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Zoologischer Anzeiger 255 (2015) 71–79

Contents lists available at ScienceDirect

Zoologischer Anzeiger

jou rn al hom epage : w ww.elsev ier .com/ locate / j cz

ntercontinental island hopping: Colonization and speciation of therasshopper genus Phaulacridium (Orthoptera: Acrididae) inustralasia

ulia Goldberg a,∗, Mary Morgan-Richards b, Steven A. Trewick b

Department for Morphology, Systematics and Evolutionary Biology, J.F. Blumenbach Institute of Zoology & Anthropology, Georg-August-Universityöttingen, GermanyEcology Group, Institute of Agriculture and Environment, Massey University, Private Bag 11-222, Palmerston North, New Zealand

r t i c l e i n f o

rticle history:eceived 24 January 2014eceived in revised form 22 February 2015ccepted 22 February 2015vailable online 24 February 2015

eywords:rthopteracrididaeustraliaew Zealandord Howe Island

a b s t r a c t

Due to their distance from the pole and extent of surrounding oceans, southern hemisphere lands werenot subjected to such severe climatic conditions in the Pleistocene as those in the northern hemisphere.Pleistocene climate cycling did however result in extensive shifts in habitat zones due to fluctuation ofrainfall and temperature. Warm and wet conditions during interglacials supported southward exten-sion of forests, whereas cooler and drier environments during glacial maxima increased the extent ofdry grassland and scrub conditions. Such fluctuations are likely to have influenced the spatial distribu-tion and evolution of the fauna of Australasia. Using data from four genes (two mitochondrial and twonuclear) the genetic structure of the trans-Tasman grasshopper genus Phaulacridium was used to inferphylogeographic patterns.

The widespread New Zealand species Phaulacridium marginale shows a phylogeographic pattern typi-cal of recent range expansion, with low genetic diversity within the species. In stark contrast, the other

hatham IslandispersalGM

New Zealand species Phaulacridium otagoense is extremely localized in two small areas in southernNew Zealand where it exhibits very high genetic diversity. The phylogeographic patterns in AustralianPhaulacridium, however, show deeper divergence within the most widespread species, than between dif-ferent species in the same area. Mismatched correlation between geographic scale and genetic diversityimply that populations’ genetics contain the signature of past species ranges.

© 2015 Elsevier GmbH. All rights reserved.

. Introduction

Since Darwin (1809–1882) and Wallace (1823–1913), a fun-amental objective for biologists has been to understand theiversity and distribution of biota. This endeavor received a sub-tantial stimulus from phylogeographic approaches (e.g., Aviset al., 1987; Avise, 2000) which continue to develop and improvee.g., Hickerson et al., 2010). An initial emphasis of phylogeographicesearch was in North America and Europe, where studies have

rovided a degree of consensus on patterns and processes (e.g.,aberlet et al., 1998; Hewitt, 1999, 2000, 2004; Schmitt, 2007).hylogeographic structuring in these landscapes has been predom-

∗ Corresponding author. Tel.: +49 551395411.E-mail address: jule.goldberg@gmail.com (J. Goldberg).

ttp://dx.doi.org/10.1016/j.jcz.2015.02.005044-5231/© 2015 Elsevier GmbH. All rights reserved.

inantly influenced by two extrinsic factors. The first is the impactof extreme climate cycling throughout the Quaternary period withextension of polar ice sheets, vast periglacial regions with per-mafrost and lower global temperatures during glacials. The secondis the largely continuous distribution of land in northern latitudesacross thousands of kilometers of longitude. The biology of tem-perate northern hemisphere regions is to a large extent the storyof dispersal and range shifting since the last glacial maximum(LGM). In the southern hemisphere, the situation is rather dif-ferent. There was no extension of polar ice sheets to continentsbeyond Antarctica, although some small adjacent islands wereaffected (McIntosh et al., 2009; Nevill et al., 2010). Furthermore,landmasses are widely spaced and separated around longitude by

wide expanses of ocean. This probably restricts dispersal of terres-trial organisms, and also ameliorates the local intensity of climatefluctuations. The Australasian region therefore provides a strong

72 J. Goldberg et al. / Zoologischer Anzeiger 255 (2015) 71–79

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ig. 1. Vegetation habitat zones in Australia and New Zealand; (A) Last glacial maxiistribution of vegetation zones in New Zealand at (i) LGM, (ii) pre-human, (iii) mo

nd intriguing contrast to most studied northern hemisphere sys-ems, as it includes land areas that differ extensively in size andegree of spatial isolation, ranging from the continent of Australia7.6 million km2) in the west to New Zealand (268.000 km2) andhe Chatham Islands (900 km2) in the east (Fig. 1).

During the LGM, extensive montane glaciers developed on theouthern Alps in South Island, New Zealand (Fig. 1A and B(i))Alloway et al., 2007). Australia on the other hand was little affectedy ice, but experienced with New Zealand a general aridificationnd cooling resulting in expansion of shrub and dry grassland andestriction of high forest (Fig. 1A) (Lancashire et al., 2002; McKinnont al., 2004; Williams et al., 2009; Nevill et al., 2010). Lowered seaevel resulted in land connection between North and South Islandsf New Zealand (Trewick and Bland, 2012) and between Tasma-ia and mainland Australia (Chappell and Shackleton, 1986), buto significant increase in the proximity of Australia, New Zealandnd Chatham Island. Climate warming in the Holocene resulted inetraction of montane glaciers and expansion of forests which cov-red about 85% of New Zealand (Fig. 1B(ii)) prior to the arrival ofumans (Trewick and Morgan-Richards, 2009). The maritime cli-ate of the low lying Chatham Islands minimized habitat variation

hrough the Pleistocene (Holt et al., 2010).New Zealand is a continental island that is an emergent part

f Zealandia, a sunken continent separated from other parts ofondwana for approximately 80 Myr (million years) (Trewick et al.,007). Its biota comprises representatives of lineages that are else-here extinct, and organisms that are more typical of an isolated

rchipelago (Daugherty et al., 1993; Goldberg et al., 2008; Wallisnd Trewick, 2009). Extensive marine inundation of Zealandia cul-inating in the late Oligocene (Cooper and Millener, 1993; Landis

t al., 2008) may explain, at least in part, the absence of manyigher-level taxa in New Zealand (Daugherty et al., 1993), radiationf others (Cooper and Cooper, 1995), and the close relationships ofhe New Zealand fauna and flora to that of Australia (Pole, 1994;napp et al., 2007; Trewick and Morgan-Richards, 2009).

Miocene New Zealand was a relatively low-lying landmass with temperate or subtropical climate (Fleming, 1979; Daugherty et al.,993; Campbell and Hutching, 2007), modified in the Plioceney climate cooling and formation of the South Island Southernlps (Batt et al., 2000; Chamberlain and Poage, 2000), and sub-equently by Pleistocene climate cycling and glaciation (Carter,

005). Ecological and molecular evidence indicates that plant andnimal inhabitants of the subalpine/alpine zone of the Southernlps radiated during this time (Buckley et al., 2001; Lockhart et al.,001; Meudt and Simpson, 2006; Goldberg et al., 2008; Wallis

(LGM), modified from Hope et al. (2004) and Alloway et al. (2007). (B) Approximatepost human settlement) New Zealand (modified from Alloway et al., 2007).

and Trewick, 2009). We infer a shift from low lying habitat dom-inated by forests to a more diverse and heterogeneous one thatprovided opportunities for both colonization and radiation in thePlio-Pleistocene. Substantial and rapid fluctuations in the distribu-tion of vegetation communities during the Pleistocene (McGlone,1985) are thought to explain the modern distribution of manycold-sensitive taxa as a result of their survival in glacial refugia(Wardle, 1963; Burrows, 1965; Dumbleton, 1969). Studies of phy-logeographic patterns in alpine and forest species (e.g., Trewick,2008; Hill et al., 2009; O’Neill et al., 2009; Goldberg et al., 2011,2014; Marske et al., 2011) show little consensus among taxa (seealso Goldberg et al., 2008; Wallis and Trewick, 2009; Trewick et al.,2011). Less is known about the response of terrestrial taxa in low-land non-forest habitat. This in part reflects a natural scarcity ofsuch open habitats in Holocene New Zealand, but also lack of clar-ity about the evolutionary association between animals and habitattypes. For instance, many insect species that can persist on scrubvegetation or are primarily soil dwellers may not require matureforest to persist in a region. Taxa in such a setting provide the mostdirect means for comparison of responses to Pleistocene climatecycling that had a major impact on the distribution of non-foresthabitats.

We focus this research on the grasshopper genus PhaulacridiumBrunner v. Wattenwyl, 1893 (Orthoptera: Acrididae) that com-prises two species in Australia [Phaulacridium vittatum (Sjöstedt,1920) and P. crassum Key, 1992], one on Lord Howe Island (P.howeanum Key, 1992) and two in New Zealand [Phaulacridiummarginale (Walker, 1870) and Phaulacridium otagoense Westermanand Ritchie, 1984] (Key, 1992). The species are relatively small(<12 mm) and inhabit native and mixed exotic grasslands (Clark,1967; Key, 1992). Phaulacridium are primarily lowland grasshop-pers occurring up to ∼1200 m above sea-level in mainlandNew Zealand (Westerman and Ritchie, 1984) but can reachhigher altitudes in equitable areas of southern New Zealand,South Australia and Tasmania (Key, 1992; Harris et al., 2013).Most Phaulacridium have reduced, non-functional wings, butmacropterous (fully-winged) individuals of P. vittatum, P. crassumand P. marginale do sporadically occur (Westerman and Ritchie,1984; Key, 1992). In general, fewer than 3% of P. crassum and10% of P. vittatum are macropterous (Key, 1992), although insome populations of P. vittatum the macropterous form can be

more abundant (Clark, 1967). Winged P. marginale are probablyquite rare (Bigelow, 1967). Hutton (1897) recorded only twowinged animals (both female) in a sample of 218 specimens, buta 2011 population in Hawkes Bay appears to have had a higher

J. Goldberg et al. / Zoologischer Anzeiger 255 (2015) 71–79 73

Fig. 2. Distribution of mtDNA COI diversity among Phaulacridium grasshoppers in Australia and New Zealand with haplotype networks for each taxon. (A) Australian andNew Zealand region with distribution of Phaulacridium taxa, with matching colored circles in corresponding haplotype networks (NZmarg = New Zealand P. marginale, NZotaAlex = New Zealand P. otagoense from Alexandra area, NZota Mack = New Zealand P. otagoense from Mackenzie area, EAhow = P. vittatum eastern Australia and P. howeanum,WAcras = P. vittatum western Australia and P. crassum). (B) Map of New Zealand showing the four main sampling areas (three in mainland New Zealand, the fourth being theChatham Islands) of P. marginale with sampling locations (colored circles); inset (i) shows haplotype network for P. marginale, with colors corresponding to sampling areasa showm in Fig.

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nd depicting the frequency of the different areas within one haplotype; inset (ii)arginale representing haplotypes according to sampling areas and colored circles

ensity of macropterous individuals (M. Lusk pers. comm.). Inustralia, P. vittatum is the most widespread species and oftenecomes abundant enough to be considered a significant pestn pasture land (Australian Department of Agriculture and Food,ttp://agspsrv34.agric.wa.gov.au/ento/pestweb/Query1 1.idc?

D=307365355), whilst P. crassum has a very narrow rangeFig. 2A). On Lord Howe Island, approximately 575 km east of

ustralia in the Tasman Sea the endemic species P. howeanums restricted to arid rock outcrops (Key, 1992). In New Zealand,. marginale is today widespread in open grasslands on thehree main islands and the smaller islands in the north. It canlso be found on the Chatham Islands, approximately 850 kmast of New Zealand in the Pacific Ocean. In contrast, P. ota-oense is confined to semi-arid parts of central Otago and centralanterbury in South Island, New Zealand (Fig. 2A). Both of theew Zealand species occupy areas that now include exoticerbs and grasses.

New Zealand Phaulacridium are thought to have been derivedrom Australian lineages. Bigelow (1967) suggested that P.

arginale either arrived in New Zealand during the last 10,000ears (i.e., after the LGM), or that it arrived earlier and persistedhrough the Pleistocene in northern refugia. Westerman and Ritchie1984) favored the latter hypothesis. They also proposed evolutionf their newly described species, P. otagoense, in situ during theleistocene. They noted as an alternative, that morphological evolu-ion of P. otagoense might have been unusually rapid and extremelyecent (∼200 ybp) in response to human modification of habitatWesterman and Ritchie, 1984), but conceded this to be unlikelyiven the extent of morphological and molecular change indi-ated by their allozyme data. Key (1992) agreed that P. otagoenseas probably derived from P. marginale during the Pleistocene,

nd proposed that, of the two, P. marginale was morphologicallyore similar to P. vittatum. Westerman and Ritchie (1984) cited

omparatively warm Pliocene climate as providing suitable condi-ions for colonization of New Zealand during that time. Lowlandrassland/scrub was rare in Holocene New Zealand but the semi-

rid environment occupied today by P. otagoense in central Southsland was likely to have been more extensive during Pleistoceneglacial’ phases (Fig. 1). These areas were not glaciated but associ-ted cooler, drier climate is thought to have resulted in expansion

s a neighbour-joining tree representing all sampled taxa, with colored labels of P. 2B(i).

of scrub/grassland (McGlone et al., 2010). Thus, P. marginale ishypothesised to have evolved from an Australian lineage, and P.otagoense to have subsequently evolved from the P. marginalelineage.

Based on four genes (two mitochondrial and two nuclear) weexamined the genetic structure of the trans-Tasman grasshoppergenus Phaulacridium to asses if its genetic diversity correlates withits current ranges and to infer phylogeographic patterns in Aus-tralasia in relation to changing environments.

2. Material and methods

2.1. Sampling and DNA extraction

Phaulacridium grasshoppers (P. vittatum, P. marginale and P. ota-goense) were collected by hand from lowland grasslands and storedin 95% Ethanol. DNA from fresh ethanol material was extractedfrom muscle tissue of one hind femur using a salting-out method(Sunnucks and Hales, 1996). Specimens from Western Australia(P. crassum) and from Lord Howe Island (P. howeanum) werepreviously pinned and dried; here an entire hind leg was usedfor DNA extraction using CTAB and phenol/chloroform extrac-tions (Trewick, 2008). Species were identified by morphologicalcharacter differences, e.g., size, sculpturing of pronotum and posi-tion of tegmina (in Australian species) following Bigelow (1967),Westerman and Ritchie (1984) and Key (1992).

2.2. Polymerase chain reaction and sequencing

Molecular analyses used primers that target the mitochondrialDNA gene cytochrome oxidase I (COI). For a subset of samplesfrom all species cytochrome oxidase II (COII) and nuclear ITSand 18S genes were also amplified. In total 95 individuals, andtwo representative outgroup taxa (Supplementary Table 1) wereemployed. Additional to 72 available sequences of COI on GenBank(JN409741–JN409816; Goldberg & Trewick, 2011) we sequenced 21

individuals for COI using primers C1-J2195 and L2-N-3014 (Simonet al., 1994). The total sampling comprised 65 specimens of P.marginale from New Zealand (including Chatham Islands), 11 P.otagoense from southern New Zealand, 15 P. vittatum from western

74 J. Goldberg et al. / Zoologischer Anzeiger 255 (2015) 71–79

Table 1DNA variation and haplotype diversity within and between regional samples of P. marginale in the New Zealand region and the two populations of P. otagoense, with thesample size for each region (n), number of observed haplotypes (Nhaps), number of unique haplotypes (Nunihaps) per population, average number of nucleotide differences(k), nucleotide diversity (� × 10−3), number of polymorphic sites (S), Tajima’s D (*P < 0.05) and haplotype diversity (h). Region abbreviations correspond to abbreviations inSupplementary Table 1 and Fig. 2B (NNZ = Northern North Island, CNZ = Central Mainland New Zealand, SNZ = Southern South Island, CHI = Chatham Islands; NZota Alex = P.otagoense population in Alexandra, Otago, NZota Mack = P. otagoense population in Mackenzie, Canterbury).

Area n Nhaps Nunihaps k � S Tajima’s D h

NNZ 16 7 4 0.358 0.81 2 −0.3817 0.342CNZ 17 4 1 0.353 0.80 3 −1.706 0.118SNZ 22 7 2 0.273 0.62 3 −2.140* 0.177CHI 10 3 0 0 0 0 −1.401 0Total population 65 12 7 0.274 0.62 7 −2.051* 0.180CHI/NNZ 0.225 0.45CHI/CNZ 0.296 0.65CHI/SNZ 0.438 0.82NNZ/CNZ 0.360 0.80CNZ/SNZ 0.308 0.70NNZ/SNZ 0.360 0.75

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nd eastern Australia, two P. crassum from western Australia, two. howeanum from Lord Howe Island and the outgroup (Minyacris)rom Australia. Twelve of these grasshoppers were sequenced for a60 bp fragment of COII using primers TL2-J-3037 and C2-N-3661Simon et al., 1994), and four taxa were sequenced for a 730 bpragment of ITS using primers ITS4 and ITS5 and a 1095 bp frag-

ent of 18S using primers 18S-S22 and 18S-A1984. PCR reactionsere performed in 10 �l volumes. The amplified products were

hecked on 1% agarose gels and purified using high pure purifi-ation columns (Roche Applied Science, Mannheim, Germany) orAP/EXO1 digest (USB Corporation) following the manufacturer’snstructions. Purified PCR products were sequenced using standardrotocols for the ABI Prism BigDye Terminator Ready Reaction KitApplied Biosystems, Mulgrave, Australia) and run on an ABI Prism77 automated sequencer (Applied Biosystems). Sequence iden-ity was confirmed by comparison with published data, checkedor nucleotide ambiguities in Sequencher 4.2 (Gene Codes Corpo-ation, Ann Arbor, MI, www.genecodes.com) and aligned by eyesing Se–Al v2.0a11 (Rambaut, 1996). Sequences newly generated

or this study have been deposited at GenBank (accession numbers:P784376–KP784415).

.3. Population and phylogenetic analyses

The program TCS 1.21 (Clement et al., 2000) was run with a5% connection limit to construct parsimony haplotype networksf Phaulacridium COI sequences to assess the geographic structur-

ng of the genus. Furthermore P. marginale haplotype frequenciesere calculated for sampled regions in New Zealand. Using a pre-

iction of spatial partitioning, mainland New Zealand was dividednto three sampled zones of similar area, using latitudinal breakshat transected the islands east to west (Fig. 2B); northern Northsland (NNZ), central New Zealand (CNZ), southern South IslandSNZ). An additional zone was designated for the Chatham IslandsCHI). These were used to test for differences in genetic compositionpanning New Zealand. Under panmixis it is expected that no sig-ificant partitioning of genetic diversity would exist (thus �ST = 0),owever, any reduction in gene flow is expected to result in geneticiversity (haplotype variation and frequency) being partitioned inpace (�ST > 0).

DnaSP v5.0 (Rozas et al., 2003) was used to calculate haplotypeiversity (h), nucleotide diversity (�, Nei, 1987) and the average

umber of nucleotide differences (k) within P. marginale popula-ions and the two distinct populations of P. otagoense. Additionally,ajima’s D statistic (Tajima, 1989) was calculated for the differ-nt regions as it provides a useful indicator for neutral markers,

95 3.50 6 −0.863 0.90567 7.79 9 −0.491 0.83300 16.36 23 0.680 0.945

such as synonymous changes within mitochondrial DNA, of popu-lation range expansion and exchange (Ray et al., 2003; Wegmannet al., 2006). Mismatch distributions for the mainland New Zealandspecies P. marginale and P. otagoense were calculated and evidenceof isolation-by-distance was sought using a Mantel test of thecorrelation of pairwise geographic distances and pairwise �ST formtDNA with 1000 permutations for mainland P. marginale popula-tion samples using IBDWS v.2 (Jensen et al., 2005).

Distance estimation and phylogenetic analyses were per-formed using PAUP* 4.0b10 (Swofford, 1998). We conductedneighbor-joining (NJ) and maximum likelihood (ML) analyses, asimplemented in PAUP* with the entire COI dataset and with a sub-set of 31 samples, respectively. We used 1000 bootstrap replicatesto test the tree topology under ML and NJ criteria.

We employed Mr. Bayes 3.1.2 (Ronquist and Huelsenbeck, 2003)to examine tree topology with a COI–COII dataset under a sixparameter model similar to that selected for ML analysis by Model-test 3.7 (Posada and Crandall, 1998). We used the GTR model withgamma-distributed rate variation across sites and a proportion ofinvariable sites. The same model was applied to the two partitions(COI and COII) with rates and nucleotide frequencies for each geneunlinked. We used four independent MCMC runs for ten milliongenerations with a burn in of 25%. Resulting posterior probabilitieson the nodes were recorded.

Data for two nuclear genes (ITS and 18S) were examined to helpresolve the phylogenetic relationships within this genus. Analyseswere run in Paup* with a subset of four species from New Zealandand Australia.

3. Results

3.1. Population analyses

Statistical parsimony networks were generated in TCS contain-ing COI sequences of all Phaulacridium species (Fig. 2A). In a sampleof 65 specimens of P. marginale from mainland New Zealand wefound 12 different COI haplotypes (Fig. 2A and B inset (i), Supple-mentary Table 1) with uncorrected genetic distances up to 0.018(1.8%). Two of these haplotypes were observed at high frequency(n = 20 (31%) and 28 (43%) (Supplementary Table 1)), and both weredistributed throughout New Zealand (Fig. 2B, inset (i), Supplemen-tary Table 1), one including samples from the Chatham Islands.

Additionally, unique haplotypes were encountered in all regionalsamples except the Chatham Islands (Fig. 2B, Table 1). Strikingly,in contrast to the situation in P. marginale, diversity in the morerange restricted P. otagoense splits into two separate haplotype

J. Goldberg et al. / Zoologischer Anzeiger 255 (2015) 71–79 75

Fig. 3. Mismatch distributions calculated for P. marginale (black) and P. otagoense (grey) from COI mtDNA illustrating the differences in genetic divergence within the twospecies. (A) Frequency distribution plot based on proportionated frequencies and pairwise genetic distances highlighting the difference in frequency distribution in the twospecies, with P. marginale mostly exhibiting low genetic distances (up to 1.5%) and P. otagoense showing the highest frequency at approximately 4% pairwise genetic distance.( e twor a wi

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B) Distribution plot based on genetic and geographic distances of individuals of thange in P. otagoense (grey triangles) and the opposite – low genetic distance within

etworks, each representing grasshoppers from one of the twomall areas in southern New Zealand where this species is foundFig. 2A). This species displays higher genetic diversity than theidespread P. marginale with nine haplotypes found in a sample of

1 grasshoppers (Fig. 2A, Table 1). Of these nine haplotypes sevenere unique within the respective population (Fig. 2B inset (i), Sup-

lementary Table 1, Table 1). The calculated uncorrected pairwiseenetic distances within this species reached 0.015 (1.5%) in thelexandra population (NZotaAlex) and 0.018 (1.8%) in the moreorthern Mackenzie population (NZotaMack) (Fig. 2A). The maxi-al genetic distance between any two P. otagoense haplotypes was

.037 (3.7%) across a spatial range of about 180 km. This is sim-lar to the maximum genetic distance between P. otagoense and. marginale (0.035/3.5%). Mismatch distributions calculated for. marginale and P. otagoense highlights the difference in geneticivergence within these two species (Fig. 3A and B).

Populations of P. marginale displayed very low levels of DNAucleotide diversity (�) within mainland New Zealand and thehatham Islands (Table 1). The sequences of the northern areaad the highest level of haplotype and nucleotide diversity.hese results of mtDNA differentiation refute the prediction of P.arginale partitioning into northern, southern and Chatham Island

opulations, but rather show panmixis in New Zealand. In theurrent data set no isolation by distance is apparent. Tajima D testsTajima, 1989) were only significant for the SNZ dataset and theotal population (Table 1). Negative Tajima’s D statistics for the dif-

species, depicting the high genetic divergence coupled with a narrow geographicder geographic distribution – in P. marginale (black dots).

ferent populations indicate an excess of sites with low frequencypolymorphisms, i.e., the population has yet to reach equilibrium(Tajima, 1989). The two P. otagoense population samples on theother hand show higher estimates of � than P. marginale popu-lations in their small modern range (Table 1), implying that thisspecies had a larger population size and range until relativelyrecently.

The Australian species P. vittatum cannot be regarded asmonophyletic. The western population of P. vittatum is sisterto the western parapatric species P. crassum (WAcras), whilethe eastern P. vittatum is sister to P. howeanum on Lord HoweIsland (EAhow; Fig. 2A). Genetic distances within the Australianhaplotype networks reached a maximum of 0.013 (1.3%) and0.006 (0.6%), respectively. In comparison, the western AustralianPhaulacridium show genetic distances of up to 0.044 (4.4%) to east-ern Australian samples, and only 0.05 (5%) to P. marginale and 0.055(5.5%) to P. otagoense from New Zealand.

3.2. Phylogenetic analyses

Phylogenetic analysis illustrates the closer relationship of thewestern Australian P. vittatum specimens to individuals of P. cras-

sum compared to eastern populations of the same species, and thecloser affinity of the P. howeanum to the eastern Australian samplesof P. vittatum (Fig. 2B inset (ii)). Bootstrap support (1000 replicates)was not high for all internal nodes, a common problem encountered

76 J. Goldberg et al. / Zoologischer Anzeiger 255 (2015) 71–79

5 changes

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S.I., Canterbury

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ig. 4. (A) COI ML tree of 31 individuals representing all species of Phaulacridiumeplicates); the tree was rooted with outgroup taxa Minyacris nana and Minyacris ohaulacridium species used in this study for COI–COII generated in Mr. Bayes with v

hen lineages are closely related (ML tree; Fig. 4A). However, P.rassum was resolved with good support as sister to western Aus-ralian P. vittatum and P. howeanum as sister to eastern Australian. vittatum.

Bayesian analysis with longer DNA sequences (COI and COII) for set of 11 specimens representing the taxonomic and geographicange of the genus returned a tree with strong support for the inter-al nodes, but little support for separation of New Zealand andustralian clades (Fig. 4B).

The two nuclear genes (ITS and 18S) provided little phylogeneticignal as the sequences were identical or differed by at most threeubstitutions (ITS had 3 variable sites; 18S sequences were identi-al). The ITS variation was consistent with the inferences of shallowvolutionary history of the genus indicated by mitochondrial DNAata (results not shown).

. Discussion

Phaulacridium is an interesting example of a trans-Tasman dis-ribution in animals as there are few instances where species of theame genus naturally occur in Australia and New Zealand. Althoughhese grasshoppers are only rarely fully winged this does not appearo have prevented their dispersal across wide expanses of land andcean. The geographic range of Phaulacridium is large by southernemisphere standards as it extends approximately 4000 km fromest to east Australia, a further 1500 km across the Tasman Sea toew Zealand and 850 km to the Chatham Islands. Within this range,ve species are recognised, but the scale of the species’ rangesiffers enormously. Three species have small geographic ranges,

. crassum near the coast in Western Australia, P. howeanum on aock outcrop on Lord Howe Island (Tasman Sea) and P. otagoense inemi-arid land in central South Island, New Zealand (Fig. 2A). Thether two species are more widespread but comprise three disjunct

in this study. Numbers on branches show bootstrap support for the nodes (1000talis from Australia; (B) Midpoint rooted phylogeny of 11 individuals representing

for Bayesian posterior probabilities mapped to the branches.

geographic populations, P. vittatum west and east of the NullarborPlain in southern Australia, and P. marginale in New Zealand. East-ern P. vitattum also extend to Tasmania (Fig. 2A), and P. marginaleto the Chatham Islands (Fig. 2A and B). These species distributionsinclude two cases of parapatry (P. crassum surrounded by P. vittatumin Western Australia and P. otagoense by P. marginale in southernNew Zealand). Apparent reproductive isolation and ecological spe-cialisation is consistent with them having speciated in response tolocal microenvironments (Westerman and Ritchie, 1984).

Phylogeographic data add to this picture, revealing that inAustralia neighboring populations are more closely related to oneanother than geographically more distant populations, even to theextent that western P. vittatum are sister to parapatric P. crassum,and eastern P. vittatum are sister to P. howeanum. Consequently P.vittatum mtDNA is paraphyletic with respect to these two species.However, the situation in New Zealand is rather different, as the twolocal endemics are, by comparison, more deeply diverged from oneanother. There is higher genetic diversity in the locally restricted P.otagoense, compared to the widespread P. marginale, and supportfor monophyly of the New Zealand taxa is rather weak. Thus themitochondrial data do not support the hypothesis that P. otagoenseis derived from P. marginale (Key, 1992; Westerman and Ritchie,1984) as it only shows that both genetic lineages of P. otagoenseare sisters to P. marginale and had to shared a common ancestorwith P. marginale at some stage. While P. marginale is widespreadtoday, population genetic analyses suggest this is the result ofrecent population expansion. Genetic diversity in this species islow, even when New Zealand and Chatham Islands samples arecompared; implying P. marginale lineages constitute a continuous

entity in New Zealand. On the other hand, high genetic diversityin P. otagoense, despite a small modern range, suggests that thisspecies had until recently a comparatively large population sizeand range. Support for this idea comes from the observation that

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he habitat/region occupied by P. otagoense today probably repre-ents the most extensive natural lowland grass/scrubland availablen New Zealand prior to human colonization. Other extensive nativerasslands are those that developed above the treeline during theleistocene (Heenan and McGlone, 2013), but these are occupied by

separate unrelated group of ‘alpine’ grasshoppers (Bigelow, 1967;rewick, 2008; Trewick and Morris, 2008). This pattern empha-izes the need to consider past climatic and geological events whennterpreting extant patterns of diversity (Graham et al., 2006).

Vegetation patterns inferred for the Pleistocene LGM in Newealand indicate that during glacials, cooling and drying in theast led to the development of extensive grass- and shrublandsMcGlone et al., 1993, 2010). An increase in grassland habitat inoncert with a westerly airflow (Sanmartin et al., 2007) may havemproved the chances of long distance dispersal and establishmentn New Zealand by Phaulacridium. The near complete coverage ofon-alpine New Zealand by forest during interglacials would, con-ersely, have severely limited habitat availability for Phaulacridium.onsequently, it is likely that the range of P. marginale was most

imited during interglacial rather than glacial episodes (in contrasto at least one alpine orthopteran; Trewick et al., 2000). Suitableatural habitat was probably limited to grassland patches asso-iated with disturbed environments around rivers and frost flats.ven by 1840 when Polynesia and European colonizers were wellstablished, it is estimated that lowland swards covered no morehan 2% of mainland New Zealand (Mark and McLennan, 2004).he pattern of the widespread New Zealand species P. marginaleoes not reveal specific locations of refugia, as has been observed inorthern hemisphere Pleistocene phylogeography, but does exhibithe typical pattern of recent range expansion, with low geneticiversity. The fact that, until the arrival of humans in New Zealand∼1000 years ago), the only “substantial area below tree lineithout complete forest cover was central Otago” (McGlone et al.,

993) suggests that this semi-arid (e.g., Walker et al., 1995) orain-shadow grassland region (McGlone, 2001) may have been theefuge for P. otagoense during interglacials, as now. Distance fromceans and presence of mountain ranges, means that grasslandabitat could have existed here since formation of the South-rn Alps in the Pliocene (i.e., before Pleistocene global climateooling). Forest clearance by humans in New Zealand resultedn the expansion of native and subsequently mixed exotic grass-and habitat was available for P. marginale to expand its range.uring the 1800s grassland expanded in mainland New Zealand

Mark and McLennan, 2004), and the process continues today (cott, 1979).

The two distinct populations of P. vittatum in eastern andestern Australia are today separated by the Nullarbor Plain, a

arge semi-arid area that stretches approximately 1200 km fromestern Australia to Southern Australia (Key, 1992). Many other

rasshoppers occupy this region including plague forming speciesf Austroicetes. Mitochondrial lineages of P. vittattum are distinctiveo west and east Australia, lack monophyly and are likely to haveeen separated at least since the LGM as vast areas of southern andestern Australia, including the area that is now the Nullarbor,

upported arid desert habitat (Hope et al., 2004; Byrne et al., 2008).e find two morphologically distinct species (P. howeanum and P.

rassum) differ from the respective P. vittatum populations by smallitochondrial DNA distances, so they are within the respectiveitochondrial clades of P. vittatum regional populations.

The New Zealand lineages might be sister to the eastern Aus-ralian lineages as suggested by NJ and ML analyses (Fig. 2B (ii);ig. 4A and B), which indicates a general west to east pattern of

ange extension for the genus. Although, the number and direc-ion of trans-Tasman dispersal events cannot be confirmed with theurrent data, the presence of the closest relative to Phaulacridiumnd a diverse non-alpine grasshopper fauna in Australia support

nzeiger 255 (2015) 71–79 77

expansion from Australia to New Zealand. Additionally, shallowmitochondrial DNA genetic distance between New Zealand andChatham Island populations and lack of unique Chatham haplo-types suggest that these islands have been recently colonized bynumerous P. marginale individuals migrating from mainland NewZealand.

The population composition of the New Zealand species is con-trary to their current range sizes, with two lineages of P. otagoenseapparently occupying distinct areas and retaining high geneticdiversity (� = 16.36) in a relatively small space, compared to a verywidespread lineage (P. marginale) with relatively low genetic diver-sity (� = 0.62) (Figs. 2 and 3; Table 1). The genetic diversity withinthe two P. otagoense populations is more or less as high as the diver-sity in the P. marginale population (Table 1). In fact the geneticdiversity is so high and the phylogenetic signal deeply structuredthat the possibility arises that the two P. otagoense lineages fromAlexandra and Mackenzie are actually two distinct biological enti-ties. Other examples of paired species endemics in this area includechafer beetles (Emerson and Wallis, 1995) and Sigaus grasshoppers(Trewick, 2008).

Phaulacridium exemplifies the manner in which phylogeogra-phy can be driven by multiple factors on different time scales. Themost important factors in geologically recent times appear to havebeen the uplift of the Southern Alps in the Pliocene (5 Ma) creat-ing new alpine habitat and rain-shadow habitat, and Pleistocene(2.4 Ma) climate cooling events restricting available habitat in NewZealand. Both factors likely favored species that are more adaptedto dry and cold environment. In Australia, Pleistocene climatecycling had similar effects on the biota but on a larger geographicscale, with locally distributed high haplotype diversity withinpopulations in many taxa (Byrne, 2008), resulting in divergent lin-eages. This structure is evident in the present sampling of AustralianPhaulacridium with deeply divergent lineages within populations.

When compared to northern hemisphere grasshoppers, thereare stark contrasts at the population and species level dependingon habitat zones occupied. In Europe low genetic diversity in thelowland Chorthippus meadow grasshopper (Lunt et al., 1998) indi-cates widespread extirpation in the LGM, followed by extensiverange expansion. Diversity within Phaulacridium is more like thatof northern lowland species, but intriguingly this pattern of rangeexpansion and speciation has operated across a region with habitatpatches widely separated by desert and ocean.

Acknowledgements

We are indebted to many people who provided speci-mens for this study and helped with collection of specimens(in alphabetical order): Nathan Curtis (Lincoln University, Canter-bury, NZ), Rowan Emberson (Lincoln University, Canterbury, NZ),Rebecca Harris (University of Tasmania, Aus), Stewart Learmouth(Department of Agriculture, WA, Aus), Simon Morris, Jacquie Recsei(ANIC, Canberra, Aus), Phil Sirvid (Te Papa, Wellington, NZ), MikeWesterman (La Trobe University, Aus) and Frank Wieland (Pfalz-museum, Bad Dürkheim, GER).

This work was supported by the Marsden Fund of the RoyalSociety of New Zealand (03-GNS-302).

Appendix A. Supplementary data

Supplementary data associated with this article can be found, inthe online version, at http://dx.doi.org/10.1016/j.jcz.2015.02.005.

References

Alloway, B.V., Lowe, D.J., Barrell, D.J.A., Newnham, R.M., Almond, P.C., Augustinus,P.C., Bertler, N.A.N., Carter, L., Litchfield, N.J., McGlone, M.S., Shulmeister, J.,

7 cher A

A

A

B

B

B

B

B

B

C

C

C

C

C

C

C

C

D

D

E

F

G

G

G

G

G

H

H

H

HH

H

8 J. Goldberg et al. / Zoologis

Vandergoes, M.J., Williams, P.W., NZ-INTIMATE members, 2007. Towards aclimate event stratigraphy for New Zealand over the past 30,000 years(NZ-INTIMATE project). J. Quat. Sci. 22, 9–35.

vise, J.C., Arnold, J., Ball, R.M., Bermingham, E., Lamb, T., Neigel, J.E., Reeb, C.A.,Saunders, N.C., 1987. Intraspecific phylogeography: the mitochondrial DNAbridge between population genetics and systematics. Annu. Rev. Ecol. Syst. 18,489–522.

vise, J.C., 2000. Phylogeography: The History and Formation of Species. HarvardUniversity Press, Cambridge.

att, G.E., Braun, J., Kohn, B.P., McDougall, I., 2000. Thermochronological analysis ofthe dynamics of the Southern Alps New Zealand. Geol. Soc. Am. Bull. 112,250–266.

igelow, R.S., 1967. The Grasshoppers of New Zealand – Their Taxonomy andDistribution. University of Canterbury Publications, Christchurch, New Zealand.

uckley, T.R., Simon, C., Chambers, G.K., 2001. Phylogeography of the New Zealandcicada Maoricicada campbelli based on mitochondrial DNA sequences: ancientclades associated with Cenozoic environmental change. Evolution 55,1395–1407.

urrows, C.J., 1965. Some discontinuous distributions of plants within NewZealand and their ecological significance. Part II: disjunctions betweenOtago-Southland and Nelson-Marlborough and related distribution pattern.Tuatara 13, 9–29.

yrne, M., 2008. Evidence for multiple refugia at different timescales duringPleistocene climatic oscillations in southern Australia inferred fromphylogeography. Quat. Sci. Rev. 27, 2576–2585.

yrne, M., Yeates, D.K., Joseph, L., Kearney, M., Bowler, J., Williams, M.A.J., Cooper,S., Donnellan, S.C., Keogh, J.S., Leys, R., Melville, J., Murphy, D.J., Porch, N.,Wyrwoll, K.-H., 2008. Birth of a biome: insights into the assembly andmaintenance of the Australian arid zone biota. Mol. Ecol. 17, 4398–4417.

ampbell, H.J., Hutching, G., 2007. In Search of Ancient New Zealand. Penguin,Auckland & GNS Science, Lower Hutt, New Zealand.

arter, R.M., 2005. A New Zealand climatic template back to c. 3.9 Ma: ODP Site1119, Canterbury Bight, south–west Pacific Ocean, and its relationship toinland successions. J. R. Soc. N. Z. 35, 9–42.

hamberlain, C.P., Poage, M.A., 2000. Reconstructing the paleotopography ofmountain belts from the isotopic composition of authigenic minerals. Geology28, 115–118.

happell, J., Shackleton, N.J., 1986. Oxygen isotopes and sea level. Nature 324,137–140.

lement, M., Posada, D., Crandall, K.A., 2000. TCS: a computer program to estimategene genealogies. Mol. Ecol. 9, 1657–1659.

lark, D.P., 1967. A population study of Phaulacridium vittatum Sjöst (Acrididae).Aust. J. Zool. 1967, 799–872.

ooper, R.A., Millener, P.R., 1993. The New Zealand biota: historical backgroundand new research. Trends Ecol. Evol. 8 (12), 429–433.

ooper, A., Cooper, R.A., 1995. The Oligocene bottleneck and New Zealand biota:genetic record of a past environmental crisis. Proc. R. Soc. London B Biol. Sci.261, 293–302.

augherty, C.H., Gibbs, G.W., Hitchmough, R.A., 1993. Mega-island ormicro-continent? New Zealand and its fauna. Trends Ecol. Evol. 8, 437–442.

umbleton, L.J., 1969. Pleistocene climates and insect distributions. N. Z. Entomol.4, 3–23.

merson, B.C., Wallis, G.P., 1995. Phylogenetic relationships of the Prodontria(Coleoptera; Scarabaeidae; subfamily Melolonthinae) derived from sequencevariation in the mitochondrial cytochrome oxidase II gene. Mol. Phylogenet.Evol. 4, 433–447.

leming, C.A., 1979. The Geological History of New Zealand and its Life. AucklandUniversity Press, Auckland, New Zealand.

oldberg, J., Trewick, S.A., Paterson, A.M., 2008. Evolution of New Zealand’sterrestrial fauna: a review of molecular evidence. Philos. Trans. R. Soc. LondonB Biol. Sci. 363, 3319–3334.

oldberg, J., Trewick, S.A., 2011. Exploring phylogeographic congruence in acontinental island system. Insects 2, 369–399.

oldberg, J., Trewick, S.A., Powlesland, R.G., 2011. Population structure andbiogeography of Hemiphaga pigeons (Aves: Columbidae) on islands in the NewZealand region. J. Biogeogr. 38, 285–298.

oldberg, J., Knapp, M., Emberson, R.M., Townsend, J.I., Trewick, S.A., 2014. Speciesradiation of carabid beetles (Broscini: Mecodema) in New Zealand. PLoS ONE 9(1), e86185.

raham, C.H., Moritz, C., Williams, S.E., 2006. Habitat history improves predictionof biodiversity in rainforest fauna. Proc. Natl. Acad. Sci. U. S. A. 103, 632–636.

arris, R.M., McQuillan, P., Hughes, L., 2013. A test of the thermal melanismhypothesis in the wingless grasshopper Phaulacridium vittatum. J. Insect Sci.13, 51.

eenan, P.B., McGlone, M., 2013. Evolution of New Zealand alpine and open-habitatplant species during the late Cenozoic. N. Z. J. Ecol. 37 (1), 105–113.

ewitt, G.M., 1999. Post-glacial re-colonization of European biota. Biol. J. Linn. Soc.London 68, 87–112.

ewitt, G.M., 2000. The genetic legacy of Quaternary ice ages. Nature 405, 907–913.ewitt, G.M., 2004. Genetic consequences of climatic oscillations in the

Quaternary. Philos. Trans. R. Soc. London B Biol. Sci. 359, 183–195.

ickerson, M.J., Carstens, B.C., Cavender-Bares, J., Crandall, K.A., Graham, C.H.,

Johnson, J.B., Rissler, L., Victoriano, P.F., Yoder, A.D., 2010. Phylogeography’spast, present, and future: 10 years after Avise, 2000. Mol. Phylogenet. Evol. 54,291–301.

nzeiger 255 (2015) 71–79

Hill, K.B.R., Simon, C., Marshall, D.C., Chambers, G.K., 2009. Surviving glacial ageswithin the biotic gap: phylogeography of the New Zealand cicada Maoricicadacampbelli. J. Biogeogr. 36, 675–692.

Holt, K.A., Wallace, R.C., Neall, V.E., Kohn, B.P., Lowe, D.J., 2010. Quaternary tephramarker beds and their potential for palaeoenvironmental reconstruction onChatham Island, east of New Zealand, southwest Pacific Ocean. J. Quat. Sci. 25,1169–1178.

Hope, G., Kershaw, A.P., van der Kaars, S., Xiangjun, S., Liew, P.-M., Heusser, L.E.,Takahara, H., McGlone, M., Miyoshi, N., Moss, P.T., 2004. History of vegetationand habitat change in the Austral–Asian region. Quat. Int. 118–119, 103–126.

Hutton, F.W., 1897. The grasshoppers and locusts of New Zealand and theKermadec Islands. Proc. N. Z. Inst. 30, 135–150.

Jensen, J.L., Bohonak, A.J., Kelley, S.T., 2005. Isolation by distance, web service. BMCGenet. 6, 13.

Key, K.H.L., 1992. Taxonomy of the genus Phaulacridium and a related new genus(Orthoptera: Acrididae). Invertebr. Taxonom. 6, 197–243.

Knapp, M., Mudaliar, R., Havell, D., Wagstaff, S.J., Lockhart, P.J., 2007. The drowningof New Zealand and the problem of Agathis. Syst. Biol. 56, 862–870.

Lancashire, A.K., Flenley, J.R., Harper, M., 2002. Late glacial beech forest: an18,000–5000-BP pollen record from Auckland, New Zealand. Global Planet.Change 33, 315–327.

Landis, C.A., Campbell, H.J., Begg, J.G., Mildenhall, D.C., Paterson, A.M., Trewick, S.A.,2008. The Waipounamu erosion surface: questioning the antiquity of the NewZealand land surface and terrestrial fauna and flora. Geol. Mag. 145,1–25.

Lockhart, P.J., McLenachan, P.A., Havell, D., Glenny, D., Huson, D., Jensen, U., 2001.Phylogeny, radiation, and transoceanic dispersal of New Zealand alpinebuttercups: molecular evidence under split decomposition. Ann. Mo. Bot. Gard.88, 458–477.

Lunt, D.H., Ibrahim, K.M., Hewitt, G.M., 1998. MtDNA phylogeography andpostglacial patterns of subdivision in the meadow grasshopper Chorthippusparallelus. Heredity 80, 633–641.

Mark, A., McLennan, B., 2004. The conservation status of New Zealand’s indigenousgrasslands. In: 16th Annual Colloquium of the Spatial Information ResearchCentre (SIRC 2004: A Spatio-temporal Workshop) 29–30 November 2004,Dunedin, New Zealand, pp. 135–137.

Marske, K.A., Leschen, R.A.B., Buckley, T.R., 2011. Reconciling phylogeography andecological niche models for New Zealand beetles: looking beyond glacialrefugia. Mol. Phylogenet. Evol. 59, 89–102.

McGlone, M.S., 1985. Plant biogeography and the Cenozoic history of New Zealand.N. Z. J. Bot. 23, 723–749.

McGlone, M.S., 2001. The origin of the indigenous grasslands of southeastern SouthIsland in relation to pre-human woody ecosystems. N. Z. J. Ecol. 25, 1–15.

McGlone, M.S., Salinger, M.J., Moar, N.T., 1993. Paleovegetation studies of NewZealand’s climate since the last glacial maximum. In: Global Climate Since theLast Glacial Maximum. University of Minnesota Press, Minneapolis, pp.294–317.

McGlone, M.S., Newnham, R.M., Moar, N.T., 2010. The vegetation cover of NewZealand during the last glacial maximum: do pollen records under-representwoody vegetation? In: Haberle, S., Stevenson, J., Prebble, M. (Eds.), AlteredEcologies: Fire, Climate and Human Influence on Terrestrial Landscapes.Australian National University EPress, Canberra.

McIntosh, P.D., Price, D.M., Slee, A.J., Eberhard, R., Barrows, T., Bottrill, R., 2009.Periglacial and Associated Deposits of Tasmania: A Review Canqua-cgrgBiennial Meeting, May (3–8). Simon Fraser University, Burnaby CampusVancouver, Canada, pp. 128, Abstracts of presented papers.

McKinnon, G.E., Jordan, G.J., Vaillancourt, R.E., Steane, D.A., Potts, B.M., 2004.Glacial refugia and reticulate evolution: the case of the Tasmanian eucalypts.Philos. Trans. R. Soc. London B Biol. Sci. 359, 275–284.

Meudt, H.M., Simpson, B.B., 2006. The biogeography of the austral, subalpine genusOurisia (Plantaginaceae) based on molecular phylogenetic evidence: SouthAmerican origin and dispersal to New Zealand and Tasmania. Biol. J. Linn. Soc.London 87, 479–513.

Nei, M., 1987. Molecular Evolutionary Genetics. Columbia University Press, NewYork.

Nevill, P.G., Bossinger, G., Ades, P.K., 2010. Phylogeography of the world’s tallestangiosperm, Eucalyptus regnans: evidence for multiple isolated Quaternaryrefugia. J. Biogeogr. 37, 179–192.

O’Neill, S.B., Buckley, T.R., Jewell, T.R., Ritchie, P.A., 2009. Phylogeographic historyof the New Zealand stick insect Niveaphasma annulata (Phasmatodea)estimated from mitochondrial and nuclear loci. Mol. Phylogenet. Evol. 53,523–536.

Pole, M., 1994. The New Zealand flora – entirely long-distance dispersal? J.Biogeogr. 21, 625–635.

Posada, D., Crandall, K.A., 1998. Modeltest: testing the model of DNA substitution.Bioinformatics 14, 817–818.

Rambaut, A., 1996. Se–Al: Sequence Alignment Editor. Available at:http://tree.bio.ed.ac.uk/software/seal/(accessed 2013).

Ray, N., Currat, M., Excoffier, L., 2003. Intra-deme molecular diversity in spatiallyexpanding populations. Mol. Biol. Evol. 20, 76–86.

Ronquist, F., Huelsenbeck, J.P., 2003. MrBayes3: Bayesian phylogenetic inference

under mixed models. Bioinformatics 19, 1572–1574.

Rozas, J., Sánchez-DelBarrio, J.C., Messequer, X., Rozas, R., 2003. DnaSP, DNApolymorphism analyses by the coalescent and other methods. Bioinformatics19, 2496–2497.

cher A

S

S

S

S

S

S

T

T

T

T

T

Zealand. Biol. J. Linn. Soc. London 21, 283–298.Williams, M., Cook, E., van der Kaars, S., Barrows, T., Shulmeister, J., Kershaw, P.,

2009. Glacial and deglacial climatic patterns in Australia and surroundingregions from 35,000 to 10,000 years ago reconstructed from terrestrial and

J. Goldberg et al. / Zoologis

anmartin, I., Wanntorp, L., Winkworth, R.C., 2007. West wind drift revisited:testing for directional dispersal in the Southern Hemisphere using event-basedtree fitting. J. Biogeogr. 34, 398–416.

chmitt, T., 2007. Molecular biogeography of Europe: Pleistocene cycles andpostglacial trends. Front. Zool. 4 (11), 1–13.

cott, D., 1979. Use and conservation of New Zealand native grasslands in 2079*. N.Z. J. Ecol. 2, 71–75.

imon, C., Frati, F., Beckenbach, A., Crespi, B., Liu, H., Flook, P., 1994. Evolution:weighting and phylogenetic utility of mitochondrial gene sequences and acompilation of conserved polymerase chain reaction primers. Ann. Entomol.Soc. Am. 87, 651–701.

unnucks, P., Hales, D.F., 1996. Numerous transposed sequences of mitochondrialcytochrome oxidase I–II in aphids of the genus Sitobion (Hemiptera:Aphididae). Mol. Biol. Evol. 13 (3), 510–524.

wofford, D.L., 1998. PAUP*: Phylogenetic Analysis Using Parsimony (*and OtherMethods) Version 4. Sinauer Associates, Sunderland, MA.

aberlet, P., Fumagalli, L., Wust-Saucy, A.-G., Cosson, J.-F., 1998. Comparativephylogeography and postglacial colonization routes in Europe. Mol. Ecol. 7,453–464.

ajima, F., 1989. Statistical method for testing the neutral mutation hypothesis byDNA polymorphism. Genetics 123, 585–595.

rewick, S., 2008. DNA barcoding is not enough: mismatch of taxonomy andgenealogy in New Zealand grasshoppers (Orthoptera: Acrididae). Cladistics 24,240–254.

rewick, S.A., Morris, S.J., 2008. Diversity and Taxonomic Status of Some New

Zealand Grasshoppers. Department of Conservation Research & DevelopmentSeries, Wellington, New Zealand, pp. 290.

rewick, S.A., Morgan-Richards, M., 2009. New Zealand biology. In: Gillespie, R.G.,Clague, D.A. (Eds.), Encyclopedia of Islands. University of California Press,Berkeley, CA, pp. 665–673.

nzeiger 255 (2015) 71–79 79

Trewick, S.A., Bland, K.J., 2012. Fire and slice: paleogeography for biogeography atNew Zealand’s North Island/South Island juncture. J. R. Soc. N. Z. 42, 153–183.

Trewick, S.A., Wallis, G.P., Morgan-Richards, M., 2000. Phylogeographical patterncorrelates with Pliocene mountain building in the alpine scree weta(Orthoptera, Anostostomatidae). Mol. Ecol. 9 (6), 657–666.

Trewick, S.A., Paterson, A.M., Campbell, H.J., 2007. Hello New Zealand. J. Biogeogr.34, 1–6.

Trewick, S.A., Wallis, G.P., Morgan-Richards, M., 2011. The invertebrate life of NewZealand: a phylogeographic approach. Insects 2, 297–325.

Walker, S., Mark, A.F., Wilson, J.B., 1995. The vegetation of flat top hill: an area ofsemi-arid grassland/shrubland in Central Otago, New Zealand. N. Z. J. Ecol. 19,175–194.

Wallis, G.P., Trewick, S.A., 2009. New Zealand phylogeography: evolution on asmall continent. Mol. Ecol. 18, 3548–3580.

Wardle, P., 1963. Evolution and distribution of the New Zealand flora: as affectedby Quaternary climates. N. Z. J. Bot. 1, 3–17.

Wegmann, D., Currat, M., Excoffier, L., 2006. Molecular diversity after rangeexpansion in heterogeneous environments. Genetics 174,2009–2020.

Westerman, M., Ritchie, J.M., 1984. The taxonomy, distribution and origins of twospecies of Phaulacridium (Orthoptera: Acrididae) in the South Island of New

near-shore proxy data. Quat. Sci. Rev. 28, 2398–2419.