Scholarly article on 2-Propanethiol, 1-(phenylamino)- 90712-75-7 from Canadian Journal of Chemistry p. 395,401

and

(Lepidoptera:

nuclear

Monophyly,

within

composition,

Saturniinae

relationships

Saturniidae):

Evidence

from

two

genes

RICHARD S.

JEROME

C.

CHARLES

MITTER,

PEIGLER

REGIER,

and TIMOTHY P. FRIEDLANDER

The

1500

lepidopterans,

of Saturniidaeor wild silk

which

systems

includesomeof the

moths,

species

have

model

provided important

approximately

of all

for studies

of

largest

ecology,

comparative

To address we

andbehavior.Such

studies

phylogeneticanalysis

wouldbenefitfroma robust

developmental

genetics,

but there has been

little

of this

framework,

this,

family.

andinternal

use nuclear

to test

genesequences

aboutthe

hypotheses

monophyly

relationships

the

of

Extending

and

geographicallywidespread

Saturniinae

644

spp.).

large

(63

subfamily

genera,

our

examinationof the

we

analyze

of Attacini,

coding sequence

from

factor-1α

and

1051 in 64

(1240nt)

elongation

dopadecarboxylase

nt)

(typically

species

four of five

in

tribes

bombycoids.

11of 16

in

and

in

Saturniinae,

Saturniini,

representing

genera

support

should

outgroups

Saturniidaeand other

The

results

a recent

includethe African

that

Saturniinae,

postulate

Oriental and Palearcticin

The

distribution,

largely

Micragonini.

alternativethat

called

or some

shown to result

Bunaeini,

supported,

thereof constituteits

own

paraphyletic

Micragonini

subgroup

subfamily(previ-

is

in

a

Saturniinae.

for

Ludiinae)

ously

Micragonini

group

with

the tribe

also African.

as is a basal

a

Saturniinae,

strongly

Monophyly

including

is

between Attacini +

split

Saturniini and

Micragonini,

strongly

+

Bunaeini

As a

to the

Africantribes

of two

Micragonini.

consequence, postulated

affinity

endemic

thus

way

Saturniini

is

paraphyletic,

Saturniini,

Madagascan

However,

rendering

rejected.

there is no

evidence either

on

of Saturniiniversus

with

strong

monophyly

paraphyly

to the

Attacini

moths and

This result reflects

relatives).

(atlas

in

respect

clearly monophyletic

weak resolution

of

Saturnini.Several

the

lower-level

groupings

generally

deeper divergences

within

Saturniiniare

'moon

tailed-hindwinged

moths'

a

lato,

corroborated,

strongly

including

that

on resinous

hostplants,

and Saturnia sensu

Actias,

Graellsia)

eight

(Argema,

specialize

consolidationof

former

small,

genera.

Jerome C.

Center for

Plant

of

Regier,

Agricultural Biotechnology,

University

Maryland

Sciences

MD

U.S.A.

Institute,

Park,

20742,

Biotechnology

er@glue.umd.edu).

Charles

Bldg., College,

(regi-

of

Plant Sciences

Mitter,

Department

MD

Entomology,University

Maryland,

Bldg.,

U.S.A.

of

Park,

20742,

College

(cm42@umail.umd.edu).

RichardS.

of the Incarnate

4301

of

Word,

Peigler,Department Biology,University

Broadway,

San

Timothy

TX

78209U.S.A.

Antonio,

(rpeigler@texas.net).

Agricultural

P.

Centerfor

&

Friedlander,

Biotechnology Department Entomology,

Plant Sciences

MD

U.S.A.

20742,

Park,

Bldg.,College

Introduction

The

interactions and

Scriber

Johnson

caterpillar ecology (e.g.,

1983),

Goldsmith & Wilkins

wild

or

of

silk

are the most

sensu stricto

Satumiidae,

moths,

Bombycoidea

1999,

see

developmental genetics

(e.g.,

and

diverse

in

family

(Minet 1994),

1995),

chapters

of about 1500

mediation of insect

behavior

consisting

which include some of the

species.

by pheromones

(e.g.,

and Baker

&

Williams

the

Riddiford &

of

Satumiids,

most

1988,

1971),

1980,

largest

provided

Vogt

Capinera

other

all

moths,

for

of

have

im-

conspicuous

model

among

subjects. Many

studies of insect/

are

portant

systems

plant

resulting hypotheses

inherently

compara-

10

tribeswithinSatumiinae.Asterisks

explicit phylogenetic

in the current

study.

1.

Figure Changingconcepts

of

identifygroupssampled

an

The nuclear

we use encode

for elon-

tive and will

require

framework for a

genes

dopa decarboxylase

genes

factor-l œ

test.

and

modem

(EF-l œ)

gation

(DDC).

vide

rigorous

role for

Examples

in the

to

been shown

have

a

include

pro-

(par-

They

previously

heterochrony

postulated

and

evolution of saturniid chorion

&

signal, singly

morphol-

strong phylogenetic

(eggshell)

a

in

at a

levels,

that subsumes those

of taxonomic

range

in and

Lepidoptera

and

combination,

1988),

ticularly)

Hatzopoulos

adaptive linkage

putative

(Regier

ogy

and

variation in larval host-

between

depths

other

divergence

and asso-

fami-

bombycoid

1984, Holloway

in this

Fried-

et

use and variation in adult

insects,

sampled

2000;

longevity

and within

plant

ciated

et al.

al.

Mitchell et

et al.

1985,

traits,

(Cho

Fang

study

among

Satumiidae

al.

lander et

lies

1987).

study

1989,

2000;

1998;

(Janzen

Regier

including

However,

there has been little modem

al. 2001).

of satumiid

Balcdzar

(but

phylogeny

& Wolfe

see, e.g., Peigler

taxon

and

of Saturni-

1997).

we extend our

Systematic background,

hypotheses

idae had been

uncertain,

sampling,

tested. - The

In this

nuclear

previous application

monophyly

as

paper

gene sequences

the

to satumiid

of

regards

particularly

phylogenet-

al.

et al.

of a robust

of the

ics

et

1998,

1998),

(Friedlander

sometimes-segregated Cercopha-

Tutt

placement

Regier

toward the

ninae

another

and

1914,

1902,

(Packard

Oxyteninae

Jordan

goal

taking

step

Forbes

for this

most

We focus on the

geo-

1923,

However,

1924, Henke 1936, Michener

family.

phylogeny

graphically

diverse

Minet

has identified

(1994)

the when Cerco-

family

are included. Lemaire

with-

nine subfamilies

assessment of their

has not

inter-relationships

and second-most-

644

1952).

seven

widespread

Saturniinae

for

(63

genera,

synapomorphies

and

subfamily,

which contains both the

saturniid

largest

Peigler

experimental

from other subfamilies and bom-

phaninae

Oxyteninae

spp.),

species

those most often

and and Minet

(1998)

recognize

the atlas

moths;

used as

1989)

models.

(e.g.,

a

in

but modem

Satumiidae,

and

their

of

monophyly

of been

Using outgroups

families,

bycoid

Saturniinae. We then

we first test the

monophyly

published.

Two of these satumiid

Satumiinae

subfamilies,

relevance to this re-

estimate

relationships among

and extend our

pre-

and

are of

tribes of Saturniinae

the

Ludiinae,

particular

as the of the former

monophyly

major

vious

to

1;

of Attacini

(Fried-

to the more

appears

to the latter

analysis

port,

hinge

generic-level

lander et al.

diverse Satumiini.

on its

relationship

1998)

(Fig.

11

Table 1.

and

of

taxa

in this

represented

study.

distribution,

Geographical

diversity,

samplingdensity higher

*

namesfollowLemaire& Minet

accordancewith

thatLudiinaeis

insideSatumiinaeas

(1998)

Subfamily

except

placed

Micragonini,

in this

3 and

(see

Fig.

in

and the results

1)

Oberprieler(1997) (see

Fig.

4).

presented

report

Tribalnameswithin

Attacinifollow

Satumiinaefollow

Generic

nameswithin Satumiiniand

Oberprieler(1997)(see

1).

Fig.

&

that

except

Graellsia is a

rather than a

within

(1994)

Oberprieler

Ndssig

genus

D'Abrera

(1996),

subgenus

Lemaire&Minet

(1998),

Actias

2 and

text).

(see

Fig.

**

Information

citedfrom

&

(1994),

Oberprieler Ndssig

(1998)

Heppner

of

and

Dictyoploca

obs.

Peiglerunpubl.

**

is listed in

the literature as a

Saturnia but is not an availablename because the

subgenus

type

is the sameas the

named

species

previously

Caligula.

12

of

Changingconcepts genera

withinSatumiini.Asterisks

in the current

identifygroupssampled study.

2.

Figure

13

Lemaire & Minet

&

Satumiini. Inclusion of Rhodinia in

our

1998,

1997).

Oberprieler

Ndssig

ar-

(1997)

sample

a test of the

advanced

sev-

by

1994,

gues

iinae

Oberprieler

permits

hypothesis,

that these

that

from the Lud-

to include Deca-

eral authors

nonetheless

are

as

see

1989),

Micragonini, expanded

of Bouvier

(Peigler

phylogenetically

the classification of Bouvier

genera

Attacini,

closest to

(1936)

Ndssig

concept

chordini

&

and Eosia

consti-

1994)

(1936;

(Oberprieler

placed

implied by

in Satumiinae:

we test the

advanced

Second,

1).

Urotini)

(formerly

Fig.

suggestion

tutes one of five tribes within

Lemaire

where-

that the

Satumiinae;

(1997)

by Oberprieler

tive,

Ceranchia

putatively primi-

Antherina and

are

phylogeneti-

tribe Bunaeini

ex-

that

& Minet

after

1994),

warrants

endemic

Madagascan genera

as,

conclude,

(Minet

Decachordini)

(1998)

in

our

(both

closer to the African

pressing previous uncertainty

their Ludiinae

sample)

than

to

(without

cally

status. Our taxon

al- other Satumiini. A test is

needed

Salassa

determine

to its own

also

to

separate subfamily

sampling

an initial

lows

molecular test of these

we lack

whether Michener's removal of

conflicting

for

renders Satumiini

(or Satumiinae)

hypotheses, although

Decachordini.

exemplars

subfamily

phyletic,

Saturniini,

para-

but

is

not

included here. Within

Salassa

Within

five tribes are

we test Michener's

of

six,

Saturniinae,

1,

recognized

of

synonymization

of which we

sampled

Bouvier

with Saturnia.

(Tab. 1,

1997).

Oberprieler

Fig.

Monophyly

eight

genera,

Bunaeini has been

1997),

questioned (Oberprieler

the

as

the

of

particularly

regards

relationship

Bunaeini to the most

Eochroa, Melanocera,

four

Materials and methods

Abbreviations. -

'higher'

taxa

primitive-looking

(e.g.,

Bunaeopsis).

EF-1

BP,

a,

bootstrap percentage;

in

we

two

Sampling

species

genera,

provide

monophy-

putatively

factor-la; DDC,

elongation

MP,

hood.

dopa decarboxylase;

maximum likeli-

an initial molecular test of the uncertain

of we lack the

maximum

ML,

parsimony;

Bunaeini,

ly

although

about which the

doubt

greatest

of Urotini also re-

as the

regards

primitive genera

has been

Taxon and

and

expressed. Monophyly

gene sampling

(1240

to 1051

experimental

length

mains

ment of Usta

in

nucleotides

EF-1 a

uncertain,

particularly

place-

Our

each)

design. -

and Parusta

and DDC

nucleotides

1997).

(Oberprieler

each)

two

Satumiidae,

(up

sequences

lacks an

from Urotini.

were obtained from 48

of

study

exemplar

species

The

Attacini

of

well estab-

of

and 14

of

Brahmaeidae,

monophyly

appears

(Tab. 2).

Sphingidae

lished

of these

were

1989).

(Peigler

have been assessed

Inter-generic relationships

Seventy-six

previously,

sequences

published

are new

Saturniidae,

mor-

while

all from

(1989),

62,

by Peigler

using

and

Friedlander et al.

(1998),

Friedlander et al.

et al.

by

2,

Regier

and

1998,

2000,

phology,

using

(Tab.

Regier

and

combined EF-1 a

for 15

DDC

al.

et

GenBank accession numbers

(2291 nt)

six

genera, plus

sequences

2001).

collection localities

all

9

are listed in

species representing

of Satumiini as

specimen

All

multiply

Tab. 2.

species

outgroups.

recovered

strongly

=

were

To

the satumiid

subfamily

(BP

sampled genera

uniquely represent

as were five of the

=

the

of

EF-1 a

100%),

level

tions of

Hemileucinae,

eight possible higher-

the

100%).

Only

Polythys-

of

sequence

DDC

ana

and the

(BP

relationships

posi-

Hyalo-

apollina

partial

sequence

and

Automeris io were combined to form a

Archaeoattacus,

Epiphora,

hybrid

ICallosamia ISamia were not

called

+

Automeris.' Simil-

phora

ported

strongly sup-

we

study

sequence

arly,

'Polythysana

=

In this

have

to

the

EF-1 cr

(BP

54-70%).

Caligula

of

uniquely represent

within the tribe

genus

increased our taxon

to determine whether

will increase.

Saturniini,

sample

sequence

node

for these

placements

and the DDC

or C.

of either

support

Monophyly

uncertain for the

Caligula japonica

were combined and called

and Ceratomia

sequence

and

are

composition

tribe,

(1936).

especially

original-

uncertain)

jonasi (origin

The Dar-

Satumiini,

largest

Bouvier

by

for Satumiini have been offered. Our

Caligula

sp.

defined

Several

were not identified

ly

of

postulates

apsa

to

specimens

because

were too

paraphyly

species

they

badly damaged.

analyzed phylo-

which includes

about two-thirds of the

Two

data sets were

sample,

genera

these. In Michener's

overlapping

tests of two of

influential revision

one for 64 taxa that included non-sat-

1,

2),

(1952)

the tribe

(Tab.

Fig.

permits

genetically,

umiid

Brahmaeidae,

(i.e.,

outgroups

Sphingidae);

taxa. The

the

to root

Tab.

of

the

the subset of 48

1,

(see

2),

other,

Fig.

formerly consisting

with

satumiid-only

designed

Rhodinia and Pararhodia was

64-taxon

was

synonymized

sample

14

Table2.

collection

and

GenBankaccessionnumbers.

localities,

Speciessampled,

15

*

EF-la

are each 1240nucleotidesin

sequences

DDC

areeach 1051

in

nucleotides

length,

unless

length.

sequences

GenBankaccessionnumber.

otherwiseindicatedwithin

afterthe

parentheses

**

Taxawith

datanew to

this

sequence

report.

Satumiidae and

a test of the

Smith et al.

mono-

There

were no indels in

of

thereby permit

1994).

Amino

any

of Satumiinae. The 48-taxon

was the

acid data

sets were

phyly

sample

sequences.

translated from nucleotide

sequences

concep-

to address

within

Satumi-

because

greater accuracy

designed

inae,

relationships

with

tually

using

Maddison &

Maddison

MacClade

1992).

3.07,

(version

potentially

most

DDC

taxa are

about a third of the

outgroup

sequence

missing

(Tab. 2)

synonymous

and because

homoplasy,

in-

across families.

Data

were

con-

EF-

analysis. - Phylogenetic

analyses

due

to

substitutions,

extended

largely

creased

ducted on the 48- and 64-taxon data sets

using

demonstrating

were

when

sampling

and DDC in

1 cr

after

combination,

from the two

Data collection and

were that the

in

not

assembly. -

Specimens

signals

genes

until

alive

frozen at

in 100%

-85°C

freezers

of

ethanol.

Department

Maryland College

conflict

Maximum

(see below).

parsimony

(MP)

Vouchers are stored in

of

at

the

of total nucleotide and amino acid data

analyses

sets were

with PAUP*4.0b2

Entomology, University

(Swofford

performed

unordered character

using

Park.

Total nucleic acids were isolated and

states and a

1998)

specif-

ic

were

the

heuristic search

100

with random

sequence-addition replicates

branch

(TBR

algorithm

sequences

chain reaction

amplified by polymerase

swapping,

EF-1 or

œ)

reverse

chain reaction

(for

by

transcrip-

(for

and conditions for

tion and then the

taxon

1985)

(typically,

swapping

values

(Felsenstein

Bootstrap

addition).

polymerase

of

were also obtained

1000

a heuristic search

with TBR branch

DDC).

Sequences

primers

using

with

nested

amplification, reamplification

primers,

have been

replications

and 10

random

and

automated

Cho et al.

isolation,

described

gel

sequencing

DDC:

sequence-addition

repli-

et cates

1995;

For

(EF-1 cr:

Friedlander et al.

Fang

the

EF-l œ,

per replication).

al.

Maximum likelihood

of com-

1997;

1998).

(ML)

analyses

between

M3 and

nucleotide data sets were

1240-basepair fragment

primers

For

bined-gene

the with

performed

rcM4 was

and

in

each case under the

DDC,

primers

PAUP*4.0b2,

fit

sequenced

analyzed.

between

simplest

statistically indistinguishable

as likelihood

1.7dF model

1051-basepair fragment

and M7.5RC was

otherwise indicated

providing

from the

and

unless

analyzed,

ratio

see also

sequenced

(Tab. 2).

optimum,

(Huelsenbeck

judged by

tests

& Rannala

1997,

A

time reversible

general

DNA

Automated

Shultz &

model

2000).

sequencer chromatograms

contiguous fragments

Regier

(general

were edited and

were

reference:

Swofford et al.

1996;

assembled

the GAP4

within the

GTR:

al.

et

1990)

was

selected.

accommodated

using

program

et al.

Rodriguez

Staden

software

were

Se-

rate variation was

(Staden

1999).

the Genetic Data

package

aligned using

Among-site

by

to a

the

of character

quences

fitting

gamma

frequency

distribution,

change

four discrete

Environment software

(version 2.2,

package

approximated by

16

A

ratio test showed that a reflected in the maximum observed

(uncorrected)

likelihood

categories.

estimated

differences across Satumiidae at the

sec-

of invariant

also

sites,

separate category

pairwise

the model's

this is called the GTR+G+I

ond codon

which is 5% for DDC but

likelihood,

(P <0.0005);

by

fit

significantly improved

position, only

1 %for EF-1 œ. Maximum differences at the third

are 55% and

for

and

DDC

model. A

time-reversible model

codon

EF-1

39%

separate general

characters were

and

position

in

which

a,

by

respectively.

the

partitioned

is

In

was also

this

from Satumiidae

position

the

by gene

explored;

When the all-nucleotide

extending

comparisons

Brahmaeidae and

to include the

called

GTR-ssr model.

outgroups

Sphin-

differences at the sec-

pairwise

data set was

ssr model

on the MP

the GTR-

maximum

tree,

a In-likelihood score

gidae,

ond codon

for

optimized

yielded

double

position approximately

11%

at

(to

approxi-

than the

to 2%

for

DDC,

while differences

1.5% lower

versus

GTR+G+I model

EF-la),

mately

Efficient

of the third codon

a

also increase for DDC

for EF-1œ. Our

(to

-18,619).

(-18,342

tree

exploration

entailed

position

remain

in likelihood

but

64%)

rate of an all-taxon data set

analysis

space

analyses

heuristic searches and

unchanged

sepa-

+

(Satumiidae

taxa)

of

(re-)

sequence

parameter

&

et al.

Brahmaeidae +

taxon

64

and a 48-

2000;

(Shultz

optimizations

Regier

Sphingidae;

Parameters fitted to the ML

were

distance matrices

data set was motivated

2000).

Satumiidae-only

topology

by

differences

used to create corrected

the

that the

pairwise

data sets

possibility

across

high pairwise

in the third

under a mini-

from 1000

mum

codon

families,

bootstrapped

particularly

could mask

model for

evolution

neighbor joining.

Average pairwise sequence

hits,

subsequent analysis by position,

multiple

inference.

undermining

phylogenetic

Both

differences were

show

base

genes frequen-

cies across Satumiidae at all codon

(P

positions

homogeneous

calculated

and

codon

by gene

PAUP*4.0b2. Base

were calculated

by position using

of terminal taxa

values from 0.58 to

further

their

With

the

1.00),

for saturniid

Brahmaeidae and

supporting

phylogenetics.

included,

frequencies

and

codon

by gene

by

position,

suitability

PAUP*4.0b2. The

also

incongruence length

as third

codon

position

Sphingidae

only

using

difference test

the

et al.

for EF -1 œbecomes

(Farris

1995),

implemented

strongly

of com-

in

<

In

0.001).

support

test

PAUP*4.0b2,

(P

partition homogeneity

was used to test for

non-homogeneous

bined

between

of total nucleotides from DDC and

tests reveal no con-

partition homogeneity

analysis

conflicting signal

Relative rates of nucleotide

character

EF-1 a,

partitions.

substitution codon

and

by gene

when

lengths

were

con-

flict in the

from DDC

signals

and

either

EF-1 œ,

inclusion of

by

estimated from

position

tree

=

with

without

(P 0.30)

sphingid outgroups.

or

(P 0.11 )

parsimony

strained to the same all-nucleotide-derived

The amount of

was estimated in PAUP*4.0b2 as the difference in

the brahmaeid and

topolo-

nucleotide

change

gy.

synonymous

and

Monophyly, composition,

phylogenetic posi-

discussion

overall tree

derived from total amino acid

from total nucleotide

In

the

results,

tion

lengths

and

when

from

of Saturniinae. -

of our

following

summarized in

3

(separately)

mapped

analysis

change

(derived

phylogenetic

Fig.

MP

onto the same

and

we make a heuristic distinction between

4,

topology

nucleotides).

of total

defined as those with

nodes,

(BP) 2:

nodes,

'strongly supported'

and the

80%,

i.e. those with BP <

other,

bootstrap percentages

more

weakly

supported

Results and discussion

EF-

80%. Low BP values can result either from low

numbers characters or from a rela-

and

œ and DDC:

of

supporting

Suitability of

Synonymous

tive abundance of

characters

conflicting

some

(or

pairwise differences,

tests.

non-synonymous changes,

base

-

and

combination).

homogeneity

frequencies,

partition

the all-

the

Within

differences

Likelihood and

of

Satumiidae,

about 92% total

of which resides at the third codon

synonymous

of character

parsimony analyses

data 64 taxa

set for

for

nucleotide

account

variation,

strongly support

of Satumiidae from the other two fam-

most

position

between

separation

ilies

Satumiidae,

=

and is

DDC

Within

are

there is

100%;

of

(BP

the two

3).

Fig.

approximately equally partitioned

sampled

and

EF-l œ.

contrast,

By

are more than threefold

non-synonymous

greater

genera Ceratocampinae

for DDC

However,

91%).

any

sampled. Similarly

strongly

no

changes

grouped

support

three subfamilies

than EF-l œ

also

et al.

for

the

(see

non-synonymous change

1998).

strong

among

Regier

Higher

grouping

weak resolu-

rates of

for DDC are also

17

3.

of maximumlikelihood

based on

topology

of total nucleotide

of EF-1 cr+

and families

Summary

Figure

analysis

sequences

subfamilies,

DDC from 64 taxa

tribes,

Brahmaeidae,

(Satumiidae,

Sphingidae).

among

Relationships

are shown with the

of

in each

listed in

after

name.

parentheses

in

consensus);

bootstrappercentages

number

terminal

the

only

species sampled

group

strict

MP

are listed

to

branches

above

(ML,

Bootstrappercentages

subtending

not

groups

ML MP

in the

or

The less

but more

likely,

(strictconsensus)

topology.

correspond

present

parentheses

of Hemileucinaeas sisterto

with

is

alsoshown.

whoserela-

groups

Satumiinae,

grouping

together

bootstrappercentages,

identify

parsimonious,

Solidlines

that are

BP values.Dashedlines

high 80%)

identifygroups

by

supported

<

The InL

the

fully

areconsidered

valuefor

resolved

is -25162.569.

topology

(BP 80%).

provisional

tionships

same three

was

of 909 nuc-

tion

obtained

leotides of the

the

subfamilies

The 64-taxon and 48-taxon

among

in an

relationships. -

ses

analy-

within

relationships

the

same

exploratory sampling

locus across 13

give essentially

Satumiinae. The two

African tribe Bunaeini are

from the

=

period

genera sampled

bombycoids

that included seven satumiids

the

et al.

Saturniinae

1998).

vis a

(BP

(Regier

strongly grouped

of

to the

all

exclusion of Satumiini.

However,

100%)

the

monophyly

Regarding

vis

the

of Bunaeini cannot be

includes a

sampling

3,

4)

Micragonini

1994)

present

definitively

Micragonini,

analysis (Fig.

monophyly

assessed until

inclusion

the

of

&

diversi-

and of

representatives

strongly

within Satumiinae

over

greater

supports

of bunaeine

ty

Urotini,

the former tribe

(Oberprieler

as a

Lemaire & Minet

Ndssig

satumiid sub-

genera,

its

to lie

within

segregation

separate

speculated

phylogenetically

1997).

thus

1999),

(Ludiinae;

family

(Oberprieler

a common African

for the

Our

corroborate a close rela-

origin

supporting

Micragonini

our

is,

analyses strongly

+

that

Bunaeini clade

of

between Attacini and

relative

This

1,

Satumiini,

=

(Tab.

is

Bunaeini,

3);

Fig.

tionship

to the other tribes

(BP

sampled

find-

of

89%).

one

exemplar

Micragonini

strongly placed

tribe

the

in

turn

sister

to the

latter

in

as

group

to Satumiinae

ing

argues strongly against

postulate

all

for

the

authors,

Saturniini,

assigned

our

amino

by

namely,

that the

1997)

sug-

Madagascan

more

both

=

and

of

nucleotides

94% under

92% )

(BP a

parsimony).

separat-

analyses

gestion

Antherina and

Ceranchia

genera

(Oberprieler

acids

With

be

(BP

may

related to the African tribe Bunaeini than

this

ed

Satumiinae is

inclusion,

very

closely

to other

strongly

from the other subfamilies sam-

=

which are

Oriental or

no

Satumiini

Satumiini,

(BP 100%)

mostly

provide

of

In

While additional

from

of

Palearctic.

our data

contrast,

sequences

Micragon-

and

strong

pled.

are

as well as

either

due

on the

postulate

ini

inclusion

Urotini

Lemaire

evidence

needed,

way

to alliance

Decachorda

& Minet

Satumiinae

as Decachordini

Rhodinia with

of

on the

or

(placed

1998),

by

paraphyly

its inclusion in Attacini

for

1989),

support

(Peigler

general

a

under

question

Ober-

&

of

for this tribe.

Thus,

ML

1994,

(Oberprieler

seems

Ndssig

strong.

monophyly

monophyletic

is

the

but

now

Satumiini

recovered

both

1997)

prieler

by

GTR+G+I and GTR-ssr

models,

(optimal)

with low

tribes in Saturniinae and their inter-

BP

Under

(17%)

of

Monophyly

support. parsimony,

18

+

48

boot-

ML-

of EF-1 cr DDC from

4. Maximumlikelihood

based on

of total nucleotide

analysis

sequences

MPstrict

branches

(ML,

topology

Figure

taxa

arelistedabove

not

consensus);

(Satumiidae

only).

subtending

in

present

Bootstrappercentages

MP

theMLor

in

to

(strictconsensus)

topology.

strappercentages parenthesescorrespond groups

below

nucleotide substitutions are shown

branches. The InL value is -

numbers

estimated

of

corresponding

are

with

bootstrappercentages,

alternative

of taxa from

summa-

18599.015.Four

Saturniini,

arrangements

together

'Actias refersto

arrangementsonly,

+

Actias +

Argema Opodiphthera.

rized

In thesealternative

that are

left).

group'

(upper

Dashed

rela-

whose

BP values.

lines

Solid lines

z 80%)

identifygroups

supportedby high

identifygroups

<

are considered

80%).

(BP

tionships

provisional

on

of Satur-

Satumiini are not

but

Lack of decisiveness

niini in our data reflects a

resolution of

monophyly

general

monophyly

monophyletic,

step beyond

lack

minimum

of

the

one

the

of

strong

gen-

requires only

3259

Four

to a

relationships among

deeper

supported

one of

steps.

weakly-to-modestly

in

are much

stronger

era

this

levels

support

alternatives

which

Satumiini,

Attacini,

tribe;

monophyletic

in Attacini

The reason for this difference

4).

with

are

Rhodinia

(Fig.

is not obvious. The maximal

groups

depicted

in

4

left).

pairwise divergences

Figure (upper

19

in

within Satumiini are

than those

within these three

the

are also

are

themselves

genera

slightly higher

tionships

ambiguous,

well

not

0.26 versus

but

so

Attacini,

mately

codon

dramatically

0.21,

EF- I a),

approxi-

at the third

across

divergences

although

(e.g.

respectively,

=

(BP 100%).

supported

and

for

position

the

basal

in

Attacini

for

within Saturniini. -

inter-

Overall,

strongly supported

0.15-0.21 at the third codon

split

(e.g.,

EF-la)

Relationships

within

Satumiini,

position

generic relationships

levels,

those in Attacini

particular-

than

recent

relatively

resolved.

than

are

those across

at

are less

actually larger

any well-sup-

loss

ly

strongly supported

deeper

within Saturniini.

of

Several

are

Thus,

4).

(Fig.

however,

ported split

due to

Satumiini cannot

signal

and saturation in

the lack of resolu-

greater divergence

easily explain

that tribe. An earlier molecular

divergences,

strongly

is 100% for the subtribe

Bootstrap support

'Actiens'of Bouvier

in

tion

&

see also

(1936;

of the

study

nucleotides

Oberprieler

and

yellow

468

taxon

of

Rhodinia),

Samia),

et

(Shimada

of the

four

2 of

al.1995),

using only

1994),

consisting

Ndssig

green

in

a

'moon moths' with tailed

in the

genera

arylphorin gene

genera

Actias,

sparse

sample -

Antheraea,

one

hindwings

and Graellsia. These three

of Saturniini

(2

Actias,

are also unusual

species

Argema,

genera

on resinous host-

2

1 of

of

and

in

Saturnia,

specializing

of Attacini

from

of

and out-

species

and

also

Sphingidae -

Australian

The

(2

genus

groups

1986).

plants (Peigler

Opodiph-

likewise feeds

resinous

so its

`moon moth'

thera

on

Bombycidae

plants,

failed to find

evidence on

of

monophyly

as the

our

data,

reasonable.

to the

sister

not

although

strong

placement

group

Satumiini.

clade

by

strongly supported,

An

alternative

is the near-simultane-

that

lineages,

implying

be

possibility

may

of basal

ous

Within the 'moon moth'

a sister

data

our

strong-

of the African

clade,

group relationship

divergence

will

be

sparse.

signal

this

generally

Arguing against

(BP 97%)

Antheraea and An-

support

ly

=

the

is

+

to Actias

which are Oriental

Graell.sia,

interpretation

strong pairing

Argema

of the two saturniine

in

In

and Holarctic

distribution.

Graellsia

turn,

genera,

included in

the aforementioned

as

the sister

is

isabellae

therina,

period

these

our current

strongly supported

species

from

et al.

In

locus

a

the latter

sampled

to

1998). contrast,

Actias,

(Regier

study

monophyletic

are never

three and

decisively

the taxon

by

genera

data

placed

species groups. Previously, Oberprieler

when

the

even

of

is reduced to

had

(1994)

synonymized

the

set,

sample

(result

Ndssig

monotypic

match that

That

not

with Actias because of evidence

Graellsia

sug-

shown).

period

deeper

plentiful

currently expanding

study

Satumiini

on

that Actias would

For

be rendered

short-tailed

green,

is,

signal

divergences

gesting

thereby

the

example,

if

be

the

is

used.

may actually

gene

for

right

paraphyletic.

are

We

the

A. luna more

Graellsia resembles

sample

period.

pale green

the

A.

which

isis,

However,

than does

brown and

has

our

yellow

Generic

tails

its

on

hindwings.

within Attarini. - Mono-

relationships

of Attacini is

straight

do

and

long,

results

for

this

syn-

not

the

corroborated

are

phyly

(BP

again strongly

support

have

necessity

therefore

=

and

a

maintained the tra-

we

100%),

from

generic relationships

onymy,

ditional

The

et

the

of Graellsia.

(Friedlander

unchanged

1998),

placement

previous study

usage

al.

with three

of

endemics Antherina and

First,

qualifications.

Madagascan

Ceranchia are

as sister

genera

to is

Loepa

hypothesis.

Epiphora,

unchanged

receives

our current

strongly supported

although

=

studies and

Their

across

close

relationship

methods,

(BP 100%).

analytical

support

a

but

from

novel,

considerably stronger

analysis

in either

only weakly supported,

with

=

ML

than under MP

one

our

(BP 93%)

analysis strong-

for the

analysis

Second,

either

Finally,

ly supports

exception,

(BP >_89%)

=

66% and

(BP

not

55%).

study

Archaeoattacus,

monophyly

large

by

in

as redefined

and diverse

Saturnia

strongly placed

to

genus

see

is now either sister

+

Michener

who

Attacus

(1952;

2),

study,

group

recovered

Fig.

The

Neori.s,

synonymized

con-

Coscinocera

lander et al.

Fried-

+

smaller

MP;

(weakly

1998)

by

major uncertainty

on which the evidence

eight genera.

or to Attacini -

recovered

cerns the

is

(Rothschildia

ML,

subgenus

as

BP

the

the 80%

indicated

4).

Eupackardia) (weakly

Fig.

conflicting,

sup-

fact

by

in

across

its inclusion

Saturnia

Third,

Samia, Callosamia,

with simi-

despite

in the

relationships

porting

that this

and

are

does not occur

optimal

which Neoris from

splits

unresolved,

Hyalophora

low levels of BP

again

support

+

placement

Copaxa,

Saturnia,

+

MP ML trees.

or

for Callosamia

rela-

larly

and

in

length

Samia

Callosamia

the other

has the

branch

longest

Hyalphora. Species

20

etic

of the nuclear

in

gene dopa decarboxylase

tribe and its

could be

spurious,

the

utility

placement

features of cocoon and larval

noctuoid moths

(Insecta:

Noctuoidea).

Lepidoptera:

morpholo-

its close rela-

although

Mol.

Farris, S.,

Evol. 15:473-486.

Phylog.

J.

and of adult

gy

wing pattern suggest

A. G. &

C.

Bult,

M.,

Källersjö,

Testingsignificance incongruence.

10:315-319.

Kluge,

to

Bouvier

1936,

Saturnia

1914,

(Packard

of

Cladis-

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Wolfe

(1995)

tics

Felsenstein,

Within

the

D'Abrera

Saturnia,

are both

1993,

1998).

J.

Confidence

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(1985)

phylogenies:

two

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strongly

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An

791.

the

Evolution39: 783-

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bootstrap.

and there is

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postu-

good

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W. T. M.

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The

New York

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Forbes,

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Lepidoptera of

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1,

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DOI: 10.1163/187631202X00028

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Article abstract of DOI:10.1163/187631202X00028

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