taxa basal to Lampyridae have flightless

females, with the single exception of Drilaster subtilis, see Tables I, and 2 in this

dissertation (Branham & Wenzel 2001) and hence could show a similar pattern to

Andersen's water striders. Due to the scattered distribution of larviform or

brachypterous females throughout the phylogeny, it is possible that a polymorphism

such as the one found in the water striders could play a role in determining which taxa

and which sex will exhibit an alate or brachypterous condition due to the occurrence

of varying amounts of paedomorphosis. The brachypterous/physogastric form,

perhaps more than the larviform female condition, appears to be associated with

habitat or ecology, as is seen in the Andersen (1993) example above. In fact, Lloyd

(1999) found varying amounts of female brachyptery in Pyropyga nigricans studied

from multiple locations across the United States, with some occurrences of

brachyptery found in several populations, which seemed to be correlated with very

restricted habitats around the edge of lakes or marshes. Lloyd suggested that these

occurrences were perhaps due to a ‘‘wing polymorphism.” One population also

showed brachyptery in both males and females (Lloyd 1999). All other known

populations o f P. nigricans are alate and capable of flight Green (1961), proposed

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. that the phenomenon of brachyptery might be associated with perm anent moisture.

Hence, like the water strider example mentioned above, the presence of a “wing

polymorphism” in fireflies appears plausible, on the basis that a segment of the

populations are fully alate and capable of flight, while others are not, and instances of

brachyptery also seem to be associated with stable and restricted habitats.

Evolution of Chemical Signals vs. Visual Signals

Here “pheromonal sexual signals” refer to long range chemical signals that are

the primary signals used in mate attraction. When the use of pheromonal sexual

signals is plotted onto the phytogeny, it occurs primarily in the basal taxa. This

phylogenetic pattern predicts at least three origins of pheromones, with at least two

losses for the family Lampyridae (Figure IS). Four possible character optimizations

exist for the evolution and toss of pheromone use within the Lampyridae. These four

schemes exist due to two equally parsimonious optimizations within each of the two

clades that contain, a) A spisom a —Pyrocoelia, and b) Macrolampis - Microphotus.

The four possible combinations of optimizations in these clades hypothesize

pheromones arising 3-5 times and then being subsequently lost 2-4 times (Figure 18).

In all optimization schemes, pheromonal sexual signals were regained in the clade

composed of Phausis and Phosphaemis , as well as in Pleotomus (Figure 18). The feet

that the basal lampyrid taxa (e.g., Brachylampis and Psilacladus), winch happen to

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. use pheromones, are also diumally active, suggests that ancestrally fireflies were

dhimaily active and employed pheromonal signals for pair formation.

Species that rely on photic signals for sexual communication are generally

restricted to the tips of the phylogenv, indicating that the use o f photic signals in

adults is derived for the family, as is nocturnal behavior. The use of bio luminescent

sexual signals has evolved at least four times in the family with at least four losses

(Figure 19). One loss of photic signaling in the adult stage seems to have occurred in

the genus A lecton, which are diurnal, brightly colored, and use pheromones to attract

mates. The second loss occurred in the genus Tenaspis, where the males of some

species possess a two-spotted photic organ on the eighth abdominal segment, which

appears to be non-functional. The third loss occurred in Macrolampis acicularis,

which is in a genus containing both luminous and non-Iuminous species.

Phosphaenus hemipterus was scored as the fourth loss. In this species, the female is

diurnal and luminous, though the photic emissions do not appear to be used as sexual

signals in this species (De Cock 2000). Therefore, the hypothesized ancestor of P.

hem ipterus used photic sexual signals, most likely in conjunction with pheromones,

and?, hem ipterus seems to have subsequently reverted to diurnal habit and the sole

use of pheromonal signals for pair formation. Treating pheromonal and photic

systems as separate characters, it appears that the combined use of pheromones with

photic signals can sometimes serve as a transition between using only pheromones to

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. using only photic signals. Both o f these signal modalities (pheromonal and photic) are

present in eight taxa scattered across the family ( Pyrocoelia, Phaenolis,

Erythrofychma, a new species, Macrolampis, Phausis, and Pleotomus,) (Figure 20).

Evolution of Photic Signaling Systems

The context as well as the manner in which photic signals are produced play

an important role in the type of signal system required to elicit courtship and pair

formation. The photic signal systems o f fireflies can be complex, with various aspects

of some systems being either retained or eliminated in the communication systems of

other species. While two general types of photic s ignal systems were noticed by early

firefly workers (Gorham 188(1, Mast 1912, McDermott 1914, and Blair 1915,1924a,

1924b), it was Lloyd (1971) who defined these two signal system types and posed a

hypothetical evolutionary scheme for their evolution.

Lloyd (1971) used “Signal System I” to represent firefly species in which one

sac, generally the female, is stationary and broadcasts a species-specific s ignal, to

which the other sex is attracted. While not explicitly stated,S ignal System I seems to

be applied only to sedentary females producing species-specific photic em ission s

rather than pheromonal signaling alone. In contrast, “S ignal System II” represented

species in which one sex, usually the flying male, broadcasts a species-specific s ignal,

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. which is then answered by a species-specific response, produced by the opposite sex.

The primary signaler is then attracted to the mate responding to the original signal. In

addition, critically timed parameters are employed by some taxa using S ignal System

Q . Lloyd (1 9 7 1 ) was also aware of signal systems that used elements of both Signal

System I and II, and he termed such systems “Compound S ignal Systems,” such as

those found in the synchronously flashing aggregations of several P teroptyx species.

In a position that seemed intermediate between Signal System E and H, Lloyd (1971)

proposed a potential transitional system, with Phausis reticulata as an example. In

this signal system, glowing females attract glowing males, and non-glowing females

also will initiate glowing in response to glowing males. After studying the behavior of

Luciola obsoleta (Olivier), Lloyd (1972b) realized that such a simple classification as

System I and II was not adequate for the larger scenario of sexual signaling and

proposed the use of the term “protocol” to replace “signal system” (Lloyd 1978). As

the old classification of signaling systems (I + H) was unsatisfactory, Lloyd (1983)

recommended that workers focus on the “key factors” of firefly mating protocols that

“emerge from modem studies of sexual selection and ecology.” Lloyd (1997)

additionally hypothesized that Signal System I and Q could have arisen multiple

times. He pointed out various components that are shared between the four signal

systems, or dropped from some, thereby highlighting the complexity of trying to

define communication protocols. Signal Systems I and Q are not supported by my

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. analysis as forming an evolutionary series, i.e., System I giving rise to System II

through an intermediate stage, such as in Phausis reticulata, and further, both appear

to have evolved more than once.

Signal System Components

In order to address signal system evolution, components should be examined,

rather than the complete, or overall communication system, because some

components may covary while others do not. In addition, because the communication

systems of some fireflies can be complex, reducing them into parts allows direct

comparisons to be made with other species by scoring the similarities and differences.

Current behavioral data permit analysis of two important components of the

signal system. Figure 21 indicates that among basal taxa, the primary signaler is a

sedentary female producing pheromonal signals. Apical to these taxa, sedentary

signaling is used by species that produce both pheromonal and photic signals. The

combined use of pheromones and photic signals in courtship appears to have evolved

at least five times. Once in Pyrocoelia, once in the common ancestor of

Erythrolychnia and Phaenolis, once in a new species from Puerto Rico that cannot be

placed with any currently described genus, once in Phausis, and once in Pleotom us

(Figures 20,21). Additionally, there seems to be a correlation between females

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. functioning as the primary signaler when the primary s ignaler is sedentary (Figure

2 1 ). ha contrast, primary signalers that produce photic s ignals while in flight appear to

be derived and strongly correlated with sign alin g males. This syndrome seems to have

evolved at least four times in the family: once in the Aspisoma - Pyractomena clade,

once in Cratomorphus, once in the Bicettonycha—Photuris - Photinus clade and

once in Robopus (Figure 2.9X

Sedentary, Synchronously Flashing Aggregations of Fireflies

In my analysis, Pteroptyx cribellata, P. tener, and P. malaccae , are the most

derived species in the Lucioline clade. These three species seem to possess a derived

and complex signal system: sedentary production of synchronous flash in g in the

context of a mating aggregation. Many of the details of what happens between the

sexes, as well as within a sex, in these mating Ieks is still far from understood and

promises to be fertile ground for future behavioral studies.

Luciola cruciata (Lloyd 1979; Ohba 1983,1984) possess a photic signaling

system that seems to be intermediate between asynchronous flash in g found in most

flashing fireflies and the true synchrony found in Pteroptyx cribellata, P. tener and P.

malaccae. Both sexes o f L. cruciata form aggregations and mate after lengthy and

complex interactions that involve males and females emitting photic signals in close

120

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. proximity (Lloyd 1979). L cruciata males emit synchronous flashes only while in

flight Sedentary females respond to these flash in g males. Upon receiving a response,

males break from the group synchrony and engage in a “male-female flash

interaction,” which ultimately leads to copulation (Ohba 1984). Yafima (1978)

determined that the female response to male signals did not involve critical timing

parameters on the part of the male. L cruciata shows little sedentary s ignaling during

courtship, though when it occurs, h is after the male has landed near the female.

Aggregations in this species occur only among females during oviposition. Females

emit unique flashes at these aggregations and tend to attract other females to an

oviposition site (Kuribayasiu 1980; Yuma 1981). Perhaps there is a benefit to having

many aposematic individuals in a restricted area in order to reinforce the association

between the aposematic signal and distastefulness. This adaptive link between

aposematism and gregarious habit is a controversial area, and some other systems also

have received special attention (Sillen-Tuilberg 1988, Maddison 1990). Luciola

cruciata is basal to Pteroptyx and may serve as an important model for how sedentary

signaling., synchrony, and the aggregation of signaling individuals evolved as

components in the sexual signal system of three Pteroptyx species, P. cribellata, P.

tener, and P. malaccae.

121

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

No adult photic organs exist in the basal species of Lampyridae. There has

been considerable confusion as to which abdom inal segments bear the adult photic

organ because a lack o f consistent use of correct terminology. In non-paedomorphic

adults (with the exception of Photinus granulatus ), the first abdominal segment is

internalized and is not visible externally (Green 1956, Lawrence & Britton 1991,

Branham & Archangelsky 2000). This situation confounds a discussion of abdominal

segment homology. As an example, when a phrase such as “seventh abdom inal

segment” was used, this could mean the seventh visible segment in the adult, or the

real seventh segment (which is the sixth visible segment in the adult) The lampyrid

taxa that use both pheromones and photic signals use only restricted regions of their

fifth, sixth, and seventh visible ventrites (true abdominal segments six, seven and

eight) for photic emission in both males and females. Species that rely solely on

photic signals for pair formation possess photic organ morphologies that use most of

the ventral surface of the fifth and sixth ventrite. The genus Robopus, however,

retains larval-type photic organs into the adult stage—the two-spot condition on the

seventh visible ventrite and produces solely photic signals.

As the most basal luminescent taxa within Lampyridae have both luminous

males and females, luminescence in adult Lampyridae seems to have arisen in adult

122

I Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. males and females at about the same time. Paedomorphic females have paired photic

spots that seem identical to larval photic organs both in structure and in types of

emission, i.e., intermittent glows (M A Branham, persormai observation). Males and

females in many genera bear only the twin-spotted photic organ on the seventh

ventrite (usually the true eighth abdominal segment) that is either non-functional or is

capable only of a very faint glow. This morphology is found throughout the lampyrid

phytogeny with the exception of the basal region, i.e., Brachylampis, Psilocladus,

E llycfm ia, etc., and is present in close to half of the taxa sampled for this analyst*;

This morphology seems to have arisen about some eight times in the family

Lampyridae, which is substantially more than the number of origins hypothesized for

any other photic organ morphology (Figure 2.10). It seems possible that the family’s

apparent predisposition toward a carry over of larval features into adults may plav a

role in the numerous independent origin s, as “larval carry-overs” of this morphology.

Photic organs on the seventh ventrite are functional in both sexes of the adults

of taxa such as Robopus, Pleotomus and Microphotus, but these organs appear not to

be functional beyond teneral adults in taxa such as Vesta, Pollaclasis, Tenaspis and

Lucidota atra. Even though the two-spot condition is functional in males of both

Pleotom us and Microphotus, it appears to have no role in courtship, though they glow

quite visibly when distressed. It is also relevant that in taxa where the two-spot

photic organ morphology is non-functional, the taxa are diumal. Yet this situation is

123

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. not restricted to the more basal regions of the phytogeny, ha addition, Phosphaenus

hem ipterus is diurnal even though the female has a functional two-spotted photic

organ from which it produces glows (in daylight) when molested or attacked by

predators (De Cock 2000).

Only three types of photic organ morphologies appear on the malers sixth

ventrite (true abdominal segment seven) in taxa used in this analysis: two luminous

spots, a luminous center-strip, and the entire ventral surface of the sixth ventrite being

luminous. When plotted on the phytogeny, the two-spot morphology arose twice,

once in Cratomorphus and once in P teroptyx. The center-strip morphology seems to

have arisen three times: once each in the generaPyrocoelia, Luciola , and

Lamprohiza. The photic organ that covers the entire ventral surface of the ventrite

appears to have arisen three times: once in the Aspisoma—Pyractomerta Clade, once

in the Bicellonycha - Photuris - Photinus Cade and once in the Luciola - Cholphotia

— Pteroptyx Clade (Figure 23).

The photic organs on male ventrite five (true abdominal segment six) in taxa

used in this analysis show four morphologies: one spot, two-spots, a centrally located

strip, or a photic organ that covers the entire surface of the ventrite. When these

photic organ morphologies are plotted on this phytogeny, the one-spot and two-spot

morphologies appear to have arisen once each, with the one-spot condition appearing

124

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. in Erythrofycfmia, and the two-spot condition arising in Cratomorphus . The center-

strip condition appears to have arisen twice, once in Pyrocoelia and once in

Lam profnza . The largest of all photic organ morphologies appearing on ventrite five,

where the entire ventral surface of the ventrite is luminous, appears to have arisen

three times, once in the Aspisoma - Pyractomena Clade, once in the Bicellonycha -

P hoturis - P hotinus Clade and once in the L uciola —Colophotia—Pteroptyx Clade

(Figure 2.12). In addition, the presence o f large photic organs covering the entire

ventrite also correlates with species that produce flashed signals. This is true of the

males in the three clades mentioned above, therefore suggesting that flashed signals

evolved at least three times in the family Lampyridae (Figure 2.12). Perhaps this

should not come as a surprise, since male photic organs used in such signal systems

are under intense sexual selection via both intra- and inter-sexual selection. At the

very least, larger photic organs in these males must increase the chance of then signal

being seen by a receptive female.

The rapid and repeated evolution of photic signals in Lampyridae may

demonstrate the proliferation of mating systems. Visual signals can be employed by

one or both sexes, with the possibility of modifying these signals immediately upon

receiving an answer.

125

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

The origin of bioluminescence in cantharoids occurred early in the

evolutionary history of the group, predating even the origin of the family lam pyridae

Cladistic analysis indicates that luminescence first appeared in larvae, probably as an

aposematic warning display, and is still found in many cantharoid larvae today. It

appears as though luminescence in adults is a carry over from larvae. Luminescence

in the adults appears to function as an aposematic warning display, which has been

co-opted in many species to serve also as a species-specific sexual signal used in

courtship.

The evolution of firefly signals is accompanied by a change from diurnal to

nocturnal behavior. The overall trend is the use of pheromones in basal species, then

pheromones used in conjunction with photic signals, then the sole use of photic

signals. Signal systems that employ only luminescence usually involve flashed

signals rather than glows, or flashed signals using critically timed signal parameters.

Not only do some components of the signal systems appear to be convergent, but so

does photic organ morphology. In addition, several correlations seem to appear across

the family. One is that primary signalers that are sedentary during signaling are

almost always females. Female primary signalers that belong to taxa appearing basal

in the family, such asBrachylampis and Psilocladtis, employ only pheromonal

signals; those in more derived taxa, such as Lam pyris and Microphotus, employ only

126

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. photic signals. A second correlation concerns species that possess large, complex

photic organs in adult males who are primary signalers. This relationship probably

reflects intense sexual selection.

I recommend that signal systems be characterized through individual

components, rather than comparing the '‘overall scheme” of various signal systems.

By comparing individual components, we can recognize and better interpret adaptive

correlations despite convergence or loss. As a result of this analysis, I demonstrate

that rapidly evolving traits can be investigated phylogenedcally.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FIGURE 17

The lampyrid clade of the strict consensus with Bremer Support values (set at a max.

Bremer value o f 5) appearing above the nodes, with the numbers below the node

indicating the number of synapomorphic characters at that node.

I Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Brachytampls sangulnicollis P silo d a d u s s p . "~~Blychnfa corrusca 3 A spisam a pulchellum _2_ 4 Aspisoma fgnitum Pyractomena ecostata 3 Pyractomena borealis Cratomorphus dlaphanus 3 i Alecton discoidalis 3 Alecton flavum £ j PyracoeSapraetexta 6 1 Pyrocoeilarufa Pyropyga nigricans Erythrolychnfa bfpartitus 3 * " Erytbrolychniaoltveri PhaenoIJs ustulata 7 Tarraspis angularfs Luddota atra L u ddota dila tlco m is Luddlrra biplagrata new species Polfacfasls bifarfa Pristo/ycus sagulatus Vesta aurantiaca Cailapisma maestra CJadodes fiabeliatus • Uacrolampis adcuiaris BlceJlonycba amoena Photurts divisa Photuris brunnipennis Phatfnus pyraUs Phatfnus Ignitus Phatfnus metearaits Robopus sp. 1 R obapus s p . 2 Ludala lateralis Ludala salomonis Luciola cruciata L udala kuroiw ae 3 p C oiopbotia s p . 1 i Pteroptyx cribelfata | P teroptyx m afaccae 2 *■ “ P tero p tyx tartar Lampmhiza splendfduia P hausis rhom bica Phosphaanus hemipterus Pteatomus nigrfpennis Pleotamus pallens Lampyris noctiluca Uicrophotus octarthrus Lampyris zankerf Uicrophotus angustus FIGURE 17

129

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FIGURE 18

The evolution o f pheiomone use in Lampyridae. Four possible optimizations exist of

seven steps each for Lampyridae when combining the optimizations (a), (a'), (b),and

(b1). The resulting topologies hypothesize from 3-5 origins^ and 2-4 losses of

pheromones.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Brachyiampis sangulnlcolHs "ftjfoc& dtB Asplsoma pulcfieiiuni Asplsoma fgnitum Pyractomena ecostata Pyractomena borealis Cratamarphus diaphanus Alecton dlscaidalis Alecton fiavum

Pyropyga nigricans Erythrolychnia blpardtus Erythrolychnia oltveri Phaendts ustulata Tanaspis angutaris Luddota atrm '• Luddota dUaticamis Luddlnablptagtata new species 1 Polladasis bifaria Prfstolycus sagulatus Vesta aurantfaca Calfopisma maestra Cladodes fJabellatvs Uacrolampis adcufaris Blceltonycha amoena Photuris dlvisa ' Photuris tvunnipennis Phatlnus pyralls Phatlnus Ignitus (b) >■»■■■»—______Phadnus —y------meteoralls------c Robopussp* T Robopus sp. 2 Ludala lateralis Ludola salomonis Ludala crudata 41- Ludala kurotwee Colophotlaep. Pterop tyx cribaUata Pteroptyx malaccae Pteroptyx tenor [ Lamprohizm splendidula Phausis rhombica ■ Phosphaenus hemiptarus Pteotomus nigripennis Pteatomus pollens Lampyris noctlluca Uicrophotus octvttinis Lampyris zenkeri Uicrophotus angustus

Asplsoma pulchaltum Uacrolampis adcularis Aspisomalgnitum Blcellonychaamoena Pyractomena ecostata Photuris dlvisa Pyractomena borealis Photuris brunnipennis Cratamarphus diaphanus Phatfnus pyralls Alecton dscoldalls Phatlnus Ignitus Alecton fiavum Phatlnus meteoralls PyrocoeOapraetaxta PyrocoeBa rufa Robopus to Uicrophotus

FIGURE 18

i Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FIGURE 19

The evolution of photic signals, produced by either sex, in Lampyridae is represented

by a single optimization of four origins and four losses.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Brachyiampis sangulnicollis Pslladadius sp Ellychnia corrusca Asplsoma puicheilum Asplsom a fgnitum Pyractomena ecostata Pyractomena borealis Cratomorphus diaphanus Alecton dscoldaSs Alecton fiavum PyrocoelfapraetBxta Pyrocoeha rvfa ——Pyropyga nigricans Erythrolychnia bipartttus Erythrolychnia oltveri PhaenoUs ustulata _ Tenaspis angutaris Luddota atra Luddota dllaticomls Ludtffna bfpiagfata new spades? Pollacfasls bifaria Pristolycus sagulatus Vesta aurantSaca . Caltopfsma maestra _ ‘ Cladodesffabellatus r j hUacrolampis adcufaris 1 Blcallonycha amoena Photuris dlvisa Photuris brunnipennis |_ _ f Phatlnus pyralls Phatlnus Ignitus mmm Phatlnus meteoralls R o b o p u ssp t Robopussp2 Ludola lateralis Ludala salomonis Ludola crudata Ludala kuroierae Colophotla sp Pteroptyx cribeUata Pteroptyx malaccae Pteroptyx taner Lamprohizm splandldula Phausis rhombica Phosphaenus hemiptarus Pfeotomusnigripennfs Pteotomus pattens Lampyris noctlluca Uicrophotus octarthrus Lampyris zenkeri Uicrophotus angustus

FIGURE 19

133

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FIGURE 20

The evolution of signaling systems that use a combination of both pheromonal and

photic signals. Five possible orgins and two losses are hypotinzed.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Brachyiampts sanguirtlcdlls Psilocladlus sp Blychnia corrusca Asplsoma pulchdlum Asplsom a fgnitum Pyractomena ecostata Pyractomena borealis Cratomorphus diaphanus Alecton dlscoldalls Alacton fiavum Pyrocoettapraetaxta Pyrocoettetuta Pyropyga nigricans Erythrolychnia blpartftus Erythrolychnia otteeri Phaenotts ustulata _ Tenaspts angutaris Luddota atra Lucldota dltaticomis Luddlna blpfagfata new spades? Pol la clasts bifaria Pristotycussagulatus Vesta atuanOaca Callopisma maestra ' Cladodesflabellatus Uacrolampis adcutaris Bicettonycha amoena Photuris tOvisa Photuris brunnlpannis Phatlnus pyralls Phatlnus Ignitus Phatlnus meteoralls R obopusspl Rabapus spZ Luclaia lateralis Ludola salomanis Luclaia cruclata Ludola kuroiwae Calophatia sp Pteroptyx cribettsta Pteroptyx malaccam Pteroptyx taner Lamprohiza sptendldula Phausis rhombica Phosphaenus hemiptarus Pteotomus nfgripennis Pfeotomus pattens Lampyris noctlluca Utcrophatus octarthrus Lampyris zenkeri Itkrophotusangustus

FIGURE 20

135

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FIGURE 21

The evolution of signal modalities, sexual signal systems, sedentary or active primary

signalers, and the sex of the primary signaler C = chemical, P = photic; I = Signal

System I, II = Signal System H; S = sedentary primary signaler, A = active primary

signaler; F = primary signaler is female, M = primary signaler is male. Representative

adult male photic organ morphologies: A. Cratomorphus diaphanous ; B. Pyrocoelia

rufar, C. Erythrolychnia oliveri', D. Bicettonycha amoena ; E. Robopus sp. #2; F.

Pteroptyx tener, G. Pleotomus pattens.

136

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. / / / / / / /// - Brachyfampis sangutnlcollls c - S F • PsilocSadus sp. C - S F Elychnia corrusca C - S F Asplsoma pulchallum p a a m Asplsoma ignitum p a A M Pyractomanaocostata PHAM Pyractomena boraatfs PHAM A. Cratomorphus daphanus P - a M Alecton d^coldaOs C - S F Alacton ffavum C - S F PyrocoaUa praatcxta C P 7 7 PyrocoaUa rufa C P I - B Pyropyga nigricans C • S F B Erythrolydmla bipartftus C P - B Eiythrotychnia olbmri C P - - B Phaenolls ustulata C P ? 7 Tenaspfs angutaris C - S F Luddota atra C - S F Luddota dUattcomls C - S F Luddlna blptagfata C - S F newspacie* CP - ? ? Pdtadads bifaria C - S F Pristotycussaguiatus C - S F Vesta aurantfaca C - S F Caffoptsmamaasba C - S P CtarioriaslhbaUatus C - S F - Uacrolampis adcdaris C - S F Blcetlonyd* amoena PHAM Photuris d visa PHAM Photuris brunnipannis PHAM d Phatlnus pyralls PHAM 'Phatlnus Ignitus PHAM 'Phatfnus metearaBs PHAM R obopussp- 1 P - A M Rbtopnip.2 P A M Ludola tataratls P - - B Ludola salomonis P - 7 ? Ludala crudata P - - B Ludola kuroiwma PHAM Colophotfasp. P A? M? Pteroptyx cribdiata P A? M? Pteroptyx mataecaa P A? M3 Pteroptyx tanar P - A? M? Lamprohiza splendduia P - - B Phausis rhombica CP? S F Phosphaonus hamlptarus C - S F a Plaotomus nigripantris C P - S F Pfeotomuspal/ens CP - S F Lampyris noctlluca P I S F Ificrophatusoctaithrus P S F Lampyris zankari P B S F Mfoophatusangustus P - S F

FIGURE 21

137

i ) Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FIGURE 22

Adult male photic organ evolution on abdominal ventrite seven (true abdominal

segment eight). Only one photic organ morphology, the two-spot condition, occurs in

this abdominal segment in male fireflies. The occurrence of the two-spot condition

(whether it appears to be functional or not) is indicated by the dashed branches of the

cladogram. This photic organ morphology is widespread throughout the family and

has evolved at least eight times. Representative adult male photic organ morphologies

include: A. Robopus sp. #2; B.Pteotomus pattens.

l Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Brachyfampls sanguinicoOls Psilocladussp. EHychrnacorrusca Asplsoma pulcheiium t^-^Aspisoirta ignitum Pyractomena acostata Pyractomena borealis Cratamarphus diaphanus Alecton discoidafls Alecton fiavum Pyrocoetiapraetexta Pyrocoeliarufa Pyropyga nigricans Erythrolychnia bipartttzjs Erythrolychnia divert Phaenolis ustulata Tenaspls angutaris =•-■5 Luddota atra Luddota dSaticomis Luddhta biplagtsta -•$ new species >= Poliadasis bifaria Pristolycus sagulatus Vesta autandaca X - - ju Cailoplsma maestra Ciadodes fiabellatus Uacrolampis adcuiaris r " Bicellonycha amoena A Photuris dlvisa ‘^=•-'2 Photuris brunnipennis Photinus pyralls Phatlnus fqnitus ~phatlnus meteoralls ’^RobopUSSp.1 * 3 3 3 Ro b o p u ssp . 2 Ludola lateralis Ludala salomonis A. Ludola crudata Ludota kurohme Colophotfm sp . Pteroptyx cribellata Ptaroptyx malaccaa Pteroptyx ten er Lamprohiza sptendUuta Phausis rhombica 3 3 Phosphaenus hemiptarus j . - - - - pteotomus nigripennis >j “ t 3 3 3 pieotamus pattens . - - j y 3 - - 3 Lampyris noctlluca uicrophotus octarthrus ~ Lampyris zenkeri 5*3Z-5 uicrophotus angustus

FIGURE 22

139

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FIGURE 23

Male photic organ evolution on abdominal ventrite six (true abdominal segment

seven). Only three types of photic organ morphologies appear in this abdominal

segment, the two-spot, center-strip and the entire ventrite photic organ morphology.

Whenever these morphologies are present, they are functional. The two-spot photic

condition appears to have evolved twice, while both the center-strip and the entire

ventrite photic organ morphology appears to have each evolved at least three times in

the seventh abdominal segment of Lampyridae. Representative adult male photic

organ morphologies: A. Cratomorphus diaphanous; B. Pyrocoelia rufar, C.

Bicettonycha amoena; D. Pteroptyx tener.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. “Brachyfampls sanguinicollis -Psilociadus sp . 'Ettychnia corrusca Asplsoma pulchellum Asplsoma Ignltum Pyractomena ecostata Pyractomena borealis i f --Cratomorphus diaphanus — ~ " Alecton discoldalls A.. ’ Alecton fiavum -Pyrocoeliapraatexta iPyrocoeBarvfa ■ Pyropyga nigricans ' Erythrolychnia bfpartltirs " Erythroiychniaoliveri ■ PhaenoUs ustulata • Tenaspis angutaris 'Lucldota atra 'Luddota dHaticomis ' Luddlna blplagfata ‘new species ' Pcliaclasis bifaria 'Pristnlycus sagulatus ' Vesta aurarrtlaca ' CaJlopisma maestra ' Cladodes ffabeSatus 8 | Uacrolampis adcufaris ■ Bicallanycha amoena Photuris dlvisa Photuris brunnipennis 1 Phatlnus pyralls Phatlnus Ignitus Phatlnus meteoralls ' Robopus s p . 1 ' Robop u s sp . 2 £Ludala lateralis = Luclaia saiomonis Ludola crudata Ludola kurotvrae Caidphotfa sp. Pteroptyx cribellata -j j F ’-'5 Pteroptyx malaccaa ^33"’■& Pteroptyx tener Lamprohizm splandldula Phausis rhombica Phosphaenus hemiptarus Pteotomus ntgripennis D. Pteotomus pallens Lampyris noctlluca 9 Uicrophotus octarthrus Lampyris zenkeri Uicrophotus an gustus

FIGURE 23

141

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. FIGURE 24

Male photic organ evolution on abdominal ventrite five (true abdominal segment six).

Whenever these morphologies are present, they are functional. The one spot, two-spot

evolved once and the center-strip photic organ morphology evolved twice in the

family Lampyridae. The photic organ morphology in which all of the surface area of

the ventrite is covered by the photic organ, appears to have evolved at least three

times in the sixth abdominal segment of Lampyridae. Representative adult male

photic organ morphologies: A. Cratomorphus diaphanous', B. P yrocoelia rufar, C.

Erythrolychnia oliverii D. B icettonycha amoena ; E. Pteroptyx tener.

142

i i Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Brachyfampis sangulnlcotlis Psilodadussp ~'EJlychnia corrusca Aspisoma pulchellum Aspisotna fgnitum Pyractomena ecostata Pyractomena borealis Cratomorphus diaphanus — ^ — Alecto n A. •A lecton fiavum 9 iPyrvcoeila praetaxta iPyrocoeHa rufa ” “ Pyropyga nigricans ss-s Erythrolychnia blpartitus r divert • Phaen oils ustulata 'Tenaspfs angutaris Luddota atra C. Luddota dltatScomls 9 Luddlna blplagiata near species!? PoHactasis bifaria Pristntycus sagulatus Vesta aurantlaca Callopisma mamstra CtadodasBabaOatus EUS r— Uacrolampis adcutaris I BIcallonycha amoena “ | Photuris dlvisa " Photuris brunnipennis Phatfnus pyralls Phatfnus Ignitus Phatfnus mataoralls T Robopus 3p t Robopus sp2 Ludala lateralis Ludola salomonls Ludala crudata Ludola kuroiwae Colophotiasp Pteroptyx cribellata Pteroptyx malaccae Pteroptyx tener Lampmhiza splendlduta N one Phausis rhombica 5 Phosphaenus hemiptarus O ne S p o t Plaotomus ntgripennis TWo S p ots Pleotomus pallens Center Strip Lampyris nactBuca Uicrophotus octarthrus All Lampyris zenkeri Uicrophotus angustus

FIGURE 24

143

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Zom, LJ*. & Carlson, A.D. (1978): Effect of mating on response o f female Photuris firefly. Anon. Behav . 26:843-847.

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. APPENDIX A. LIST OF TAXA USED IN THE ANALYSIS Material studied was borrowed from the following institutions: California Academy o f Sciences, San Francisco, CA [CASC]; Field Museum of Natural History, Chicago, IL [FMNH]; Florida State Collection of Arthropods, Gainesville, FL [FSCA]; Collection of author [MABC]; Museum of Comparative Zoology, Harvard University, Cambridge, MA [MCZC]; Ohio State University Collection, The Ohio State University, Columbus, OH [OSUC]; Snow Entomological Museum Collection, University of Kansas, Lawrence, KS [SEMC]; Museum of Zoology, University o f Michigan, Aim Arbor, MI [UMMZ]; National Museum of Natural History, Smithsonian Institution, Washington, D.C.[USNM].

List of Species Studied: The higher classification used here is based on Lawrence & Newton (1995), and in the case of Rhagophthalmidae. Wittmer and Ohba (1994).

Plastoceridae Plastocerus (=Ceroplatus) angulosus (Germar) [FMNHI Omalisidae Omalisus (=Omafystis, Homaltsus) fontisbellaguei (Fourar.) 1785 [FMNH] O. sangum ipenm s Cast. 1840 [FMNH] Drilidae Drihis concolor Ahr. 1812 [FMNH] D. jlavescens G_A. Olivier 1790 [USNM] Selasia sp. [FMNH] Omethidae Matheteinae Mathetens theveneti LeConte 1874 [CASC] Gmgtymocladus luteicollis Van Dyke 1918 [CASC] Omethinae Omethes marginatus LeConte 1861 [CASC] Blalchleya gracilis Blatchley 1910 [OSUC] Phengodidae

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Phengodinae Cenophengus pallidus Schaeffer 1904 fFSCA] Phrixothrix reducticomis Whtmer 1963 [UMMZ] Pseudophengodes pulchella Guer 1843 [USNM]

Zarhipis integripennis LeConte 1874 [MABC], [UMMZ] Rhagophthalminae Dioptoma adamsi Pascoe 1860 [USNM] Diplocladon sp. [CASC]

Rhagnphthalmfriaft Rhagophthalmus ohbai Wittmer and Ohba 1994 [MABC] Rhagophthalmus sp. [SEMC], [CASC] Telegusidae Pseudotelegeusis sp. [SEMC] Telegeusis nubifer Martin 1931 [SEMC] Lycidae Calochrominae Calochromus perfacetus (Say) 1825 [OSUC] Lycinae Calopteron discrepans (Newman) 1838 [OSUC] Celetes basalts LeConte 1851 [OSUC] Erotmae Dictyoptera aurora (Herbst) 1789 [OSUC] Cantharidae Cantharinae Cultelhmguis ingenuus LeConte 1881 [OSUC] Silinae Discodon bipunctatum Schaeffer 1908 [OSUC] Maithininae Malthinus occipitalis LeConte 1851 [OSUC] Chanfingnathfnaft Trypherusfrisoni Fender I960 [OSUC] Lampyridae Pterotmae Pterotus obscuripermis LeConte 1859 [UMMZ] Cyphonocerinae Pottaclasis bifaria (Say) 1835 [MCZC], [FSCA] Ototretinae Brachylampis sangumicollis Van Dyke 1939 [CASC] Drilaster subtilis (E. Olivier) 1908 [CASC] D riliaster sp. [MABC] Harmatelia bilinea Walker 1858 [CASC] Stenocladius sp. [MABC]

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Amydetmae Cladodes flabeUatus Solier 1849 [CASC] Psilocladus sp. [MABC] Vesta aurantiaca E. Olivier 1886 [USNM] Lampyrinae Alecton discoidalis Laporte 1833 [MCZC]

A flavum Leng and Mutchler 1922 [MCZC] Aspisoma igrntum (Linnaeus) 1767 [OSUC] A ptdchettum (Gorham) 1880 [FSCA] Callopisma maestra Mutchler 1923 [CASC] Cratomorphus diaphamis (Germar) 1824 [USNM] Ellychnia comisca (Linnaeus) 1767 [MABC] Erythrofychma bipartitus (E. Olivier) 1912 [FSCA] £ olivieri Leng and Mutchler 1922 [FSCA] Lamprohiza splendidula (Linnaeus) 1767 [CASC] Lampyris noctiluca Linnaeus 1767 [CASC], [FSCA] L zenkeri Germar 1817 [FMNH] Lucidina biplagiata (Motschulsky) 1866 [MABC] Lucidota atra (GA . Olivier) 1790 [MABC] L dilaticomis (Motschulsky) 1854 [FMNH] Macrolampis acicularis (E. Olivier) 1907 [CASC] Microphotus angustus LeConte 1874 [FMNH] M octarthrus Fall 1912 [MABC] New Species [MABC] Phaenolis ustulata Gorham 1880 [FSCA] Phausis rhombica Fender 1962 [MABC] Photinus ignitus Fall 1927 [OSUC] P. meteoralis (Gorham) 1881 [CASC] P. p yra lis (Linnaeus) 1767 [MABC] Phosphaenus hemipterus (Fourcroy) 1785 [CASC] Pleotom us m gripen n is LeConte 1885 P. pattens LeConte 1 8 6 6[MABC] Pristofycus sagulatus Gorham 1883 [CASC] Pyractomena ecostata (LeConte) 1878 [FSCA]

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. P. borealis (Randall) 1828 [MABC] Pyrocoelia praetexta E. Olivier 1911 [MABC] P. rufa E. Olivier 1886 [MABC] Pyropyga nigricans (Say) 1823 [MABC], [CASC] Robopus sp. #/ [MABC] Robopus sp. #2 [MABC]

Tenaspis angularis (Gorham) 1880 [CASC], [MCZC] Luciolinae Colophotia sp. [MABC] Luciola cruciata Motschulsky 1854 [MABC] L kuroiwae Matsumura 1918 [MABC] L lateralis Motschulsky 1860 [CASC] L salomonis (E. Olivier) 1911 [CASC] Pteroptyx cribellata (E. Olivier) 1891 [MABC], [UMMZ] P: malaccae (Gorham) 1880 [MABC] P. tener E. Olivier 1907 [MABC] Photurinae BiceUonycha amoena Gorham 1880 [FSCA] Photuris brunnipennis Jacq.-Duv. 1856 [OSUC] P. divisa LeConte 1852 [MABC]

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

CHARACTERS AND CHARACTER STATES Multistate characters treated as ordered are specified below. Values for Consistency Index (C J.) and Retention Index (RJ-) fin: each character in the analysis as they appear on the consensus tree are indicated after the last character state (CJ., RJ.). The character- taxon matrix is presented in Appendix 3. The morphological terminology of Lawrence & Britton (1991) and Snodgrass (1993) was used. Wing venation scheme follows that used in Kukalova-Peck & Lawrence (1993).

0. Headposition : 0-exposed; I-partially exposed; 2-covered. (C J. 0.18, RX 0.72)

1. Head shape: O-deflexed between eyes; 1-partially deflexed; 2-not deflexed. (C.1.0.8, R J. 0.52)

2. Antennal insertions (ordered): 0-widely separated; 1-moderately approximate; 2- approximate. (CJ. 0.8, RJ. 0.61)

3. Antennal sockets : 0-prominent; I-flush. (CJ. 0.5, RJ. 033 )

4. Number segments (antennomeres) in male antennae (ordered): 0-eight; 1-ten; 2- eleven; 3-twelve; 4-thirteen. (C J. 0.44, RJ. 0.50)

5. A ntennal seg. #5(flagellomere M) (ordered): 0-short, 1-same as #4; 2-Iong. (C J. 0.7, R J. 0.52)

6. Antennalfeatures (general): 0-filiform; I-serrate; 2-flahelIate; 3-pectinate; 4- bipectinate. (C J. 035, RJ. 0.63)

7. Distal antennal flagellomeres (ordered): 0-Ionger than wide; 1-about as long as wide; 2-much wider than long. (CJ. 0.14, RJ. 0.29)

8. Basal antennalflagellomere/s: 0-not symmetrical with apical flagellomeres; I- symetrical with apical flagellomeres. (CJ. 0.14, RJ. 0.53)

9. Distal margins o f flagellomeres: 0-straight; 1-concave. (CJ. 0.11, RX 0.46)

10. Distal margin o fantennal flagellomeres: 0-approximating proximal margin in width; 2-wider than proximal margin. (CJ. 0.7, RJ. 038)

1 6 7

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 11. Antennalflagellomere #2 (ordered): 0-not compressed; 1-slightly compressed; 2- greatly compressed (CJ. 0.8, RJ. 0.60)

12. Lateral margins ofthe distal antennalflagellomeresz Q-parallel; I-non-parallel. (CJ. 0.5, RJ. 0.52)

13. Antennal lobes producedfrom (ordered): 0-basal region of flagellomere; 1-medial region of flagellomere; 2-apicaI region of flagellomere. (C J. 0.25, RJ. 0.62)

14. Number o felongated antennal lobes per segment. 0-one lobe; 1-two lobes. (C J. 0.25, R J. 0.62)

15. A ntennal lobes: 0-compressed; l-not compressed. (CJ. 0J20, RJ. 0.33)

16. Length o f antennal lobes (ordered): 0-less than length o f flagellomere; 1- approximating length of flagellomere; 2-greater than length of flagellomere. (C J. 0.50, RJ. 0.66)

17. Antennal lobe/flagetlomere juncture : 0-broad; 1-narrow (CJ. 0.16, RJ. 0.44)

18. Antennal lobes : 0-not bearing a sensory depression at apex; I-bearing a sensory depression at apex (C J. 1.0, R J. 1.0)

19. M andibles (ordered): 0-prominent; 1-normal sized; 2-reduced; 3-very reduced. (CJ. 0.14, RJ. 0.68)

20. Mandible tooth : 0-absent; I-present (CJ. 1.0, RJ. 1.0)

21. Mandible -width. 0-stout; 1-slender. (CJ. 0.12, RJ. 0.68)

22. M andible shape: 0-apices acute (inside angle <90 degrees); 1-apices non-acute(inside angle near 180 degrees). (CJ. 0.5, RJ. 0.56)

23. Mandible type: 0-normal type (arcuate, regularly narrowing to tips); 1-specialized type (tips slender and glabrous with discontinuous curvature). (C.1.0.25, RJ. 0.84)

24. Hypomera: 0-not exten d in g to anterior edge of pronotai shield; I-narrowly extending to anterior edge of pronotai shield; 2-broadly extending to anterior edge of pronotai shield; 3-Iacking. (CX 030, RJ. 0.82)

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 25. Hypomera space around head (side view)' O-head (eyes) not able to retract between hypomera; 1-head (eyes) partially enclosed (up to half width of eyes); 2-head (eyes) retractable (less than half eye width exposed). (CJ. 0.11, R J. 0.65)

26. Maxillary palpi'. 0-filiform; I-clavate compressed; 2-cIavate; 3-modified. (CJ. 0.12, R J.0.57)

27. Maxillary palp apical seg. 0-filiform; I-securiform; 2-eIongate; 3-greatly elongate and flattened; 4-conicai. (CJ. 0.4, RJ. 0.5)

28. Labial palpi'. 0-filiform; l-clavate compressed; 2-clavate; 3-modified. (C J. 0.16, RJ. 0.44)

29. Labial palp apical seg: 0-filiform; I-securiform; 2-elongate; 3-greatly elongate and flattened. (C J. 0.20, RJ. 0.52)

30. Eyes: 0-oval; 1-emarginate. (C J. 1.0, R J. 1.0)

31. Eyes postero-ventrally (ordered): 0-seperated; 1-approximate; 2-condguous. (CJ. 0.25, R J. 0.66)

32. Pronotum border. 0-smooth; 1-margined; 2-expIanate. (C J. 0.22, R J. 0.46)

33. Hind angles ofpronotum: 0-truncate (juncture between lateral and hind margin = 90 degrees); 1-acute (juncture <90 degrees); 2-lateraIly expanded (juncture >90 degrees); 3-notched (juncture <90 degrees due to deep notch in hind margin). (CJ. 0.13, RJ. 038)

34. Overall surface area ofhypomeron (ordered): 0-absent; 1-small; 2-large/broad. (CJ. 0.22, RJ. 0.58)

35. Scutellum shape: 0-distinct; l-poorly developed. (CJ. 0.25, RJ. 0.40)

36. Scutellum : 0-membranous, 1-sclerotized. (CJ. 1.0, RJ. 1.0)

37. P rostem um (ordered): 0-small; I-medium; 2-Iarge. (CJ. 0.22, RX 0.53)

38. Mesostemum (cmt. margin): 0-straight; 1-emarginate. (CJ. 0.50, RJ. 0.75)

39. Mesal margins o f metepistenur. 0-sigmoid; 1-straight or nearly so. (C J. 033, RJ. 0.83)

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 40. Anterior coxae (ordered): O-contiguous; 1-nearly contiguous; 2-separate at base. (CX 0.33, R J. 0.42)

41. Anterior coxal shape: O-conical; l-subconical; 2-trianguIar; 3-broad; 4-buIbous. (C J. 0.19, R J. 0.54)

42. Middle coxae (ordered): O-contiguous; 1-nearly contiguous; 2-separate. (C.1.022, R J. 0.63)

43. H ind coxae (ordered): O-contiguous; 1-nearly contiguous; 2-separate. (C.1.0.09, RJ. 033)

44. Hind coxae/femoral plates (ordered)- 0-plates obsolete; l-< length of coxae; 2-entire length of coxae. (CJ. 0.16, RJ. 0.74)

45. Trochanter attachment to femora : 0-oblique; I-very oblique; 2-interstitial. (CJ. 0.22. R J. 0.82)

46. Middle trochantinsr. 0-setiferous; 1-glabrous. (CJ. 0.10, RJ. 0.47)

47. Femora'. 0-sIender, 1-normal; 2-Qattened; 3-swollen. (CJ. 022, RJ. 0.56)

48. Tibiae'. 0-slender; 1-normal; 2-flattened; 3-swollen. (CJ. 0.14, RJ. 0.50)

49. Tibial spurs (ordered): 0-absent; 1-small; 2-well developed. (CJ. 0.06, RJ. 0.42)

50. Hind tarsal segment one: 0-normal; 1-elongate. (C J. 022, RJ. 0.53)

51. Tarsal segment three-. 0-simple; l-Iobed beneath. (CJ. 025, RJ. 0.66)

52. Tarsal segmentfour. 0-simple; l-Iobed beneath. (CJ. 025, RJ. 0.0)

53. Claws: 0-snnpIe; 1-cleft (CJ. 0.50, RJ. 0.66)

54. Male elytra (ordered): 0-fiiIIy covering abdomen; 1-somewhat reduced; 2-greatly reduced. (C J. 028, RJ. 0.54)

55. Elytral surface: 0-slight punctures with no costae; I-slight punctures with longitudinal costae; 2-deep window shaped punctures with longitudinal costae; 3- coarse punctures with no costae; 4-slightly coarse punctures with longitudinal costae. (CJ. 026, RJ. 034)

1 7 0

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 56. Elytral epipleuralfold (ordered): 0-absent; 1-narrow; 2-broad at base. (C J. 0.22, RJ. 0.84)

57. Abdominal ventrite #(includingpygidiian) (ordered): 0-six visible; I-seven visible; 2- eight visible. (C J. 020, RJ. 0.72)

58. Male ninth abdominal tergite : 0-not emarginate behind; 1-emarginate behind. (C J. 0.11, R J. 0.75)

59. Setae on claws' 0-absent; I-present. (CJ. 0.50, RJ. 0.85) 60. Abdominal segment 6, shape o fphotic organ/sr. 0-two-spots; I-one spot; 2-all; 3- center-strip; 4-none. (C J. 0.57, RJ. 0.85)

61. Abdominal segment 7t shape o f photic organ/s: 0-two-spots; 2-strip; 3-all; 4-none. (CJ. 037, RJ. 0.73)

62. Abdominal segment 8, photic organs 0-absent; I-present (C J. 0.08, RJ. 0.47)

63. Pairedphotic organs on segments 1 - 7 : 0-absent; 1-present (C J. 0.50, RJ. 0.0)

64. Wing vein r3: 0-absent; 1-present. (CJ. 0.08, RJ. 0.26)

65. Wing vein r4 (ordered): 0-absent; l-partial; 2-compIete. (CJ. 0.09, R J. 0.42)

66. Wing Radial Cell: 0-open; 1-closed; 2-not present (CJ. 0.13, RJ. 0.23)

67. Wing vein MP3: 0-contacting MPl+2; l-not contacting MP1+2. (CJ. 0.07. RJ. 027)

68. Wing Ist Cubito-Anal Cell: 0-absent; 1-present (C J. 0.16, RJ. 0.54)

69. Wing 2”^ Cubito-Anal Cell: 0-absent; 1-present (CJ. 0.05, RJ. 0.55)

70. Wing CttAl(cross-vein): 0-absent; l-partial; 2-compIete. (CJ. 0.11, RJ. 030)

71. Wing CitAl vein intersecting MP vein: 0-above fork (MP3a&MP3b); I-at fork 2- below fork; 3-other (no fork present). (CJ. 0.16, RJ. 021)

72. Wing CuA2(cross vein): 0-absent; l-partial; 2-complete. (CJ. 023, RJ. 0.67)

73. WmgAA3+4 vein: 0-absent; 1-present (C J. 0.08, RX 037)

171

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

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sp,

sp. sp,

sp.

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Plaatocerua anguloaua Selaaia Prilua flaveacenaPrilua concolor Pterotua obaouripennia Harmatelia bilinea Trypherua friaoni Rhagophthalmua Drilaater aubtllia Plplocladon D1 op coma adamal coma op D1 Omallaua Omallaua fonciabellaguel Oinallsua aanguinipennla Rhagophthalmua Rhagophthalmua ohba Caloohromua perfacetua Pictyoptera aurora placodon bipunctatum Calopteron diaorepana Celecea baaalia APPENDIX C. APPENDIX MORPHOLOGICAL CHARACTER MATRIX Character Number 567890123456789012345678901234567890123456789012345678901234567890123 01234 Telegeuaia nubifer pulchellaPaeudophengodea Paeudotelegeuaia Phrlxothrlx raductlcornia Omethea marglnatuaOmethea Blatohleya graciliaGlnglymocladua lutelcollia P rila a te r C ultellunguia Ingenuua Character Number (10) 1 2 3 Stenocladlua Cenophengua palliaCenophengua u a Mathetaua thevenetl Malthlnua occipltalla

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sp.

ustuJata ustuJata integripennis integripennis atra atra Psiiocladus Pyractomena Pyractomena ecostata Pyrocoelia praetentaPyrocoelia rufa Pyractomena Pyractomena borealis Earhipia Braahyiampia aanguinicoilis Eiiychnia corrusca Pyropyga Pyropyga nigriaana Cratomorphua Cratomorphua diaphanua Phaenolia Tenaapla angularia Character Number (10) Number (10) Character Aapiaoma Aapiaoma puichelium Character Number Character Aapiaoma Aapiaoma ignitum Lucidota Lucldina biplagiata Pollaolaaia bifaria Aleoton dlacoidalia Erythrolyohnia bipartitua luddota dilaticornia Priatolycua aagulatua Vesta aurantiaca Photinus pyraila Alecton flavum Erythrolyohnia oliveri apecieaNew Calloplama maeatra Photinus ignituaPhotinus meteoraiia Cladodea Cladodea fla b e lla tu s Macrolampis acicularis

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sp. sp. sp2 sp2 Photuris brunnlpennis Bicellonycha amoena Photuris divisa Robopus spiRobopus Robopus Luciola lateralisLuciolais Salomon Luciola oruciata Pteroptyx crlbellataPteroptyx malaccae Luciola kuroiwae Ptecoptyx tener Character Number (10) Number (10) Character Pleotomus pallens Phosphaenus Phosphaenus hemlpterus Pleotomus nigripennis Colophotia Character Number Character Phausis rhomblca Lampyris noctiluca Lamprohisa splendidula Lampyris senkeri Microphotus angustus Microphotus octarthrus 33

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