242 Bull.Br.arachnol.Soc.(2002)12(5),242–244 12S DNA sequence data confirm the separation of Material and methods Alopecosa barbipes and Alopecosa accentuata AmalespecimenofA.barbipeswascollectedbyCJV (Araneae, Lycosidae) and M. A. Hudson on 6 October 1999 in grass at Redgrave and Lopham Fen, Suffolk, (Grid ref. TM Cor J. Vink and Anthony D. Mitchell 0479, 52)23*N, 01)00*E). The specimen was stored in Ecology&EntomologyGroup, 95% ethanol and washed in sterile, deionised, distilled POBox84,LincolnUniversity, water before DNA extraction. DNA was extracted by Canterbury,NewZealand homogenising one each of legs III and IV using a proteinase-K digestion and high salt precipitation Summary method (White et al., 1990). A 263 base pair segment Phylogenetic analyses of DNA sequence data from the from the 12S rDNA was amplified and sequenced from 12S ribosomal subunit supported the relationship of A. barbipes following the methods in Zehethofer & AlopecosaaccentuataassisterspeciestoA.barbipes.Other Sturmbauer (1998). The sequence data have been sub- Alopecosaspecies,althoughmoremorphologicallydistinct, mitted to GenBank (Benson et al., 2000) (accession werefoundtohavefarlessvariationintheirsequencedata. number AY028420). Sequence data of the six Alopecosa species studied by Introduction Zehethofer&Sturmbauer(1998),andTrochosaterricola The Lycosidae is a morphologically conservative Thorell, 1856, were obtained from GenBank (accession family. In some cases species are separated by slight numbers AJ008022, AJ008024, AJ008025, AJ008026, morphologicaldifferences,whichhavebecomeapparent AJ008027, AJ008030, AJ008017 respectively). only after behavioural and/or ecological observations. Phylogenetic analyses were performed on the six This has been the case with species in genera such as Alopecosa sequences generated by Zehethofer & Alopecosa Simon, 1885 (Kronestedt, 1990), Schizocosa Sturmbauer(1998)plusA.barbipes.Itshouldbepointed Chamberlin, 1904 (Stratton, 1991) and Pardosa C. L. out that for this analysis we did not attempt to test the Koch, 1847 (Kronestedt, 1992, 1999). monophylyofAlopecosa,butsimplyrootedtheresulting Dahlem et al. (1987) studied the courtship behaviour trees on T. terricola, since Zehethofer & Sturmbauer of Alopecosa spp. and found differences between (1998) found this species to be distinct, but closely Alopecosa barbipes (Sundevall, 1833) and A. accentuata related to Alopecosa. The data were analysed with (Latreille,1817).Subsequently,Cordes&vonHelversen PAUP* (Swofford, 2001) using parsimony (an exhaus- (1990)notedmorphological,behavioural,andecological tivesearch)andmaximumlikelihood(aheuristicsearch differencesbetweenthespeciesandremovedA.barbipes with TBR branch swapping and 10 random sequence from synonymy with A. accentuata. Alopecosa barbipes additions). The general time reversible model (Yang, is found in regions with an oceanic climate, adults mate 1994) was used to estimate the maximum likelihood in autumn and often in the following spring, and males tree. The rate and among-site heterogeneity parameters haveanexaggeratedcourtshippostureandhaveablack wereestimatedinthesearch.Supportfortheparsimony tibial hair-brush on the front legs (Cordes & von tree was assessed by bootstrap analysis (Felsenstein, Helversen,1990).Alopecosaaccentuataisfoundinparts 1985). of Central Europe with a warm, continental climate, adults mate in spring, the courtship posture of males is Results less distinctive and they have no black tibial hair-brush on the front legs (Cordes & von Helversen, 1990). The Maximum likelihood analysis resulted in two trees, validity of A. barbipes as a species distinct from A. one of which is shown here (Fig. 1). The second tree accentuata is now generally accepted (e.g. Merrett & differed only in the placement of A. inquilina, which Millidge, 1992; Roberts, 1993). came out in a trichotomy with a second clade of A. 12S DNA sequences have recently been used to infer accentuataandA.barbipesandtheremainingAlopecosa Europeanlycosidphylogeneticrelationships(Zehethofer speciesinathirdclade.Theparsimonyanalysisresulted & Sturmbauer, 1998), which included sequences from in one most parsimonious tree that had an identical Alopecosa accentuata, A. cuneata (Clerck, 1757), A. topology to the maximum likelihood tree shown in pulverulenta (Clerck, 1757), A. taeniata (C. L. Koch, Fig. 1. Bootstrap analysis (1000 replicates) showed 1835),A.trabalis(Clerck,1757)andA.inquilina(Clerck, goodsupport(89%)forthecladecontainingA.cuneata, 1757). Zehethofer & Sturmbauer (1998) found that 12S A. pulverulenta, A. taeniata and A. trabalis, and some was especially suitable for resolving relationships support (44%) for the clade containing A. barbipes, between more distantly related taxa. In this study, a A. accentuata and A. inquilina. Alopecosa barbipes and sectionof12SmitochondrialDNAfromA.barbipeswas A. accentuata were found to be sister species (bootstrap sequenced and compared with sequences from other value of 44%). The sequences of A. barbipes (4 unique species of Alopecosa, including A. accentuata. Our nucleotide sites) and A. accentuata (six unique objectives were to see whether sequence data supported nucleotide sites) differed by nine transitions and one the separation of A. barbipes and A. accentuata and transversion. Within the seven Alopecosa species how these species were related to other species of examined there were 29 variable sites and these are Alopecosa. shown in Fig. 1. C.Vink&A.D.Mitchell 243 Discussion otherAlopecosaspecieswasunresolvedinZehethofer& Sturmbauer (1998), but the addition of A. barbipes in The ten base pair difference between the same 12S this study provides some resolution. Alopecosa barbipes mitochondrial subunit sequence from A. barbipes and is also shown to be sister to A. accentuata. A. accentuata supports the morphological, behavioural The genus Alopecosa contains more than 130 species and ecological differences found by Cordes & von (Platnick, 2000) and a very small proportion of this Helversen (1990). In contrast, A. pulverulenta and A. genus has been sequenced. However, this study demon- trabalis, which are morphologically distinct from each strates that morphological and genetic distinctiveness other (in both male and female genitalia) (Heimer are not strictly correlated. & Nentwig, 1991), have identical 12S sequences (Zehethofer & Sturmbauer, 1998). The clade containing A.cuneata,A.pulverulenta,A.taeniataandA.trabalisis Acknowledgements part of the ‘‘pulverulenta group’’ (Lugetti & Tongiorgi, Thanks to Marie Hudson for helping collect the 1969). There are only seven variable sites within this specimen of A. barbipes and to Peter Hudson for the clade, so it would appear that A. barbipes and A. trip to Redgrave and Lopham Fen. We thank Adrian accentuatahavebeenseparatespeciesforatleastaslong Patersonforhelpwiththephylogeneticanalysisandfor as the divergence of the species in the ‘‘pulverulenta comments on the manuscript. We also thank Rowan group’’. The section of mtDNA sequenced has been Emberson for comments on the manuscript. Thanks to foundtobehighlyconservedamongmanytaxa(Kocher Torbjörn Kronestedt for sending a copy of Cordes & et al., 1989), so sequencing of other gene regions (e.g. von Helversen (1990) and helpful advice on European COI (see Garb, 1999), ND1 (see Hedin, 1997)) may lycosids. more fully distinguish between closely related species such as A. pulverulenta and A. trabalis. The topology of the clade containing four species of References the ‘‘pulverulenta group’’ in the parsimony analysis and BENSON, D. A., KARSCH-MIZRACHI, I., LIPMAN, D. J., the maximum likelihood analysis agrees with the tree OSTELL, J., RAPP, B. A. & WHEELER, D. L. 2000: produced by Zehethofer & Sturmbauer (1998). 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MERRETT,P.&MILLIDGE,A.F.1992:Amendmentstothecheck Fig.1: MaximumlikelihoodtreeofAlopecosaspp.Numberswithout listofBritishspiders.Bull.Br.arachnol.Soc.9:4–9. bracketsarebootstrapvalues(1000replicates)fromtheparsi- PLATNICK, N. I. 2000: The world spider catalog. http:// monytreewiththesametopology.Numbersinbracketsshow research.amnh.org/entomology/spiders/catalog81–87/NewYork, numbersofapomorphies(nucleotidechanges). TheAmericanMuseumofNaturalHistory. 244 12SDNAsequenceofAlopecosabarbipes ROBERTS,M.J.1993:AppendixtoThespidersofGreatBritainand & T. J. White (eds.), PCR protocols: a guide to methods and Ireland.Colchester,HarleyBooks. applications:315–322.SanDiego,AcademicPress. STRATTON, G. E. 1991: A new species of wolf spider, Schizocosa YANG, Z. 1994: Maximum likelihood phylogenetic estimation from stridulans(Araneae,Lycosidae).J.Arachnol.19:29–39. DNA sequences with variable rates over sites: approximate SWOFFORD, D. L. 2001: PAUP*: phylogenetic analysis using methods.J.mol.Evol.39:306–314. parsimony (and other methods), version 4.0b5. Sunderland, ZEHETHOFER, K. & STURMBAUER, C. 1998: Phylogenetic Massachusetts,SinauerAssociates. relationships of Central European wolf spiders (Araneae: WHITE,T.J.,BRUNS,T.,LEE,S.&TAYLOR,J.1990:Amplifi- Lycosidae)inferredfrom12SribosomalDNAsequences.Mol. cation and direct sequencing of fungal ribosomal RNA genes phylogenet.Evol.10:391–398. forphylogenetics.InM.A.Innis,D.H.Gelfand,J.J.Sninsky Bull.Br.arachnol.Soc.(2002)12(5),244–248 Opopaea fosuma, n. sp. from Sumatra, Indonesia Diagnosis: Males of Opopaea fosuma, n. sp. are (Araneae, Oonopidae) separated from those of most other Opopaea species by the structure of the palp. The cymbium and bulb are Matthias Burger, Wolfgang Nentwig only partly fused in O. fosuma, n. sp. whereas they are ZoologicalInstitute,UniversityofBern, completely fused in most other Opopaea species, and Baltzerstrasse6,CH-3012Bern,Switzerland the embolus is long and slender in O. fosuma, n. sp. The species is similar to the Micronesian O. foveolata and Roewer, 1963, where, according to the original description, the cymbium and bulb of the male palp Christian Kropf are also only partly fused (Roewer, 1963: fig. 6g). NaturalHistoryMuseum, Males of O. fosuma, n. sp. differ from those of Bernastrasse15,CH-3005Bern, O. foveolata, however, by their long and slender Switzerland embolus, whereas it is rather short and blunt in O. foveolata. Summary Male: Measurements (n=1): Prosoma length 0.59, width0.48,height0.23.Opisthosomalength0.67,width Anewspeciesofoonopid,Opopaeafosuma,isdescribed from both sexes collected in the Kerinci Seblat National 0.42, height 0.41. Appendages: ParkinCentralSumatra.Possiblerelationshipswithother speciesofOpopaeaandthestructuresofthefemalegenitalia Fe Pa Ti Mt Ta Total andtheembolusarediscussed. LegI 0.37 0.23 0.26 0.20 0.15 1.21 LegII 0.32 0.20 0.22 0.19 0.15 1.08 LegIII 0.29 0.15 0.16 0.17 0.15 0.92 Introduction LegIV 0.41 0.22 0.30 0.26 0.20 1.39 Palp 0.12 0.16 0.06 — 0.16 0.50 The oonopid genus Opopaea was described by Simon in 1891. The male palp of the type species, Colour (alcohol-preserved material): Prosoma and O. deserticola, is notable for a characteristically chelicerae orange; legs and palps light orange. enlarged patella (Simon, 1891). The following descrip- Opisthosoma:ventralscutumorange;dorsalscutumand tion of an Indonesian species of the genus Opopaea sclerite that partially surrounds spinnerets light orange; is based on material collected in the Kerinci Seblat spinnerets pale yellow; soft areas white. Carapace: National Park in Central Sumatra and provided for Ovoid; narrowed in eye region (Fig. 1), slightly ascend- description by Dr C. Deeleman. All measurements are ing behind PME, then almost horizontal and posterior in mm. 1/3 steeply descending (Fig. 2). Tiny hairs forming U-shaped band; three areas of tubercles near lateral borders; additional hair-bearing tubercles along lateral Opopaea fosuma Burger, n. sp. (Figs. 1–24) borders, some in posterior area (Fig. 1). Eyes: Six; Types: Male holotype, Kerinci Seblat National Park, almost circular, with roughly same diameter; AME 800m a.s.l., Central Sumatra, Indonesia, 21–30 July lacking;eyegroupoccupiesslightlymorethan1/2width 1988, leaf litter, Suh. Djojosudharmo leg., deposited in of head; posterior row slightly recurved (Fig. 1); PME NMBE (Natural History Museum, Bern, Switzerland). almostcontiguous;ALEseparatedfromPMEandfrom Paratypes, leg. Suh. Djojosudharmo at type locality: PLE by 1/2 and from each other by one diam. of ALE; 1X 2Y (NMBE), 2X 2Y (Coll. Deeleman, Ossendrecht, PLEseparatedfromPMEby1/2diam.ofPLE(Fig.3). The Netherlands). Exact position and size of eyes somewhat variable. Etymology: The specific name is an abbreviation of Clypeus (Fig. 3): Two diam. of ALE high; slightly ‘‘found in Sumatra’’. extended between chelicerae in middle. Sternum and