Phylogenomics of peacock spiders and their kin (Salticidae: Maratus), with implications for the evolution of male courtship displays
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Biological Journal of the Linnean Society, 2021, XX, 1–24. With 6 figures.
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Phylogenomics of peacock spiders and their kin
(Salticidae: Maratus), with implications for the evolution
of male courtship displays
MADELINE B. GIRARD1, DAMIAN O. ELIAS1,*, , GUILHERME AZEVEDO2, KE BI3,
MICHAEL M. KASUMOVIC4, , JULIANNE M. WALDOCK5, ERICA BREE ROSENBLUM1
and MARSHAL HEDIN2
1
Department of Environmental Science, Policy and Management, University of California, Berkeley,
Berkeley, CA 94720-3114, USA
2
Department of Biology, San Diego State University, San Diego, CA 92182-4614, USA
3
Museum of Vertebrate Zoology, University of California, Berkeley, Berkeley, CA 94720-3160, USA
4
Ecology & Evolution Research Centre, School of Biological, Earth & Environmental Sciences, UNSW,
Sydney, 2052, NSW, Australia
5
Collections and Research, Western Australian Museum, 49 Kew Street, Welshpool, 6106, Western
Australia, Australia
Received 7 July 2020; revised 16 September 2020; accepted for publication 22 September 2020
Understanding diversity has been a pursuit in evolutionary biology since its inception. A challenge arises when sexual
selection has played a role in diversification. Questions of what constitutes a ‘species’, homoplasy vs. synapomorphy,
and whether sexually selected traits show phylogenetic signal have hampered work on many systems. Peacock
spiders are famous for sexually selected male courtship dances and peacock-like abdominal ornamentation. This
lineage of jumping spiders currently includes over 90 species classified into two genera, Maratus and Saratus. Most
Maratus species have been placed into groups based on secondary sexual characters, but evolutionary relationships
remain unresolved. Here we assess relationships in peacock spiders using phylogenomic data (ultraconserved
elements and RAD-sequencing). Analyses reveal that Maratus and the related genus Saitis are paraphyletic. Many,
but not all, morphological groups within a ‘core Maratus’ clade are recovered as genetic clades but we find evidence
for undocumented speciation. Based on original observations of male courtship, our comparative analyses suggest
that courtship behaviour and peacock-like abdominal ornamentation have evolved sequentially, with some traits
inherited from ancestors and others evolving repeatedly and independently from ‘simple’ forms. Our results have
important implications for the taxonomy of these spiders, and provide a much-needed evolutionary framework for
comparative studies of the evolution of sexual signal characters.
ADDITIONAL KEYWORDS: Australia – character evolution – multimodal signals – rapid evolution – sexual
selection – taxonomy – ultraconserved elements.
INTRODUCTION emerged from many of these studies is that sexually
selected traits sometimes show low phylogenetic signal
Sexual selection has driven the evolution of a
(Kusmierski et al., 1997; Gleason & Ritchie, 1998;
spectacular diversity of colours, sounds, smells, shapes
Ohmer et al., 2009; Puniamoorthy et al., 2009; Hosner
and forms across the animal kingdom, from darters
& Moyle, 2012; Hebets et al., 2013; Owens et al., 2020),
(Hulse et al., 2020) to bower birds (Frith & Frith, 2004)
often in the traits that are the most obvious to human
to mantis shrimps (Porter et al., 2010) to treehoppers
(Rodriguez et al., 2004). One common theme that has observers (i.e. bower bird plumage and bowers, hind
wings in swallowtails, tyrant flycatcher plumage,
frog calls, etc.). It thus follows that for comparative
*Corresponding author. E-mail: doelias@berkeley.edu studies, particularly those focused on the evolution
© 2021 The Linnean Society of London, Biological Journal of the Linnean Society, 2021, XX, 1–24 1
2 M. B. GIRARD ET AL.
of potentially homoplastic sexually selected traits, 2020). Sexual dimorphism is profound in this group,
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species hypotheses and phylogenetic relationships with cryptic adult females contrasting with highly
should be based on independent evidence (e.g. decorated adult males. Male Maratus are now famous
phylogenomic data). for their elaborate visual and vibrational courtship
Peacock spiders of the genus Maratus Karsch displays (Girard et al., 2011; Otto & Hill, 2011; Girard
1878 comprise a diverse clade of jumping spiders & Endler, 2014; Girard et al., 2018; Otto & Hill, 2019a;
(Fig. 1; Supporting Information, Fig. S1) distributed Wilts et al., 2020), facilitated by conspicuous behaviour
predominately in eastern and western Australia, with in which they raise their often colourful abdomens
over 90 described species (Otto & Hill, 2019a; World vertically and along with elongated, brush-adorned,
Spider Catalog, 2020) and active on-going species third legs perform a vigorous courtship display. Some
discovery (e.g. Schubert & Whyte, 2019; Schubert, male peacock spiders have cuticular flaps that wrap
Figure 1. Peacock spiders in courtship posture and variation in fan morphology. A, Maratus sarahae – elliptical fan. B,
M. calcitrans – no fan. C, M. mungaich – elliptical fan. D, M. pardus – elliptical fan. E, M. amabilis – elliptical fan. F,
M. vespa –posterior lobed fan. G, Saratus hesperus – no fan. H, M. flavus – no fan. I, M. rainbowi – round fan. J, M. plumosus
– posterior lobed fan. K, M. harrisi – lobed fan. L, M. anomalus – no fan. M, M. jactatus – round fan. N, M. volans – elliptical
fan. O, M. linnaei – no fan. P, M. clupeatus – elliptical fan. Q, M. cinerus – no fan. R, M. personatus – no fan. S, M. elephans
– elliptical fan. T, M. madelineae – elliptical fan. U, M. proszynskii – no fan. V, M. australis – lobed fan. W, M. spicatus – no
fan. X, M. vespertilio – lobed fan. Y, M. pavonis – round fan. Z, M. speciosus – no fan. Latitude and longitude data for all
specimens can be found in Supporting Information, Table S1.
© 2021 The Linnean Society of London, Biological Journal of the Linnean Society, 2021, XX, 1–24
PEACOCK SPIDER PHYLOGENOMICS 3
beneath the abdomen (Dunn, 1947; Waldock, 1993; The genus Maratus included fewer than ten
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Hill, 2009), unfurled during the male courtship display, described species in 2008 (Waldock, 2008). The
revealing remarkable patterns of scale pigmentation description of new Maratus species has exploded over
and structural colours (Stavenga et al., 2016; Hsiung recent years, as interest in these spiders has increased,
et al., 2017, 2019; Wilts et al., 2020; Fig. 1; Fig. S1; Table and macrophotography and video of live specimens
S1). The extensive radiation of peacock spider courtship has facilitated the discovery of complex male courtship
ornaments and displays is perhaps comparable to ornaments and behaviours (Otto & Hill, 2019a;
that of the better-known birds of paradise (Cooper & Schubert, 2020). Most peacock spider species have
Forshaw, 1977; Nunn & Cracraft, 1996; Ligon et al., been placed into complexes of related species (species
2018), but these spiders have remained obscure groups) based on male morphology and/or display
until recently because of their small size. Despite characters (Otto & Hill, 2019a). As of early 2020,
tremendous sexual signal diversity seen in this group, 16 species groups have been recognized (Table 1).
the evolutionary patterns and processes of ornament Relationships within and among groups have not been
evolution remain mostly unstudied due to a lack of independently tested using other types of evidence,
formal phylogenetic study and resolution. and resolution within these groups is mostly lacking.
Previous molecular systematics research has placed Some species are not currently placed into species
Maratus within a clade of mostly Australasian salticids groups (Table 1) and it remains unclear whether these
in the Saitis Clade (Fig. 2A; Zhang & Maddison, 2013), taxa are truly phylogenetically isolated, or are nested
within the tribe Euophryini (Maddison, 2015; Zhang & cryptically within described groups. Also, some species
Maddison, 2015). Zhang & Maddison (2015) considered appear to share morphological features of multiple
the multiple genera in the Saitis Clade (Maratus, groups (Schubert, 2020), blurring species group limits.
Saratus Otto & Hill 2017a, Hypoblemum Peckham & Similar to arguments above for generic-level diagnoses
Peckham 1885, Jotus Koch 1881, Saitis Simon 1876, and because species groups are defined mostly on male
etc.) as so closely related as to be possibly congeneric. courtship morphologies and behaviours (but see Baehr
Conversely, other authors have maintained the above & Whyte, 2016), current phylogenetic hypotheses do
taxa as separate genera (Otto & Hill, 2012a, 2019a; not present an independent framework to understand
Otto & Hill, 2017a; Prószyński et al., 2018; Baehr potential homoplasy in these same character systems
et al., 2019). A hypothesized sub-lineage in this clade (Maddison & Hedin, 2003).
includes the ‘Maratus group’ (Otto & Hill, 2012a, Of the now more than 90 accepted species in Maratus,
2019a; Otto et al., 2019), including Maratus, Saratus, no species hypotheses have been independently tested
‘Lycidas’ (now mostly synonymized with Maratus) and using molecular evidence. There are many species
Hypoblemum. Otto et al. (2019) suggested that the two known only from single (type) locations, and these could
species of Hypoblemum are sister to peacock spiders, represent divergent populations of otherwise known
including Maratus and Saratus (Fig. 2B). Saratus is species (i.e. geographical variation). Some ‘varieties’
monotypic, with male and female genitalia that differ have been described within wide-ranging species [e.g.
from Maratus, but with somatic morphology, male M. pavonis (Dunn 1947); Hill & Otto, 2011; Otto &
coloration and male display behaviours otherwise Hill, 2012a; Baehr & Whyte, 2016], possibly indicating
similar to the latter (Otto & Hill, 2017a). undetected speciation. Some of these wide-ranging
Except for the Sanger-sequencing-based studies species have conspicuously disjunct distributions
of Zhang & Maddison (2013, 2015), relationships with populations along the eastern seaboard and in
within the Saitis Clade have never been tested via south-western Australia, but populations are absent
phylogenetic analysis, although authors have inferred from the more xeric temperate arid zone [e.g. eastern
relationships based on character argumentation vs. western M. pavonis, M. vespertilio (Simon 1901)].
(Fig. 2B). A unique aspect of generic-level diagnoses Finally, researchers have discussed possible evidence
in this clade is that many are based mostly or entirely for gene flow across species boundaries (Otto & Hill,
on male courtship morphology and behaviour (Otto & 2014b, 2016a; Schubert, 2020), again blurring species
Hill, 2012a, 2017a; Prószyński et al., 2018; Baehr et al., limits, and testable using genetic data.
2019; Otto et al., 2019), rather than the traditional We collected ultraconserved element (UCE) and
(for salticid systematics) male and/or female genitalic RAD-sequencing (RAD-seq) phylogenomic data to
morphology (see Zhang & Maddison, 2015). It address relationships in the Saitis Clade, emphasizing
remains to be seen whether independent evidence Maratus relationships. Our sample includes type
(e.g. molecular evidence) supports taxa based on male species for core genera [Jotus auripes L. Koch 1881,
courtship characters, or rather, if such characters Saitis barbipes (Simon 1868), Hypoblemum griseum
might be subject to higher levels of homoplasy, calling (Keyserling 1882), ‘Lycidas’ anomalus (=Maratus
into question current generic-level diagnostic traits anomalus) (Karsch 1878), Saratus hesperus Otto &
(see also Zhang & Maddison, 2015). Hill 2017a, and Maratus amabilis (Karsch 1878)]. For
© 2021 The Linnean Society of London, Biological Journal of the Linnean Society, 2021, XX, 1–24
4 M. B. GIRARD ET AL.
Maileus cf. fuscus (Malaysia)_JXZ098 A) Sanger sequence data -
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Saitis barbipes (SPAIN)_JXZ147 Zhang & Maddison 2013, fig 1
cf Saitis sp. (WA)_JXZ091 Saitis mutans
Maratus rainbowi (WA)_JXZ160
Maratus sp. (soAU)_JXZ159
Lycidas cf. karschi (NSW)_JXZ157 Hypoblemmum scutulatum
Hypoblemum cf. albovittatum (soAU)_JXZ164 Hypoblemmum scutulatum?
Hypoblemmum sp. (NSW)_JXZ085 Hypoblemmum scutulatum
Lycidas cf. griseus (NSW)_JXZ092 Hypoblemmum griseum?
Lycidas cf. vittatus (QL)_JXZ146
XZ1
uncertain
cf. Jotus sp. (QL)_JXZ155
Prostheclina sp. (NSW)__JXZ095
Jotus auripes (QL)_JXZ090
Servaea vestita
B) Compilation - multiple sources, see legend
Saitis
Saitis
(5 species, S. barbipes type)
image
ima
iim
m
maage
male leg III long, w/ “bottle brush”
slender unmodifed leg I
flexibile pedicel
MF elevation of abdomen
Hypoblemum scutulatum
Hypoblemum griseum
heavy outer embolar ring
image
iima
im
m
maage
“Maratus Hypoblemum
group”
??
Maratus
M abdomen with image
iima
im
m
maage
dorsal plate or scute
Barraina
(monotypic, B. anfracta type) M palp, Saratus hesperus
F epigynum unique
Maileus
(monotypic, M. fuscus type) Maratus
Jotus
(8 species, Jotus
J. auripes type)
carapace with lateral stripes image
ima
iim
m
ma
mag
age
abdomen with lateral bands
male leg I setal brush
combined conductor +
embolus of male palp
metatarsus leg I fringe in males
Prostheclina
(7 species, P. pallida type)
Figure 2. Overview of the phylogenetic structure of the Saitis clade. A, results of Zhang & Maddison (2013: fig. 1), based
on maximum-likelihood analysis of four concatenated genes. Annotation of uncertain taxon names reflects UCE by-catch
phylogenetic results (see Supporting Information, Fig. S3). B, hypothesized phylogenetic structure of the Saitis Clade from
multiple sources (Richardson & Żabka, 2007; Otto & Hill, 2012a, 2016a, 2017a; Prószyński et al., 2018; Otto et al., 2019;
Baehr et al., 2019), with partial list of supporting characters for genera and generic groupings.
© 2021 The Linnean Society of London, Biological Journal of the Linnean Society, 2021, XX, 1–24
Table 1. Maratus species groups, following Otto and Hill (2019a), and results presented here
Groups (no. of species) Otto & Hill (2019a) Otto & Hill (2019a) species (taxa Taxa sampled in this study RAD results
characters; see also Baehr & described after 2019 denoted as such)
Whyte (2016)
fimbriatus group (4) Wheel-rim-shaped embolus; M. fimbriatus Otto & Hill 2016a, M. licunxini M. speculifer, M. ‘flame’, M. ‘carmel’
legs I extended in courtship Baehr & Whyte 2016, M. speculifer (Simon
1909), M. volpei Schubert 2020
chrysomelas group (3) Apex of the embolus with con- M. chrysomelas (Simon 1909), M. kiwirrkurra M. chrysomelas, M. nigromaculatus
vergent inner and outer Baehr & Whyte 2016, M. nigromaculatus
edge (Keyserling 1883)
spicatus group (4) Conical proximal tegulum, M. nimbus Otto & Hill 2017b, M. purcellae M. nimbus, M. purcellae, M. robinsoni, spicatus group intermixed
etc.; probably related to Otto & Hill 2013a, M. robinsoni Otto & Hill M. spicatus with chrysomelas group
chrysomelas group 2012a, M. spicatus Otto & Hill 2012a
pavonis group (8) Target on fan, sometimes ob- M. leo Otto & Hill 2014b, M. literatus Otto & M. cf. leo, M. leo, M. literatus,
scured; zigzag ridges on em- Hill 2014b, M. maritimus Otto & Hill 2014b, M. maritimus, M. pavonis,
bolic disc M. montanus Otto & Hill 2014b, M. pavonis M. rainbowi, M. watagansi
(Dunn 1947), M. rainbowi (Roewer
1951), M. sylvestris Otto & Hill 2019c,
M. watagansi Otto & Hill 2013b
anomalus group (11) Bifurcated apex of the outer M. albus Otto & Hill 2016b, M. anomalus M. albus, M. anomalus, M. aurantius, Includes M. sceletus from
ring of embolus (Karsch 1878), M. aurantius Otto & Hill M. cinereus, M. michaelorum, calcitrans group
2017a, M. cinereus Otto & Hill 2017a, M. neptunus, M. cf. neptunus,
M. inaquosus Schubert 2020, M. julianneae M. vultus
Baehr & Whyte 2016, M. kochi (Żabka
1987), M. lentus Otto & Hill 2017a,
M. michaelorum Baehr & Whyte 2016,
M. neptunus Otto & Hill 2017a, M. vultus
Otto & Hill 2016b
vespertilio group (2) M. sapphirus Otto & Hill 2017b, M. vespertilio M. vespertilio
(Simon 1901)
velutinus group (2) Squamous setae on abdomen M. proszynskii Waldock 2015, M. velutinus M. proszynskii, M. velutinus
Otto & Hill 2012a
harrisi group (3) Lobate flaps on fan M. constellatus Schubert 2020, M. harrisi Otto M. harrisi
© 2021 The Linnean Society of London, Biological Journal of the Linnean Society, 2021, XX, 1–24
& Hill 2011, M. lobatus Otto & Hill 2016b
calcitrans group (7) Males with asymmetric dis- M. calcitrans Otto & Hill 2012b, M. digitatus M. calcitrans, M. digitatus, M. eliasi, M. plumosus, M. cf. plumosus
play, most inflating spin- Otto & Hill 2012b, M. eliasi Baehr & Whyte M. jactatus, M. ottoi, M. plumosus, near tasmanicus group,
nerets 2016, M. jactatus Otto & Hill 2015a, M. ottoi M. cf. plumosus, M. sceletus M. sceletus in anomalus
Baehr & Whyte 2016, M. plumosus Otto & group
Hill 2013b, M. sceletus Otto & Hill 2015a
volans group (3) Large fringed fans with dis- M. elephans Otto & Hill 2015b, M. pardus M. elephans, M. pardus, M. volans M. pardus in expanded
tinctive figures Otto & Hill 2014c, M. volans (O. Pickard- mungaich (*from
Cambridge 1874) south-west Australia)
PEACOCK SPIDER PHYLOGENOMICS
5
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Table 1. Continued
Groups (no. of species) Otto & Hill (2019a) Otto & Hill (2019a) species (taxa Taxa sampled in this study RAD results
characters; see also Baehr & described after 2019 denoted as such)
Whyte (2016)
tasmanicus group (3) Triangular fan with lobes, M. australis Otto & Hill 2016b, M occasus M. australis, M. tasmanicus M. australis in expanded
black spots Schubert 2019b, M. tasmanicus Otto & Hill mungaich (*from
2013b south-west Australia)
mungaich group (10) Wide fans with red scales; M. avibus Otto & Hill 2014a, M. bubo Otto M. avibus, M. caeruleus, M. madelineae, Not monophyletic, part of ex-
south-west Australia & Hill 2016b, M. caeruleus Waldock 2013, M. mungaich, M. sarahae panded mungaich
M. B. GIRARD ET AL.
M. gemmifer Otto & Hill 2017c, M. hortorum
Waldock 2014, M. karrie Waldock 2013,
M. madelineae Waldock 2014, M. melindae
Waldock 2013, M. mungaich Waldock 1995,
M. sarahae Waldock 2013
flavus group (4) south-west Australia endemic, M. boranup Otto & Hill 2018a, M. felinus M. flavus Part of expanded mungaich
related to linnaei and vespa Schubert 2019, M. flavus Otto & Hill 2018a,
groups? M. suae Schubert 2020
linnaei group (4) Rotating abdomen during M. cuspis Otto & Hill 2019c, M. electricus Otto M. linnaei Part of expanded mungaich
courtship; south-west Aus- & Hill 2017c, M. laurenae Schubert 2020,
tralia; related to vespa M. linnaei Waldock 2008
group?
personatus group (2) south-west Australia M. banyowla Otto & Hill 2019b, M. personatus M. personatus Part of expanded mungaich
Otto & Hill 2015c
vespa group (10) Lobate flaps with lateral orna- M. aquilus Schubert 2019, M. azureus Schu- M. vespa Part of expanded mungaich
ments; south-west Australia, bert 2020, M. combustus Schubert 2019,
related to linnaei group? M. cristatus Otto & Hill 2017c, M. icarus
Otto & Hill 2019c, M. noggerup Schu-
bert 2020, M. tortus Otto & Hill 2018b,
M. unicup Otto & Hill 2018b, M. vespa Otto
& Hill 2016b
Taxa not placed into spe- M. amabilis (Karsch 1878), M. clupeatus Otto M. amabilis, M. clupeatus, M. speciosus M. clupeatus in ex-
cies groups (6) & Hill 2014d, M. sagittus Schubert & Whyte panded mugaich (*from
2019, M. speciosus (O. Pickard-Cambridge, south-west Australia)
1874), M. tessellatus (south-west Australia)
Otto & Hill 2016b, M. trigonus (south-west
Australia) Otto & Hill 2017c
© 2021 The Linnean Society of London, Biological Journal of the Linnean Society, 2021, XX, 1–24
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peacock spiders (Maratus and Saratus), we generated (Supporting Information, Table S1). We also included
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data for a representative sample of species, including the more divergent genus Servaea Simon 1888 to root
species from all 16 currently recognized species phylogenies (data shared by W. Maddison), following
groups (Table 1). Our phylogenomic results reveal the results of Zhang & Maddison (2013, 2015).
that both Saitis and Maratus, as currently recognized, We lacked samples of Barraina Richardson 2013,
are paraphyletic taxa. Most described species are Maileus Peckham & Peckham 1907 and Prostheclina
recovered as exclusive genetic groupings, but we find Keyserling 1882, all hypothesized members of the
evidence for undetected speciation and we present Saitis Clade (Zhang & Maddison (2013, 2015). We used
data for three probable new species. Phylogenetic the MYbaits Spider v.1 kit (Arbor Biosciences, Ann
comparative analysis of male visual and vibratory Arbor, MI, USA; Kulkarni et al., 2020) to capture UCE
signals reveals a combination of synapomorphic and loci, using methods of library preparation as in recent
homoplastic signal. Overall, our research establishes a publications (Starrett et al., 2017; Hedin et al., 2019).
phylogenetic framework for future studies of peacock Sequencing was conducted on an Illumina HiSeq400 at
spider morphology, behaviour and evolution. the U.C. Davis Genome Center with 150-bp paired-end
reads. Sequence reads were trimmed and assembled
using TRINITY v.2.1.1. (Grabherr et al., 2011)
using default settings (trimmomatic = full_cleanup,
MATERIALS AND METHODS
kmer = 25), then processed using Phyluce (Faircloth,
For a majority of specimens, genomic DNA was 2016). Assembled contigs were matched to probes
extracted from legs or whole spiders using a QIAamp using standard minimum coverage and minimum
DNA mini kit (Qiagen, Valencia, CA, USA). Saitis identity values (80_80). UCE loci were aligned with
barbipes samples used in UCE experiments were MAFFT (Katoh & Standley, 2013) and trimmed with
extracted using a Qiagen DNeasy blood and tissue kit. Gblocks (Castresana, 2000; Talavera & Castresana,
2007) using strict settings (--b1 0.5 --b2 0.85 --b3 4
--b4 8). Resulting Phyluce alignments with at least
Taxonomic correction 70% sample occupancy were imported into Geneious
The name Maratus rainbowi (Roewer, 1951) is used 11.0.4 (Biomatters, Auckland, New Zealand), where all
in this study, in accordance with the World Spider individual alignments were visually inspected.
Catalogue (2020), replacing the use of Maratus Maximum-likelihood (ML) phylogenetic analyses
splendens common in other catalogues (Otto & Hill, were conducted using IQ-TREE v.2.0-rc2 (Nguyen
2019a). The name Attus splendens was assigned et al., 2015). Initial partitions corresponded to
to a North American salticid species by Peckham individual loci, and ModelFinder (Kalyaanamoorthy
& Peckham (1883), which is now considered to be a et al., 2017) was then used to find best-fit models
junior synonym of Habronattus decorus (Blackwall, and merge partitions (-s -p -m TESTMERGE
1846). Rainbow (1896) also established a species -rcluster 10); the relaxed hierarchical clustering
that he called Attus splendens for a new species from algorithm (Lanfear et al., 2014) was used to reduce
Australia, overlooking the prior usage by Peckham & computational burden. Support was assessed via 1000
Peckham (1883). Attus splendens Rainbow, 1896 was ultrafast bootstrap replicates (Hoang et al., 2018).
transferred to Saitis by Simon (1901) and to Maratus An SVDQuartets analysis (Chifman & Kubatko,
by Żabka (1991). In the meantime, Roewer (1951) 2014; Chifman & Kubatko, 2015) was conducted on
recognized that Attus splendens Peckham & Peckham, a concatenated matrix using PAUP* 4.0a (Swofford,
1883 and Attus splendens Rainbow, 1896 were primary 1988), implementing the multispecies coalescent tree
homonyms, and provided the replacement name model with exhaustive quartets sampling and 1000
Saitis rainbowi Roewer, 1951. With the transfer of all bootstrap replicates.
Australian species of Saitis to Maratus, this species is Using IQ-TREE 2 we calculated gene (gCF) and
correctly known as Maratus rainbowi (Roewer, 1951). site concordance (sCF) factors. For every branch of a
This usage is in accordance with the International reference tree, gCF can be defined as the percentage of
Code of Zoological Nomenclature (1999). ‘decisive’ gene trees containing that branch, while sCF
can be defined as the percentage of decisive sites (in
an alignment) supporting a branch (Minh et al., 2020).
UCE data The latter support metric is particularly useful when
Specimens of Jotus auripes, Saitis barbipes, Saitis individual gene trees are uncertain. Because ML and
mutans Otto & Hill 2012a, Saitis virgatus Otto & Hill SVDQuartets UCE tree topologies agreed (see below),
2012a, Hypoblemum griseum, H. scutulatum L. Koch we used the partitioned ML tree as a reference for CF
1881, Saratus hesperus, and a subsample of seven calculations, with individual gene trees calculated as
Maratus species were included in UCE experiments default in IQ-TREE 2 (-S command).
© 2021 The Linnean Society of London, Biological Journal of the Linnean Society, 2021, XX, 1–248 M. B. GIRARD ET AL.
UCE by-catch taxa (rather than all taxa), this enabled us to extract
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We assembled ribosomal 28S and mitochondrial more loci per clade.
16S_ND1 ‘by-catch’ using standard BLAST searches
in Geneious 11.0.4 (Biomatters). Specifically, we
RAD analysis – all taxa
used published sequences to query UCE TRINITY
assemblies using blastn (maximum e-value of 1e-5). A custom script was used to process RAD data (available
We then stitched together UCE by-catch data with at https://github.com/CGRL-QB3-UCBerkeley/RAD),
published sequences from Zhang & Maddison (2013, calling various external programs. This pipeline has
2015), aligned matrices manually, and conducted been used in several other recent publications (Krohn
ML analyses using IQ-TREE 2 (-s, -B 1000, -bnni). et al., 2018; Maas et al., 2018; Klonoski et al., 2019).
Our primary interest here was to potentially resolve Exact duplicates were removed using Super Deduper
uncertain taxon identities from Zhang & Maddison (https://github.com/dstreett/Super-Deduper). Raw
(2013) (e.g. see Fig. 2A), while also including Maileus reads were filtered using Cutadapt (Martin 2011)
and Prostheclina, not sampled in other matrices. and Trimmomatic (Bolger et al., 2014) to trim adapter
contaminations and low-quality reads. The resulting
cleaned forward reads of each individual were first
clustered using Cd-hit (Li & Godzik, 2006; Fu et al.,
RAD data collection 2012), and only clusters with at least two reads
Specimens of Jotus auripes, Saitis mutans, S. supported were kept. To remove potential paralogues,
virgatus, Hypoblemum griseum, H. scutulatum and Blastn (Altschul et al., 1990) was used to compare
Saratus hesperus were included as ‘outgroup taxa’ clustered loci against themselves, and remove any
(Supporting Information, Table S1). Lacking Servaea locus that matched to a locus other than itself. The
RAD data, we used UCE topology results to root RAD resulting RAD loci from each individual were then
trees. Specimens representing 48 described Maratus combined for all individuals and the resulting marker
species were included, based on 2013–2015 collections sets served as a master reference. We then used Blastn
from eastern and western Australia (ACT, NSW, QLD, (Altschul et al., 1990) to compare markers from each
SA, TAS, VIC, WA; Table 1; Appendix S1). One to three individual to the master reference and only kept
individuals per species were sampled per location, and those that had unique hits. These uniquely matched
for more broadly distributed species we attempted to markers from each individual served as a reference
sample populations spanning known geographical for that individual. Cleaned sequence reads from each
ranges. Our taxon sample included five novel forms individual were aligned to its own reference using
(Maratus cf. neptunus, M. ‘carmel’, M. ‘flame’, M. cf. Novoalign (http://www.novocraft.com) and reads that
leo, M. cf. plumosus), representing potentially new mapped uniquely to the reference were kept. Picard
species. (http://picard.sourceforge.net) was used to add read
We generated RAD libraries using the protocol of groups and GATK (McKenna et al., 2010) to perform
Ali et al. (2016) without completing the targeted bait realignment around indels. SAMtools/BCFtools and
capture step and using Pippin Prep (Sage Science, ‘vcfutils.pl vcf2fq’ implemented in SAMTools (Li et al.,
Beverley, MA, USA) instead of beads to size-select 2009) were used to generate individual consensus
fragments between 250 and 600 bp. Each of two sequences by calling genotypes and incorporating
96-sample libraries was sequenced on an Illumina ambiguous sites. We kept a consensus base only when
HiSeq 4000 lane at the U.C. Davis Genome Center its depth was at least 3× or above and retained loci
with 150-bp paired-end reads. Raw fastq reads were that contained no more than 80% missing data. We
de-multiplexed allowing one barcode mismatch. also masked sites within 5-bp windows around indels.
De-multiplexed reads were removed if the expected We converted the resulting consensus fastq sequence
cut site (also one mismatch allowed) was not found at file to fasta format using Seqtk (https://github.com/
the beginning of the 5′-end of sequences. lh3/seqtk). Using MAFFT (Katoh & Standley, 2013),
Two separate sets of analytical pipelines and final filtered loci from each individual were aligned by
downstream analyses were conducted on RAD data: comparing against the master reference. Ambiguously
(1) a custom pipeline was used to extract a set of loci aligned regions were then trimmed using Trimal
conserved across all taxa – this locus set was used to (Capella-Gutierrez et al., 2009). To avoid excess
reconstruct a phylogeny for Maratus and ‘outgroups’; missing data, we removed alignments if more than
(2) based on the ‘all taxa’ RAD results, the ipyrad 90% missing data were present in at least 30% of
pipeline (Eaton et al., 2017) was used to assemble samples.
locus sets for well-supported species groups (or clades Using a subsample of the entire available sample
comprising multiple species groups). Because ipyrad (152 specimens), we performed several all-taxa
analyses were focused on more closely related sets of phylogenomic analyses. Species trees were inferred
© 2021 The Linnean Society of London, Biological Journal of the Linnean Society, 2021, XX, 1–24PEACOCK SPIDER PHYLOGENOMICS 9
using the multispecies coalescent model implemented S2. Character scorings for Saitis barbipes were
Downloaded from https://academic.oup.com/biolinnean/advance-article/doi/10.1093/biolinnean/blaa165/6126965 by University of California, Berkeley user on 05 February 2021
in SVDQuartets. Originally developed for unlinked extracted from other sources (Hill, 2009; https://
single nucleotide polymorphism (SNP) data, where www.youtube.com/watch?v=wQT2bHTOdwo); some
each site has an independent genealogy drawn from scorings for Hypoblemum scutulatum were taken from
a coalescent model, the method has also been shown the courtship description of Otto et al. (2019). The
to perform well for linked SNP data (Chifman & characters presented here are not intended to capture
Kubatko, 2014), as used here. SVDQuartets was the full diversity or complexity of courtship characters
used to construct a ‘lineage’ tree, relating individual seen within Maratus, but rather represent a set of
sequences. We also used a taxon partition to assign scoreable traits with hypothesized deeper homology
individual samples to species (partition = species); for across all genera included.
M. pavonis, which includes multiple well-supported
sub-lineages (see Results below), each sub-lineage
here was treated as a ‘species’. For both lineage and Character evolution
partitioned analyses we evaluated all possible quartets For tree-based character evolution analyses we used
(evalq = all, quartet assembly algorithm = QFM), pruned UCE and all-taxa RAD matrices, removing
and conducted a non-parametric bootstrap analysis, duplicate specimens/populations for individual
resampling with replacement both loci and sites species, while retaining the most data-rich samples
within loci (bootstrap = multilocus, loci = combined, (summarized using AMAS; Borowiec, 2016). We then
nreps = 100). IQ-TREE was also used to reconstruct conducted partitioned and unpartitioned ML analyses
an ML tree from a concatenated matrix, with a single of these matrices, as outlined above. Mesquite v.3.6
best-fit model automatically chosen by ModelFinder, (build 917) (Maddison & Maddison, 2018) was used to
and support assessed via 1000 ultrafast bootstrap reconstruct ancestral states for courtship phenotypes,
replicates (-s, -B 1000, -bnni). scored as alternative discrete states (Supporting
Information, Table S3). ML character reconstructions
were conducted using the one-parameter Markov
RAD analysis – species groups k-state model (Lewis, 2001).
Based on RAD phylogenomic results from all-
taxa analyses (see below), additional analyses
were conducted for members of four well-
RESULTS
supported clades (FC = fimbriatus + chrysomelas,
pavonis, anomalus, MTVC = expanded Voucher specimen data and relevant summary values
mungaich + tasmanicus + volans + calcitrans). Here, are presented in Supporting Information, Table S1.
forward read (R1) RAD data were de novo assembled Raw UCE and RAD data have been submitted to the
using ipyrad v.0.7.30 (Eaton & Overcast, 2016), with Short Read Archive (BioProjects PRJNA667490 and
the following settings adjusted from default: max_ PRJNA665271), and all data matrices and resulting
Indels_locus = 4, clust_threshold = 0.85 (within and tree files referenced below are available at Dryad
across), min_samples_locus = 5; see Eaton et al., 2017). (https://doi.org/10.5061/dryad.9p8cz8wdp).
We used a phylogenetic invariants analysis (Lake,
1987; Chifman & Kubatko, 2015) on linked SNPs to
infer quartet trees and a species tree using tetrad UCE data
v.0.7.30, part of the ipyrad.analysis toolkit, sampling The final primary 70% occupancy UCE matrix
all quartets with 100 bootstrap replicates. included 472 loci, with a combined alignment length
of 282 674 bp and 39 224 parsimony-informative sites.
ML and SVDquartets tree topologies were identical,
Courtship characters and both were well supported, with almost all nodes
Adult male visual and vibrational courtship signals receiving maximum support (Fig. 3; Supporting
were characterized using video and laser vibrometer Information, Fig. S2). Both gene (gCF) and site
recordings (see Girard et al., 2011 for a detailed concordance (sCF) factors were relatively high, except
description of methods). From these recordings we for a node uniting Saitis mutans with ‘Maratus
scored nine characters for all taxa, based on original group’ genera.
observations: (1) lateral fan flaps, (2) overall fan shape, The genus Saitis is not recovered as monophyletic
(3) raises fan, (4) third leg use, (5) white brushes, (6) (Fig. 3). Saitis virgatus is sister to Jotus auripes,
elongated spinneret display, (7) vibratory display, (8) possibly misplaced in Saitis. Saitis mutans and Saitis
vigorous tapping and (9) pre-mount display. Detailed barbipes are also not recovered together; instead they
character descriptions and scorings can be found are found to be successive sister species to ‘Maratus
in Supporting Information, Appendix S1 and Table group’ taxa (Hypoblemum, Maratus, Saratus; Fig. 2B).
© 2021 The Linnean Society of London, Biological Journal of the Linnean Society, 2021, XX, 1–2410 M. B. GIRARD ET AL.
Saitis_virgatus_160
98.8/98.4
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Saitis_virgatus_159
79.2/59.4
Jotus_auripes_162
98.5/97.2
Jotus_auripes_161
Saitis_barbipes_SDSUG3313
99.7/98.5
Saitis_barbipes_SDSUG3314
gCF/sCF
Saitis_mutans_178
ML/SVD bootstraps 99.4/98.7
(100 if not shown)
Saitis_mutans_180
73.6/57.5
Hypoblemum_scutulatum_166
93.9/84.5
Hypoblemum_griseum_d529
Hypoblemum
86.4/86.8 Hypoblemum_griseum_168 Clade
32.6/35 53.8/50.5
55.1/43
Hypoblemum_griseum_167
95/96
M_chrysomelas_24c
98.3/89.3
Maratus_flame_18c
58.1/52.8 Maratus_rainbowi_84
83.4/81.5
Maratus_western_pavonis_44
Saratus_52c
89.6/94.9
67/67.1
Saratus_51c
‘core Maratus’
45.4/45.8
0.02 Saratus_145
53.8/53.9
Maratus_volans_58c
65.4/74.6 Maratus_cf_plumosus_41c
26.9/41.2
Maratus_madelineae_120
85/80
Servaea_inanca_JXZ093
Figure 3. UCE maximum-likelihood phylogeny, with concordance factor (CF) values. All bootstrap values = 100, unless
otherwise indicated. Bootstrap values from SVDQuartets analysis are also shown for nodes with values 7000 bp); these were trimmed to match the Sanger
to this entire clade, also recovered in RAD analyses data. Based on phylogenetic placement in gene trees,
(see below), as the Hypoblemum clade. A second we tentatively resolve the identity of five samples
primary branch includes other Maratus species, with from Zhang & Maddison (2013, 2015) (see Fig. 2A;
Saratus nested within this larger clade. We hereafter Supporting Information, Fig. S2). Two Hypoblemum
refer to this clade as ‘core Maratus’ (also recovered in samples have discordant 28S vs. 16S_ND1 placements;
RAD analyses, see below). we leave these identified to genus only. Prostheclina
© 2021 The Linnean Society of London, Biological Journal of the Linnean Society, 2021, XX, 1–24PEACOCK SPIDER PHYLOGENOMICS 11
appears allied to Jotus (e.g. Fig. 2B), with Maileus we recovered 513 loci (alignment length = 59 997 bp,
Downloaded from https://academic.oup.com/biolinnean/advance-article/doi/10.1093/biolinnean/blaa165/6126965 by University of California, Berkeley user on 05 February 2021
surprisingly nested, although some nodes are poorly 11 290 SNPs, 7196 parsimony-informative sites) for
supported for these individual gene trees (Supporting the sample including Jotus, Saitis, Hypoblemum,
Information, Fig. S3). Saratus and Maratus.
Following UCE results (see above), we rooted
phylogenies using Jotus auripes + Saitis virgatus. With
RAD analysis – all taxa this root placement, neither Saitis nor Maratus were
Thirty-four samples were removed from downstream recovered as monophyletic in unpartitioned ML (Fig. 4;
analysis because of low raw read counts. Of retained Supporting Information, Fig. S3) or SVDQuartets
samples, 152 were included in all-taxa phylogenomic (Supporting Information, Figs S4, S5) analyses. One
analysis. Using the custom pipeline, the number of primary branch includes members of the fimbriatus,
filtered reads for these samples ranged from 101 077 chrysomelas and spicatus morphological species groups
to 3623 248, with an average coverage of individual as defined by Otto & Hill (2019a; Table 1). As for UCE
RAD loci of 12× (range from 3.0 to 61.1; Supporting data, members of this lineage occur on a conspicuously
Information, Table S1). We recovered between 9400 long branch. Members of the chrysomelas group are
and 27 893 loci per sample, but the number of shared intermixed with species of the spicatus group; because
loci across the entire taxon sample was lower, probably the former species were described first, we hereafter
due to the deep divergences among genera. In total refer to this entire larger clade as the chrysomelas
Saitis_mutans_178
Saitis_mutans_180
Hypoblemum_grisuem_167
Hypoblemum_griseum_168
Hypoblemum_scutulatum_166 carmel_SpringHIllSA_12C
carmel_SpringHIllSA_93C
flame_LakeGairdnerSA_18C
FC
Hypoblemum Clade flame_LakeGairdnerSA_95C
speculifer_TriggWA_121
fimbriatus chrysomelas_BordertownSA_17C
chrysomelas_BordertownSA_22C
chrysomelas_CarnavonQLD_24C
chrysomelas_TregoleQLD_19C
spicatus_KoondoolaWA_116
spicatus_KoondoolaWA_71
spicatus_KoondoolaWA_53
nigromaculatus_BoondallQLD_65C
nigromaculatus_BoondallQLD_66C
nimbus_LakeEyreSA_13C
nimbus_LakeEyreSA_16C
nimbus_LakeEyreSA_87C
chrysomelas purcellae_CanberraACT_179
purcellae_CanberraACT_38C
purcellae_CanberraACT_9C
robinsoni_NewcastleNSW_131
robinsoni_NewcastleNSW_170
rainbowi_LaneCoveNSW_132 robinsoni_NewcastleNSW_172
rainbowi_LaneCoveNSW_84
maritimus_EsperanceWA_146
western_pavonis_HeardsmanWA_141
western_pavonis_TakalarupWA_88
western_pavonis_WalpoleWA_44
western_pavonis_HeardsmanWA_90
cf_leo_WaterfallSA_73C
leo_DarlingtonSA_25C
leo_DarlingtonSA_7C
leo_DarlingtonSA_85C
literatus_BooroopkiVIC_71C pavonis
cf_leo_WaterfallSA_74C
literatus_NhilSwampVIC_72C
cf_leo_WaterfallSA_75C
SE_pavonis_MaquarieTAS_37C
SE_pavonis_OrfordTAS_35C
SE_pavonis_OrfordTAS_36C
SE_pavonis_MelbourneVIC_139
SE_pavonis_MelbourneVIC_129
ACT_pavonis_NamadgiACT_32C
ACT_pavonis_NamadgiACT_33C
ACT_pavonis_NamadgiACT_34C
watagansi_WatagansNSW_169
albus_FlindersSA_21C
albus_FlindersSA_2C
albus_FlindersSA_8C
albus_FlindersSA_90C
albus_PtAdelaideSA_29C
vultus_StrathdownieVIC_11C
anomalus
vultus_StrathdownieVIC_15C
anomalus_DunmoreQLD_81
aurantius_OrangeNSW_62C
neptunus_ButterwickNSW_173
aurantius_OrangeNSW_63C
‘core Maratus’ aurantius_OrangeNSW_76C
cfneptunus_CassilisNSW_47C
neptunus_ButterwickNSW_171
neptunus_ButterwickNSW_174
cinereus_StanthropeQLD_55C
cinereus_StanthropeQLD_56C
cinereus_StanthropeQLD_84C
michaelorum_CarnavonQLD_1C
michaelorum_CarnavonQLD_4C
sceletus_IslaGorgeQLD_61C
sceletus_WondulQLD_137
Saratus anomalus_NewcastleNSW_67
Saratus_CanberraACT_145
sceletus_WondulQLD_75
Saratus_NamadgiACT_51C
Saratus_NamadgiACT_52C
amabilis_CowanNSW_30C
amabilis_KaputarNSW_123
amabilis_KaputarNSW_68
amabilis_MtBarneyQLD_27C
amabilis
amabilis_MtBarneyQLD_88C
vespertillio_NgarkatSA_43C
vespertillio_LittleDesVIC_46C vespertilio
= bp > 95 vespertillio_MajuraACT_45C
vespertillio_MajuraACT_44C
speciosus_TriggWA_64
speciosus
speciosus_TriggWA_86
= bp > 90 proszynskii_GoulburnNSW_3C
proszynskii_NamadgiACT_143
proszynskii_MtWilliamTAS_89C velutinus
proszynskii_NamadgiACT_103
velutinus_KuringgaiNSW_133
harrisi_WadabilligaNSW_14C
harrisi_WadabilligaNSW_28C
harrisi
87 avibus_CapeAridWA_92C
caeruleus_MiddleIslWA_10C
caeruleus_MiddleIslWA_6C
pardus_CapeLeGrandeWA_157
clupeatus_GnangaraWA_126
clupeatus_GnangaraWA_96
flavus_YalgorupWA__124
linnaei_TwoPeoplesWA_112 expanded
65 linnaei_TwoPeoplesWA_87
linnaei_TwoPeoplesWA_63
madelineae_DardanupWA_120
mungaich
madelineae_DardanupWA_57
madelineae_DardanupWA_69
sarahae_BluffsKnollWA_114
sarahae_BluffsKnollWA_85
mungaich_MtDaleWA_128
mungaich_MtDaleWA_52
vespa_lakeJasperWA_70C
personatus_CapeRicheWA_154
australis_EsperanceWA_153
tasmanicus_MelbourneVIC_144
tasmanicus_MtWilliamTAS_48C
tasmanicus
tasmanicus_MtWilliamTAS_50C
tasmanicus_PiccaninnieSA_49C (grade)
tasmanicus_PiccaninnieSA_82C
cf_plumosus_GleneigVIC_39C
cf_plumosus_GleneigVIC_40C
cf_plumosus_GleneigVIC_41C
cf_plumosus_HeathertonVIC_101
cf_plumosus_HeathertonVIC_102
plumosus_KuringgaiNSW_108
plumosus_GoldburnRiverNSW_130
volans_BlackdownQLD_58C
volans_BlackdownQLD_96C
volans_LaneCoveQLD_94
volans_MtBarneyQLD_54C
elephans_BowlingAlleyNSW_151
volans
0.0050 MTVC elephans_BowlingAlleyNSW_74
elephans_DungowanNSW_5C
digitatus_KaputarNSW_125
digitatus_KaputarNSW_142
eliasi_NugaNugaQLD_80C
eliasi_TregoleQLD_81C
eliasi_NugaNugaQLD_97C
ottoi_VenmanQLD_59C
ottoi_VenmanQLD_67C
calcitrans
ottoi_VenmanQLD_68C
calcitrans_BlackMtnACT_72
jactatus_WondulQLD_118
jactatus_WondulQLD_46
jactatus_WondulQLD_69C
Jotus_auripes_161
Jotus_auripes_162C
Saitis_virgatus_159
Saitis_virgatus_160
100
Figure 4. RAD all-taxa maximum-likelihood phylogeny. Bootstrap values below 90 are not shown for most nodes. Placement
of Maratus sceletus, M. pardus and M. plumosus to the original groups of Otto & Hill (2019a) is designated with arrows.
© 2021 The Linnean Society of London, Biological Journal of the Linnean Society, 2021, XX, 1–2412 M. B. GIRARD ET AL.
group (redefined). Many species in the above three 2014b, M. neptunus Otto & Hill 2017a, M. aurantius
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groups were previously placed in the genus ‘Lycidas’ Otto & Hill 2017a]. The only clear case of strongly
(but not including the type species of ‘Lycidas’). supported non-monophyly involves the geographically
Consistent with UCE data, fimbriatus + chrysomelas widespread M. pavonis. In both tetrad and ML results,
(FC) clades are recovered as sister to Hypoblemum, M. pavonis specimens from Western Australia are more
and together comprise the Hypoblemum clade. We closely related to Western Australia M. maritimus Otto
discuss below possible morphological and behavioural & Hill 2014b, with eastern M. pavonis populations
synapomorphies that unite members of this clade. phylogenetically closer to geographically adjacent
A second primary Maratus branch includes all other M. literatus and M. leo (Figs 4, 5). This result supports
Maratus taxa (‘core Maratus’), with Saratus samples the contention that M. pavonis currently includes
nested within this larger clade (Fig. 4; Supporting several distinct species, as supported by both genitalic
Information, Figs S4, S5). The ‘core Maratus’ clade (Baehr & Whyte, 2016) and male ornamentation
includes the anomalus species group, including both characters (Hill & Otto, 2011; Otto & Hill, 2019a).
the type species of ‘Lycidas’, and M. amabilis, the type Introgression with geographical neighbours might also
species of the genus Maratus. Many clades within be driving this pattern, but if this were true we might
‘core Maratus’ are mostly concordant with the species expect more lineage non-monophyly (e.g. Western
group divisions of Otto & Hill (2019a), as based on Australia M. pavonis intermixed with Western
morphological/behavioural similarities. Exceptions Australia M. maritimus, etc.).
include a monophyletic anomalus group, with the Multiple analyses support the hypothesis that three
inclusion of M. sceletus Otto & Hill 2015a (from the novel forms included here represent undescribed
calcitrans group). Maratus plumosus Otto & Hill species (M. ‘carmel’, M. ‘flame’, M. cf. plumosus).
2013b, a hypothesized (but divergent) member of the Two forms (M. cf. neptunus, M. cf. leo) might best be
calcitrans group, also falls outside of this group. The interpreted as geographical variants (Figs 4, 5), but
tasmanicus group is not recovered as monophyletic, larger sample sizes and formal integrative species
but instead represents a grade sister to the ‘expanded delimitation analyses are needed to rigorously test
mungaich’ (EM) clade. The large, poorly resolved EM these species hypotheses.
clade includes species from several groups as follows:
M. clupeatus Otto & Hill 2014d (not a straggler),
M. pardus Otto & Hill 2014c (from the volans group), Character evolution
M. australis Otto & Hill 2016b (from the tasmanicus We lacked some character data for Saitis barbipes
group), and single sampled species from the flavus, and Hypoblemum (Supporting Information, Table
linnaei, personatus and vespa groups. The EM clade S2), causing ambiguity in certain ancestral character
is further discussed below, but this grouping is reconstructions. For the sparse UCE taxon sample that
biogeographically compelling, as all included taxa are importantly includes Saitis barbipes, we reconstruct
endemic to south-west Australia. the common ancestor of the entire Saitis Clade as
males using third legs during courtship (character 4,
Fig. S6), and with vibrations produced during a pre-
RAD analysis – species groups mount display (character 8, Fig. S7; see Girard et al.,
Tetrad results and locus statistics for the FC, pavonis, 2011), with reversals observed for both characters.
anomalus and MTVC clades are shown in Figure 5. The common ancestor of the Maratus group is
Interspecific relationships in tetrad analyses are unambiguously reconstructed as with males raising
generally better supported than in ML or SVDQuartets their abdomen during courtship (character 1, Fig. S6),
analysis, perhaps reflecting the greater number of consistent with the hypothesis of Otto & Hill (2012a;
loci recovered for each group. An exception is the EM Fig. 2B). Ancestral state reconstructions for other
clade – although tetrad analyses included more SNPs characters are shown in Figures S6 and S7.
and parsimony-informative sites than in all-taxa Ancestral state reconstructions for the denser RAD
ML analyses, relationships are conspicuously poorly taxon sample that included the full Maratus sample
supported in both (Figs 4, 5). (but lacked Saitis barbipes) are shown in Supporting
Almost all a priori species are supported as Information, Figures S8–S15. These reconstructions
monophyletic on tetrad trees (Fig. 5). One caveat of suggest the evolution of a strong white brush on the
this claim is that several species with multiple samples male third leg evolving at the base of ‘core Maratus’
only include specimens from the same geographical (character 5, Fig. S11). Two characters suggest that
location. Five species not recovered as monophyletic the pre-mount display and pre-mount vibrations may
on ML trees are recovered as exclusive groupings on be unique in the Hypoblemum clade (characters 8 and
tetrad trees [M. chrysomelas (Simon 1909), M. leo 9, Figs S14, S15), predicting that missing data for
Otto & Hill 2014b + M. cf. leo, M. literatus Otto & Hill Hypoblemum species will conform to the FC clade. The
© 2021 The Linnean Society of London, Biological Journal of the Linnean Society, 2021, XX, 1–24PEACOCK SPIDER PHYLOGENOMICS 13
84
flame_SA_95C FC Clade
Downloaded from https://academic.oup.com/biolinnean/advance-article/doi/10.1093/biolinnean/blaa165/6126965 by University of California, Berkeley user on 05 February 2021
100
flame_SA_26C
flame_SA_18C 5381 loci,
100 carmel_SA_93C 63102 SNPs
83
89
carmel_SA_23C 17622 PIs
100
carmel_SA_12C
speculifer_WA_121
chrysomelas_SA_20C
43 chrysomelas_QLD_24C
46
chrysomelas_QLD_19C
47
chrysomelas_SA_22C
90 68
chrysomelas_SA_17C
spicatus_WA_71
95 spicatus_WA_116
40
74
spicatus_WA_53
nigromaculatus_QLD_64C
96 97 nigromaculatus_QLD_65C
90 nigromaculatus_QLD_66C
nimbus_SA_16C
100 nimbus_SA_87C
100 43
nimbus_SA_13C
purcellae_ACT_9C
100 purcellae_ACT_38C
71 albus_SA_2C
100
purcellae_ACT_179 albus_SA_8C anomalus
59
robinsoni_NSW_172 76 albus_SA_21C
100 robinsoni_NSW_131 100 67
52 albus_SA_90C 11609 loci,
robinsoni_NSW_170 albus_SA_29C
100 115608 SNPs
vultus_VIC_11C
100 33606 PIs
vultus_VIC_15C
anomalus_NSW_67
100 anomalus_QLD_81
cfneptunus_NSW_47C
47 neptunus_NSW_173
98 neptunus_NSW_174
100 45 neptunus_NSW_171
pavonis 100 aurantius_NSW_76C
85 aurantius_NSW_63C
9055 loci, 47 aurantius_NSW_62C
100
103477 SNPs cinereus_QLD_84C
rainbowi_NSW_132 26175 PIs 83 cinereus_QLD_56C
100 rainbowi_NSW_84 50
72 cinereus_QLD_55C
western_pavonis_WA_141 100 michaelorum_QLD_1C
68 western_pavonis_WA_90 92
65 michaelorum_QLD_4C
western_pavonis_WA_107 sceletus_QLD_75
100 51
western_pavonis_WA_88 sceletus_QLD_137
99 88 western_pavonis_WA_44 100
sceletus_QLD_61C
maritimus_WA_146
SE_pavonis_VIC_139
90 SE_pavonis_VIC_129
93
65 SE_pavonis_TAS_37C
46 avibus_WA_92C
79 SE_pavonis_TAS_35C
SE_pavonis_TAS_36C 88 caeruleus_WA_10C
98 caeruleus_WA_6C
100 leo_SA_85C
leo_SA_25C sarahae_WA_114
96 100 sarahae_WA_85
84 leo_SA_7C 53
vespa_WA_70C
55 cf_leo_SA_75C 22 linnaei_WA_112
36 cf_leo_SA_73C 94 linnaei_WA_63
63 45 50
cf_leo_SA_74C 52 36 linnaei_WA_87
ACT_pavonis_ACT_32C madelineae_WA_69
62 madelineae_WA_57
55 ACT_pavonis_ACT_33C 71
53 34 17 madelineae_WA_120
51 ACT_pavonis_ACT_34C EM flavus_WA__124
literatus_VIC_71C 78 34 mungaich_WA_52
57 83 mungaich_WA_128
literatus_VIC_72C clupeatus_WA_126
watagansi_NSW_169 57 79 clupeatus_WA_96
personatus_WA_154
australis_WA_153
44 pardus_WA_157
cf_plumosus_VIC_41C
68 46 cf_plumosus_VIC_39C
48 cf_plumosus_VIC_101
93 62 cf_plumosus_VIC_102
cf_plumosus_VIC_40C
77 tasmanicus_VIC_144
85 tasmanicus_TAS_48C
95 tasmanicus_TAS_50C
91 83
26
93
tasmanicus_SA_82C
tasmanicus_SA_49C
plumosus_NSW_108
49 plumosus_NSW_130
elephans_NSW_5C
99 elephans_NSW_74
99 elephans_NSW_151
97 volans_QLD_94 MTVC
55 volans_QLD_54C
100 volans_QLD_96C 13264 loci,
100 volans_QLD_58C
jactatus_QLD_118 152942 SNPs
100 jactatus_QLD_69C 40981 PIs
82 80
jactatus_QLD_46
calcitrans_BlackMtnACT_72
eliasi_QLD_81C
91 100 eliasi_QLD_97C
94 eliasi_QLD_80C
100
digitatus_NSW_142
94 100 digitatus_NSW_125
ottoi_QLD_59C
99 ottoi_QLD_67C
38 ottoi_QLD_68C
Figure 5. RAD tetrad results for the FC, pavonis, anomalus and MTVC clades.
© 2021 The Linnean Society of London, Biological Journal of the Linnean Society, 2021, XX, 1–2414 M. B. GIRARD ET AL.
‘core Maratus’
Downloaded from https://academic.oup.com/biolinnean/advance-article/doi/10.1093/biolinnean/blaa165/6126965 by University of California, Berkeley user on 05 February 2021
MTVC
expanded mungaich
elephans (2)
avibus (2)
pavonis anomalus
volans (2)
caeruleus (2)
vultus (1)
watagansi
Saratus
rainbowi (1)
albus
flavus
calcitrans
plumosus (4)
amabilis (2)
Hypoblemum Clade
clupeatus (2)
anomalus
maritimus
jactatus (1)
cf plumosus (4)
chrysomelas
speciosus
madelineae (2)
western pavonis (1)
robinsoni
H griseum
sceletus
calcitrans
tasmanicus (2)
purcellae
vespertillio (3)
ACT pavonis (1))
H scutulatum
linnaei
cinereus
outgroups
nimbus
ottoi
Saitis mutans
michaelorum
SE pavonis (1)
vespa (4)
australis (3)
fimbriatus
chrysomelas nigromaculatus
velutinus
digitatus (3)
speculifer
proszynskii
sarahae (2)
Saitis virgatus
cf neptunus
neptunus +
personatus
literatus (1)
flame
eliasi (5)
5
mungaich (2)
Jotus auripes
aurantius
leo + cf leo
pardus (2)
harrisi (3)
spicatus
carmel
1 1 1 1 1 1 1 2 3 3 1 3 5 2 2 4 4 2 3 2 2 2 2 2 2 2 4
Character 3:
overall fan shape
0=no fan
1=round
2=elliptical
3=lobed
4=lobed(post)
5=diamond
Figure 6. Maximum-likelihood ancestral character reconstruction for fan shape, on the skeletal RAD taxon sample
phylogeny. Character states are noted on the phylogeny, and are associated with taxon names; all taxa with character state
0 (=no fan) are left unlabelled.
evolution of fan shape diversity across Maratus group the abdomen during courtship are ancestral to the
members is illustrated in Figure 6. ‘core Maratus’ clade. Other traits, particularly fan
shape, show evidence of multiple independent gains
and losses throughout what is currently recognized
as Maratus.
DISCUSSION
Our phylogenomic results reveal that both Saitis and
Maratus, as currently recognized, are paraphyletic Saitis Clade
taxa. In Maratus, some species are more closely Zhang & Maddison (2015) define the Saitis Clade as
related to Hypoblemum taxa. A ‘core Maratus’ including multiple genera (Saitis, Maratus, Saratus,
clade includes several well-supported sub-clades Jotus, Hypoblemum, Prostheclina, Maileus, Barraina
that mostly correspond to morphological species and possibly Margaromma), united in sharing several
groups, although some hypothesized groupings were male genitalic features, including a lamella on the
not supported. Saratus is nested within the ‘core tegular shoulder distinctive for euophryine spiders.
Maratus’ clade. Based on this phylogenomic tree Zhang & Maddison (2015) recognized the striking
topology, some traits such as using third legs and variation in male courtship ornamentation in the
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