Extant primitively segmented spiders have recently

Downloaded from http://rspb.royalsocietypublishing.org/ on May 6, 2015
rspb.royalsocietypublishing.org
Research
Cite this article: Xu X et al. 2015 Extant
primitively segmented spiders have recently
diversified from an ancient lineage.
Proc. R. Soc. B 282: 20142486.
http://dx.doi.org/10.1098/rspb.2014.2486
Received: 13 October 2014
Accepted: 14 April 2015
Subject Areas:
evolution, ecology, taxonomy and systematics
Keywords:
living fossils, genetic diversity, plesiomorphies,
vicariance, dispersal, ancestral areas
Authors for correspondence:
Matjazˇ Kuntner
e-mail: [email protected]
Daiqin Li
e-mail: [email protected]
Extant primitively segmented spiders
have recently diversified from an
ancient lineage
Xin Xu1, Fengxiang Liu1, Ren-Chung Cheng3, Jian Chen1, Xiang Xu5,
Zhisheng Zhang6, Hirotsugu Ono7, Dinh Sac Pham8, Y. Norma-Rashid 9,
Miquel A. Arnedo10, Matjazˇ Kuntner1,3,4 and Daiqin Li1,2
1
Centre for Behavioural Ecology and Evolution (CBEE), College of Life Sciences, Hubei University, Wuhan,
People’s Republic of China
2
Department of Biological Sciences, National University of Singapore, Singapore, Singapore
3
Evolutionary Zoology Laboratory, Biological Institute ZRC SAZU, Ljubljana, Slovenia
4
Department of Entomology, National Museum of Natural History, Smithsonian Institution, Washington, DC, USA
5
College of Life Sciences, Hunan Normal University, Changsha, Hunan, People’s Republic of China
6
Key Laboratory of Eco-environments in Three Gorges Reservoir Region (Ministry of Education),
School of Life Science, Southwest University, Chongqing, People’s Republic of China
7
Department of Zoology, National Museum of Nature and Science, 4-1-1 Amakubo, Tsukuba-shi,
Ibaraki-ken 305-0005, Japan
8
Institute of Ecology and Biological Resources (IEBR), Vietnamese Academy of Science and Technology (VAST),
18 Hoang Quoc Viet, Cau Giay, Hanoi, Vietnam
9
Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur 50603, Malaysia
10
Institut de Recerca de la Biodiversitat, Departament de Biologia Animal, Universitat de Barcelona,
Avinguda Diagonal 643, Barcelona 08028, Spain
Living fossils are lineages that have retained plesiomorphic traits through long
time periods. It is expected that such lineages have both originated and diversified long ago. Such expectations have recently been challenged in some
textbook examples of living fossils, notably in extant cycads and coelacanths.
Using a phylogenetic approach, we tested the patterns of the origin and diversification of liphistiid spiders, a clade of spiders considered to be living fossils
due to their retention of arachnid plesiomorphies and their exclusive grouping
in Mesothelae, an ancient clade sister to all modern spiders. Facilitated by original sampling throughout their Asian range, we here provide the phylogenetic
framework necessary for reconstructing liphistiid biogeographic history. All
phylogenetic analyses support the monophyly of Liphistiidae and of eight
genera. As the fossil evidence supports a Carboniferous Euramerican origin
of Mesothelae, our dating analyses postulate a long eastward over-land dispersal towards the Asian origin of Liphistiidae during the Palaeogene (39–58 Ma).
Contrary to expectations, diversification within extant liphistiid genera is
relatively recent, in the Neogene and Late Palaeogene (4–24 Ma). While no
over-water dispersal events are needed to explain their evolutionary history,
the history of liphistiid spiders has the potential to play prominently in vicariant
biogeographic studies.
1. Introduction
Electronic supplementary material is available
at http://dx.doi.org/10.1098/rspb.2014.2486 or
via http://rspb.royalsocietypublishing.org.
Living fossils are those lineages that were once more diverse than today, or that
have survived over long evolutionary timescales with little morphological
change [1]. Textbook examples include plants such as the Gingko tree and the
cycads, and animals such as coelacanth fishes and horseshoe crabs. These
living fossils are hypothesized to exhibit ancient roots and early diversification
patterns [2,3]. However, such assertions are rarely tested within a time-calibrated
phylogenetic framework. The latest dating analyses of extant cycads and coelacanths have discovered that although these lineages are ancient, their extant
species are a surprisingly recent radiation [1,4]. Within arachnids, liphistiid spiders are widely recognized as ‘living fossils’ [5] as they are the only surviving
& 2015 The Author(s) Published by the Royal Society. All rights reserved.
Downloaded from http://rspb.royalsocietypublishing.org/ on May 6, 2015
Here we use molecular data from our original sampling
to test the monophyly of Liphistiidae and its genera. We
then devise a time-calibrated phylogeny to elucidate the
origin of the stem group Mesothelae and its major clades.
We use these data to examine the biogeographic history of
these spiders, and, given their sedentary lifestyle and long
evolutionary history, propose that the lineage is an ideal
model for reconstructing past events. Finally, we test the
assumption that present-day liphistiids exhibit not only early
origin but also diversification patterns, as would be expected
from their labelling as living fossils.
The greater detailed methods are described in the electronic supplementary material.
(a) Material and phylogenetic inference
Our study primarily focused on heptathelines (the liphistiids of
East Asia), and we added two species of liphistiines from Laos
and Malaysia (figure 1e). Our ingroup sampling thus had a
total of 124 liphistiid specimens from 105 localities (figure 1e).
We sampled mygalomorph outgroups of the families Atypidae
and Ctenizidae and augmented original data with sequences
from GenBank (electronic supplementary material, table S1).
We performed parsimony [34], maximum-likelihood (ML) [35]
and Bayesian (BI) [36] phylogenetic analyses using nucleotide
data from five genes (CO1, 16S, 28S, H3, ITS2) (electronic supplementary material, table S2) and selecting the best fit DNA
substitution model for each of the four partitions [37].
(b) Calibrations and divergence dating
To estimate divergence times within Liphistiidae, we devised a
matrix whose taxon sample maximized fossil calibration points.
Analyses conducted using only fossil-based time constraints
yielded extremely low estimates for mtDNA substitution rates,
more than twice lower than any value reported in the literature
[38 – 41]. Therefore, we conducted additional analyses with
normal distribution on the mtDNA rate prior based on information available in the literature. We linked the mitochondrial
genes (CO1 and 16S) in a single clock, assigning the substitution
rate parameter a normal prior with a mean 0.017 and standard
deviation 0.0045, as recently estimated for other spider lineages
[42]. For the nuclear genes (28S, H3 and ITS2) and to speed-up
analyses, we restricted the parameter space by setting diffuse,
uniform priors to the starting value of 0.00115, with the minimum and maximum bounds set to 0.0001 and 0.0115. These
values are based on the universal substitution rate proposed
for arthropod mtDNA [43], and were selected under the assumption that the nuclear genes are about one order of magnitude
slower than mitochondrial (starting value) and generally no
nuclear protein coding gene will show higher rates than the
mitochondrial genes (upper bound) [44].
Based on the jModeltest results, we used the substitution
models GTR þ I þ G for CO1, 16S, 28S and ITS2 and TN93 þ
I þ G for H3, and base frequencies were set to estimate except
for H3 and ITS2 which were set to equal. A starting tree including the time constraints was generated with starttree (http://
bodegaphylo.wikispot.org/starttree_program) and incorporated
to the analysis by modifying the xml file. All analyses were
run assuming a lognormal relaxed clock for each gene partition
and a Yule tree prior. We specified an exponential prior for the
yule.birthRate with a mean of 0.05, estimated using the pyule
tool (available at https://github.com/joaks1/pyule). For each
analysis, three independent chains were run for 50 million
Proc. R. Soc. B 282: 20142486
2. Material and methods
2
rspb.royalsocietypublishing.org
group of the suborder Mesothelae that have retained spider
plesiomorphies such as abdominal tergites and spinnerets
located in the middle of the abdominal venter (figure 1a,b)
[6–9]. Because of this plesiomorphic morphology, the family
and genera of living liphistiid spiders native only to parts of
Southeast and East Asia are assumed to be ancient and exhibit early diversification patterns. This assumption, however,
remains untested.
The family Liphistiidae consists of 89 extant species-level
taxa currently grouped in three genera, and displays an interesting geographical distribution confined to Southeast and
East Asia [10]. The group is a species-poor, but seemingly
ancient lineage of relatively large, dispersal-limited, unusually long-lived (5 –18 years), ground-dwelling spiders that
build trapdoor burrows (figure 1c,d) [8,9,11]. Since their discovery [12], classical authors [13] divide Liphistiidae into
two subfamilies, Liphistiinae with a single genus, Liphistius
and 50 species, and Heptathelinae with 39 species within
two genera, Heptathela and Ryuthela [10]. Liphistiines are geographically separated from heptathelines, Liphistius occurring
in Southeast Asia, and Heptathela and Ryuthela confined to
East Asia (figure 1e).
As the sister clade to all other known spiders, Mesothelae
should play prominently in any attempts at reconstructing the
arachnid tree of life and testing alternative biogeographic
hypotheses. Surprisingly, however, the research on Mesothelae
and Liphistiidae has thus far been devoid of comprehensive
phylogenies. Although liphistiid exemplars are routinely used
in family level or higher spider phylogenetic research, these
works either use solely morphological data [7–9,14–17], or
where molecular data are used, employ a single or at the
most two representatives of the family [18–21] except for a
recent study reporting the molecular phylogeny for the genus
Ryuthela [22]. To date, no molecular phylogeny has been
available for the family and for other genera.
Representing an ancient lineage with a geographically
restricted distribution and very likely limited colonization
ability, the biogeography of liphistiids is of great interest. It
is, therefore, surprising how little we know about the origin
of Mesothelae (the basal-most branch of all spiders) and
Liphistiidae (the only Mesothelae family with extant species),
and how geological and biological processes in the Earth’s
history might have affected them. The extant Mesothelae
are only distributed in East and Southeast Asia, from where
no Palaeozoic arachnids are known, and the only fossil
species is the Late Carboniferous Palaeothele montceauensis
[23], known from the part of Euramerica that corresponds
to today’s France [23,24]. At that time in the Earth’s history
(295 Ma), those regions where Mesothelae are found today
were on landmasses isolated from Pangea [25–32]. Recent
authors [13,33] hypothesized that Mesothelae originated in
Euramerica prior to its integration with Pangea, and that
during the Late Triassic the ancestor of Liphistiidae colonized
East and Southeast Asia from Euramerica over one of several
possible landmasses. The occurrence of Mesothelae on the
islands of southern Japan (Kyushu and Ryukyu Archipelago)
was explained both through vicariant origin in the Tertiary
when the Japanese island arc separated from mainland Asia
[33], or alternatively, as a consequence of dispersal events
over-land bridges from east China during the Pleistocene
[9]. Considering the acute absence of a liphistiid phylogeny,
none of the above biogeographic hypotheses have been
empirically tested.
Downloaded from http://rspb.royalsocietypublishing.org/ on May 6, 2015
(b)
(a)
3
(c)
rspb.royalsocietypublishing.org
ST
(d)
SE
Proc. R. Soc. B 282: 20142486
BK
S
TG
(e)
Hebei
Shanxi
Shandong
Shannxi
D
Henan
B
Hubei
Sichuan
Chongqing
F
Japan
Zhejiang
Jiangxi
Hunan
Guizhou
Fujian
Yunnan
G
E
Hong Kong
Laos Vietnam
Hainan
C
Heptathela
Liphistius
Ganthela
Qiongthela
Ryuthela
A
Sinothela
Songthela
Vinathela
500 km
Malaysia
Figure 1. Phenotypical plesiomorphies of liphistiid spiders, and a map with the sampling localities. (a) Dorsal view of Ryuthela nishihirai; (b) ventral view of
R. nishihirai; (c) trapdoor of Liphistius with radiating signal lines; (d ) heptatheline trapdoor (note absence of signal lines). BK, book lungs; S, spinnerets; SE, sternite,
ST, sternum; TG, tergite. Scale bars, 2 mm; (e) map with the sampling localities. Colours, corresponding with those in figure 3, represent eight liphistiid genera.
Letters denote areas for biogeographic reconstruction: A: Southeast Asia; B: Kyushu; C: Hainan; D: North China (north of the Yangtze River); E: East China (Fujian,
Hubei, Hunan, Hong Kong, Jiangxi and Zhejiang); F: West China and North Vietnam (Chongqing, Guizhou, Hubei (Enshi, Jianshi and Lichuan), Hunan (Cili,
Fenghuang and Zhangjiajie), Sichuan and Yunnan); G: Ryukyu archipelago. (Online version in colour.)
Downloaded from http://rspb.royalsocietypublishing.org/ on May 6, 2015
(c) Biogeographical analyses
3. Results
(a) Phylogenetic reconstruction
All analyses, BI, parsimony and ML under the four partition
schemes consistently recover Liphistiidae monophyly with
high support values ( posterior probability, hereafter PP, ML
bootstrap support, BS and parsimony jackknife, JK, all reaching
100%), and all analyses converged on similar within-family
topologies (figure 2). The monophyly of Liphistius, in our taxonomic sample represented by exemplars from Malaysia and
Laos, is well supported (PP ¼ 1, JK ¼ 100 and BS ¼ 100) and
this clade was always sister to heptathelines, a clade uniting
all remaining liphistiids (figure 2). The samples from Japan
fit into two reciprocally monophyletic and highly supported
clades (figure 2), one including all species of Ryuthela from
Ryukyu Island and southern part of Okinawa, and the other
clade consisting of some but not all Heptathela (those from
Ryukyu Island and Kyushu; figure 2). The fact that all other
Heptathela species, those from the Asian mainland, fall into
other clades makes Heptathela paraphyletic. In an accompanying paper [51], we revise the genus level taxonomy based on
the phylogenetic evidence (this paper) and morphological
and natural history diagnoses. Here, we refer to those taxonomic decisions by treating Liphistius Schio¨dte, 1849 and
Ryuthela Haupt, 1983 as valid genera, and Heptathela sensu
lato as six genera (nomenclature formalized in [51]): Ganthela,
Heptathela sensu stricto, Qiongthela, Sinothela Haupt, 2003,
Songthela Ono, 2000, Vinathela Ono, 2000.
(b) Divergence dating and biogeographical
reconstructions
Results of the analyses with and without the informed
prior in the mtDNA substitution rate resulted in markedly
different time frames of diversification for liphistiids (electronic
supplementary material, table S3 and figure S1; figure 3a).
Informing the substitution rate resulted in a compression of the
liphistiid clade towards the present, yielding dates about onethird younger than the fossil, unconstrained analysis (electronic
supplementary material, table S3). The estimated rate of substitution (ucld.mean) of the mtDNA under the unconstrained
4
Proc. R. Soc. B 282: 20142486
To infer biogeographical events, we defined seven discrete
geographical areas (figure 1e) and reconstructed ancestral distributions on a simplified matrix using three alternative methods in
RASP v. 3.0 [47,48]: (i) statistical dispersal – vicariance analysis
(S-DIVA; [47]), with a maximum of three areas per node; (ii) BI
binary MCMC analysis (BBM; [47,49]), with 5000 cycles using
10 chains, sampling every 100 cycles, allowing for a maximum
of three areas, giving a ‘custom’ root distribution, using the
JC þ G substitution model; and (iii) dispersal – extinction –
cladogenesis analysis (DEC; [50]), setting the maximum areas
to three [48], with an unconstrained model allowing for equal
rates of dispersal (1.0) between areas at any time.
fossil analysis yielded very low values, about twice lower than
those reported in the literature [38–41] (electronic supplementary
material, table S4). In addition, some of the parameter values
reported for the unconstrained fossil analysis (coefficient of variation of the 28S and ITS2) were higher than those recommended
in the literature (greater than 1) (electronic supplementary
material, table S4). On the other hand, increasing the time
range of the maximum bound had little to no effect on the results
(electronic supplementary material, tables S3 and S4, column
rates and rates (20%)).
The time-calibrated phylogenetic analyses based on fossil
(exponential prior with 95% of the posterior density falls
within the range 10% older than the minimum bound) and
on substitution rates estimated the basal-most spider divergence between the Mesothelae (treated as a stem group)
and the Opisthothelae (represented by Mygalomorphae
and Araneomorphae) at 297.6 Ma (295–306 Ma) (figure 3a).
Given the estimated age of the extant mygalomorphs and araneomorphs at 245.3 Ma (240–259 Ma) and 280.8 (262–297 Ma),
respectively, and a long, 250 million year Mesothelae branch,
the Palaeogene origin of the family Liphistiidae estimated at
48 Ma (39–58 Ma) is relatively recent. Heptathelinae origin is
estimated at 32.9 Ma (28–39 Ma). The origins of all liphistiid
genera fall into the Late Palaeogene and Neogene (4–36 Ma).
Within Heptathelinae, the clade uniting eastern island
Oligocene taxa Heptathela (9.5; 7–12 Ma), Qiongthela (6.4;
4–10 Ma) and Ryuthela (7.5; 5–10 Ma) originated at 30.6 Ma
(25–36 Ma). The clade of a comparable Oligocene age (27.9;
23–33 Ma) unites the mainland genera Ganthela (15.4;
11–20 Ma), Sinothela (17.7; 14–22 Ma), Songthela (12.4;
10–15 Ma) and Vinathela (16.2; 13–20 Ma). The extant diversity
of the Heptathelinae genera can be traced to the Miocene
(approx. 4–22 Ma), while Liphistius may have diversified
slightly earlier (17.3; 11–24 Ma).
Ancestral area reconstructions indicate a number of likely
dispersal and vicariant events (electronic supplementary
material, figure S2 and table S5). None of postulated dispersal
events in the liphistiid biogeographic history, however, need
to invoke any over-water dispersal. The ancestral area for liphistiids is as likely to be Southeast as East Asia, which since that
geological time (approx. 48 Ma) continued to be a single land
mass. The heptatheline ancestors occupied the areas corresponding to today’s Hainan, East China or Ryukyu archipelago, at
the time (approx. 33 Ma) all part of continental Asia. Likewise,
the ancestors of today’s island taxa (Heptathela s.s., Ryuthela,
Qiongthela) occupied either Hainan or Ryukyu archipelago
before they were islands (approx. 31 Ma). These three genera
diversified vicariantly, before or when their ancestral areas
became continental islands, i.e. when Kyushu separated from
the Asian continent with the opening of the Japanese sea
approximately 15 Ma [53–55], and when Ryukyu archipelago
was gradually isolated from Asia approximately 10–5.3 Ma
[56–61]. On the mainland, the separation of the lineages
Sinothela and Ganthela (approx. 20; 16–25 Ma) corresponds
well to the timing of the Yangtze River origin approximately
23 Ma [55,62]. This suggests that their vicariant origin on each
side of the river is still mirrored in today’s distributions.
Similarly, Vinathela and Songthela have never in their 23 Ma
(19–28 Ma) history left the areas E and F (figure 1e) over Yangtze
to the north or over the ocean to the south and east, and neither
has Ganthela in approximately 15 Ma (15–20 Ma). Similarly, the
approximately 6 Ma (3–10 Ma) evolution of Qiongthela invokes
no over-water dispersals from Hainan.
rspb.royalsocietypublishing.org
generations. Chain convergence and correct mixing of each
MCMC chain was assessed with TRACER v. 1.6 [45]. Ten
per cent of the first generation of each chain was removed as
burnin and the remaining values were combined into a single
file with the help of LogCombiner [46] and consensus trees
were obtained with TreeAnnotator [46].
Downloaded from http://rspb.royalsocietypublishing.org/ on May 6, 2015
Atypus heterothecus
Atypus baotianmanensis
Atypus yajuni 01
Atypus yajuni 02
0.1
Figure 2. Summary phylogenetic results. The topology is from the partitioned BI analysis of the full matrix. Black stars indicate solid branch supports, i.e. above the
threshold in more than half of the 12 analyses using BI inference, ML and parsimony (thresholds of posterior probabilities . 0.95, boostrap . 70%, jackknife . 70%).
(Online version in colour.)
Proc. R. Soc. B 282: 20142486
Latouchia sp
Bothriocyrtum californicum
Stasimopus mandelai
Cyrtocarenum grajum
Cyrtocarenum cunicularium
Cteniza moggridgei
Cteniza sauvagesi
Latouchia typica 03
Latouchia formosensis smithi
Latouchia typica 01
Latouchia swinhoei 04
Conothele taiwanensis 06
Conothele taiwanensis 07
Conothele sp
Ummidia sp1
Ummidia sp4
Ummidia algarve
Ummidia picea
Ummidia aedificatoria
Ummidia sp3
Ummidia algeriana
Ummidia sp2
Malaysia 01
Malaysia 02
Etulaos 3256
Liphistius
Etulaos 3257
Fanelaos 3265
Fanelaos 3266
AharenTokashiki 2409
DarumaKumejima 2387
Hirakuboishigaki 3222
Iriomotejima 3200
Asato 2542
Ryuthela
Shuri 2302
Kisenbaru 2477
Yamazato 2538
MtOtowa 2536
MtNago 2447
HenokoDam 2468
Taira 2333
MtkankaeYakushima 3490
IwatoMiyazaki 3442
TakeyamaOita 3418
ShiroyamaKagoshima 3349
LakemiikeMiyazaki 3468
HonjoMiyazaki 3449
HonjoMiyazaki 3462
TakachihokawaraKagoshima 3472
HitoyoshiKumamoto 3360
HanaokaKumamoto 3391
MukoyamaMiyazaki 3435
GinamaDam 2432
KojimaTokunoshima 3339
Heptathela
NazeAmami 3278
SumiyoAmami 3287
UkenAmami 3297
YamatoAmami 3305
MikyoTokunoshima 3316
MikyoTokunoshima 3333
TokuwaseTokunoshima 3323
Shuri 2309
Tokashiki 2417
MtKubayamaiheya 2481
MtOtowa 2535
TaihoDam 2441
Yona 2322
Untenport 2522
Taira 2328
Yofuke 2458
Jianfeng 2106
Baisha 2087
Wuzhishan 2108
Qiongthela
Bawangling 1001
Jianfeng 2098
Jianfeng 2017
JiAn 3540
Jinggangshan 3517
Wangjiangshan 3159
Ganthela
Qingyuanshan 2287
Xianyou 3151
Yundingshan 3135
Luotian 2079
Badong 2140
Yichang 2272
Yiyuan 2045
Zhangqiu 2034
Luquan 1217
Sinothela
Yongnian 1214
Dengfeng 2030
Yuncheng 1234
Heyang 1242
Gaoling 1255
Jingyang 1270
Hongkong 2152
Jinggangshan 3501
JiAn 3519
MtYuelu 1037
Wugaishan 3559
Yizhang 3556
Vinathela
LucNam 3075
TamDao 3021
CucPhuong 3007
LaoCai 3047
Yenbai 3060
Geleshan 2177
Banzigou 2165
Jinyunshan 2189
Wanzhou 2256
Wangerbao 2268
Zhangjiajie 1168
Enshi 2127
Jianshi 2137
Lichuan 2110
MtYuelu 1019
Xiujian 1131
Hangzhou 3166
Xianning 2056
Zhangjiajie 1190A
MtYuelu 1021
Gouloufeng 1098
Gouloufeng 1097
Hengshan 1039
Cili 1194
Zhangjiajie 1170
Songthela
Fenghuang 1153
Fenghuang 1156
Chengxi 1144
Xiujian 1137
Wuzhufeng 2232
Dali 3082
Kunming 3119
Sapa 3031
Eshan 3090
Yuanjiang 3275
Mojiang 3270
WesternHill 3106
Jiangan 2202
Wuzhufeng 2215
Hengshan 1054
Hengshan 1055
Xiazhou 1127
Yanhe 2013
Jiangan 2201
Xiujian 1130
Geleshan 2175
Nanshan 2237
rspb.royalsocietypublishing.org
Cyclocosmia ricketti 08
5
Downloaded from http://rspb.royalsocietypublishing.org/ on May 6, 2015
(a)
(b)
280.9
245.3
Mygalomorphs
Euramerica
North China
‘Stepp
ing-on
Middl
e Eas
**
9.5
t’
North China
Sibumasu
Heptathela
6
Hebei
23.4 **
Shanxi
5
**
7.5
30.6 *
7
4
** 6.4
Ryuthela
Qiongthela
8
297.6
Qia
Lh ngtan
asa
g
Hunan
**
Ganthela
15 Sinothela
**17.3
350
300
Carboniferous
Permian
Palaeozoic
II
III
250
200
Triassic
V
IV
150
Jurassic
Mesozoic
100
Cretaceous
Malaysia
Heptathela
Liphistius
Liphistius
Ganthela
2
Qiongthela
VII IX
VI VIII
50
0
P Eoc O Mi P PH
Palaeogene Neo
Borneo
500 km
Sumatra
Ryuthela
Sinothela
Songthela
Vinathela
Cenozoic
Figure 3. Divergence time estimates and geographical distribution of liphistiid genera. (a) Chronogram from BEAST with inferred node ages in million years and
95% confidence intervals (bars), estimated using fossil calibrations and informed priors on the mtDNA substitution rates. The arrow indicates the fossil age of
Palaeothele montceauensis but does not imply direct ancestry. Black numbered circles represent the major nodes matching those in electronic supplementary
material, figure S2. Triangles with Roman numbers mark relevant geological events that may have affected the dispersal routes and diversification of mesothelid
and liphistiid lineages. I, Pangea started to form; II, Pangea started to break-up and Sibumasu terrane contacted with Eurasia; III, Sibumasu terrane accreted to the
Indochina, South China and North China plates; IV, North China plate connected to Eurasia; V, India collided with Eurasia; VI, the Yangtze River formed; VII, Japanese
islands separated from East Asia; VIII, Taiwan formed; IX, Hainan Island separated from the mainland. (b) General distribution maps of the eight genera delimited in
this study, and the demarcation of main continental plates and terranes that formed Southeast and East Asia (in part from [52]). Inset map shows the two most
plausible of the three mutually exclusive hypotheses explaining the eastward dispersal routes of liphistiid ancestors (simplified on today’s land masses, but see text
for details). WB, West Burma. (Online version in colour.)
4. Discussion
Bristowe [5] referred to Liphistiidae as a ‘family of living
fossil spiders’ [5] because they ‘retained many characters
from their ancestors and resemble fossil spiders of the
Carboniferous period’. Our extensive fieldwork over their present range (figure 1e) discovered numerous new localities
and species and, albeit emphasizing heptathelines, secured a
taxon sample needed for the first comprehensive specieslevel phylogeny of Liphistiidae. Our phylogenetic (figures 2
and 3a) and biogeographic (electronic supplementary
material, figure S2) analyses enabled rigorous testing of the
monophyly of the family, its two subfamilies, its genera and
species groups and the timing of the major divergences
and the ancestral ranges. Interesting strong evolutionary and
biogeographic patterns emerge. While extant liphistiids
indeed possess arachnid plesiomorphies (figure 1a,b) not
shared with any other spiders, and the Mesothelae lineage
traces back as far as the Carboniferous, the origin of Liphistiidae and its diversification are surprisingly much more recent
than once thought, repeating the patterns recently detected
also in other textbook living fossils [1,4]. We trace the origin
of the family in Southeast or East Asia to the Late Palaeogene
and Eocene (39– 58 Ma), and the origins of all eight genera to
the Late Oligocene and Neogene (figure 3a; electronic supplementary material, figure S1 and table S3). However,
liphistiids do belong to a very long evolutionary branch that
connects them with the only mesothelean fossil known from
Eurasia (figure 3a), and consequently, some of their
phenotypic traits resemble the hypothetically ancestral spiders. In the current absence of any additional fossil data, this
long phylogenetic branch suggests that mesothelean evolution
must account for a very slow eastward move with possibly
numerous extinction events. Below, we discuss three possible
scenarios and suggest that the eventual discovery of new
mesothelean fossils may support one over the other, depending on its location. Finally, we align liphistiid natural history,
in particular their sedentary lifestyle (figure 1c,d), to the emerging phylogenetic and biogeographic patterns to reveal that
they apparently lack over-water dispersal ability, and this
makes them ideal biogeographic models.
(a) Phylogeny and dating analysis
We provided the first phylogenetic hypothesis for the family
Liphistiidae, with a particular focus on the genera and species
groups within Heptathelinae. We used exemplars obtained
through our several years of sampling throughout their
range, obtained data on five mitochondrial and nuclear markers, and analysed them along with numerous mygalomorph
outgroups using three phylogenetic approaches (BI inference,
ML and parsimony) under four partition schemes. The results
(figures 2 and 3a) have converged on the following general
patterns: (i) the monophyly of the family Liphistiidae is well
supported; (ii) the genus Liphistius is the sister group to
Heptathelinae, the latter containing all other liphistiids;
(iii) within Heptathelinae, Ryuthela is monophyletic but the
genus ‘Heptathela’ s.l. as currently known is paraphyletic.
Proc. R. Soc. B 282: 20142486
14
Cambodia
Indochina
Sibumasu
Vinathela
13 **
I
Hong Kong
Laos
Hainan
12
20.2
**
17.7
Ryukyu Islands
Taiwan
Thailand
**
* * 16.2
27.9
15.4
Fujian
Vietnam
Myanmar
10
9
Liangxi
South China
** 22.9
1
Zhejiang
Guizhou
WB
48 **
Kyushu
Chongqing
Songthela
11
Shandong
Henan
Hubei
Sichuan
Yunnan
32.9 * **
12.4
3
Shannxi
rspb.royalsocietypublishing.org
**PP > 95%
* PP > 50%
6
‘Silk road’
Araneomorphs
291
Downloaded from http://rspb.royalsocietypublishing.org/ on May 6, 2015
Mesothelid spiders are sedentary, dispersal-limited spiders
whose evolutionary history dates back to the very origin of
spiders in the Late Carboniferous. They have therefore already
attracted considerable biogeographical interest [9,33,64–66],
but our view is that they should play a more prominent role
in future reconstructions of biogeographical histories. As all
prior hypotheses have been devoid of molecular data, the
here presented phylogenetic framework and the inferred
chronogram (figures 2 and 3a) shed new light on the origin
and biogeography of the group. Interestingly, the reconstructed ancestral areas (electronic supplementary material,
figure S2) and dispersal versus vicariant events (electronic supplementary material, table S5) do not require any explanations
of over-water dispersal in liphistiid biogeographic history.
Apparently, effective water barriers include not only bodies
of ocean, but also major rivers, notably Yangtze.
The available fossil evidence supports the ‘Euramerican
origin hypothesis’ for Mesothelae (figure 3b inset; [13,33]).
Our various dating analyses agree that the spider origin, and
thus the basal-most split between Mesothelae and Opisthothele
was in the Late Carboniferous and Early Permian, approximately 295–320 Ma (figure 3a; electronic supplementary
7
Proc. R. Soc. B 282: 20142486
(b) Historical biogeography
material figure S1 and table S3). During the Late Carboniferous
and Permian, the Laurentia and the Baltica plates together
formed the equatorially situated Euramerica [28,28,31,67].
It was there that the only known Mesothelae fossil lived
around 295 Ma [23,24]. By contrast, the regions inhabited
today by extant liphistiids (Southeast and East Asia) correspond to isolated microplates of that time, and are not
known to contain any Palaeozoic arachnid fossils. Therefore,
all evidence points to the origin of Mesothelae in Euramerica.
The long evolutionary history from the Mesothelae origin
in Euramerica to modern time is estimated at about 298 Ma.
Prior to our dating analysis, the node Liphistiidae was traditionally viewed as ‘primitive’ and ancient, but in fact our dating
analysis places its origin at ‘only’ approximately 48 Ma, and
therefore, recovers an extraordinarily long (approx. 250 Ma)
branch between the time of the mesothelean fossil (295 Ma)
and the origin of Liphistiidae. Even the alternative analysis
that only used fossil data unconstrained for mtDNA substitution rates uncovered a relatively recent origin of Liphistiidae
at approximately 124 Ma, implying a minimum estimate of
the mesothelean branch of 177 Ma. This means that liphistiid
ancestors must have arrived east over Laurasia (or Gondwana,
see below) over a long geological time.
The available palaeogeographical and palaeoclimatological data [28,29,31,52,67–71] allow for three possibilities of
such a long journey over land from Laurasia to Asia (figure
3b; see electronic supplementary material): (i) the liphistiid
ancestors may have taken a southern route along the northern margins of Gondwana, to which Euramerica connected
during the Late Carboniferous [33], then drifted north
towards Southeast Eurasia on Sibumasu, i.e. eastern Burma,
western Thailand, northwestern peninsular Malaysia and a
part of Sumatra [68 –70,72 –74]. We name this scenario the
‘Out of Gondwana’ hypothesis; (ii) they may have taken a
route along the southern margin of Laurasia by stepping
onto the Cimmerian continent strip [33]. This is the ‘Stepping-on Middle East’ hypothesis (figure 3b, inset); and
finally (iii) they may have travelled through northern Laurasia and on to the North China plate after it had connected to
Laurasia during the Mid-Jurassic. We name this the ‘Silk
road’ hypothesis (figure 3b, inset). Wherever the over-land
dispersal took the liphistiid ancestors during this long evolutionary time, they (along with any branches they may
have formed) must have repeatedly gone extinct along the
way. Finding new fossil Mesothelae representatives from
the Late Carboniferous to Eocene is therefore expected, but
depending on the route taken, these fossils could be on
either of the continental masses of Gondwana, Central and
North Asia, Sibumasu or China. See electronic supplementary material for discussion on plausibility of these
hypotheses and their predictions.
Liphistiidae diverged in the Palaeogene and diversified
during the Neogene (figure 3a; electronic supplementary
material figure S1 and table S3). The estimated diversification
times and ancestral area reconstructions (figure 3a; electronic
supplementary material figures S1 and S2, tables S3 and S5)
suggest that the origins of Heptathela s.s. Qiongthela, and
Ryuthela, the three genera exclusively inhabiting the southwestern Japanese islands and Hainan Island, predate the break-up
of the islands form the mainland estimated at 15 Ma for Japan
and approximately 2–2.5 Ma for Hainan [53,55,56,75–77]. Our
results thus support Schwendinger’s hypothesis [33], and
refute the explanation of liphistiids dispersing onto Japanese
rspb.royalsocietypublishing.org
Liphistiid monophyly has never been questioned, and the
clade has been recovered in all morphological and molecular
analyses that focused on higher level phylogenetics [7–9,
14,15,17–20,63]. However, these studies only included a
small subset of liphistiid diversity, typically one Liphistius
and/or one Heptathela [7,8,18 –20]. Our study is the first to
test and confirm liphistiid monophyly with a comprehensive
taxon sampling and using molecular data.
Two distinct clades can be identified within Liphistiidae:
Liphistiinae and Heptathelinae. Liphistiinae contains only
Liphistius whose monophyly has not been disputed [9,13].
Our dating analysis estimates the crown group Liphistius to
be approximately 17 Ma, but its stem may be as old as
approximately 48 Ma. Its sister lineage, Heptathelinae,
which contains all other liphistiids from China, Japan and
Vietnam, originated during the Eocene/Oligocene. Our
results unequivocally confirm the monophyly of Ryuthela in
agreement with morphological [9,13] and preliminary molecular datasets [22]. However, all phylogenetic evidence
refutes the monophyly of ‘Heptathela’ s.l., contradicting most
recent taxonomic literature [13,64].
The monophyly of each of the mainland Chinese genera
Songthela, Vinathela, Sinothela, and in part, Ganthela is well
supported in the phylogeny, and also by the dating analysis
and morphological diagnostics [51]. The exception is the suboptimal clade support for Ganthela in the parsimony analyses,
and poorly resolved relationships within Songthela in most
analyses. The apparent conflict in the data may be due to
inadequate taxonomic sampling in south and southwest
China. The position of the genus Qiongthela from Hainan
remains unresolved. With relatively low support, the phylogenetic analyses place it as sister to all mainland taxa
(figure 2), but the dating analysis places it as sister to the
doublet Ryuthela and Heptathela s.s. the remaining two island
genera (figure 3a; electronic supplementary material, table S5).
More sampling and data are needed to resolve this topological
instability for the three genera that may be crucial for the
understanding of liphistiid island biogeographic patterns.
Downloaded from http://rspb.royalsocietypublishing.org/ on May 6, 2015
Our results combined with the fossil record indicate that
Mesothelae originated in Euramerica in the Carboniferous
and Permian boundary, and suggest that the period that followed was marked by a long eastward over-land dispersal of
liphistiid ancestors towards the origin of Liphistiidae in
Southeast or East Asia. The notable pattern seen is an up to
250-Ma-long lag between the stem of Mesothelae and the
origin of extant Liphistiidae, combined with the occurrence
of likely extinction events during the evolutionary history
of the family. Since their origin in the Palaeogene Asia is relatively recent, we argue that liphistiids are modern spiders
that happened to have retained certain characters considered
plesiomorphic within spiders. As such, their labelling as
living fossils is accurate from the phenotypical stance, however, their diversification is much more recent than expected.
Organismal dispersal abilities profoundly affect their
diversification patterns [83] and biogeographic work benefits
from identifying such lineages that due to their ancient origins
and limited dispersal abilities retain their genetic imprint and
biogeographic signal [84]. The family Liphistiidae should join
Data accessibility. DNA sequences can be accessed via GenBank
(KP229800-KP230400); phylogenetic matrices can be accessed via
Dryad: http://dx.doi.org/10.5061/dryad.b8d6m; and all other datasets supporting this study are made available as part of the electronic
supplementary material.
Acknowledgements. We thank numerous people for local assistance, help
and advice: Gary Ades, Paul Crow, Yorkie Wong and Zoie Wong in
Hong Kong, Xianjin Peng, Luyu Wang, Bo Wu, Chengqiong Wu,
Tingbang Yang and Zizhong Yang in China, Chu Thi Thao and
Neuyen Thi Dinh in Vietnam, Zolta´n Korso´s, Mamoru Toda and Bo
Wu in Japan. We thank Ingi Agnarsson, Danwei Huang, Wei Shong
Hwang, Nina Vidergar and Matjazˇ Gregoricˇ for kind help and/or
advice on molecular resources and data analyses, Peter Ja¨ger for providing information on collection locality in Laos, Gonzalo Giribet for
commenting on an early draft, and Reta Mehta as well as two anonymous reviewers for their insightful comments. Finally, we thank the
staff of the Centre for Behavioural Ecology and Evolution (Hubei
University) and the Behavioural Ecology and Sociobiology Lab
(National University of Singapore) for support, in particular, Zhanqi
Chen, Seok Ping Goh, Xiaoguo Jiao, Hongze Li, Jie Liu, Yu Peng,
Xiaoyan Wang, Chen Xu, Long Yu and Zengtao Zhang.
Funding statement. This work was supported in part by the NSFC (grant
no. 31272324) and the Singapore Ministry of Education (AcRF Tier 1
grant no. R-154-000-591-112) grants to D.L., and by the Slovenian
Research Agency grant nos. P1-10236 and MU-PROM/12-001 to M.K.
Authors’ contributions. D.L., M.K. and X.X. designed this study; D.L.,
M.K., F.X.L., H.O., D.S.P., Y.N-R., X.X., X.X. and Z.S.Z. conducted
fieldwork; and X.X., R.-C.C., M.A.A., D.L. and M.K. performed
data analyses. All the authors contributed to the writing, revising
of the manuscript and gave final approval for publication.
References
1.
2.
3.
4.
5.
6.
Nagalingum NS, Marshall CR, Quental TB, Rai HS,
Little DP, Mathews S. 2011 Recent synchronous
radiation of a living fossil. Science 334, 796 –799.
(doi:10.1126/science.1209926)
Norstog K, Nicholls TJ. 1997 The biology of the
cycads. Ithaca, NY: Comstock Pub. Associates.
Watson J, Cusack HA. 2005 Cycadales of the
English Wealden. Palaeontogr. Soc. Monogr. 622,
1– 189.
Inoue JG, Miya M, Venkatesh B, Nishida M. 2005
The mitochondrial genome of Indonesian coelacanth
Latimeria menadoensis (Sarcopterygii:
Coelacanthiformes) and divergence time estimation
between the two coelacanths. Gene 349, 227–235.
(doi:10.1016/j.gene.2005.01.008)
Bristowe WS. 1975 A family of living fossil spiders.
Endeavour 34, 115– 117. (doi:10.1016/0160-9327
(75)90130-1)
Pocock RI. 1892 XXXVIII. - Liphistius and its bearing
upon the classification of spiders. Ann. Mag. Nat
Hist. Ser. 6 10, 306–314. (doi:10.1080/
00222939208677416)
7.
8.
9.
10.
11.
12.
13.
Platnick NI, Gertsch WJ. 1976 The suborders of
spiders: a cladistic analysis (Arachnida, Araneae).
Am. Mus. Nov. 2607, 1–15.
Coddington JA, Levi HW. 1991 Systematics and
evolution of spiders (Araneae). Annu. Rev. Ecol. Syst.
22, 565– 592. (doi:10.1146/annurev.es.22.110191.
003025)
Haupt J. 2003 The Mesothelaea—monograph of an
exceptional group of spiders (Araneae: Mesothelae)
(morphology, behaviour, ecology, taxonomy,
distribution and phylogeny). Zoologica 154, 1–102.
World Spider Catalog. 2015 World spider catalog. version
16. Bern, Switzerland: Natural History Museum. See
http://wsc.nmbe.ch (accessed 13 April 2015).
Bristowe WS. 1976 A contribution to the knowledge
of liphistiid spiders. J. Zool. Lond. 178, 1 –6.
(doi:10.1111/j.1469-7998.1976.tb02260.x)
Schio¨dte JC. 1849 Om en afigende sloegt af
spindlernes orden. Naturhistorisk Tidsskr. 2,
617 –624.
Schwendinger PJ, Ono H. 2011 On two Heptathela
species from southern Vietnam, with a discussion
14.
15.
16.
17.
18.
of copulatory organs and systematics of the
Liphistiidae (Araneae: Mesothelae). Rev. Suisse Zool.
118, 599–637.
Haupt J. 1983 Vergleichende Morphologie der
Genitalorgane und Phylogenie der liphistiomorphen
Webspinnen (Araneae: Mesothelae). I. Revision der
bisher bekannten Arten. Z. Zool. Syst. Evol. Forsch.
21, 275 –293. (doi:10.1111/j.1439-0469.1983.
tb00296.x)
Haupt J. 1984 Comportement sexuel, morphologie
ge´nitale et phylogene`se des araigne´es
liphistiomorphes. Rev. Arachnol. 5, 161 –168.
Platnick N, Goloboff PA. 1985 On the monophyly
of the spider suborder Mesothelae (Arachnida:
Araneae). New York Entomol. Soc. 93, 1265–1270.
Kraus O, Kraus M. 1993 Divergent transformation of
chelicerae and original arrangement of eyes in
spiders (Arachnida, Araneae). Mem. Queensl. Mus.
33, 579 –584.
Agnarsson I, Coddington JA, Kuntner M. 2013
Systematics—progress in the study of spider
diversity and evolution. In Spider research in the
8
Proc. R. Soc. B 282: 20142486
5. Conclusion
other prominent litter or soil-dwelling spiders [40,85–88] and
be considered an excellent model for biogeographic research
on a par with selected few phylogenetically old terrestrial
arthropod lineages, such as harvestmen [89], scorpions [90],
centipedes [91] and velvet worms [84]. Both the reconstructed
biogeographic history of Liphistiidae and their natural history
namely suggest that these spiders are very poor and slow
dispersers that, rather than ever move over-water or over
short-lived land bridges, prefer to maintain their restricted
ranges, or ride vicariantly on drifting continents.
rspb.royalsocietypublishing.org
islands over land bridges during the Pleistocene [9]. Likewise,
the occurrence of Qiongthela on Hainan Island requires no ad
hoc over-water dispersal events.
We can now also explain the absence of liphistiids from
Taiwan. The island namely formed de novo about 9 Ma
during the collision of the Philippine Sea plate with the
Eurasian plate [78]. In accordance with their sedentary terrestrial life history and an over 298 Ma long evolutionary history
of slow, over-land dispersal, the best explanation at hand is
that liphistiids have never set their tarsi on Taiwan, not
even during relatively short-lived land bridges that formed
between the southern Japanese islands and the Asian
mainland during the Pleistocene glacial cycles [79–82].
Downloaded from http://rspb.royalsocietypublishing.org/ on May 6, 2015
20.
22.
23.
24.
25.
26.
27.
28.
29.
30.
31.
32.
33.
34.
35.
37.
38.
39.
40.
41.
42.
43.
44.
45.
46.
47.
48.
49. Nylander JAA, Olsson U, Alstrom P, Sanmartı`n I.
2008 Accounting for phylogenetic uncertainty in
biogeography: a Bayesian approach to dispersal –
vicariance analysis of the thrushes (Aves: Turdus).
Syst. Biol. 57, 257–268. (doi:10.1080/1063515
0802044003)
50. Ree RH, Smith SA. 2008 Maximum likelihood
inference of geographic range evolution by dispersal
local extinction and cladogenesis. Syst. Biol. 57,
4–14. (doi:10.1080/10635150701883881)
51. Xu X, Liu FX, Chen J, Ono H, Li D, Kuntner M. 2015 A
genus level taxonomic revision of primitively
segmented spiders (Mesothelae: Liphistiidae). Zookeys
488, 121–151. (doi:10.3897/zookeys.488.8726)
52. Metcalfe I. 1998 Palaeozoic and Mesozoic geological
evolution of the SE Asian region: multidisciplinary
constraints and implications for biogeography. In
Biogeography and geological evolution of SE Asia
(eds R Hall, JD Holloway), pp. 25 –41. Leiden,
The Netherlands: Backhuys Publishers.
53. Otofuji Y, Matsuda T, Nohda S. 1985 Opening mode
of the Japan Sea inferred from paleomagnetism of
the Japan arc. Nature 317, 603– 604. (doi:10.1038/
317603a0)
54. Otofuji Y, Itaya T, Matsuda T. 1991 Rapid rotation of
Southwest Japan: paleomagnetism and K-Ar ages of
Miocene volcanic rocks of southwest Japan.
Geophys. J. Intern. 105, 397–405. (doi:10.1111/j.
1365-246X.1991.tb06721.x)
55. Yin A. 2010 Cenozoic tectonic evolution of Asia: a
preliminary synthesis. Tectonophysics 488,
293–325. (doi:10.1016/j.tecto.2009.06.002)
56. Kimura M. 2000 Paleogeography of the Ryukyu
Islands. Tropics 10, 5 –24. (doi:10.3759/tropics.10.5)
57. Kimura M. 2002 The formation age of the Ryukyu
Arc and migration of biota to the arc. Naha,
Okinawa: Okinawa Times Co.
58. Kimura M. 2003 Land connections between Eurasian
continent and Japanese islands—related to human
migration. Mig. Diff. 4, 14– 33.
59. Ota H. 1998 Geographic patterns of endemism and
speciation in amphibians and reptiles of the Ryukyu
Archipelago, Japan, with special reference to their
paleogeographical implications. Res. Popul. Ecol. 40,
189–204. (doi:10.1007/BF02763404)
60. Ota H. 2000 The current geographic faunal patterns
of reptiles and amphibians of the Ryukyu
Archipelago and adjacent regions. Tropics 10,
51– 62. (doi:10.3759/tropics.10.51)
61. Otsuka H, Takahashi A. 2000 Pleistocene vertebrate
faunas in the Ryukyu Islands: their migration and
extinction. Tropics 10, 25– 40. (doi:10.3759/tropics.
10.25)
62. Zheng HB, Clift PD, Wang P, Tada R, Jia JT, He MY,
Jourdan F. 2013 Pre-Miocene birth of the Yantze
River. Proc. Natl Acad. Sci. USA 110, 7556 –7561.
(doi:10.1073/pnas.1216241110)
63. Bond JE, Hendrixson BE, Hamilton CA, Hedin M.
2012 A reconsideration of the classification of the
spider infraorder Mygalomorphae (Arachnida:
Araneae) based on three nuclear genes and
morphology. PLoS ONE 7, e38753. (doi:10.1371/
journal.pone.0038753)
9
Proc. R. Soc. B 282: 20142486
21.
36.
sequence datasets under the maximum likelihood
criterion. PhD thesis, The University of Texas
at Austin.
Ronquist F et al. 2012 MrBayes 3.2: efficient
Bayesian phylogenetic inference and model choice
across a large model space. Syst. Biol. 61, 539– 542.
(doi:10.1093/sysbio/sys029)
Darriba D, Taboada GL, Doallo R, Posada D. 2012
jModelTest 2: more models, new heuristics and
parallel computing. Nat. Methods 9, 772. (doi:10.
1038/nmeth.2109)
Bond J, Hedin M, Ramirez M, Opell B. 2001 Deep
molecular divergence in the absence of
morphological and ecological change in the
Californian coastal dune endemic trapdoor spider
Aptostichus simus. Mol. Evol. 10, 899–910. (doi:10.
1046/j.1365-294X.2001.01233.x)
Macı´as-Herna´ndez NE, Oromı´ P, Arnedo MA. 2010
Integrative taxonomy uncovers hidden species
diversity in woodlouse hunter spiders (Araneae,
Dysderidae) endemic to the Macaronesian
archipelagos. Syst. Biod. 8, 531–553. (doi:10.1080/
14772000.2010.535865)
Bidegaray-Batista L, Arnedo MA. 2011 Gone
with the plate: the opening of the Western
Mediterranean basin drove the diversification of
ground-dweller spiders. BMC Evol. Biol. 11, 317.
(doi:10.1186/1471-2148-11-317)
Planas E, Ribera C. 2014 Uncovering overlooked
island diversity: colonization and diversification of
the medically important spider genus Loxosceles
(Arachnida: Sicariidae) on the Canary Islands.
J. Biogeogr. 41, 1255 –1266. (doi:10.1111/
jbi.12321)
Bidegaray-Batista L, Ferra´ndez MA´, Arnedo MA. 2014
Winter is coming: Miocene and Quaternary climatic shifts
shaped the diversification of Western-Mediterranean
Harpactocrates (Araneae, Dysderidae) spiders. Cladistics
30, 428–446. (doi:10.1111/cla.12054)
Brower AVZ. 1994 Rapid morphological radiation and
convergence among races of the butterfly Heliconius
erato inferred from patterns of mitochondrial DNA
evolution. Proc. Natl Acad. Sci. USA 91, 6491–6495.
(doi:10.1073/pnas.91.14.6491)
Opatova V, Arnedo MA. 2014 From Gondwana to
Europe: inferring the origins of Mediterranean
Macrothele spiders (Araneae, Hexathelidae) and the
limits of the family Hexathelidae. Invert. Syst. 28,
361 –374.
Drummond AJ, Rambaut A. 2007 BEAST: Bayesian
evolutionary analysis by sampling trees. BMC Evol.
Biol. 7, 214. (doi:10.1186/1471-2148-7-214)
Drummond AJ, Suchard MA, Xie D, Rambaut A.
2012 Bayesian phylogenetics with BEAUti and the
BEAST 1.7. Mol. Biol. Evol. 29, 1969 –1973. (doi:10.
1093/molbev/mss075)
Yu Y, Harris AJ, He XJ. 2010 S-DIVA (Statistical
Dispersal-Vicariance Analysis): a tool for inferring
biogeographic histories. Mol. Phylogenet. Evol. 56,
848 –850. (doi:10.1016/j.ympev.2010.04.011)
Yu Y, Harris AJ, He XJ. 2014 RASP (reconstruct
ancestral state in phylogenies) 3.0. See http://mnh.
scu.edu.cn/soft/blog/RASP.
rspb.royalsocietypublishing.org
19.
21st century (ed. D Penney), pp. 58 –111.
Manchester, UK: Siri Scientific Press.
Bond JE, Garrison NL, Hamilton CA, Godwin RL,
Hedin M, Agnarsson I. 2014 Phylogenomics resolves
a spider backbone phylogeny and rejects a
prevailing paradigm for orb web evolution. Curr.
Biol. 24, 1765 –1771. (doi:10.1016/j.cub.2014.
06.034)
Ferna´ndez R, Hormiga G, Giribet G. 2014
Phylogenomic analysis of spiders reveals
nonmonophyly of orb weavers. Curr. Biol. 24,
1772–1777. (doi:10.1016/j.cub.2014.06.035)
Sharma PP, Kaluziak ST, Pe´rez-Porro AR, Gonza´lez VL,
Hormiga G, Wheeler WC, Giribet G. 2014
Phylogenomic interrogation of Arachnida reveals
systemic conflicts in phylogenetic signal. Mol. Biol.
Evol. 31, 2963–2984. (doi:10.1093/molbev/msu235)
Tanikawa A. 2013 Phylogeny and genetic variation
in the spiders of the genus Ryuthela (Araneae:
Liphistiidae). Acta Arachnol. 62, 41 –49. (doi:10.
2476/asjaa.62.41)
Selden PA. 1996 Fossil mesothele spiders. Nature
379, 498–499. (doi:10.1038/379498b0)
Selden PA. 1996 First fossil mesothele spider, from
the Carboniferous of France. Rev. Suisse Zool. 2,
585–596.
McLoughlin S. 2001 The breakup history of
Gondwana and its impact on pre-Cenozoic floristic
provincialism. Aust. J. Bot. 49, 271 –300. (doi:10.
1071/BT00023)
Scotese CR. 2002 Plate tectonic maps and
continental drift animations. PALEOMAP Project.
See www.scotese.com/earth.htm (accessed
11 October 2014).
Rogers JJW, Santosh M. 2003 Supercontinents in
earth history. Gondwana Res. 6, 375. (doi:10.1016/
S1342-937X(05)70993-X)
Metcalfe I. 2002 Permian tectonic framework and
palaeogeography of SE Asia. J. Asian Earth Sci. 20,
551–566. (doi:10.1016/S1367-9120(02)00022-6)
Metcalfe I. 2011 Tectonic framework and
Phanerozoic evolution of Sundaland. Gondwana Res.
19, 3–21. (doi:10.1016/j.gr.2010.02.016)
Seton M et al. 2012 Global continental and
ocean basin reconstructions since 200 Ma. Earth
Sci. Rev. 113, 212– 270. (doi:10.1016/j.earscirev.
2012.03.002)
Metcalfe I. 2013 Gondwana dispersion and Asian
accretion: tectonic and palaeogeographic evolution
of eastern Tethys. J. Asian Earth Sci. 66, 1–33.
(doi:10.1016/j.jseaes.2012.12.020)
Torsvik TH, Cocks LRM. 2013 Gondwana from top
to base in space and time. Gondwana Res. 24,
999–1030. (doi:10.1016/j.gr.2013.06.012)
Schwendinger PJ. 2009 Liphistius thaleri, a new
mesothelid spider species from southern Thailand
(Araneae, Liphistiidae). Contrib. Nat. Hist. 12,
1253 –1268.
Goloboff PA, Farris JS, Nixon KC. 2008 TNT, a free
program for phylogenetic analysis. Cladistics 24,
774–786. (doi:10.1111/j.1096-0031.2008.00217.x)
Zwickl DJ. 2006 Genetic algorithm approaches for
the phylogenetic analysis of large biological
Downloaded from http://rspb.royalsocietypublishing.org/ on May 6, 2015
75.
76.
77.
79.
80.
81.
82.
83.
84.
85. Hendrixson B, Bond J. 2007 Molecular phylogeny
and biogeography of an ancient Holarctic lineage of
mygalomorph spiders (Araneae: Antrodiaetidae:
Antrodiaetus). Mol. Phylogenet. Evol. 42, 738 –755.
(doi:10.1016/j.ympev.2006.09.010)
86. Hedin M, Starrett J, Hayashi C. 2013 Crossing the
uncrossable: novel trans-valley biogeographic
patterns revealed in the genetic history of lowdispersal mygalomorph spiders (Antrodiaetidae,
Antrodiaetus) from California. Mol. Ecol. 22,
508–526. (doi:10.1111/mec.12130)
87. Opatova V, Bond JE, Arnedo MA. 2013 Ancient
origins of the Mediterranean trap-door spiders of
the family Ctenizidae (Araneae, Mygalomorphae).
Mol. Phylogenet. Evol. 69, 1135–1145. (doi:10.
1016/j.ympev.2013.08.002)
88. Wood HM, Matzke NJ, Gillespie RG, Griswold CE.
2013 Treating fossils as terminal taxa in divergence
time estimation reveals ancient vicariance patterns
in the palpimanoid spiders. Syst. Biol. 62,
264–284. (doi:10.1093/sysbio/sys092)
89. Giribet G, Sharma PP, Benavides LR, Boyer SL,
Clouse RM, de Bivort BL, Kawauchi GY, Murienne J,
Schwendinger PJ. 2012 Evolutionary and
biogeographic history of the harvestman suborder
Cyphophthalmi (Arachnida, Opiliones)—an ancient
and global group of arachnids. Biol. J. Linn. Soc.
105, 92– 130. (doi:10.1111/j.1095-8312.2011.
01774.x)
90. Bryson Jr RW, Savary WE, Prendini L. 2013
Biogeography of scorpions in the Pseudouroctonus
minimus complex (Vaejovidae) from south-western
North America: implications of ecological
specialization for pre-Quaternary diversification.
J. Biogeogr. 40, 1850 –1860.
91. Giribet G, Edgecombe GD. 2012 Reevaluating the
arthropod tree of life. Annu. Rev. Entomol. 57,
167–186. (doi:10.1146/annurev-ento-120710-100659)
10
Proc. R. Soc. B 282: 20142486
78.
harvestmen (Opiliones, Cyphophthalmi, Stylocellidae)
in Southeast Asia. J. Biogeogr. 37, 1114–1130.
(doi:10.1111/j.1365-2699.2010.02274.x)
Zeng ZX, Zeng XZ. 1989 Physical geography of
Hainan Island. Beijing, China: Science Press.
Zhao HT, Wang LR, Yuan JY. 2007 Origin and time
of Qiongzhou Strait. Marine Geol. Quat. Geol. 27,
33 –40.
Huang Y, Guo X, Ho SY, Shi H, Li J, Li J, Cai B, Wang
Y. 2013 Diversification and demography of the
oriental garden lizard (Calotes versicolor) on Hainan
Island and the adjacent mainland. PLoS ONE 8,
e64754. (doi:10.1371/journal.pone.0064754)
Sibuet JC, Hsu SK. 2004 How was Taiwan created?
Tectonophysics 379, 159–181. (doi:10.1016/j.tecto.
2003.10.022)
Wageman JM, Hilde TMC, Emery KO. 1970 Structural
framework of East China Sea and Yellow Sea. Am.
Assoc. Pet. Geol. Bull. 54, 1611–1643.
Ye XQ. 1982 On the formation and development
of the geology and geomorphology of Taiwan.
J. Central China Teachers College 1982, 83 –89.
Juan VCC. 1986 Thermal-tectonic evolution of the
Yellow sea and East China Sea—Implication for
transformation of continental to oceanic crust and
marginal basin formation. Tectonophysics 125,
231 –244. (doi:10.1016/0040-1951(86)90016-8)
Teng LS. 1992 Geotectonic evolution of Tertiary
continental margin basins of Taiwan. Petrol. Geol.
Taiwan 27, 1 –19.
Agnarsson I, Cheng RC, Kuntner M. 2014 A multiclade test supports the intermediate dispersal
model of biogeography. PLoS ONE 9, e86780.
(doi:10.1371/journal.pone.0086780)
Murienne J, Daniels SR, Buckley TR, Mayer G, Giribet
G. 2014 A living fossil tale of Pangaean
biogeography. Proc. R. Soc. B 281, 20132648.
(doi:10.1098/rspb.2013.2648)
rspb.royalsocietypublishing.org
64. Ono H. 2000 Zoogeographic and taxonomic notes
on spiders of the subfamily Heptathelinae (Araneae,
Mesothelae, Liphistiidae). Mem. Natl Sci. Mus. 33,
145–151.
65. Paik K. 1953 A study on the geographical
distribution of Heptathela kimurai Kishida. Acta
Arachnol. 13, 53 –68. (doi:10.2476/asjaa.13.63)
66. Haupt J. 2003 Zoogeography in southern Japan as
revealed by ground-living arachnids. Rev. Suisse
Zool. 110, 133–139.
67. Cox CB, Moore PD. 2005 Biogeography: an ecological
and evolutionary approach, 7th edn. Malden, MA:
Blackwell Publishing.
68. Ali JR, Aitchison JC. 2005 Greater India. Earth
Sci. Rev. 72, 169–188. (doi:10.1016/j.earscirev.
2005.07.005)
69. Ali JR, Aitchison JC. 2006 Positioning Paleogene
Eurasia problem: solution for 60 –50 Ma and
broader tectonic implications. Earth Planet. Sci. Lett.
251, 148–155. (doi:10.1016/j.epsl.2006.09.003)
70. Ali JR, Aitchison JC. 2008 Gondwana to Asia: plate
tectonics, paleogeography and the biological
connectivity of the Indian sub-continent from the
Middle Jurassic through latest Eocene
(166–35 Ma). Earth Sci. Rev. 88, 145– 166. (doi:10.
1016/j.earscirev.2008.01.007)
71. Torsvik TH, Cocks LRM. 2004 Earth geography from
400 to 250 Ma: a palaeomagnetic, faunal and facies
review. J. Geol. Soc. Lond. 161, 555–572. (doi:10.
1144/0016-764903-098)
72. Golonka J, Krobicki M, Pajak J, Giang NV, Zuchiewicz
W. 2006 Phanerozoic palaeogeography of Southeast
Asia. Geolines 20, 40 –43.
73. Golonka J. 2007 Phanerozoic paleoenvironment and
paleolithofacies maps. Mesozoic. Geologia 33,
211–264.
74. Clouse RM, Giribet G. 2010 When Thailand was an
island—the phylogeny and biogeography of mite