systematics and ecology of anachipteria geminus sp. nov ...publish.uwo.ca/~zlindo/publications1/my...

Systematics and ecology of Anachipteria geminussp. nov. (Acari: Oribatida: Achipteriidae) from

arboreal lichens in western North America

Zoë Lindo1

Department of Biology, University of Victoria, P.O. Box 3020, Station CSC,Victoria, British Columbia, Canada V8W 3N5

Marilyn ClaytonForest Biodiversity Network, Pacific Forestry Centre, Canadian Forest Service,

Natural Resources Canada, Victoria, British Columbia, Canada V8Z 1M5

Valerie M. Behan-PelletierBiodiversity Program, Research Branch, Agriculture and Agri-Food Canada,

K.W. Neatby Building, Ottawa, Ontario, Canada K1A 0C6

Abstract—We present the systematics and ecology of a new species of arboreal oribatid mite inthe family Achipteriidae, Anachipteria geminus sp. nov., a dominant arthropod in canopy lichensassociated with western hemlock (Tsuga heterophylla (Raf.) Sarg. (Pinaceae)) and Pacific silverfir (Abies amabilis (Dougl. ex Loud.) Dougl. ex Forbes (Pinaceae)) in the coniferous temperateand montane forests of the Pacific Northwest of North America. The species is described on thebasis of the morphology of all active instars plus molecular sequence data for the D3 region ofthe 28s rRNA (D3-28s) and mitochondrial cytochrome oxidase 1 (CO1) genes. Anachipteriageminus is the dominant oribatid mite in foliose lichens in the upper canopies of Pacific montaneconifer trees. Specimens were found in 100% of canopy lichens sampled, and abundances canreach 1450 individuals / 100 g dry mass of lichen. Analysis of the population structure indicatesthat A. geminus has seasonally stable populations consistent with overlapping generations.Anachipteria geminus expresses morphological character states that require expanding the diag-noses of the genus Anachipteria and the family Achipteriidae. A revised diagnosis and morpho-logical key to species of Anachipteria in Canada are given.

Résumé—Nous présentons la systématique et l’écologie d’une nouvelle espèce d’oribatide arbori-cole dans la famille Achipteriidae, Anachipteria geminus sp. nov., un arthropode dominant dans leslichens de la canopée associés à la pruche de l’ouest (Tsuga heterophylla (Raf.) Sarg. (Pinaceae))et au sapin gracieux (Abies amabilis (Dougl. ex Loud.) Dougl. ex Forbes (Pinaceae)) dans les forêtsde conifères tempérées de montagne du Pacifique au Nord-ouest de l’Amérique du Nord. L’espèceest décrite sur la base de la morphologie de toutes les stades actifs, plus les données d’ordre molé-culaires pour la région D3 de 28 rRNA (D3-28) et mitochondrial cytochrome oxidase 1 (CO1).Anachipteria geminus est l’oribatide dominant dans les lichens foliose des canopées supérieuresdes conifères de montagne du Pacifique au Nord-ouest. Les exemplaires ont été trouvés dans 100 %des lichens des canopées échantillonnés et la densité peut atteindre 1450 individus / 100 g demasse sec du lichen. L’analyse de la structure démographique indique que A. geminus a des popu-lations saisonnièrement stables en accord avec les générations chevauchantes. Anachipteria gemi-nus exprime des états de caractères morphologiques qui exigent le développement de lesdiagnostiques du genre Anachipteria et de la famille Achipteriidae. Une diagnostique révisée et uneclé morphologique aux espèces d’Anachipteria au Canada sont données.

556Lindo et al. Introduction

The genus Anachipteria Grandjean (Acari:Oribatida: Achipteriidae) is known from the

Holarctic, Oriental, and Neotropical regions,with most species described from the centraland eastern United States of America, centralEurope, and Japan. The genus includes 45

Can. Entomol. 140: 539–556 (2008) © 2008 Entomological Society of Canada

539

Received 20 January 2008. Accepted 11 April 2008.

1Corresponding author (e-mail: [email protected]).

named species of which 9 are known fromNorth America (Marshall et al. 1987; Subías2007). Four species of Anachipteria were re-corded previously from Canada: Anachipteriaacuta (Ewing) and A. australoides Jacot fromBritish Columbia, A. howardi (Berlese) fromnorth-central Canada, and A. magnilamellata(Ewing) from eastern Canada (Behan-Pelletierand Eamer 2004). Members of this genus aretypically collected from forest-floor organicsoils, but recent studies have shown some spe-cies to be arboreal (Lindo and Winchester 2006;Root et al. 2007a).

Oribatid mites are known from many arborealhabitats, including the bark and trunk of trees(Nicolai 1993), leaf domatia and stems (Walterand O’Dowd 1995), moss, lichen, and othercorticolous epiphytic cover (Seyd and Seaward1984; André 1985; Root et al. 2007b), and in ac-cumulations of organic matter known as suspendedsoils (Wunderle 1992; Behan-Pelletier et al. 1993;Lindo and Winchester 2007). Among Oribatida,representatives of the families Camisiidae,Ceratozetidae, Scheloribatidae, and Mycobatidaeare usually the numerically dominant mites in thecanopy of old-growth Pacific Northwest forests(Behan-Pelletier and Walter 2000).

In this paper we describe the first species inthe family Achipteriidae that is known solelyfrom arboreal habitats in the Pacific Northwest.Anachipteria geminus sp. nov. is described onthe basis of specimens collected from arboreallichens growing on western hemlock (Tsugaheterophylla (Raf.) Sarg. (Pinaceae)) and Pa-cific silver fir (Abies amabilis (Dougl. exLoud.) Dougl. ex Forbes (Pinaceae)) trees incoastal Oregon and British Columbia. In addi-tion to describing the external morphology ofall active stages, we provide molecular se-quence data for the D3 region of the 28s rRNA(D3-28s) and mitochondrial cytochromeoxidase 1 (CO1) genes, and ecological data.Anachipteria geminus expresses morphologicalcharacter states that require expansion of the di-agnoses of Anachipteria and Achipteriidae. Wediscuss the morphology of this species in thecontext of its genus, family, and superfamily,and provide a key to adults of the five speciesof Anachipteria now known from Canada.

Materials and methods

Species descriptionMorphological terminology is mostly that of

F. Grandjean (for general references see Travé

and Vachon (1975), for leg setal nomenclaturesee Norton (1977), and for an overview seeWeigmann (2006)). The following conventionsfor measurements and description are used: to-tal length, distance measured dorsally from thetip of the rostrum to the posterior edge of thenotogaster, on specimens in lactic acid in cavityslides; total width, distance measured at thewidest part of the notogaster, perpendicular tothe circumgastric scissure on specimens in lac-tic acid in cavity slides; and setae, length mea-sured on dissected slide-mounted specimens:ro, rostral setae; le, lamellar setae; in,interlamellar setae; ex, exobothridial setae; bo,bothridial seta; 1x, 2x, 3x, and 4x, coxisternalsetae; g, genital seta; ag, aggenital seta; ad,adanal seta; an, anal seta. The unideficience no-menclature is used for the notogastral setation.Setal measurements are averaged from 10 spec-imens.

Specimens for scanning electron microscopywere stored in 70% ethanol, dehydrated in etha-nol and acetone, critical-point-dried, mountedon aluminum stubs with double-sided tape, andcoated with gold–palladium.

Molecular sequencing techniquesA single live individual of A. geminus was se-

quenced for two genes: the D3 region of the 28srRNA (D3-28s) and mitochondrial CO1. Forcomparison, a single specimen of each of the fol-lowing seven other oribatid mite species was se-quenced (specimen provenance is given inTable 1): Achipteria curta Aoki, A. acuta,Anachipteria sp., and a species of DentachipteriaNevin (Achipteriidae); Ceratozetes pacificusBehan-Pelletier (Ceratozetidae); and one specieseach of the genera Scheloribates Berlese(Scheloribatidae) and Tegoribates Ewing(Tegoribatidae). We extracted total genomic DNAof each specimen with DNeasy Tissue Kit(QIAGEN), using protocols therein, with 50 μLelution buffer. Primers used in the polymerasechain reaction (PCR) amplification cycle for theD3-28s gene, which are given in Maraun et al.(2004), were diluted to 50 μmol. Five replicateamplifications per sample were performed in50 μL total volume containing 5 μL 2.5 mmolMgCl2 buffer solution, 1 μL each primer, 1 μLDNA, and 25 μL 2× Readymix Taq PCR reactionstock (QIAGEN). PCR consisted of an initial de-naturation step at 95 °C for 3 min, followed by 35amplification cycles (94 °C for 30 s, 54 °C for45 s, 72 °C for 1 min), ending with a 7-min elon-gation step at 72 °C.

© 2008 Entomological Society of Canada

540 Can. Entomol. Vol. 140, 2008

The primer pair LCO1490 (5′-GGTCAA-CAAATCATAAAGATATTGG-3′) and HCO2198(5′-TAAACTTCAGGGTGACCAAAAAATCA-′3 ) (Folmer et al. 1994) diluted to 50 μmol was

used in the PCR amplification cycle for CO1.Amplifications were performed in 50 μL totalvolume containing 5 μL 2.5 mmol MgCl2 buffersolution, 5 μL each primer, 1 μL DNA, and 25μL 2× Readymix Taq PCR reaction stock(QIAGEN). PCR consisted of an initial denatur-ation step at 95 °C for 15 min followed by 36amplification cycles (94 °C for 30 s, 47 °C for60 s, 72 °C for 60 s), ending with a 10-minelongation step at 72 °C based on protocolsgiven in Heethoff et al. (2007).

PCR products were purified using the QIAquickPCR purification kit (QIAGEN). Purified DNAwas run on a 1% agarose gel at 100 V for 1 h toestimate the concentration of DNA in the puri-fied products. Products estimated at >5 ngDNA/μL were prepared for cloning. Ligationsteps followed protocols of Promega (PromegaCorporation, Madison Wisconsin) usingpGem plasmid vector. Transformation followedInvitrogen protocols (Invitrogen Canada Inc.,Burlington, Ontario) whereby 2 μL ligation wasadded to 50 μL competent cells (Esche-richia coli) incubated on ice for 30 min, fol-lowed by heat shock for 45 s at 42 °C. Twohundred and fifty microlitres SOC media was

added to each sample, which was incubated for1 h at 37 °C while rotating. Final solutions wereplated on LB medium plates (with ampicillin, X-Gal, and IPTG) using 50 μL transformed prod-uct. Plates were incubated overnight at 37 °C.Five colonies for each sample for each specieswere selected by blue/white screening followedby growth in 1.5 mL LB media at 37 °C, whileshaking, for 24 h. Plasmid preparation was car-ried out with the QIAprep Spin Miniprep Kit(QIAGEN) and final DNA concentrations wereestimated by measuring sample absorbance at260 nm. Approximately 500 ng DNA was usedin the restriction digest with 4 μL 10× BSA,10× Nebuffer3, NCo1, and Pst1 (New EnglandBiolabs, Inc., Pickering, Ontario). Restriction di-gests were performed at 37 °C for 3 h and runon 1% agarose gels to assess whether D3-28srRNA or CO1 mtDNA inserts were present inthe final samples. The sample with the highestconcentrations of oribatid mite D3-28s rRNAand CO1 mtDNA for each species was sent tothe University of Victoria sequencing labora-tory for sequencing. All sequences were con-firmed to be of oribatid mite origin bycomparisons with known sequences inGenBank using the BLASTn search algorithm(Altschul et al. 1997). Sequences were alignedand compared using ClustalW (Thompson et al.1994).


Lindo et al. 541

GenBankaccession No. Gene* Locality Habitat notes

Tegoribates sp. EF989716 D3-28s (353) Sidney Inlet, ClayoquotSound, Vancouver Island

Forest floor underSitka spruce

Achipteria curta EF989724 D3-28s (355) Walbran Valley,Vancouver Island

Forest floor underwestern redcedar

Anachipteria acuta EF989720 D3-28s (355) Walbran Valley,Vancouver Island

Suspended soil inwestern redcedar

Anachipteriageminus sp. nov.

EF989717EF989722

D3-28s (355)CO1 (709)

Walbran Valley,Vancouver Island


Anachipteria sp. EF989725 D3-28s (355) Sidney Inlet, ClayoquotSound, Vancouver Island

Forest floor underSitka spruce

Dentachipteria sp. EF989718 D3-28s (356) Walbran Valley,Vancouver Island

Forest floor underwestern redcedar

Ceratozetes pacificus EF989719EF989723

D3-28s (355)CO1 (709)

Walbran Valley,Vancouver Island


Scheloribates sp. EF989721 D3-28s (353) Walbran Valley,Vancouver Island


*The number in parentheses is the number of nucleotide base pairs in the sequence.

Table 1. List of specimens used for DNA sequencing, with GenBank accession numbers for nucleotidesequences, and collection localities and habitat notes.

Sampling to obtain ecological dataEcological data were generated from sam-

ples obtained in 1996 at the Montane Alterna-tive Silvicultural Systems (M.A.S.S.) site situatedin the Montane Moist Maritime Coastal West-ern Hemlock biogeoclimatic variant (Greenand Klinka 1994) on Vancouver Island, BritishColumbia. Plots within the M.A.S.S. site areeither (i) old-growth coniferous forest domi-nated by western hemlock (200–800 years old)and Pacific silver fir (generally <250 yearsold) or (ii) shelterwood, which is an experi-mental selection harvest system derived fromold growth, in which 30% of trees (represent-ing the entire stand profile) remain after har-vesting.

Three replicate subsamples from each of twohabitat substrates (branch tips and foliose li-chen scrapings) were collected within the uppercanopy (~40 m) of six Pacific silver fir and sixwestern hemlock trees in both an old-growthand a shelterwood plot at the M.A.S.S. site (12trees were sampled, for a total of 72 samplesper collection period, i.e., 216 samples in total).The same trees were resampled over three col-lection periods in 1996: period 1: 25–28 May,while snow was still on the ground; period 3:29 July – 2 August; and period 6: 4–6 Novem-ber, before the first snowfall. Single rope climb-ing techniques were used to access the canopy(Barker and Standridge 2002). Lichen samples(approximately 25 g) were hand-picked andbagged and branch samples (approximately50 cm lengths, including stem, needles, andcones) were bagged and cut in the canopy andlowered to the ground. Specimens were extractedfrom lichen and branch samples using a sodiumhydroxide wash (Clayton and Humble 1999) andstored in 70% ethanol. All data are standardizedas number of specimens per 100 g dry mass(DM) of substrate material (branch, lichen). Data(standardized abundances of adult, immature, andtotal A. geminus) were analysed using a repeated-measures nested analysis of variance.

The specimens of Acari belonged to the fol-lowing collections:

CNC Canadian National Collection of Insects,Arachnids, and Nematodes, Agricultureand Agri-Food Canada, Ottawa, Ontario,Canada

PFC Pacific Forestry Centre, Canadian For-estry Service, Natural Resources Canada,Victoria, British Columbia, Canada

ZLC Personal collection of Zoë Lindo

Anachipteria geminus sp. nov.(Figs. 1–30)

Material examinedHolotype: Adult female. CANADA. British

Columbia: Vancouver Island: Campbell River,M.A.S.S. site, 49°50′53′′N, 125°26′27′′W, ele-vation 740–850 m, 28.iv.1996 (L.M. Humbleand N.N. Winchester), from foliose lichens onwestern hemlock; stored in 70% ethanol anddeposited in CNC, type No. 23577. Para-types: Twenty with same data as holotype,slide-mounted in Hoyer’s medium, deposited inCNC, PFC, and ZLC. British Columbia: Van-couver Island: Mt. Cain, 50°13′N, 126°18′W,5.vi.1996 (L.M. Humble and N.N. Winchester),from foliose lichen on Pacific silver fir;M.A.S.S. site, 49°55′N, 125°25′W, 28.iv.1996(L.M. Humble and N.N. Winchester), from folioselichen on Pacific silver fir; Upper CarmanahValley, 48°44′N, 124°37′W, 30.viii.1991 (N.N.Winchester), 2 from moss in canopy of Sitkaspruce (Picea sitchensis (Bong.) Carr. (Pinaceae);Upper Walbran Valley, 48°39′N, 124°35′W,10.i.2005 (Z. Lindo), 2 from litter traps in west-ern redcedar (Thuja plicata Donn ex D. Don(Cupressaceae)) canopy; Victoria, 48°45′N,123°37′W, 14.ii.2007 (Z. Lindo), 3 from folioselichen on Oregon white oak (Quercus garryanaDougl. ex Hook. (Fagaceae)). United States ofAmerica. Oregon: Curry Co., Siskiyou Na-tional Forest, Butler Bar Campground on ElkRiver Road, 42°43′N, 124°16′W, 18.iii.2004 (V.Behan-Pelletier), 1 from moss and lichens fromdeciduous shrub on bank of river.

EtymologyThe specific epithet geminus refers to the

closely adjacent setae 3c and 3d positioned lat-erally in the coxisternal region.

DiagnosisAdultTotal length 533–615 μm, with character

states of Achipteriidae (Grandjean 1954) andcharacter states of Anachipteria as describedbelow. Lamellae about 146 μm long to lateraltip of cusps. Lamellar cusps widely spaced, dis-tance between lamellae at base of cusps 36 μm.Notogastral porose areas small but clearly visi-ble. Coxisternal setal formula 3–1–4–3. Inser-tions of coxisternal setae 3c and 3d closelyadjacent, positioned laterally on coxisternalprotuberance ventrally at base of pedotectum II.



DescriptionAdult (Figs. 1–21)Measurements. Mean total length: females

(n = 13) 587 μm (range 549–615 μm); males(n = 7) 562 μm (range 533–607 μm) (Figs. 1, 3).Mean notogastral width: females (n = 13)399 μm (range 369–418 μm); males (n = 7) 386 μm(range 349–426 μm). Integument. Notogasterand ventral plate weakly foveate (Figs. 3, 5, 15);with minute microtubercles, particularly evidentin lenticular region. Microtubercles formingshort striae on abaxial surface of femora I–IVand trochanters III and IV. Granularcerotegument clearly evident at base of prodor-sum (Figs. 7, 19) and on mentum (Fig. 9).Prodorsum. Rostrum rounded in outline, anteri-orly with distinct rounded longitudinal crest(Fig. 8). Setae ro 89 μm long (range 85–95 μm),acuminate, strongly barbed on lateral edge, curv-ing medially, and extending well beyond rostrum(Fig. 12). Lamellae about 146 μm long (range141–156 μm) to lateral tip of cusps. Lamellarcusps about 64 μm long laterally (range 62–66 μm), about 42 μm long medially (range 38–45 μm), with sharp dens laterally, roundeddistally (Figs. 1, 7). Translamella absent, dis-tance between lamellae at base of cusps 36 μm.Setae le about 97 μm long (range 80–107 μm),barbed, inserted medially on ventral side of lam-ellar cusps (Fig. 8) and extending anteriorly wellbeyond rostrum (Fig. 7). Setae in lightly barbed,140 μm long (range 131–148 μm), extending an-teriorly beyond tips of lamellae (Fig. 7). Mutualdistance between individual setae in each pairro–ro, le–le, and in–in, about 123, 43, and 83 μm,respectively. Setae ex thin and barbed, about50 μm long (range 47–55 μm), inconspicuous(Fig. 19). Bothridial setae 55–65 μm long (mean60.5 μm), curved medially, clavate; head abouttwo-thirds to three-quarters total length (Fig. 19).Lateral aspect of prodorsum. Tutoria broad,with undulating teeth distally (Figs. 8, 10);appearing pointed in dorsal aspect, owing to atapering, thickened dorsal margin (Fig. 7) ex-tending well anterior of insertion of setae ro.Genal tooth rounded to sharply pointed distally,with genal notch extending posterior to insertionof seta ro (Fig. 8). Pedotectum I well developed,fully covering acetabulum I (Fig. 7). Porose or-gans Am and Ah present, Am porose area diffuse;Ah (subalar vesicle) well delineated, dorsal toacetabulum III, consistently 33 μm in diameterand covered with fine cerotegument. Porose areaAl absent. Notogaster. Longer than wide, ratio1.2:1; hysterosoma often fattened with 2–4 eggs

of considerable size (about 258 μm long). Noto-gastral setae thick, barbed, with setae c2 longest,mean 57 μm (range 47–62 μm). Setae la, lm, andlp about 46, 34, and 32 μm long, respectively.Setae of h series usually about 29–33 μm long(range 23–35 μm). Setae of p series smaller thanother notogastral setae, about 23 μm long (range16–29 μm). Lateral edge of pteromorphs smooth,without striations (Fig. 6). Lenticular region sub-triangular with diffuse margins, but weakly visi-ble and lacking true lenticulus. Porose areas smallbut clearly visible (A3 hard to discern in somespecimens). Diameter of Aa, A1, A2, and A3, 6,7, 5, and 5 μm, respectively. Distance betweenAa and A1 about 166 μm. Normal five pairs oflyrifissures present, all about 12 μm long, vary-ing in orientation: ia at ventral edge of ptero-morph, im about 60 μm posterior of Aa, ih andips very close to each other (25 μm apart) (Fig. 16)anterior of A2, ip posteromedial of A3. Ventralregion. Coxisternal setae barbed, relatively long;formula (epimeres I–IV) 3–1–4–3 (Figs. 2, 4).Setae increasing in length laterally with 1a, 2a,3a, and 4a about 37 μm long and 1b and 3branging from 49 to 78 μm long. Setae 1c, 97 μmlong. Coxisternal setae 3c and 3d, 86 and 75 μmlong, respectively, insertions closely adjacent,positioned laterally on coxisternal protuberanceventrally at base of pedotectum II (Figs. 4, 13,14). Coxisternal setae 4c very thin, about 35 μmlong, only visible in dissected specimens. Sixpairs of genital setae ranging in length from 23to 41 μm, with longest anteriorly (Fig. 15).Aggenital setae about 51 μm long, lightlybarbed. Two pairs of anal setae about 22 μmlong, barbed; three pairs of adanal setae lightlybarbed, 25 μm long. Lyrifissure iad 15 μm long,paraanal, anterior of ad3 (Fig. 2). Gnathosoma.Subcapitular mentum without tectum (Fig. 9);setae h, m, and a about 43, 37, and 35 μm long,respectively, all barbed (Fig. 9). Cheliceraechelate–dentate (Figs. 11, 12). Axillary sacculeabsent from base of palp. Legs. Ratio of legIV length to body length ~0.6:1. Approximatelengths of leg segments (femur, genu, tibia, tar-sus (μm): I: 98, 34, 54, 67; II: 88, 32, 52, 63; III:75, 31, 58, 63; IV: 60, 45, 68, 81. Pretarsushetero-tridactylous with large smooth empodialclaw and finely barbed thin lateral claws (Fig. 18).Setation (I–IV, with number of solenidia inparentheses): trochanters 1–1–2–1; femora 5–5–3–2; genua 3(1)–3(1)–1(1)–2; tibiae 4(2)–4(1)–3(1)–3(1); tarsi 19(2)–15(2)–15–12; setalhomologies are indicated in Table 2. Genu I withventral keel (Fig. 21). Seta l′′ on tibia and genu I


Lindo et al. 543

and II strongly thickened (Figs. 18, 21). Seta son tarsus II thick, pectinate, with six or sevenlong barbs (Fig. 20). Ovipositor (Fig. 17). Un-paired dorsal-lobe seta ψ1 approximately 51 μmand seta ψ2 about 20 μm in length. Pairedventral-lobe setae τ1 longest at 56 μm, τ2, τ3, and

τ4, 31, 24, and 24 μm long, respectively. Threepairs of coronal setae, about 14 μm long.

Immatures (Figs. 22–30)Measurements. Mean total length: trito-

nymphs (n = 6) 514 μm (range 459–582 μm);deutonymphs (n = 4) 418 μm (range 385–



Figs. 1, 2. Anachipteria geminus, adult female: 1, dorsal aspect; 2, ventral aspect. Legs removed. Fordescriptions of setae see Materials and methods. Scale bars = 100 μm.

443 μm); protonymphs (n = 6) 358 μm (range320–377 μm); larva (n = 7) 268 μm (range 230–303 μm); egg (n = 7) 258 μm (range 230–287 μm). Tritonymph. Integument plicate withtransverse and longitudinal folds (Fig. 22).Prodorsum. Rostrum rounded. Integument mod-erately plicate with curved plicae extending frombase of bothridia to setae le. Setae in, le, and ro

short, 12, 14, and 21 μm in length, respectively.Bothridial setae 52 μm long, with stalk 23 μmlong and clavate barbed head 29 μm long.Bothridia directed laterally. Gastronotic region.Integument finely porose. Transverse plicae in4–6 rows on anterior of notogaster. Setae of cseries increasing in length medially, c1 23 μmlong, with dense short barbs. Setae la and lm


Lindo et al. 545

Figs. 3–6. Anachipteria geminus, scanning electron microscopic images of adult: 3, habitus, dorsal aspect;4, habitus ventral aspect; the arrow indicates coxisternal setae 3c and 3d; 5, habitus, anterodorsal aspect;6, habitus, lateral aspect; the arrow indicates the pteromorph. Scale bars = 100 μm.



Figs. 7–12. Anachipteria geminus, scanning electron microscopic images of adult: 7, prodorsum in dorsalaspect, interlamellar seta (in), lamellar seta (le), tutorium (TU), pedotectum I (Pd); 8, prodorsum, anterioraspect, lamellar seta (le), rostral crest (a), genal tooth (b), tutoria (TU); 9, gnathosome, ventral aspect, setae a,m, and h; 10, prodorsum, lateral aspect, showing thickened dorsal margin of tutorium (TU); 11, detail ofmouthparts, dorsal aspect; palptarsus (palp) with closely adjacent eupathidium acm and solenidion, tip ofchelicerae (ch), rutella (RU); 12, detail of mouthparts, anterior aspect; rutella (RU), chelicerae (ch), rostralsetae (ro). Scale bars = 10 μm (Figs. 7–9, 11–12) and 100 μm (Fig. 10).


Lindo et al. 547

Figs. 13–17. Anachipteria geminus, scanning electron microscopic images of adult: 13, left side of theproterosoma, ventral aspect, showing position of coxisternal setae 3c and 3d; 14, closeup of coxisternal setae3c and 3d, orientation as in 13; 15, genital region, showing ventral foveation; 16, notogaster in vicinity of setah3, dorsal aspect of left side, showing lyrifissures ih and ips, opisthonotal gland (gla), notogastral seta h3;17, extended ovipositor, lateral aspect. Scale bars = 10 μm (Figs. 13–16) and 100 μm (Fig. 17).

subequal in length to c1, about 23 μm long,curved, with dense short barbs. Setae da and dmcurved, about two-thirds length of la. Allnotogastral setae slightly curved with barbed totuberculate dorsal edge. Setae dp and h seriesabout 33 μm long, clubbed with thick barbs dor-sally (Fig. 25). Ventral. Cerotegument in poste-rior region with pitted or foveate appearance.Epimeral border II notably long, thin, straight,and clearly delineated. Genital setae thin,straight, and apparently smooth. Legs. Monodac-tylous with minute barbs dorsally.

Other developmental instars (Figs. 23–30)Prodorsum. Bothridial setae similar to trito-

nymph in all instars. Gastronotic region. Integ-ument finely porose. Notogastral setae similarto tritonymph in all instars. Ventral region.Coxisternal and anogenital setation for develop-mental instars given in Table 3. Paraproctal atri-chosy present to deutonymph. Legs. All tarsiwith small pulvillus. Ontogeny of setae and

solenidia given in Table 2. Solenidia short, iso-diametric on genua and tibiae. Seta d absentunilaterally from tibia IV of one tritonymph.

Genetic analysisEight oribatid mite species were successfully

sequenced for the D3 region of the 28s rRNAgene (Table 1). Among Achipteriidae, these in-cluded A. geminus, and A. acuta from suspendedsoil at the type locality of A. geminus; an undes-cribed species of Anachipteria from a forest-floor site on Vancouver Island; and one specieseach of Achipteria and Dentachipteria from for-est floor at the type locality of A. geminus. Alsoincluded was a species of Tegoribates from aforest-floor site on Vancouver Island, andC. pacificus and a species of Scheloribates fromthe type locality of A. geminus. All sequencesare available in GenBank under accession num-bers EF989716–EF989725. BLASTn confirmed thatall sequences were of oribatid mite origin.



Trochanter Femur Genu Tibia Tarsus

Leg ILarva — d b ′′v (l) σ (l) v′ φ1 (ft) (pl) (tc) (p) (u) s (a) (pv) e, ω1

Protonymph — — — — ω2

Deutonymph — (l) — φ2 —Tritonymph v′ v′ v′ — (it)Adult — — — ′′v v′

Leg IILarva — d b ′′v (l) σ l′ v′ φ (p) (tc) (ft) (u) s (a) (pv) ω1

Protonymph — — — — —Deutonymph — (l) — ′′l ω2

Tritonymph v′ — v′ ′′v (it)Adult — v′ — — —

Leg IIILarva — d ev′ l′ σ v′ φ (p) (tc) (ft) (u) s (a) (pv)Protonymph — — — — —Deutonymph v′ l′ — l′ —Tritonymph l′ — — ′′v (it)Adult — — — — —

Leg IVLarva — — — — —Protonymph — — — — (p) f ′′t (u) (pv)Deutonymph — d ev′ d l′ v′ d φ (tc) (a) sTritonymph v′ — — l′ v′′ —Adult — — — [d] —

Note: Setae (Roman) and solenidia (Greek) are listed opposite the instar in which they first appear. Pairs of setae areshown in parentheses and setal loss in square brackets.

Table 2. Development of leg setiform organs in Anachipteria geminus.


Lindo et al. 549

Figs. 18–21. Anachipteria geminus, scanning electron microscopic images of adult: 18, leg I, anterodorsalaspect, showing setae l ′′ of genu and tibia; 19, left proximal region of prodorsum, lateral aspect, showingbothridial setae (bo) and exobothridial seta (ex); 20, leg II tarsus, anteroventral aspect, showing seta s; 21, legI trochanter, genu with ventral keel (a) and seta l ′′ indicated. Scale bars = 10 μm.

Only A. geminus and C. pacificus were suc-cessfully sequenced for CO1, preventing the useof this gene to help resolve relationships. WithinAchipteriidae, A. acuta, Anachipteria sp., andA. curta formed a group. Dentachipteria sp. andA. geminus grouped closer to Tegoribates sp. thanto other Achipteriidae, with A. geminus moresimilar (96%) to Dentachipteria sp. than to otherspecies in the Achipteriidae–Tegoribatidae group(Table 4). The D3-28s sequence of A. geminuswas also 96% similar to that of C. pacificus, sug-gesting additional genetic divergence of this spe-cies from other members of Achipteriidae.

EcologyAdults and immatures of A. geminus were

found on 100% of the lichen samples (n = 107),but on only 9.4% of branch samples (n = 106)collected from the M.A.S.S. site. Whether thispreference relates to favourable complexity of thelichen substrate as habitat or to the use of lichenas a food source is not known, since food boluseswere not studied. In lichen samples collectedfrom the M.A.S.S. site, A. geminus was the domi-nant oribatid species among adults, constituting28% of the total number of adult individuals col-lected. The average abundances of both adult andimmature stages of A. geminus were significantlygreater in lichen samples than on branch samples(adults: lichen 375.1 / 100 g DM, branches 0.2 /

100 g DM (F8,7 = 4.432, P = 0.032); immatures:lichen 427.5 / 100 g DM, branches 0.6 / 100 gDM (F8,7 = 4.689, P = 0.028); total: lichen 802.6 /100 g DM, branches 0.8 / 100 g DM (F8,7 =6.357, P = 0.012)). Abundances of adults and im-mature stages were consistent throughout thethree seasonal sampling periods, and tree species(Pacific silver fir, western hemlock) showed littleeffect on A. geminus abundance (Fig. 31). Themaximum number of individual A. geminus spec-imens found in one sample was 1462/100 g DMlichen (adults and immatures combined).

Discussion

SystematicsAnachipteria Grandjean, 1932, p. 301.Type species: Anachipteria deficiens Grand-

jean, 1932, p. 301.

DiagnosisCharacter states of Anachipteria were previ-

ously given by Grandjean (1932), Shaldybina(1975), Pérez-Iñigo (1993), and Weigmann(2006). Adults of this genus are unique amongAchipteriidae in having the following combina-tion of character states. Lamellae large, eithermeeting medially with small translamella or wellseparated; if meeting medially, occluding mostof anterior part of prodorsum. Pteromorphs lacking



Fig. 22. Anachipteria geminus, tritonymph, dorsal aspect. Scale bar = 100 μm.


Lindo et al. 551

Figs. 23–30. Anachipteria geminus, scanning electron microscopic images of immature stages: 23, nymph–nymph moult, lateral aspect; 24, egg–larva hatching, anterodorsal aspect; 25, nymph, detail of unidentifiedposterior opisthosomal seta, dorsal aspect; 26, larva, lateral aspect; 27, nymph, dorsolateral aspect; 28,tritonymph–adult moult, dorsal aspect; 29, larva, prodorsum, dorsal aspect, showing setae in; 30, nymph,prodorsum, dorsal aspect showing setae in. Scale bars = 100 μm (Figs. 24–24, 26–28) and 10 μm (Figs. 25,29–30).

knife-like humeral projection. Octotaxic systemexpressed as four pairs of porose areas or sac-cules. Axillary saccule of gnathosoma absent.Genu IV concave dorsally and segment longerthan genu III. Coxisternal setation either 3–1–3–3 or 3–1–4–3. Humerosejugal porose organAh with form of dome-shaped subalar vesicle.Postanal porose area absent. Leg tarsi tridactylous.

Placement of A. geminus in Anachipteria isbased on character states of both adults and im-mature stages (Shaldybina 1975; Seniczak 1977;Pérez-Iñigo 1993; Root et al. 2007a). However,we expand the diagnosis of Anachipteria to in-clude other morphological character states pres-ent in A. geminus. In particular, the widelyspaced lamellae and coxisternal setation in adultA. geminus, 3–1–4–3, is unique in the genus.Additionally, the coxisternal setation of allknown Achipteriidae is 3–1–3–3 where thischaracter state has been examined (Travé 1960;Seniczak and Seniczak 2007). Just as Weigmann(2006) and Root et al. (2007a) argued that thepresence of saccules on Anachipteria dubiaWeigmann and A. sacculifera Root et al. did notwarrant proposing a separate genus, we do notthink that the narrow lamellae or addition or po-sition of coxisternal seta 3d in A. geminus justifyseparate generic status.

Adults of A. geminus bear a superficial simi-larity to species of Hypozetes Balogh (Tegori-batidae) in the lateral position of lamellae onthe prodorsum. However, adults of A. geminusand other species of Anachipteria differ fromthose of Hypozetes and other members of theTegoribatidae by the absence of a postanal porosearea and axillary saccule of the subcapitulum(Behan-Pelletier 2001). Immatures of species ofAnachipteria, including A. geminus, lack com-panion seta d on tibiae I–III in all instars, asynapomorphy shared with Ceratozetoidea,Phenopelopoidea, and many Licneremaeoidea.By contrast, this seta is present in immatures ofHypozetes and other known Tegoribatidae(Behan-Pelletier 2001).



Ana

chip

teri

age

min

usA

nach

ipte

ria

acut

aA

nach

ipte

ria

sp.

Ach

ipte

ria

curt

aD

enta

chip

teri

asp

.Te

gori

bate

ssp

.C

erat

ozot

espa

cifi

cus

A.

gem

inus

100

A.

acut

a95

100

Ana

chip

teri

asp

.95

9910

0A

.cu

rta

9599

9810

0D

enta

chip

teri

asp

.96

9897

9710

0Te

gori

bate

ssp

.95

9897

9797

100

Cer

atoz

otes

paci

ficus

9696

9696

9695

100

Sche

lori

bate

ssp

.91

9190

9092

8992

Tab

le4.

Clu

stal

Wpe

rcen

tsi

mil

arit

yin

D3-

28S

rRN

Ase

quen

ceal

ignm

ents

for

Ana

chip

teri

age

min

usw

ith

thos

eof

othe

rsp

ecie

sof

Ori

bati

da.

Larva Protonymph Deutonymph Tritonymph Adult

Coxisternal 3–1–2–0 3–1–2–1 3–1–2–2 3–1–3–3 3–1–4–3Genital 0 1 3 5 6Aggenital 0 0 1 1 1Anal 0 0 0 2 2Adanal 0 0 3 3 3

Table 3. Development of coxisternal and anogenital chaetotaxy in Anachipteria geminus.

However, our D3-28s sequence data suggestthat the relationship between members ofAchipteriidae and Tegoribatidae (Table 4) is lessclearly resolved than morphology indicates.Family-level distinction is low, so separation ofmembers of these two families is likely recent.Although the D3 region sequence for A. geminuswas different from that for other members of theAchipteriidae and Tegoribates sp., it was moresimilar to the sequence for Dentachipteria sp.than that for Anachipteria acuta (Table 4). Maraunet al. (2004) also had difficulties in placing twospecies of Achipteriidae within a greater oribatidmite phylogeny using the D3-28s region. Theyfound that Achipteriidae did not group withinthe poronotic Brachypylina, except under condi-tions of maximum parsimony. However, usingthe ribosomal 18S region and the nuclear elon-gation 1 alpha gene, Achipteria coleoptrata (L.)clustered with the poronotic brachypyline higherplicates (Domes et al. 2007). These results sug-gest that using the D3 region is insufficient to re-solve relationships between Achipteriidae and

Tegoribatidae, and that further morphologicaland molecular studies of members of thissuperfamily are warranted.

EcologyThe habitat of A. geminus mimics that of

A. sacculifera, recently described from arboreallichens in New York State (Root et al. 2007a).Whereas A. geminus is the numerically dominantoribatid mite in canopy lichens of western hem-lock and Pacific silver fir, A. sacculifera was thenumerically dominant oribatid mite on foliose li-chens on the crown branches of sugar maple(Acer saccharum Marsh. (Aceraceae)) trees inNew York State (Root et al. 2007b). The structureof the arboreal oribatid community of a congenerin the Pacific Northwest, bigleaf maple (Acermacrophyllum Pursh (Aceraceae)), has not beenstudied. The abundances of adults and immatureinstars of A. geminus throughout the active grow-ing season indicate that the populations studiedreproduce continuously throughout the season.

Key to adults of described species of Anachipteria known from Canada

1. Mentum of subcapitulum with tectum. . . . . . . . . . . . A. australoides Jacot, 1938(British Columbia)

— Mentum of subcapitulum without tectum . . . . . . . . . . . . . . . . . . . . . . 2

2. Lamellae widely spaced on prodorsum; coxisternal setation 3–1–4–3, with insertions ofcoxisternal setae 3c and 3d closely adjacent, positioned laterally on coxisternal protuberance

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A. geminus sp. nov.(British Columbia)

— Lamellae meeting or closely adjacent medially; coxisternal setation 3–1–3–3 . . . . . . . 3

3. Bothridial setae short, subcapitate; lamellae closely adjacent, but not meeting medially;notogastral porose areas about 13 μm in diameter; all notogastral setae long, subequal in length. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A. acuta (Ewing, 1918)

(Oregon, Alberta, British Columbia)— Bothridial setae long, clavolanceolate, directed medially; lamellae meeting medially; notogastral

porose area Aa larger than 15 μm in diameter; notogastral setae c and la longer than othernotogastral setae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4

4. Anterior margin of lamellar cusps truncated anteriorly, lateral dens absent or very short . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . A. howardi (Berlese, 1908)

(Missouri, Nunavut, Northwest Territories, Manitoba, also Europe and Far East)— Anterior margin of lamellar cusps oblique, with lateral dens strongly acute . . . . . . .

. . . . . . . . . . . . . . . . . . . . . . . . . A. magnilamellata (Ewing, 1909)(Ontario, New Brunswick)


Lindo et al. 553

Acknowledgements

We thank Roy Norton for helpful commentson the manuscript. Canopy arthropod research atthe M.A.S.S. research site was supported byfunding from the Canadian Forest Service, Natu-ral Resources Canada, and research awards fromForest Renewal British Columbia (PA96580-RE)and the Science Council of British Columbia(IA-97/98-006). We thank K. Jordan for canopysampling and D. Bennett, S. Curry, L. Richmond,

P. Bowering, and W. Halstrom for sorting andwashing branch samples. Molecular sequenceswere generated with the help of R. Roy andM. Hill. The Natural Sciences and EngineeringResearch Council of Canada provided financialsupport (CGS-D-332720-06) for Z.L.

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