Phylogenetic position and composition of Zygiellinae and … · 2019-01-27 · Phylogenetic position and composition of Zygiellinae and Caerostris, with new insight into orb-web evolution

Phylogenetic position and composition of Zygiellinaeand Caerostris, with new insight into orb-web evolutionand gigantism

MATJAŽ GREGORIC1,2*, INGI AGNARSSON3,4, TODD A. BLACKLEDGE2 andMATJAŽ KUNTNER1,4,5

1Institute of Biology, Scientific Research Centre, Slovenian Academy of Sciences and Arts, Novi trg 2,P. O. Box 306, SI-1001 Ljubljana, Slovenia2Integrated Bioscience Program, Department of Biology, University of Akron, Akron, OH 44325-3908,USA3Department of Biology, University of Vermont, Burlington, VT, USA4Department of Entomology, National Museum of Natural History, Smithsonian Institution,Washington, DC, USA5Centre for Behavioural Ecology and Evolution, College of Life Sciences, Hubei University, Wuhan,Hubei, China

Received 15 October 2014; revised 27 March 2015; accepted for publication 27 March 2015

Orb-weaving spiders are good objects for evolutionary research, but phylogenetic relationships among and withinorb-weaving lineages are poorly understood. Here we present the first species-level molecular phylogeny that in-cludes the enigmatic orb weavers ‘Zygiellidae’ and Caerostris. Zygiellidae is interesting for the evolution of thesector web, and Caerostris is noteworthy for web gigantism and extraordinary silk biomechanics. We assembled amolecular data set using mitochondrial (COI, 16S) and nuclear (H3, 18S, 28S, ITS2) gene fragments for 112 orbicularianexemplars, focusing on taxa with diverse web architecture and size. We show that ‘Zygiellidae’ contains the HolarcticZygiella genus group (Leviellus, Parazygiella, Stroemiellus, and Zygiella) and the Australasian Phonognatha andDeliochus. As this clade is placed with Araneidae in all analyses we treat it as a subfamily, Zygiellinae. Using thenew phylogeny, we show that the sector web evolved eight times, and coevolved with the silk tube retreat, butthat these features are not zygielline synapomorphies. Zygiellinae, Caerostris, and some other araneids form abasal grade of araneids that differ from ‘classical’ araneids in web-building and preying behaviour. We also confirmthat Caerostris represents the most striking case of spider-web gigantism.

© 2015 The Linnean Society of London, Zoological Journal of the Linnean Society, 2015, 175, 225–243.doi: 10.1111/zoj.12281

ADDITIONAL KEYWORDS: bark spider – Caerostris – sector web – web building – web gigantism – Zygiellidae– Zygiellinae.

INTRODUCTION

The spider group Orbiculariae is defined by the originof the orb web, and contains all orb-weaving taxa andtheir relatives. These include several radiations of taxawith diverse alternative web architecture (sheet, cob,

and tent webs) and taxa that have reduced webs intotriangles or bolas, or those that have even aban-doned web-building altogether (Griswold et al., 1998;Blackledge et al., 2009; Hormiga & Griswold, 2014).The classical Orbiculariae contains over 12 000 de-scribed species (Griswold et al., 1998; Blackledge et al.,2009; World Spider Catalog, 2015); however, themonophyly of Orbiculariae has been controversial inthe past decade (Hausdorf, 1999; Ayoub et al., 2007;*Corresponding author. E-mail: [email protected]

bs_bs_banner

Zoological Journal of the Linnean Society, 2015, 175, 225–243. With 4 figures

© 2015 The Linnean Society of London, Zoological Journal of the Linnean Society, 2015, 175, 225–243 225

mailto:[email protected]

Blackledge et al., 2009; Agnarsson, Coddington &Kuntner, 2013; Hormiga & Griswold, 2014), and recentrobust molecular evidence places the diverse clade ofmostly cursorial spiders, the ‘RTA-clade’, withinOrbiculariae (Bond et al., 2014; Fernández, Hormiga& Giribet, 2014), suggesting that the majority of spiderdiversity, some 30 000 species, are descendants of orb-weaving ancestors. Many orb weavers are routinely usedas models in studies of development, functional mor-phology and physiology, adaptive evolution, sexual se-lection, sexual size dimorphism, phylogeography,evolutionary ecology, and other fields (Coddington, 1994;Bond & Opell, 1998; Gillespie, 2004; Blackledge,Kuntner & Agnarsson, 2011; Herberstein & Wignall,2011). Furthermore, orb webs are directly measur-able ‘extended phenotypes’ of behaviour, and are thusideal objects for addressing a variety of topics likepredator–prey interactions (e.g. Eberhard, 1986; Craig,Weber & Bernard, 1996; Opell & Schwend, 2007),producer–consumer interactions (e.g. Robinson &Robinson, 1973; Rypistra, 1981; Elgar, 1989; Higgins& Buskirk, 1998; Agnarsson, 2003, 2011), behav-ioural plasticity (e.g. Eberhard, 1990; Watanabe, 2000;Blamires, 2010), resource use (Blackledge & Gillespie,2004; Gillespie, 2004), evolution of web-related behav-iour (e.g. Peters, 1937; Witt, Reed & Peakall, 1968;Eberhard, 1982; Vollrath & Selden, 2007; Kuntner &Agnarsson, 2009; Blackledge et al., 2012a), and otherbehavioural questions (e.g. Eberhard, 1982, 2007;Kuntner, Gregoric & Li, 2010; Gregoric et al., 2013).Moreover, because orb webs are made with nature’stoughest biomaterial and covered with glue, orb weaversare central objects of biomaterial research (e.g. Hayashi& Lewis, 2000; Agnarsson et al., 2009; Swanson et al.,2009; Vollrath & Porter, 2009; Agnarsson, Kuntner &Blackledge, 2010; Sensenig, Agnarsson & Blackledge,2010; Sahni, Blackledge & Dhinojwala, 2011; Blackledgeet al., 2012b; Vasanthavada et al., 2012; Grawe, Wolff& Gorb, 2014; Stellwagen, Opell & Short, 2014). Hence,a solid phylogenetic understanding of orbicularianspiders would benefit all of these biological disci-plines. Yet phylogenetic relationships among and withinmost orb-weaving families are poorly understood(Lopardo, Giribet & Hormiga, 2011; Agnarsson et al.,2013; Hormiga & Griswold, 2014), rendering evolu-tionary research on orb weavers less robust.

The majority of studies focusing on the above topicschoose orb webs as objects of study because of theirsimple architecture, superior material properties, andlarge diversity in size, shape, and resource use(Blackledge et al., 2011; Herberstein & Wignall, 2011).Most spiders building orb webs are grouped into thefamily Araneidae, the third largest spider family, con-taining more than 3000 species (World Spider Catalog,2015). Although sharing primitive orb web-building an-cestors, araneid spiders have greatly diversified mor-

phologically, behaviourally, and genetically (Dimitrovet al., 2012). Because the diversification of orb weaversis closely linked with the evolution of traits associat-ed with orb-web biology (Blackledge et al., 2011), wefocus on two araneid groups that have controversialsystematics and are interesting models for several evo-lutionary questions, including web biology: the generaZygiella Pickard-Cambridge, 1902, in the broad sense(hereafter termed Zygiella s.l.), and Caerostris Thorell,1868. Our aim is to build a more robust tool for evo-lutionary research through better-resolved relation-ships among orb-weaving groups.

Zygiella s.l. (containing Leviellus, Parazygiella,Stroemiellus, and Zygiella) spiders were among the firstobjects of studies focusing on the architecture, func-tion, and building of orb webs (e.g. Wiehle, 1927, 1929;Peters, 1937; Witt et al., 1968), and are still widely usedas objects of studies focusing on predator–prey inter-actions, sexual selection, behavioural plasticity, and web-building behaviour and physiology (e.g. Zschokke &Vollrath, 1995; Venner et al., 2000, 2003; Thevenard,Leborgne & Pasquet, 2004; Venner & Casas, 2005;Bel-Venner & Venner, 2006; Bel-Venner et al., 2008;Mayntz, Toft & Vollrath, 2009). The genus Zygiella s.l.contains mostly widespread Palaearctic species that typi-cally possess a characteristic orb-web feature: a spiral-free sector in the upper part of the orb with a signalline leading to the tubular silk retreat (Levi, 1974;Gregoric, Kostanjšek & Kuntner, 2010; Fig. 1).Zygiella s.l. is currently catalogued in Araneidae (WorldSpider Catalog, 2015), but was transferred between thefamilies Tetragnathidae (Levi, 1980; Heimer & Nentwig,1991; Wunderlich, 2004) and Araneidae (Levy, 1987;Roberts, 1995; Scharff & Coddington, 1997) in the past,and several species were transferred to other genera.Recently, Wunderlich (2004) split Zygiella s.l. into foursmaller genera and proposed them to belong toZygiellidae, uniting taxa that build a characteristic sectorweb (Fig. 1). According to Wunderlich (2004), this groupconsists of Leviellus, Parazygiella, the monotypicStroemiellus, Zygiella in the strict sense (hereafterZygiella), and Chrysometa, traditionally a tetragnathid.Despite the lack of phylogenetic evidence the new generaare catalogued (World Spider Catalog, 2015), but thefamily ‘Zygiellidae’ is generally not accepted (Hormiga& Griswold, 2014).

Several recent molecular phylogenies of araneoidspiders cast new doubt on the araneid affinity ofZygiella s.l. (Alvarez-Padilla et al., 2009; Sensenig et al.,2010; Dimitrov & Hormiga, 2011; Agnarsson et al., 2012;Blackledge et al., 2012a; Dimitrov et al., 2012; Kuntneret al., 2013). These analyses include only two Zygiellaspecies, Zygiella x-notata (Clerck, 1757) and/or Zygiellaatrica (C.L. Koch, 1845), and no representatives of theother three genera, and they establish a clade thatunites Zygiella with the Australasian leaf-curling

226 M. GREGORIC ET AL.

© 2015 The Linnean Society of London, Zoological Journal of the Linnean Society, 2015, 175, 225–243

araneids Deliochus and Phonognatha, with both generapreviously considered nephilines (Hormiga, Eberhard& Coddington, 1995; but see Kuntner, Coddington &Hormiga, 2008). They ambiguously place this cladeeither as sister to all other araneids or as sister to thefamily Nephilidae. Depending on how this ambiguity

is resolved adjustments to the classification may benecessary, with one option being to adopt a familyranking for ‘Zygiellidae’.

Until recently, bark spiders of the genus Caerostriswere grossly understudied ecologically, behaviourally,and systematically; however, this enigmatic genus

A B C D

E F G H

I J K

Figure 1. Webs of Zygiellinae, other sector-web taxa, and Caerostris: A, Zygiella x-notata, Slovenia; B, Deliochus sp.,Brisbane, Australia; C, Phonognatha graeffei, Australia; D, undetermined araneid from Halmahera, Indonesia; E, Araneusmitificus, Singapore; F, Dolicognatha sp., Yunnan, China; G, Acusilas coccineus, Gombok, Malaysia; H, Milonia sp., Yunnan,China; I, Guizygiella sp., Yunnan, China; J, Caerostris darwini webs bridging river in Ranomafana, Madagascar; K, C. darwiniweb over small stream in Andasibe-Mantadia, Madagascar.

ORB WEB EVOLUTION AND GIGANTISM 227


contains diverse, large orb weavers that are wide-spread throughout the Old World tropics (Grasshoff,1984), and exhibit peculiar web biology (Kuntner &Agnarsson, 2010). Caerostris spiders build the largestorb webs using the toughest silk (Agnarsson et al., 2010;Gregoric et al., 2011a), and some species even use aunique habitat by suspending their webs above largebodies of water, using a unique set of web-building be-haviours (Kuntner & Agnarsson, 2010; Gregoric et al.,2011b). Because of their gigantic webs, reaching 2 min diameter (Gregoric et al., 2011a), and made of ex-tremely tough silk, Caerostris spiders have the poten-tial to become models for biomaterial research.Additionally, the genus Caerostris represents a prom-ising object for the research of sexual size dimor-phism, and its correlated behaviours (Kuntner et al.,2008, 2013; Kuntner & Agnarsson, 2010).

With an outdated taxonomy (Grasshoff, 1984),however, and with the near absence of phylogeneticdata, bark spider systematics is largely unknown andconflicted. Whereas only 12 Caerostris species are de-scribed, a much larger diversity is expected to exist,e.g. in Madagascar (Kuntner & Agnarsson, 2010;Gregoric et al., 2011a). Morphological data placedCaerostris deep within araneids (Scharff & Coddington,1997), or as sister to Argiope (Kuntner et al., 2008),but molecular data instead recovered it as sister to allother araneids, excluding ‘zygiellids’ (Sensenig et al.,2010; Kuntner et al., 2013). For this reason, any con-sideration of Caerostris systematics depends upon arobust analysis of Zygiella s.l.

We provide a phylogenetic analysis of orbicularianfamilies, emphasizing orb-web building taxa to test themonophyly and phylogenetic placement of Zygiella s.l.(and its four genera) and Caerostris, and use that phy-logeny to understand the macroevolution of web sizeand the architectural traits that characterize these twogroups. Using two mitochondrial and four nuclear genefragments, our molecular analysis includes all fourZygiella s.l. genera, several former and potential zygiellidgenera, as well as several Caerostris species with anextensive out-group selection. We use the new phy-logeny to test whether the sector web indeed repre-sents a zygielline synapomorphy, to investigate theevolutionary interplay of the sector web and its po-tential functional correlate, an off-web silk tube retreat,to explore the evolution of web gigantism, and toexamine behavioural traits that may support the place-ment of Zygiellinae and Caerostris as early splits fromthe main Araneidae branch.

MATERIAL AND METHODSTAXONOMIC SAMPLING

As in-group taxa, we included all four Zygiella s.l. generawith at least two species per genus (except for the

monotypic Stroemiellus), summing up to nine of the17 described Zygiella s.l. species and two undescribedspecies. We also included Phonognatha and Deliochus,the two genera consistently recovered as sister toZygiella in recent analyses (Alvarez-Padilla et al., 2009;Sensenig et al., 2010; Dimitrov et al., 2012; Kuntneret al., 2013), as well as the tetragnathid Chrysometaalboguttata (O. Pickard-Cambridge, 1889), a memberof Zygiellidae sensu Wunderlich (2004). To inferthe phylogenetic position of Caerostris, we includedsix of the 12 currently described species, which wesampled in Southeast Asia, continental Africa, andMadagascar.

As out-group taxa, we included several species for-merly included in Zygiella s.l., such as the monotypicaraneid Yaginumia (Archer, 1960) and several speciesof the supposedly tetragnathid genus Guizygiella (Zhu,Kim & Song, 1997). Additionally, we included severalspecies with web biology resembling ‘zygiellids’, e.g.the leaf-curling Acusilas (Murphy & Murphy, 1983;Birkedal Schmidt & Scharff, 2008), as well as the sectorweb-building Milonia, Araneus mitificus (Simon, 1886),and several other araneids and tetragnathids. Becauseprevious molecular analyses (Sensenig et al., 2010;Dimitrov et al., 2012; Kuntner et al., 2013) infer bothCaerostris and Zygiella to be at the base of Araneidae,or to be associated with Nephilidae, we also includedtaxa that previous phylogenetic studies recovered atthese basal araneid nodes, e.g. Oarces, Gnolus, andMicrathena (Sensenig et al., 2010; Dimitrov et al., 2012),and some former or potential zygiellids currently be-longing to Tetragnathidae, as well as several othertetragnathid genera. In addition to our taxon choicethat is biased towards families that buildorb webs (especially Araneidae, Nephilidae, andTetragnathidae), we included all other orbicularian fami-lies, except Synaphridae, Sinopimoidae, andMicropholcommatidae. Altogether, our data set includ-ed 112 species from 75 genera and 18 families (Ap-pendices S1 and S4). We rooted the trees with the non-araneoid orbicularian Deinopis (Deinopidae).

MOLECULAR PROCEDURES

We isolated DNA from leg muscles using the DNeasyBlood and Tissue Kit (QIAGEN), following the proto-col for mammals. We amplified two mitochondrial [COIand 16S] and four nuclear [H3, 18S, 28S, ITS2] genefragments, which are among the standard genes ofchoice in spider phylogenetics. All polymerase chainreactions (PCRs) had a total volume of 25 μL and con-sisted of 13.1 μL of double-distilled H2O, 5 μL of 5× PCRbuffer ‘GoTaqFlexi’ (Promega), 2.25 μL of MgCl2 (25 mM;Promega), 0.15 μL of ‘5U GoTaqFlexi Polymerase’(Promega), 2.5 μL of ‘dNTP Mix’ (2 μM each; Biotools),0.5 μL of each forward and reverse 20-μM primers, and



1.5 μL of DNA. We included 0.15 μL of bovine serumalbumin (10 mg mL−1; Promega) to some reactions, anddecreased the H2O volume accordingly. We performedthe PCR amplifications using a ‘2720 Thermal Cycler’(Applied Biosystems) and a ‘Mastercycler® ep’(Eppendorf). Primer details and the different PCRprotocols with varying annealing temperatures and cyclesettings are summarized in Appendix S5.

PHYLOGENETIC INFERENCE

We aligned protein-coding genes (COI and H3) usingClustalW alignment, and checked for stop codons tovalidate sequences and alignments. Because non proteincoding gene fragments (16S, 18S, 28S, and ITS2) showunequal lengths, we aligned them with the online versionof the automatic aligner MAFFT 6 (http://mafft.cbrc.jp/alignment/server/; Katoh & Standley, 2013), using in-formation on the secondary structure of RNA duringthe alignment process (Q-INS-i strategy) and with othervalues set to default. The alignments of the ribosomalgenes contained highly unequal distributions of indels.We thus used GBLOCKS 0.91b (http://molevol.cmima.csic.es/castresana/Gblocks_server.html) to elimi-nate poorly aligned positions and divergent regions ofthe alignment in order to make our data set more suit-able for phylogenetic analyses (Talavera & Castresana,2007). We set the options to be less stringent, allowingsmaller final blocks, gap positions within final blocks,and less strict flanking positions. Using MESQUITE 2.75(Maddison & Maddison, 2013), we concatenated genefragments into two matrices, one with the full 5533 bpof data, and another containing ribosomal genes trimmedusing Gblocks, summing to 3875 bp of data.

We conducted both Bayesian and maximum-likelihood(ML) analyses for different partition schemes. We usedunlinked models for each gene of both the full andGblocks-trimmed data sets (the ‘full gene partition’ and‘gblocks gene partition’, respectively). In Bayesian analy-ses, we also used unlinked models for each gene andcodon position in protein-coding genes, of both the fulland Gblocks-trimmed data sets (‘full codon partition’and ‘gblocks codon partition’, respectively). We usedjModel test 2.1.3 (Darriba et al., 2012), implementingthe Akaike information criterion, to statistically selectthe best-fitting models of nucleotide substitutions.

We conducted Bayesian analyses using MrBayes 3.1.2(Ronquist & Huelsenbeck, 2003) run remotely at theCIPRES Science Gateway (Miller, Pfeiffer & Schwartz,2010). We performed two independent runs with foursimultaneous Markov chain Monte Carlo (MCMC) chains,each with random starting trees, running for a total of50 million generations. We discarded 30% of genera-tions as burn-in. We conducted ML analyses usingGARLI 2.0 (Zwickl, 2006). For every partition scheme,we first ran two search replicates, used stepwise-

addition starting topologies, and used automated stop-ping criteria. Using the best trees obtained by thesesearch replicates, we then ran 100 bootstrap replicatesunder the same conditions.

EVOLUTIONARY ANALYSES

Of the 112 terminals in our analyses, 80 represent taxathat build orb webs, for which we coded selected weband size traits. Using discrete variables, we coded thesector web as 0 (no sector web) or 1 (sector web), andthe silk tube retreat as 0 (no silk tube retreat) or 1(silk tube retreat). Any retreat shaped as a silken tube,even if inside a rolled leaf or another object, was char-acterized as a silk tube retreat. Simple off-web re-treats were not characterized as such. FollowingEberhard (1982), Hormiga et al. (1995), and Kuntneret al. (2008), we coded the following characters: build-ing of radial threads as 0 (cut and reeled single radii),1 (no cut and reel double radii), 2 (built as in Caerostris,0 and 1 in same web), or 3 (built as in Nephilidae,two single radii built with one trip from web hub toframe and back); building of the web hub as 0 (hubcentre left intact), 1 (hub centre bitten out), 2(hub centre bitten out and sealed back), 3 (entire hubremoved); attack behaviour as 0 (biting and then wrap-ping prey) or 1 (wrapping and then biting prey). Weused continuous variables for web size (calculated as:web width × web height × π) and spider size (length offirst leg patella + tibia). Actual web measurements aremissing from most of the literature, but based on ourown field data and literature records (Eberhard, 1987;Hajer & Rehakova, 2003; Ott & Brescovit, 2003;Ramirez, Lopardo & Platnick, 2004; Hormiga,Alvarez-Padilla & Benjamin, 2007; Brescovit & Lopardo,2008; Lopardo & Hormiga, 2008; Miller, Griswold &Yin, 2009; Lin & Li, 2013) we succeeded in gatheringboth size measures for 61 of the 80 terminals. To scoreabsolute web size for more terminals, we used de-scriptive literature measures for web size and addi-tionally categorized web size with discrete variablesas 0 (small web < 0.1 m2), 1 (0.1 m2 < mediumweb < 0.5 m2), and 2 (large web > 0.5 m2).

In MESQUITE 2.75 (Maddison & Maddison, 2013),we employed Felsenstein’s independent contrasts(Felsenstein, 1985) through the PDAP package (Midford,Garland & Maddison, 2002) to analyse correlationsamong continuous variables. We used stochastic Bayes-ian correlation analysis in SIMMAP 1.0 to analyse thecoevolution of discrete variables (Bollback, 2006).Because it is logically expected that larger spiders buildlarger webs, we regressed web size to spider size andused the standardized residuals as a measure of rela-tive web size. We plotted these relative measures onour preferred phylogeny, and included a scatter plotshowing the standard deviation of mean orbicularian



http://mafft.cbrc.jp/alignment/server/

http://mafft.cbrc.jp/alignment/server/

http://molevol.cmima.csic.es/castresana/Gblocks_server.html

http://molevol.cmima.csic.es/castresana/Gblocks_server.html

web size to illustrate the disproportionately large orsmall webs in certain terminals.

RESULTS

Bayesian and ML analyses, under the four differentpartition schemes, yielded similar topologies regard-ing the position of Zygiellinae and Caerostris (Fig. 2;Appendix S2). All analyses recovered a strongly sup-ported clade including the Zygiella genus group (Zygiella,Leviellus, Parazygiella, and Stroemiellus). This cladewas sister to a well-supported clade with the two Aus-tralian genera (Phonognatha + Deliochus), and togeth-er these groups excluded other potential zygiellid genera(Chrysometa, Yaginumia, and Guizygiella); however, thegenera Zygiella, Parazygiella, and Phonognatha werenot recovered as monophyletic in all analyses. Zygiellanearctica Gertsch, 1964 from North America groupedwith Parazygiella, whereas Parazygiella sp. A fromTaiwan was ambiguously placed in either Parazygiellaor Zygiella. The monotypic Stroemiellus was mostly re-covered as sister to Leviellus. We summarized theZygiellinae taxonomic emendations in Appendix 1.

All analyses strongly supported a monophyleticZygiellinae + Araneidae + Nephilidae, but the place-ment of Zygiellinae within this clade remains some-what ambiguous: whereas most analyses recovered itas sister to all other araneids, the gblocks partitionleft an unresolved trichotomy. All analyses strongly sup-ported monophyletic Nephilidae and Araneidae ex-cluding Zygiellinae (Fig. 2; Appendix S2). Other taxawith a sector web, tubular silk retreat, or a leaf-retreat in web were not recovered as close relativesto zygiellines, being within Araneidae (Acusilas, Milonia,and Yaginumia + Guizygiella) or Tetragnathidae(Chrysometa alboguttata; Figs 2, 3).

Although all analyses strongly support the monophyleticCaerostris within araneids (excluding Zygiellinae; Fig. 2;Appendix S2), its precise placement is ambiguous, withunstable relationships between Caerostris and otherbasal araneid lineages, e.g. Oarces + Gnolus, Micrathena,and an undescribed araneid (code ARA019). The cladecontaining all the remaining araneids also received strongsupport, but the topology within it changed among thepartition schemes.

Our results suggest that the sector web evolved in-dependently in the Zygiella genus group, twice inTetragnathidae, and at least five times in Araneidae(Fig. 3). The sector web correlates strongly with thesilken tube retreat (P < 0.001; Fig. 3). As expected, andshown only for a small number of spider taxa (Senseniget al., 2010), web size coevolved with body size (P < 0.001,R = 0.687; Appendix S3). Thus, in absolute terms,medium-sized webs (> 0.1 m2) evolved in large spiderspecies of the families Tetragnathidae, Nephilidae, andseveral times in Araneidae (Appendix S3); however, cor-rected for spider size, Caerostris and some Nephila builddisproportionately large webs, whereas the araneidPoltys builds disproportionately small webs (Fig. 4). Thewebs of Caerostris and nephilids are outliers in termsof exceptional web size (> 0.5 m2) in absolute terms(Appendix S3).

DISCUSSIONPHYLOGENETIC PLACEMENT AND GENEALOGY

OF ZYGIELLINAE

Recent molecular phylogenies of araneoid spiders con-sistently recover a zygielline clade uniting Zygiella withPhonognatha and Deliochus, but contradict each otherin placing such a clade as sister to Araneidae(Alvarez-Padilla et al., 2009; Dimitrov et al., 2012;Blackledge et al., 2012a), Nephilidae (Blackledge et al.,2009; Kuntner et al., 2013), or Araneidae + Nephilidae(Sensenig et al., 2010; Agnarsson et al., 2012); however,all of these studies strongly support the monophylyof a group containing Nephilidae, Zygiellinae, and otherAraneidae. As these recent phylogenetic studies eachaddressed a different question, none aimed to resolvethe relationships between zygiellines, araneids, andnephilids, mostly including one or two Zygiella species(Z. x-notata and/or Z. atrica) and nephilid species, andfew atypical araneids. In addition to an extensive out-group sample, our analyses included all four Zygiella s.l.genera, several other former and potential zygiellids,all five nephilid genera, several atypical araneids, andtetragnathids that resemble zygiellids in web biology.Our results support the clade consisting of Zygiellinae,Araneidae, and Nephilidae, and also support the place-ment of Zygiellinae as sister to other Araneidae, rather

▶Figure 2. Summary orbicularian phylogeny. The topology is from the Bayesian analysis under the ‘full data’ scheme,and partitioned ‘by gene’. Bayesian inference (BI) posterior probabilities and maximum-likelihood (ML) bootstrap valuesat nodes: high support (PB > 0.95, ML boot > 0.75) is marked with green; a recovered clade, but below the high supportthreshold, is marked with grey; and a clade that was not recovered is marked with red. Apart from Zygiellinae, otherAraneidae, Nephilidae, and Tetragnathidae families are labelled using the following codes: ANA, Anapidae; CYA, Cyatholipidae;DEI, Deinopidae; HOL, Holarchaeidae; LIN, Linyphiidae; MAL, Malkaridae; MIC, Micropholcommatidae; MIM, Mimetidae;MYS, Mysmenidae; NES, Nesticidae; NIC, Nicodamidae; PIM, Pimoidae; SYM, Symphytognathidae; SYN, Synotaxidae;THD, Theridiidae; THS, Theridiosomatidae; ULO, Uloboridae.





Figure 3. Evolution of behavioural traits in Orbiculariae plotted on the preferred topology (see Fig. 2). Above, evolutionof the sector web and silk tube retreat; below, evolution of radius building, hub building, and attack behaviour.



than Nephilidae or Nephilidae + Araneidae (Fig. 2). Interms of classification, our results do not require adopt-ing a family rank for ‘Zygiellidae’; a simpler solutionis to treat Zygiellinae as an Araneidae subfamily, aswe do henceforth (Appendix 1). The placement ofZygiellinae as sister to Tetragnathidae sensu Wunderlich(2004) is clearly refuted, as is the inclusion of thetetragnathid Chrysometa in that clade (Fig. 2; e.g.Hormiga et al., 1995; Alvarez-Padilla et al., 2009;Dimitrov et al., 2012). Within Zygiellinae, Leviellus ismonophyletic and sister to the monotypic Stroemiellus(Fig. 2), Zygiella is paraphyletic, and most analyses alsorecover a paraphyletic Parazygiella. Furthermore, thesupposedly tetragnathid Guizygiella and the araneid

Yaginumia entirely consist of former species of Zygiella,and were regarded as related to Zygiella by Zhu et al.(1997) and Wunderlich (2004), based on verbal argu-mentation rather than formal analyses. Our resultsstrongly support Guizygiella as araneid rather thantetragnathid, with the monotypic Yaginumia as its sister(Fig. 2). Our results further indicate a possible groupconsisting of Yaginumia + Guizygiella and Milonia, bothcontaining sector web-building species.

PHYLOGENETIC PLACEMENT OF CAEROSTRIS

Like some other molecular studies, our results supportthe basal position of Caerostris within Araneidae

Figure 4. Relative web size (standardized residuals of web size regressed on spider size) of Orbiculariae plotted on thepreferred topology (see Fig. 2). Dashed lines in the scatter plot represent the standard deviation for mean relative websize.



(excluding Zygiellinae; Sensenig et al., 2010; Kuntneret al., 2013), contrasting with morphological phylogeniesthat placed them with gasteracanthines (Scharff &Coddington, 1997) or argiopines (Kuntner et al., 2008);however, based on our results, the exact positionof Caerostris is ambiguous. Depending on the parti-tion scheme used in our analyses, Caerostris,Oarces + Gnolus, Micrathena, and an undescribedaraneid are generally recovered as a grade leading toall other araneids, but the relationships among themare not resolved. These ‘basal araneids’ are all mor-phologically distinct from other araneids (Scharff &Coddington, 1997), and with the exception of Micrathena,little is known about their biology. For example, vir-tually nothing is known about the cursorial Oarces andGnolus, which have been placed in Mimetidae basedon several morphological features (Platnick & Shadab,1993), but recent molecular studies place them at basalnodes of Araneidae (Dimitrov et al., 2012; this study).Caerostris morphologically differs from typical araneids,e.g. by the flattened tibiae and metatarsi, modifiedclypeus, abdominal sigillae, and macrosetae on femur IV(Scharff & Coddington, 1997; Smith, 2006; Kuntneret al., 2008).

CLASSIFICATION IMPLICATIONS

The classification implications of our study mainly delveinto the systematics of the Zygiella genus group, anddelimitation of the subfamily Zygiellinae (see Appen-dix 1 for details). Similar to Zygiellinae, Nephilidaewas transferred between Araneidae and Tetragnathidaein the past, and based on molecular data, could thusbe regarded araneids in the broadest sense. Nephilidaeis morphologically and behaviourally well-defined anddistinctively different from other Araneidae, however,and thus remains ranked as family (Kuntner et al., 2008,2013; World Spider Catalog, 2015). Further classifi-cation implications deal with the definition of Araneidae.Whereas some araneid groups are seemingly welldefined, e.g. Araneinae and Argiopinae (hereafter termed‘classical araneids’), synapomorphies of the diverseAraneidae have been elusive, and the composition ofthe family is a phylogenetic problem (Scharff &Coddington, 1997; Kuntner et al., 2013). We show thatseveral araneid groups, especially among lineages atbasal nodes, behaviourally differ from ‘classical’ araneids;however, phylogenetic ambiguity suggests that furtherdata are necessary before a stable classification ofAraneidae and relatives can be proposed.

EVOLUTION OF THE SECTOR WEB AND SILK

TUBE RETREAT

The sector web is a proposed synapomorphy and di-agnostic character of Zygiellinae (Wunderlich 2004).

We here confirm that the sector web-buildingChrysometa is a tetragnathid, and that the sector webevolved several times in other groups, including clas-sical araneids and tetragnathids (Fig. 3; Alvarez-Padillaet al., 2009). Furthermore, the sector web defines asubclade of Zygiellinae, as the Australasian zygiellinegenera Deliochus and Phonognatha do not build sectorwebs (Hormiga et al., 1995; Kuntner et al., 2008). Allzygiellines use some variation of silk tube retreats, beit a silken tube above the web in Zygiella s.l. (Gregoricet al., 2010), a silken tube with a leaf next to the webin Deliochus, or a rolled-leaf retreat inside the web inPhonognatha (Kuntner et al., 2008). The sector web andsilk tube retreat coevolve (Fig. 3), as would be expect-ed because such a retreat form enables the spiders toquickly charge towards the capture areas of the webin order to subdue prey. Similar retreats evolvedconvergently within orb weavers several times, indi-cating their adaptiveness. For example, they are foundin the araneids Acusilas (leaf retreat inside web;Kuntner et al., 2008; Murphy & Murphy, 1983), inMilonia and Guizygiella (silken tube and sector webin both genera), in Singa and Perilla (silk tube retreatinside a rolled grass stem; Kuntner et al., 2008; Gregoric,Kuntner & Blackledge, 2015), and silk tube retreatsare also found in the nephilids Nephilengys andNephilingis (Kuntner et al., 2008, 2013).

ORB WEB-RELATED TRAITS IN ZYGIELLINAE

Although not documented for all genera, at least somezygiellines seem to exhibit several atypical araneid be-haviours (Fig. 3). Zygiella s.l. builds double radial threadsin their orb webs (Hormiga et al., 1995; Gregoric, et al.,2015), a feature typical of uloborid webs (Eberhard,1982). Araneids, nephilids, and tetragnathids typical-ly build single radial threads, but differ in the detailsof construction behaviour (Eberhard, 1982), but Kuntneret al. (2008) considered the unique nephilid radii to bedoubled. Furthermore, Phonognatha and Deliochus leavethe web hub intact after orb construction (characterstate unknown for Zygiella s.l.), as do uloborids andnephilids (Eberhard, 1982; Hormiga et al., 1995; Kuntneret al., 2008), whereas tetragnathids bite it out andaraneids bite it out and seal the hole back up (Eberhard,1982). Phonognatha leaves the temporary spiral in thefinished orb, as do nephilids, whereas most other orbweavers, including Zygiella s.l. and Deliochus, removeit when building the sticky spiral (Kuntner et al., 2008).No zygiellines ‘decorate’ their webs with stabilimentaor detritus, as do some nephilids, araneids, and uloborids(e.g. Nephila, Argiope, Cyclosa, and Uloborus; Eberhard,1982; Kuntner et al., 2008). All zygiellines attack theirprey by biting first and then wrapping the prey(Kuntner et al., 2008; Gregoric et al., 2010), likenephilids, and unlike tetragnathids and most araneids



(Eberhard, 1982; Kuntner et al., 2008). Zygiellines hidein the retreat during the day, and they do not shaketheir body or switch sides of the web when threat-ened (Kuntner et al., 2008; Gregoric et al., 2010), asdo some araneids (e.g. Argiope and Azilia) and nephilids(e.g. Nephilengys, Nephila, and Clitaetra; Kuntner et al.,2008), but instead run to the retreat or jump off theweb.

ORB WEB-RELATED TRAITS IN CAEROSTRIS

Similar to zygiellines, Caerostris also exhibits special-ized orb web architecture and building behaviour.Whereas the behaviours reported here are document-ed only for Caerostris darwini Kuntner & Agnarsson,2010, preliminary data on three additional Caerostrisspecies (Caerostris extrusa Butler, 1882 and twoundescribed species) indicate that the behaviours aresynapomorphic for the genus (Gregoric, unpubl. data).Caerostris deviates in early orb web construction fromall other known orb weavers: it employs almost no website exploration, builds no secondary web frames, andconstructs the entire orb below the initial bridge line,in contrast to other orb weavers that extensively exploretheir web sites, typically construct secondary webframes, and build the orb around the initial bridge line(Gregoric et al., 2011b). Furthermore, Caerostris buildsorb webs that contain two types of radial threads, singleradial threads in the upper half and doubled radialthreads in the lower half of the orb, and this behav-iour is unique (Gregoric et al., 2011b). Moreover,Caerostris spiders sometimes build weak stabilimentaand never ‘decorate’ webs with detritus (Gregoric et al.,2011a). Some Caerostris species are nocturnal and hide,mimicking bark, during the day, but do not build re-treats, whereas other species never leave the web hub.They do not shake their body or switch sides of theweb when threatened, but instead run to the edge ofthe web or jump off of the web (Gregoric, pers. observ.).They attack their prey by biting first and then wrap-ping the prey, and carry all but the largest prey backto the hub in their massive chelicerae, or lift the largestprey to the hub still attached to other web parts(Gregoric et al., 2011a). Other orb weavers typically hangall but the smallest prey to their spinnerets and inthis way carry the wrapped prey back to the hub (Foelix,2011).

WEB GIGANTISM

Large webs evolved several times in tetragnathids,nephilids, and araneids, simply because larger spiderspecies generally build larger webs (Sensenig et al., 2010;Appendix S3); however, true web gigantism, i.e. thebuilding of disproportionately large webs, has not yetbeen studied, and has evolved only in Caerostris and

in some Nephila (Fig. 3). Unlike the large-bodiednephilid spiders like Nephila and Nephilingis (Kuntner& Coddington, 2009), Caerostris are not exception-ally large spiders, but are instead similar in size toaraneid genera like Argiope, Araneus, Parawixia, andNeoscona, etc. (Sensenig et al., 2010). Interestingly,Nephila and Caerostris represent web architectures thatshow opposite strategies in the compensatory evolu-tion of web performance, where the quality of silk tradesagainst web architecture and the volume of silk used(Sensenig et al., 2010). Namely, Nephila builds websusing silk threads of average quality (low quality cor-rected for spider size), but these threads densely coverthe capture area of the web (Kuntner et al., 2008;Sensenig et al., 2010), such that Nephila webs arecapable of stopping and retaining even vertebrate prey(Sensenig et al., 2010; Nyffeler & Knornschild, 2013).Caerostris on the other hand, builds sparse webs(Sensenig et al., 2010; Gregoric et al., 2011a), but becauseof the extremely tough silk, the stopping potential ofthe web is comparable with that of nephilid and thebest-performing araneid webs (Sensenig et al., 2010).

CONCLUSIONS

We provide the first species-level molecular phylogeniesof the taxonomically controversial and/or understud-ied Zygiellinae and Caerostris. First, we have shownthat the subfamily Zygiellinae contains the HolarcticZygiella s.l., and the Australasian Phonognatha andDeliochus, and is likely to be sister to all other araneids,whereas Caerostris is a basal araneid genus, not amember of Argiopinae or Araneinae. Second, despiteextensive in-group and out-group taxon sampling, thesomewhat ambiguously supported phylogenetic posi-tions of Zygiellinae and Caerostris confirms that theusual selection of genetic markers is insufficient forunambiguously resolved phylogenetic relationshipsbetween spider groups at the family level; however,we believe that our results represent important pro-gress towards resolving phylogenetic relationships atbasal nodes of Araneidae. Third, the sector web is nota zygiellid synapomorphy, but evolved several timesindependently, coevolving with the silk tube retreat.Fourth, phylogenetic exclusivity seems to reflect be-havioural differences in zygiellines and ‘basal araneids’.Fifth, true web gigantism evolved in Caerostris andto a lesser extent in Nephila, and the two genera exhibitopposite strategies in the compensatory evolution ofweb performance.

ACKNOWLEDGEMENTS

We thank Ren-Chung Cheng, Shakira G. QiñonesLebron, Heine C. Kiesbüy, Tjaša Lokovšek, LauraMay-Collado, Yadira Ortiz, and Joel Duff for their



laboratory, logistic, and analytic help. We thank KlemenCandek, Jan Dolansky, Petr Dolejš, Jason Dunlop,Charles Haddad, Peter Jäger, Rudy Jocqué, RokKostanjšek, Daiqin Li, Wenjin Gan, Liu Shengjie, YuriMarušik, Honore Rabarison, Sahondra LalaoRahanitriniaina, Jutta Schneider, Mark Townley,Shichang Zhang and MICET and ICTE crews formuseum loans, fresh material and help in the field.We also thank B.A. Huber for a useful review of thearticle. This contribution was funded by the SlovenianResearch Agency (grants P1-0236 and J1-2063), theUnited States National Science Foundation (IOS-0745379), and the National Geographic Society (grant8655-09).

REFERENCES

Agnarsson I. 2003. Spider webs as habitat patches – the dis-tribution of kleptoparasites (Argyrodes, theridiidae) amonghost webs (Nephila, tetragnathidae). Journal of Arachnol-ogy 31: 344–349.

Agnarsson I. 2011. Habitat patch size and isolation as pre-dictors of occupancy and number of argyrodine spiderkleptoparasites in Nephila webs. Die Naturwissenschaften 98:163–167.

Agnarsson I, Coddington JA, Kuntner M. 2013. System-atics: progress in the study of spider diversity and evolu-tion. In: Penney D, ed. Spider research in the 21st century:trends and perspectives. Rochdale, UK: Siri Scientific Press,58–111.

Agnarsson I, Dhinojwala A, Sahni V, Blackledge TA. 2009.Spider silk as a novel high performance biomimetic muscledriven by humidity. Journal of Experimental Biology 212:1989–1993.

Agnarsson I, Gregoric M, Blackledge TA, Kuntner M. 2012.The phylogenetic placement of Psechridae within Entelegynaeand the convergent origin of orb-like spider webs. Journalof Zoological Systematics and Evolutionary Research 51: 100–106.

Agnarsson I, Kuntner M, Blackledge TA. 2010.Bioprospecting finds the toughest biological material: extraor-dinary silk from a giant riverine orb spider. PLoS ONE 5:e11234.

Alvarez-Padilla F, Dimitrov D, Giribet G, Hormiga G. 2009.Phylogenetic relationships of the spider family Tetragnathidae(Araneae, Araneoidea) based on morphological and DNA se-quence data. Cladistics 25: 109–146.

Archer AF. 1951. Studies in the orbweaving spiders(Argiopidae). 1. American Museum Novitates 1487: 1–52.

Archer AF. 1960. A new genus of Argiopidae from Japan. ActaArachnologica Tokyo 17: 13–14.

Ausserer A. 1871. Neue Radspinnen. Verhandlungen derKaiserlich-Königlichen Zoologisch-Botanischen Gesellschaft inWien 21: 815–832.

Ayoub NA, Garb JE, Hedin M, Hayashi CY. 2007. Utilityof the nuclear protein-coding gene, elongation factor-1 gamma(EF-1 gamma), for spider systematics, emphasizing family

level relationships of tarantulas and their kin (Araneae:Mygalomorphae). Molecular Phylogenetics and Evolution 42:394–409.

Bakhvalov VF. 1974. Identification key of the spider familyAraneidae from Kirgizia. Entomologicheskie Issledovaniya vKirgizii 9: 101–112.

Bel-Venner MC, Dray S, Allaine D, Menu F, Venner S.2008. Unexpected male choosiness for mates in a spider.Proceedings of the Royal Society B-Biological Sciences 275:77–82.

Bel-Venner MC, Venner S. 2006. Mate-guarding strategiesand male competitive ability in an orb-weaving spider: resultsfrom a field study. Animal Behaviour 71: 1315–1322.

Birkedal Schmidt J, Scharff N. 2008. A taxonomic revi-sion of the orb-weaving spider genus Acusilas Simon, 1895(Araneae, Araneidae). Insect Systematics & Evolution 39:1–38.

Blackledge TA, Gillespie RG. 2004. Convergent evolutionof behavior in an adaptive radiation of Hawaiian web-building spiders. Proceedings of the National Academy of Sci-ences of the United States of America 101: 16228–16233.

Blackledge TA, Kuntner M, Agnarsson I. 2011. The formand function of spider orb webs: evolution from silk to eco-systems. In: Casas J, ed. Advances in insect physiology, Vol41: spider physiology and behaviour – behaviour. Burling-ton, Elsevier, 175–262.

Blackledge TA, Kuntner M, Marhabaie M, Leeper TC,Agnarsson I. 2012a. Biomaterial evolution parallels behavioralinnovation in the origin of orb-like spider webs. ScientificReports 2: 833.

Blackledge TA, Perez-Rigueiro J, Plaza GR, Perea B,Navarro A, Guinea GV, Elices M. 2012b. Sequential originin the high performance properties of orb spider dragline silk.Scientific Reports 2: 782.

Blackledge TA, Scharff N, Coddington JA, Szuts T, WenzelJW, Hayashi CY, Agnarsson I. 2009. Reconstructing webevolution and spider diversification in the molecular era. Pro-ceedings of the National Academy of Sciences of the UnitedStates of America 106: 5229–5234.

Blamires SJ. 2010. Plasticity in extended phenotypes: orb webarchitectural responses to variations in prey parameters.Journal of Experimental Biology 213: 3207–3212.

Bollback JP. 2006. SIMMAP: stochastic character mappingof discrete traits on phylogenies. BMC Bioinformatics 7: 88.

Bond JE, Garrison NL, Hamilton CA, Godwin RL, HedinMC, Agnarsson I. 2014. Phylogenomics resolves a spiderbackbone phylogeny and rejects a prevailing paradigm fororb web evolution. Current Biology 24: 1–7.

Bond JE, Opell BD. 1998. Testing adaptive radiation and keyinnovation hypotheses in spiders. Evolution 52: 403–414.

Brescovit AD, Lopardo L. 2008. The first record on the spidergenus Trogloneta Simon in the southern hemisphere (Araneae,Mysmenidae), with descriptions of three new species fromBrazil and remarks on the morphology. Acta Zoologica 89:93–106.

Clerck C. 1757. Svenska spindlar, uti sina hufvud-slågter indeltesamt under några och sextio särskildte arter beskrefne ochmed illuminerade figurer uplyste. Stockholmiae.



Coddington JA. 1994. The roles of homology and conver-gence in studies of adaptation. In: Eggleton PV, Vane-Wright R, eds. Phylogenetics and ecology. London: The LinneanSociety of London, 53–78.

Craig CL, Weber RS, Bernard GD. 1996. Evolution ofpredator-prey systems: spider foraging plasticity in re-sponse to the visual ecology of prey. American Naturalist 147:205–229.

Darriba D, Taboada GL, Doallo R, Posada D. 2012.jModelTest 2: more models, new heuristics and parallel com-puting. Nature Methods 9: 772–772.

Dimitrov D, Hormiga G. 2011. An extraordinary new genusof spiders from Western Australia with an expandedhypothesis on the phylogeny of Tetragnathidae (Araneae).Zoological Journal of the Linnean Society 161: 735–768.

Dimitrov D, Lopardo L, Giribet G, Arnedo MA, Alvarez-Padilla F, Hormiga G. 2012. Tangled in a sparse spiderweb: single origin of orb weavers and their spinning workunravelled by denser taxonomic sampling. Proceedings of theRoyal Society B-Biological Sciences 279: 1341–1350.

Eberhard WG. 1982. Behavioral characters for the higher clas-sification of orb-weaving spiders. Evolution 36: 1067–1095.

Eberhard WG. 1986. Effects of orb-web geometry on prey inter-ception and retention. In: Shear WA, ed. Spiders, webs, be-havior and evolution. Stanford: Stanford University Press,70–100.

Eberhard WG. 1987. Web-building behavior of Anapid,Symphytognathid and Mysmenid spiders. Journal of Arach-nology 14: 339–356.

Eberhard WG. 1990. Function and phylogeny of spider webs.Annual Review of Ecology and Systematics 21: 341–372.

Eberhard WG. 2007. Miniaturized orb-weaving spiders:behavioural precision is not limited by small size.Proceedings of the Royal Society B-Biological Sciences 274:2203–2209.

Elgar MA. 1989. Kleptoparasitism – a cost of aggregating foran orb-weaving spider. Animal Behaviour 37: 1052–1055.

Felsenstein J. 1985. Phylogenies and the comparative method.American Naturalist 125: 1–15.

Fernández R, Hormiga G, Giribet G. 2014. Phylogenomicanalysis of spiders reveals nonmonophyly of orb weavers.Current Biology 24: 1–6.

Foelix RF. 2011. Biology of spiders. Oxford: Oxford Univer-sity Press.

Franganillo BP. 1909. Arañas de la familia de los argiópidos,observada junto á la desembocadura del Miño. Act. Actas yMemorias del Congreso de Naturaleza de España. 1: 185–189.

Gertsch WJ. 1964. The spider genus Zygiella in North America(Araneae, Argiopidae). American Museum Novitates 2188:1–21.

Gillespie R. 2004. Community assembly through adaptiveradiation in Hawaiian spiders. Science 303: 356–359.

Grasshoff M. 1984. Die Radnetzspinnen-Gattung Caerostris(Arachnida: Araneae). Revue Zoologique Africaine 98: 725–765.

Grawe I, Wolff JO, Gorb SN. 2014. Composition and substrate-dependent strength of the silken attachment discs in spiders.

Journal of the Royal Society Interface 11: doi: 10.1098/rsif.2014.0477.

Gregoric M, Agnarsson I, Blackledge TA, Kuntner M.2011a. Darwin’s bark spider: giant prey in giant orb webs(Caerostris darwini, Araneae: Araneidae)? Journal of Arach-nology 39: 287–295.

Gregoric M, Agnarsson I, Blackledge TA, Kuntner M.2011b. How did the spider cross the river? Behavioral ad-aptations for river-bridging webs in Caerostris darwini(Araneae: Araneidae). PLoS ONE 6: e26847.

Gregoric M, Kiesbuy HC, Lebron SGQ, Rozman A,Agnarsson I, Kuntner M. 2013. Optimal foraging, notbiogenetic law, predicts spider orb web allometry. DieNaturwissenschaften 100: 263–268.

Gregoric M, Kostanjšek R, Kuntner M. 2010. Orb web fea-tures as taxonomic characters in Zygiella s.l. (Araneae,Araneidae). Journal of Arachnology 38: 319–327.

Gregoric M, Kuntner M, Blackledge TA. 2015. Does bodysize predict foraging effort? Patterns of material invest-ment in spider orb webs. Journal of Zoology 296: 67–78.

Griswold CE, Coddington JA, Hormiga G, Scharff N. 1998.Phylogeny of the orb-web building spiders (Araneae,Orbiculariae: Deinopoidea, Araneoidea). Zoological Journalof the Linnean Society 123: 1–99.

Hajer J, Rehakova D. 2003. Spinning activity of the spiderTrogloneta granulum (Araneae, Mysmenidae): web, cocoon,cocoon handling behaviour, draglines and attachment discs.Zoology 106: 223–231.

Hausdorf B. 1999. Molecular phylogeny of araneomorph spiders.Journal of Evolutionary Biology 12: 980–985.

Hayashi CY, Lewis RV. 2000. Molecular architecture and evo-lution of a modular spider silk protein gene. Science 287:1477–1479.

Heimer S, Nentwig W. 1991. Spinnen Mitteleuropas. EinBestimmungsbuch. Berlin: Blackwell Wissenschafts-Verlag.

Herberstein M, Wignall A. 2011. Introduction: spider biology.In: Herberstein M, ed. Spider behaviour: flexibility and ver-satility. Cambridge: Cambridge University Press, 1–30.

Higgins LE, Buskirk RE. 1998. Spider-web kleptoparasitesas a model for studying producer-consumer interactions.Behavioral Ecology 9: 384–387.

Hormiga G, Alvarez-Padilla F, Benjamin SP. 2007. Firstrecords of extant Hispaniolan spiders of the familiesMysmenidae, Symphytognathidae, and Ochyroceratidae(Araneae), including a new species of Ochyrocera. Ameri-can Museum Novitates 3577: 1–21.

Hormiga G, Eberhard WG, Coddington JA. 1995. Web-construction behavior in australian Phonognatha and thephylogeny of nephiline and tetragnathid spiders (Araneae,Tetragnathidae). Australian Journal of Zoology 43: 313–364.

Hormiga G, Griswold CE. 2014. Systematics, phylogeny, andevolution of orb-weaving spiders. Annual Review of Ento-mology 59: 487–512.

Katoh K, Standley DM. 2013. MAFFT multiple sequencealignment software version 7: improvements in perfor-mance and usability. Molecular Biology and Evolution 30:772–780.



Koch CL. 1834. Arachniden. In: Herrich-Schäffer GAW, ed.Deutschlands Insekten. Heft: 122–127.

Koch CL. 1839. Die Arachniden. Nürnberg: Funfter Band, 125–158, Sechster Band, 1–156, Siebenter Band, 1–106.

Koch CL. 1845. Die Arachniden. Nürnberg: Zwolfter Band,1–166.

Kulczynski W. 1885. Araneae in Camtschadalia a Dre B.Dybowski collectae. Pamietnik Akademji umiejetnosci wKrakow wydzial matematyczno-przyrodniczy 11: 1–60.

Kuntner M, Agnarsson I. 2009. Phylogeny accuratelypredicts behaviour in Indian Ocean Clitaetra spiders(Araneae: Nephilidae). Invertebrate Systematics 23: 193–204.

Kuntner M, Agnarsson I. 2010. Darwin’s bark spider: webgigantism in a new species of bark spiders from Madagas-car (Araneidae: Caerostris). Journal of Arachnology 38:346–356.

Kuntner M, Arnedo MA, Trontelj P, Lokovšek T,Agnarsson I. 2013. A molecular phylogeny of nephilid spiders:evolutionary history of a model lineage. Molecular Phylogeneticsand Evolution 69: 961–979.

Kuntner M, Coddington JA. 2009. Discovery of the largestorbweaving spider species: the evolution of gigantism inNephila. PLoS ONE 4: e7516.

Kuntner M, Coddington JA, Hormiga G. 2008. Phylogenyof extant nephilid orb-weaving spiders (Araneae, Nephilidae):testing morphological and ethological homologies. Cladistics24: 147–217.

Kuntner M, Gregoric M, Li DQ. 2010. Mass predicts webasymmetry in Nephila spiders. Die Naturwissenschaften 97:1097–1105.

Levi HW. 1974. The orb-weaver genus Zygiella (Araneae:Araneidae). Bulletin of the Museum of Comparative Zoology146: 267–290.

Levi HW. 1980. The orb-weaver genus Mecynogea, the sub-family Metinae and the genera Pachygnatha, Glenognathaand Azilia of the subfamily Tetragnathinae north of Mexico(Araneae: Araneidae). Bulletin of the Museum of Compara-tive Zoology 149: 1–74.

Levy G. 1987. Spiders of the genera Araniella, Zygiella, Zillaand Mangora (Araneae, Araneidae) from Israel, with noteson Metellina species from Lebanon. Zoologica Scripta 16: 243–257.

Lin Y, Li S. 2013. Two new species of the genera Mysmenaand Trogloneta (Mysmenidae, Araneae) from SouthwesternChina. ZooKeys 303: 33–51.

Lopardo L, Giribet G, Hormiga G. 2011. Morphology to therescue: molecular data and the signal of morphological char-acters in combined phylogenetic analyses-a case study frommysmenid spiders (Araneae, Mysmenidae), with commentson the evolution of web architecture. Cladistics 27: 278–330.

Lopardo L, Hormiga G. 2008. Phylogenetic placement of theTasmanian spider Acrobleps hygrophilus (Araneae, Anapidae)with comments on the evolution of the capture web inAraneoidea. Cladistics 24: 1–33.

Maddison WP, Maddison DR. 2013. Mesquite: a modularsystem for evolutionary analysis. Available at: http://mesquiteproject.org

Mayntz D, Toft S, Vollrath F. 2009. Nutrient balance affectsforaging behaviour of a trap-building predator. Biology Letters5: 735–738.

Menge A. 1866. Preussische Spinnen. Erste Abtheilung.Schriften der Naturforschenden Gesellschaft in Danzig (N.F.)1: 1–152.

Midford PE, Garland T, Maddison WP. 2002. PDAP:PDTREEpackage for Mesquite, version 1.00.

Miller JA, Griswold CE, Yin CM. 2009. The symphytognathoidspiders of the Gaoligongshan, Yunnan, China (Araneae,Araneoidea): systematics and diversity of micro-orbweavers.ZooKeys 11: 9–195.

Miller MA, Pfeiffer W, Schwartz T. 2010. Creating theCIPRES Science Gateway for inference of large phylogenetictrees. Proceedings of the Gateway Computing Environ-ments Workshop (GCE), 14 Nov. 2010, New Orleans, LA: 1–8.

Murphy J, Murphy F. 1983. The orb weaver genus Acusilas(Araneae, Araneidae). Bulletin of the British ArachnologicalSociety 6: 115–123.

Nentwig W, Blick T, Gloor D, Hänggi A, Kropf C.2014. Araneae: spiders of Europe. Available at: http://www.araneae.unibe.ch

Nyffeler M, Knornschild M. 2013. Bat predation by spiders.PLoS ONE 8: e58120.

Opell BD, Schwend HS. 2007. The effect of insect surfacefeatures on the adhesion of viscous capture threads spun byorb-weaving spiders. Journal of Experimental Biology 210:2352–2360.

Ott R, Brescovit AD. 2003. Description of the females of Anapiscastilla and Anapisona bordeaux (Araneae, Anapidae). Journalof Arachnology 31: 340–343.

Peters HM. 1937. Studien am Netz der Kreuzspinne (Araneadiadema.). 1. Die Grundstruktur des Netzes und Beziehungenzum Bauplan des Spinnenkorpers. Zeitschrift für Morphologieund Ökologie der Tiere 33: 128–150.

Pickard-Cambridge FO. 1872. General list of thespiders of Palestine and Syria, with descriptions ofnumerous new species, and characters of two new genera.Proceedings of the Zoological Society of London 1871: 212–354.

Pickard-Cambridge FO. 1902. A revision of the genera ofAraneae or spiders with reference to their type species. Annalsand Magazine of Natural History 9: 5–20.

Platnick NI, Shadab MU. 1993. A review of the pirate spiders(Araneae, Mimetidae) of Chile. American Museum Novitates3074: 1–30.

Ramirez MJ, Lopardo L, Platnick NI. 2004. Notes on Chileananapids and their webs. American Museum Novitates 3428:2–16.

Roberts MJ. 1995. Spiders of Britain & Northern Europe.London: HarperCollins Publishers.

Robinson MH, Robinson B. 1973. Ecology and behaviorof the giant wood spider Nephila maculata (Fabr.) in NewGuinea. Smithsonian Contributions to Zoology 149: 1–73.

Ronquist F, Huelsenbeck JP. 2003. MrBayes 3: Bayesianphylogenetic inference under mixed models. Bioinformatics19: 1572–1574.



http://mesquiteproject.org

http://mesquiteproject.org

http://www.araneae.unibe.ch

http://www.araneae.unibe.ch

Rypistra AL. 1981. The effect of kleptoparasitism on prey con-sumption and web relocation in a Peruvian population of thespider Nephila clavipes. Oikos 37: 179–182.

Sahni V, Blackledge TA, Dhinojwala A. 2011. A review onspider silk adhesion. Journal of Adhesion 87: 595–614.

Scharff N, Coddington JA. 1997. A phylogenetic analysis ofthe orb-weaving spider family Araneidae (Arachnida, Araneae).Zoological Journal of the Linnean Society 120: 355–434.

Schmidt G. 1968. Zur Spinnenfauna von Teneriffa. ZoologischeBeiträge (N.F.) 14: 387–425.

Sensenig A, Agnarsson I, Blackledge TA. 2010. Behav-ioural and biomaterial coevolution in spider orb webs. Journalof Evolutionary Biology 23: 1839–1856.

Simon E. 1886. Arachnides recueillis par M. A. Pavie (sous-chef du service des postes au Cambodge) dans le royaumede Siam, au Cambodge et en Cochinchine. Actes de la SociétéLinnéenne de Bordeaux 40: 137–166.

Simon E. 1889. Arachnidae transcaspicae ab ill. Dr. G. Radde,Dr. A. Walter et A. Conchin inventae (annis 1886–1887).Verhandlungen der Kaiserlich-Königlichen Zoologisch-Botanischen Gesellschaft in Wien 39: 373–386.

Simon E. 1894. Histoire naturelle des araignées. Paris: 1,489–760.

Smith HM. 2006. A revision of the genus Poltys in Austral-asia (Araneae: Araneidae). Records of the Australian Museum58: 43–96.

Stellwagen SD, Opell BD, Short KG. 2014. Temperature me-diates the effect of humidity on the viscoelasticity of glyco-protein glue within the droplets of an orb-weaving spider’sprey capture threads. Journal of Experimental Biology 217:1563–1569.

Sundevall CJ. 1833. Svenska spindlarnes beskrifning.Fortsättning och slut. Bihang till Kongliga Svenska Vetenskaps-Akademiens Handlingar 1832: 172–272.

Swanson BO, Anderson SP, DiGiovine C, Ross RN, DorseyJP. 2009. The evolution of complex biomaterial perfor-mance: the case of spider silk. Integrative and ComparativeBiology 49: 21–31.

Talavera G, Castresana J. 2007. Improvement of phylogeniesafter removing divergent and ambiguously aligned blocks fromprotein sequence alignments. Systematic Biology 56: 564–577.

Thevenard L, Leborgne R, Pasquet A. 2004. Web-buildingmanagement in an orb-weaving spider, Zygiella x-notata: in-fluence of prey and conspecifics. Comptes Rendus Biologies327: 84–92.

Thorell T. 1870. Remarks on synonyms of European spiders.Part I. Uppsala: 1–96.

Vasanthavada K, Hu X, Tuton-Blasingame T, Hsia Y,Sampath S, Pacheco R, Freeark J, Falick AM, Tang S,Fong J, Kohler K, La Mattina-Hawkins C, Vierra C. 2012.Spider Glue Proteins Have Distinct Architectures Com-pared with Traditional Spidroin Family Members. Journalof Biological Chemistry 287: 35986–35999.

Venner S, Bel-Venner MC, Pasquet A, Leborgne R. 2003.Body-mass-dependent cost of web-building behavior in an orbweaving spider, Zygiella x-notata. Die Naturwissenschaften90: 269–272.

Venner S, Casas J. 2005. Spider webs designed for rare butlife-saving catches. Proceedings of the Royal Society B-BiologicalSciences 272: 1587–1592.

Venner S, Pasquet A, Leborgne R. 2000. Web-building be-haviour in the orb-weaving spider Zygiella x-notata: influ-ence of experience. Animal Behaviour 59: 603–611.

Vollrath F, Porter D. 2009. Silks as ancient models for modernpolymers. Polymer 50: 5623–5632.

Vollrath F, Selden P. 2007. The role of behavior in the evo-lution of spiders, silks, and webs. Annual Review of EcologyEvolution and Systematics 38: 819–846.

Watanabe T. 2000. Web tuning of an orb-web spider, Octonobasybotides, regulates prey-catching behaviour. Proceedings ofthe Royal Society of London Series B-Biological Sciences 267:565–569.

Wiehle H. 1927. Beiträge zur Kenntnis des Radnetzbaues derEpeiriden, Tetragnathiden und Uloboriden. Zeitschrift fürMorphologie und Ökologie der Tiere 8: 468–537.

Wiehle H. 1929. Weitere Beiträge zur Biologie der Araneen,insbesondere zur Kenntnis des Radnetzbaues. Zeitschrift fürMorphologie und Ökologie der Tiere 15: 262–308.

Witt PN, Reed CF, Peakall DB. 1968. A spider’s web: prob-lems in regulatory biology. New York: Springer Verlag.

World Spider Catalog. 2015. World spider catalog. Bern:Natural History Museum. Available at: http://wsc.nmbe.ch

Workman T, Workman ME. 1894. Malaysian spiders. Belfast:9–24.

Wunderlich J. 2004. Fossil spiders in amber and copal. Con-clusions, revisions, new taxa and family diagnoses of fossiland extant taxa. Hirschberg-Leutershausen: J. Wunderlich.

Zawadsky AM. 1902. Materialy k faounié i biology Paoukow(Araneina) Zakawkazia. Izwestia Imperatorskago ObtchiestwaLioubitelei Iestwoznania, Antropology i Etnografy, Sostoiaschagopri Imperatorskom Moskowskom Ouniwersitete 98: 1–5.

Zhu MS, Kim JP, Song DX. 1997. On three new genera andfour new species of the family Tetragnathidae (Araneae) fromChina. Korean Arachnology 13: 1–10.

Zschokke S, Vollrath F. 1995. Web construction patterns ina range of orb-weaving spiders (Araneae). European Journalof Entomology 92: 523–541.

Zwickl DJ. 2006. Genetic algorithm approaches for thephylogenetic analysis of large biological sequence datasetsunder the maximum likelihood criterion. PhD Dissertation,The University of Texas at Austin.

APPENDIX 1TAXONOMIC EMENDATIONS

Family Araneidae Clerck, 1757Subfamily Zygiellinae Wunderlich, 2004.

RemarksThe Zygiella genus group was transferred between thefamilies Tetragnathidae (Levi, 1980; Heimer & Nentwig,1991; Wunderlich, 2004) and Araneidae (Levy, 1987;Roberts, 1995; Scharff & Coddington, 1997) in thepast. The genera Deliochus and Phonognatha were



http://wsc.nmbe.ch

transferred from Tetragnathidae to Araneidae (Kuntner,Coddington & Hormiga, 2008). The monophyly ofZygiellinae, uniting the above genera, and exclusiveof other members of Araneidae, is highly supported byBayesian and ML analyses (PB = 0.89–1.00, MLboot = 0.82–0.92; Fig. 2, Appendix S2). Molecular datastrongly support Parazygiella and Stroemiellus as newsynonymies of Zygiella and Leviellus, respectively (Fig. 2;Appendix S2).

DiagnosisSpecies of Zygiellinae differ from other araneids by theaggregate spigots being apart from the flagelliform spigots,rather than embracing them (Hormiga, Eberhard &Coddington, 1995; Kuntner et al., 2008; Alvarez-Padillaet al., 2009), and by employing doubled rather thansingle radial threads in the web. Species ofDeliochus + Phonognatha differ from other araneids bythe tarsus IV median claw being shorter than the pairedmain claw, by the spermathecae being lobed rather thanspherical or oval, and by the absence of the sustentaculumand light pigmented pattern on female venter (Kuntneret al., 2008). Species of Leviellus + Zygiella differ fromother araneids by the paracymbium being complex inshape, rather than a short basal structure, by the paracy-mbium base being less sclerotized than the cymbium,and by the tegulum being of the same size or longerthan subtegulum in ectal view (Hormiga et al., 1995;Kuntner et al., 2008; Alvarez-Padilla et al.,2009).

RemarksSpecies of Deliochus + Phonognatha differ from thoseof Leviellus + Zygiella by the tarsus IV median clawbeing shorter than the paired main claw, by the absenceof the sustentaculum, the presence of paired whitedots around spinnerets, and a light pigmented patternon female venter, by the lobed rather than sphericalspermathecae, the presence of embolic plugs, by theembolus of medium length (0.5–1.5 cymbium length),rather than short (<0.5 cymbium length), andby the paracymbium base being as sclerotized asthe cymbium (Hormiga et al., 1995; Kuntner et al.,2008; Alvarez-Padilla et al., 2009; Gregoric unpubl.data). Species of Leviellus + Zygiella all build aweb with a spiral-free sector, whereas species ofDeliochus + Phonognatha do not.

CompositionFollowing the revised phylogeny, the subfamilyincludes the genera: Deliochus Simon, 1894;distribution, Australia; Leviellus Wunderlich,2004; distribution, Holarctic; Phonognatha Simon, 1894;distribution, Australia, and southern and easternAsia; Zygiella Pickard-Cambridge, 1902; distribution,Holarctic.

LEVIELLUS WUNDERLICH, 2004

Stroemiellus Wunderlich, 2004 syn. nov., description ofStroemiellus stroemi (= Leviellus stroemi).

RemarksBecause Stroemiellus as a monotypic genus does notcontain grouping information, we synonymizeStroemiellus Wunderlich, 2004 with LeviellusWunderlich, 2004. Leviellus sp. (code ZYG311; Appen-dix S1) from our data set is likely to represent a ju-venile female Zygiella poriensis Levy, 1987, collectednear the type locality of the species. Based on the resultsof our analyses and the genital morphology ofZ. poriensis (epigynal scape), we transfer the speciesto Leviellus.

Type speciesZygiella kochi Thorell, 1870.

DiagnosisSpecies of Leviellus differ from Zygiella by the malepalpal embolus tip being flat rather than cylindrical,and by the presence of female epigynal scape.

DescriptionFemales small to large in size (3–10 mm body length),males same size (Levi, 1974). Female epigynum witha scape, male embolus tip flat (Levi, 1974: 71–72, 81–83, 85–86, 88–91, 93–94, 96–101, 104–110; Levy, 1987:17–25, 30–33). Carapace light brown, head region slight-ly darker, often outlined in black, chelicerae dark brown.Sternum dark brown with a light-brown median band.Abdomen grey to brown, dorsally with a wave-edgeddarker area that sometimes contains a whitish medianband. Legs light brown to brown, annulated dark brownto black (Gregoric, pers. observ.; Nentwig et al., 2014).Orb webs with a spiral-free sector. During the day, thespiders hide off of the web in a tubular silk retreat,connected to the web hub with a signal line (Levi, 1974;Gregoric, Kostanjšek & Kuntner, 2010).

CompositionFollowing the revised phylogeny here, the genus in-cludes: Leviellus caspica (Simon, 1889), for synony-mies and diagnosis, see Levi (1974) and World SpiderCatalog (2015); distribution, Central Asia; Leviellusinconveniens (Pickard-Cambridge, 1872), for synony-mies and diagnosis, see Levi (1974) and World SpiderCatalog (2015); distribution, Lebanon, Israel; Levielluskochi (Thorell, 1870), for synonymies and diagnosis,see Levi (1974) and World Spider Catalog (2015); dis-tribution, Southern Europe, North Africa, Central Asia;Leviellus poriensis (Levy, 1987) comb. nov., for syn-onymies and diagnosis, see Levy (1987) and WorldSpider Catalog (2015); distribution, Israel; Leviellus



stroemi (Thorell, 1870) comb. nov., for synonymies anddiagnosis, see Levi (1974) and World Spider Catalog(2015); distribution, Palaearctic; Leviellus thorelli(Ausserer, 1871), for synonymies and diagnosis, see Levi(1974) and World Spider Catalog (2015); distribution,Europe.

ZYGIELLA PICKARD-CAMBRIDGE, 1902

Araneus Clerck, 1757, description of Araneus x-notatus(= Zygiella x-notata); Epeira Sundevall, 1833, descrip-tion of Epeira calophylla (= Zygiella x-notata); ZillaC.L. Koch, 1839, description of Zilla calophylla(= Zygiella x-notata); Eucharia C.L. Koch, 1845, de-scription of Eucharia atrica (= Zygiella atrica); ZygiaMenge, 1866, description of Zygia atrica (= Zygiellaatrica); Parazygiella Wunderlich, 2004 syn. nov., de-scription of Parazygiella montana (= Zygiella montana).

RemarksBecause the mutually paraphyletic Parazygiella andZygiella together form a well-supported clade, wesynonymize Parazygiella Wunderlich, 2004 with ZygiellaPickard-Cambridge, 1902. Based on morphology, Levi(1974) doubted that Zygiella calyptrata (Workman &Workman, 1894) is related to other Zygiella species.Although our analyses did not include Z. calyptrata,this species might be related to Yaginumia (see Ap-pendix 2), based on its distribution.

Type speciesEucharia atrica C.L. Koch, 1845.

DiagnosisSpecies of Zygiella differ from Leviellus by the malepalpal embolus tip being cylindrical rather than flat,and by the absence of a female epigynal scape.

DescriptionFemales small to medium in size (3–6.5 mm bodylength), males same size (Levi, 1974). Female epigynumwithout a scape, male embolus tip cylindrical (Levi,1974: 2–7, 9–11, 13–25, 28–39, 51–56, 60–64, 67–69,73–78). Carapace yellowish brown to light brown, headregion much darker to only slightly darker, many timesonly with a median black band, chelicerae dark brown.Sternum dark brown, mostly with a light-brown medianband. Abdomen yellowish brown to greyish brown, some-times with silver pigment spots, dorsally with a wave-

edged darker area that is sometimes darker only atthe edges. Legs yellowish brown to brown, lightlyannulated dark brown (own data; Nentwig et al., 2014).Orb webs with a spiral-free sector. During the day, thespiders hide off of the web in a tubular silk retreat,connected to the web hub by a signal line (Levi, 1974;Gregoric et al., 2010).

CompositionFollowing the revised phylogeny here, the genus in-cludes: Zygiella atrica (C.L. Koch, 1845), for synony-mies and diagnosis, see Levi (1974) and World SpiderCatalog (2015); distribution, Central and NorthernEurope, and Russia. Introduced to Canada and USA;Zygiella calyptrata (Workman & Workman, 1894), forsynonymies and diagnosis, see Levi (1974) and WorldSpider Catalog (2015); distribution, China, Myanmar,Malaysia; Zygiella carpenteri (Archer, 1951) comb. nov.,for synonymies and diagnosis, see Levi (1974) and WorldSpider Catalog (2015); distribution, USA; Zygiella dispar(Kulczynski, 1885) comb. nov.; for synonymies and di-agnosis, see Levi (1974) and World Spider Catalog (2015);distribution, Holarctic; Zygiella keyserlingi (Ausserer,1871), for synonymies and diagnosis, see Levi (1974)and World Spider Catalog (2015); distribution, South-ern Europe, Ukraine; Zygiella kirgisica Bakhvalov, 1974,for synonymies and diagnosis, see Bakhalov (1974) andWorld Spider Catalog (2015); distribution, Kyr-gyzstan; Zygiella minima Schmidt, 1968, for synony-mies and diagnosis, see Levi (1974) and World SpiderCatalog (2015); distribution, Canary Islands; Zygiellamontana (C.L. Koch, 1834) comb. nov., for synonymiesand diagnosis, see Levi (1974) and World Spider Catalog(2015); distribution, Palaearctic; Zygiella nearcticaGertsch, 1964, for synonymies and diagnosis, see Gertsch(1964) and World Spider Catalog (2015); distribution,Alaska, Canada, and USA; Zygiella pulcherrima(Zawadsky, 1902), for synonymies and diagnosis, seeZawadsky (1902) and World Spider Catalog (2015); dis-tribution, Russia; Zygiella x-notata (Clerck, 1757), forsynonymies and diagnosis, see Levi (1974) and WorldSpider Catalog (2015); distribution, Holarctic, Neotropical;Zygiella x-notata chelata (Franganillo, 1909), for syn-onymies and diagnosis, see Franganillo (1909) and WorldSpider Catalog (2015); distribution, Portugal; Zygiellax-notata percechelata (Franganillo, 1909), for synony-mies and diagnosis, see Franganillo (1909) and WorldSpider Catalog (2015); distribution, Portugal.



APPENDIX 2

Collection details of the specimens sequenced here. Unless otherwise stated, all specimen are deposited at the NationalMuseum of Natural History, Smithsonian Institution, Washington DC, USA

Family Genus Species Collection details

Araneidae Acusilas coccineus 1 female, China, Yunnan, Xishuangbanna, N21.936107 E101.254153, i. 2011, GregoricM., Kuntner M.

Araneidae Acusilas malaccensis 1 female, China, Yunnan, Xishuangbanna, N21.936107 E101.254153, i. 2011, GregoricM., Kuntner M.

Araneidae Araneus diadematus Slovenija, Razvanje, N46.532199 E15.641761, 15.x.2006, Gregoric M.Araneidae Araneus mitificus 1 female, Singapore, Bukit Timah, N1.348376 E103.777479, Kuntner M., Gregoric M.Araneidae Argiope lobata 1 male, Spain, Schneider J.Araneidae Caerostris cowani 1 female, Madagascar, Ranomafana, N21.256514 E47.437372, 19.iii.2010, Agnarsson

I., Kuntner M., Gregoric M.Araneidae Caerostris darwini 1 female, Madagascar, Madraka, N18.912647 E47.892627, 2.iii.2010, Agnarsson I.,

Kuntner M., Gregoric M.Araneidae Caerostris extrusa 1 female, Madagascar, Ranomafana, N21.256514 E47.437372, 22.iii.2010, Agnarsson

I., Kuntner M., Gregoric M.Araneidae Caerostris mitralis 1 female, Madagascar, Montagne d’Ambre, N12.4713 E49.21283, 17.xii.2005, Wood H.,

Raholiarisendra H., Rabemahafaly J.Araneidae Caerostris sexcuspidata 1 female, RS. Africa, Hogsback, N32.595483 E26.931567, 27.iii.2011, Haddad C.Araneidae Caerostris sumatrana 1 female, China, Yunnan, Xishuangbanna, Baka, N21.713675 E100.783023, 6.i.2011,

Gregoric M., Kuntner M.Araneidae Cyclosa conica 1 female, Switzerland, Gasteretahl, N46.44572 E7.74133, 7.vii.2011, Kuntner M.,

Gregoric M., Candek K.Araneidae Cyrtophora unicolor 1 female, Taiwan, New Taipei City, Wulai TonHo Cycing. 22.vi.2011, N24.850000

E121.616667, Cheng R.C.Araneidae Deliochus sp. B 1 female, Australia, NSW, Hornsby, N33.700004 E151.083328, 26.i.2006, Smith H.Araneidae Deliochus sp. E 1 female, Australia, NSW, Hornsby, N33.700004 E151.083328, 25.ii.2006, Smith H.Araneidae Eriowixia sp. 1 subadult female, China, Yunnan, Xishuangbanna, Baka, N21.713675 E100.783023,

i.2011, Gregoric M., Kuntner M.Araneidae Leviellus inconveniens 1 penultimate female, Israel, Tel Dan Reserve, N33.245980 E35.648300, 13.ix.2011,

Gregoric M., Cheng RC.Araneidae Leviellus thorelli 2 females, Macedonia, Ohrid, N41.113604 E20.793403, 22.x.2010, Gregoric M.Araneidae Leviellus sp. 1 penultimate male, Israel, Kineret, N32.780511 E35.419334, 16.ix.2011, Gregoric M.,

Cheng RC.Araneidae Milonia sp. A 1 female, China, Yunnan, Xishuangbana, Mengla, N21.611695 E101.583285, 1.i.2011,

Gregoric M., Kuntner M.Araneidae Milonia sp. H 1 female, Malaysia, Gombak, N3.324850 E101.752917, 6.i.2010, Gregoric M., Kuntner

M.Araneidae Neoscona sp. 1 subadult female, Yunnan, China, Xishuangbanna, N21.936107 E101.254153, i. 2011,

Gregoric M., Kuntner M.Araneidae Parazygiella montana 3 females, Slovenia, Goreljek, N46.32805833 E13.97433611, 16.x.2007, Gregoric M.Araneidae Parazygiella sp. A 2 females, Taiwan, Taichung, Hoping, N24.277540 E120.946086, Kuntner M.Araneidae Phonognatha sp. 1 female, Australia, Cape Arid NP, N33.666813 E123.347819, 1.i.2008, Frarnenau

V.W., Thomas M.L.Araneidae Poltys sp. A 1 female, China, Yunnan, Xishuangbanna, Baka, N21.713675 E100.783023, 8.i.2011,

Kuntner M., Gregoric M.Araneidae Poltys sp. E 1 female, Madagascar, Mantadia, N18.783784 E48.427617, 28.ii.2010, Agnarsson I.,

Kuntner M., Gregoric M.Araneidae Stroemiellus stroemi 4 females, Slovenia, Obrež, N46.40616667 E16.24122222, 7.x.2006, Gregoric M.Araneidae Yaginumia sia 1 female, 1 male, China, Hainan, Zhang S.Araneidae Zygiella atrica 1 females, 2 males, Czech Republic, Pardubice, N50.034303 E15.781696, 24.ix.2009,

Gregoric M.Araneidae Zygiella keyserlingi 3 females, Macedonia, Ohrid, N41.111951 E20.795727, 22.x.2010, Gregoric M.Araneidae Zygiella x-notata 3 females, Montenegro, Ulcinj, N41.930781 E19.214045, 21.x.2010, Gregoric M.Araneidae ‘halmahera’ 3 females, Indonesia, Halmahera, N1.828370 E127.816271, 6.vii.2007, Kuntner M.,

Gregoric M.Araneidae sp. ‘ORB019’ 1 female, 1 male, Madagascar, Ranomafana, N21.256514 E47.437372, 19.iii.2010,

Agnarsson I., Kuntner M., Gregoric M.



Appendix 2 Continued

Family Genus Species Collection details

Araneidae sp. ‘ORB021’ 1 female, Kent Ridge Park, Singapore, N1.286950 E103.788575, Kuntner M., GregoricM.

Araneidae sp. ‘ORB B’ 1 female, 1 male, China, Yunnan, Xishuangbanna, N21.936107 E101.254153, i. 2011,Gregoric M., Kuntner M.

Araneidae sp. ‘ORB D’ 1 female, China, Hainan, Ledong, N18.744921 E108.864331, vi.2010, Kuntner M.Araneidae sp. ‘ORB E’ 1 female, China, Hainan, Ledong, N18.744921 E108.864331, vi.2010, Kuntner M.Araneidae sp. ‘ORB F’ 1 female, China, Hainan, Ledong, N18.744921 E108.864331, vi.2010, Kuntner M.Deinopidae Deinopis sp. 1 female, Australia, Cape Arid NP, N33.666813 E123.347819, 1.i.2008, Frarnenau

V.W., Thomas M.L.Linyphiidae Linyphia triangularis 1 female, Slovenia, Tabor, N46.202998 E14.972155, 31.vii.2007, Gregoric M.Linyphiidae Meioneta rurestris 1 male, Slovenia, Sekirišce, N45.86315 E14.53671, 23.vi.2011, Candek K.Linyphiidae Neriene radiata 1 female, Switzerland, Grison Alps, N46.68081 E9.65574, 15.vii.2011, Kuntner M.,

Gregoric M., Candek K.Nephilidae Herennia etruscilla 1 female, Indonesia, Java, Cibodas, N6.738857 E107.011287, 19.vii.2007, Kuntner M.,

Gregoric M.Nephilidae Nephila clavipes 1 female, French Guiana, 27.xi.2005, Kuntner M.Nephilidae Nephilingis livida 1 female, Mayotte, 8.iv.2008, Kuntner M., Agnarsson I.Tetragnathidae Dolicognatha sp. 1 female, China, Yunnan, Xishuangbanna, Baka, N21.713675 E100.783023, i.2011,

Kuntner M., Gregoric M.Tetragnathidae Guizygiella guangxiensis 1 female, China, Yunnan, Xishuangbanna, N21.936107 E101.254153, i. 2011, Gregoric

M., Kuntner M.Tetragnathidae Guizygiella nadleri 1 female, 1 male, China, Yunnan, Xishuangbanna, N21.936107 E101.254153, i. 2011,

Gregoric M., Kuntner M.Tetragnathidae Guizygiella salta 1 female, China, Hainan, Ledong, N18.744921 E108.864331, vi.2010, Kuntner M.Tetragnathidae Guizygiella sp. B 1 female, China, Wuhan, N30.559600 E114.317600, 5.vi.2010, Kuntner M.Tetragnathidae Guizygiella sp. D 1 female, China, Hainan, Ledong, N18.744921 E108.864331, vi.2010, Kuntner M.Tetragnathidae Guizygiella sp. F 1 female, China, Yunnan, Xishuangbanna, N21.936107 E101.254153, i. 2011, Gregoric

M., Kuntner M.Tetragnathidae Guizygiella sp. G 1 female, China, Yunnan, Xishuangbanna, N21.936107 E101.254153, i. 2011, Gregoric

M., Kuntner M.Tetragnathidae Metellina merianae 1 female, Slovenia, Biš, N46.53738 E15.89630, 22.vii.2011, Kostanjšek R.Theridiidae Steatoda bipunctata 1 female, Slovenia, Spodnje Praprece, N46.161820 E14.693580, 1.xii.2007, Kuntner

M.Theridiidae Theridion varians 1 female, Slovenia, Primostek, N45.62986 E15.29969, 25.viii.2010, Kuntner M.,

Gregoric M., Lokovšek T.

SUPPORTING INFORMATION

Additional supporting information may be found in the online version of this article at the publisher’s web-site:

Appendix S1. Taxonomic and gene information of the 112 terminals used in our analyses, with GenBank ac-cession numbers and proportion of missing nucleotide data.Appendix S2. Phylogenies recovered by the different partition and coding schemes.Appendix S3. Coevolution of spider size (length of first leg patella + tibia) and web size (m2), and absoluteweb size plotted on the preferred topology (see Fig. 2) as discrete variables.Appendix S4. Web and behavioural character information of the 112 terminals used in our analyses.Appendix S5. Primer details and PCR protocols with varying annealing temperatures and cycle settings foreach gene fragment.



Documents

Phylogenetic position and composition of Zygiellinae and … · 2019-01-27 · Phylogenetic position and composition of Zygiellinae and Caerostris, with new insight into orb-web evolution