Variation in Male and Female Genitalia Among Ten Species of North American Anthocoris (Hemiptera: Heteroptera: Anthocoridae)

SYSTEMATICS Variation in Male and Female Genitalia Among Ten Species of North American Anthocoris (Hemiptera: Heteroptera: Anthocoridae)

1,2 1 DAVID R. HORTON AND TAMERA M. LEWIS

USDAÐARS, 5230 Konnowac Pass Road, Wapato, WA 98951

Ann. Entomol. Soc. Am. 104(6): 1260Ð1278 (2011); DOI: http://dx.doi.org/10.1603/AN11087 ABSTRACT We compared morphology of internal reproductive anatomy and genitalia among 10 species of North American Anthocoris (Hemiptera: Heteroptera: Anthocoridae). Reproductive structures of males, including internal reproductive organs (testes, seminal vesicles, ejaculatory bulb, phallus), the paramere, and the endosoma, were found to vary in size, shape, and appearance among the 10 species, although with similarities among species within some groupings. All species except Anthocoris bakeri Poppius have two testicular follicles per testis; A. bakeri has Þve to seven follicles per testis. In seven of the 10 species, a longitudinal groove was found to be present on the paramere; we believe this groove functions to guide the maleÕs intromittent organ as it enters the female during copulation. Variation among species in morphology of the maleÕs inßated endosoma included differences in length, presence or absence of looping, and in presence of spinulate, dentate, or tuberculate projections. The summary presented here is the Þrst description of the endosoma for any species of Anthocoris. We observed substantial variation among species in length of the femaleÕs copulatory tube (i.e., the organ which receives the maleÕs intromittent organ), although again with some similarities among certain groupings of species. Variation in length of the femaleÕs copulatory tube parallels variation in length of the maleÕs endosoma, which suggests that morphology of genitalia within this genus has coevolved between sexes.

KEY WORDS paragenital system, true bugs, sexual selection, internal reproductive anatomy

The genus Anthocoris Falle´n, 1814 (Hemiptera: Het- in classiÞcation of some taxa within this family (Ke and eroptera: Anthocoridae) includes Ϸ70 described spe- Bu 2005, 2007). Recently, Ke and Bu (2007) used cies (Ke and Bu 2007) distributed primarily in the features of the femaleÕs copulatory tube (see below) Palearctic and Nearctic regions (Pe´ricart 1972, 1996). to deÞne species groupings within the worldwide These insects associate often with trees and shrubs fauna of Anthocoris. The results supplemented similar (Anderson 1962, Horton and Lewis 2000, Horton et al. attempts at subdividing Anthocoris made previously 2004) and may be important sources of biological (Hill 1957, Pe´ricart 1972, Bu and Zheng 2001). Here, control in perennial crops and woodlands (Herard we examine male and female reproductive anatomy 1986, Lattin 1999). The North American (north of and genitalia of 10 species of North American Antho- Mexico) fauna of Anthocoris is composed of 11 de- coris. We show that the North American species of scribed species, including two species introduced into Anthocoris exhibit extensive variation in size and shape North America from the Old World (Henry 1988, Maw of genitalia but Þnd that certain species cluster into et al. 2000, Horton and Lewis 2009). The fauna also relatively well-deÞned groups based upon features of includes an unknown number of undescribed cryptic the genitalia (as in Ke and Bu 2007). Lastly, we show species very similar in appearance to Anthocoris mus- that variation among species in characteristics of the culus (Say, 1832) and Anthocoris antevolens White, maleÕs intromittent organ is accompanied by similar 1879 (Horton et al. 2007, 2008), as well as an unde- changes in morphology of the femaleÕs copulatory scribed species similar in appearance to Anthocoris tube, which suggests that male and female genitalia in tomentosus Pe´ricart (TML unpublished data). these species have coevolved. IdentiÞcation of species within the Anthocoridae is often based extensively on characteristics of the genitalia, particularly the male genitalia (Pericart 1972, ´ Characteristics of Genitalia of Anthocoris Kelton 1978, Bu and Zheng 2001), although traits associated with the female genitalia also may be useful We provide here a brief description of genitalic morphology and mechanics for Anthocoris. Terminol- 1 The authors contributed equally to this study. ogy follows Dupuis (1970), Carayon (1972), Pe´ricart 2 Corresponding author, e-mail: [email protected]. (1972), and Schuh and Slater (1995). November 2011 HORTON AND LEWIS:GENITALIA OF NORTH AMERICAN Anthocoris 1261

sperm pouch

st IV "trunk" of st V conductive tissue

intersegmental st VI copulatory tube membrane

tVII

t VIII

Fig. 1. Female genitalia. Location of copulatory tube and sperm pouch of Anthocoris (illustration is for A. antevolens). ModiÞed from Þg. 1 in Ke and Bu (2007).

Female. The Anthocoridae and related taxa such as at the base of the sperm pouch (Fig. 1). Inseminated the bedbugs (Cimicidae) have evolved several unique sperm move from the sperm pouch into the conduc- reproductive structures associated with extragenital tive tissues, and eventually arrive at the base of the (“traumatic”) insemination (Carayon 1966, 1977). Ex- ovaries (Carayon 1953). Eggs are fertilized in the tragenital insemination in the Anthocoridae (and ovaries before the chorion has been formed and be- Cimicidae) has led to the evolution in females of fore the eggs have moved into the oviduct. structures collectively referred to as the spermalege Comments About Terminology. Terminology used (Carayon 1966). The spermalege consists of two struc- here to describe genitalia of female Anthocoris differs tures, a swelling, thickening, or invagination of the from that in a previous review (Ke and Bu 2007), integument which acts to guide the maleÕs intromit- which could lead to some confusion. Ke and Bu (2007) tent organ to a speciÞc insemination site (ectosper- referred to the copulatory tube as comprising two malege); and, a structure to receive the sperm parts, a basal duct (equivalent to the copulatory tube, (mesospermalege). In many Anthocoridae, the ecto- as described by Carayon 1953) and an apical sac spermalege has evolved into a specialized copulatory (equivalent to the sperm pouch, as described by Car- tube (“tube copulateur,” Carayon 1953, p. 1206) that ayon 1953). That is, the copulatory tube in Ke and Bu Carayon (1966) considered to represent the most de- (2007) includes the entire copulatory organ (ectosper- rived form of the ectospermalege. The distal end of the malege and mesospermalege), rather than just the ec- copulatory tube of Anthocoridae ends at the meso- tospermalege. We follow terminology in Carayon spermalege (Carayon 1966), referred to as the sperm (1953), and consider the copulatory tube and sperm pouch or spermatic pouch (“poche spermatique,” Car- pouch to be two separate structures. Carayon (1966) (p. ayon 1953, p. 1206). The copulatory tube of Anthocoris 131; 1977: p. 358Ð359, 384) has shown that the copulatory opens to the exterior of the female on her ventral side tube and sperm pouch arise from different anatomical between abdominal segments seven and eight (Fig. 1). structures during the insectÕs development. The structure is membranous, tubular, and often with Male. Two structures associated with the genitalia noticeable pleating of the outer wall. The copulatory of male Anthocoris are of particular importance with tube extends from its basal opening between sterna 7 and respect to mating: the paramere (clasper) and the 8, forward to abdominal segments 4 or 5 (Ke and Bu phallus. The ninth abdominal segment (genital cap- 2007) where the tube meets her sperm pouch (Fig. 1). sule or pygophore) houses the genitalia (Fig. 2A and When in place within the female, the copulatory tube B). There is a relatively pronounced groove along the may exhibit noticeable coiling or twisting (Ke and Bu left border of the capsule; this groove houses the left 2007). paramere (Fig. 2B). The genitalia of males are asym- The sperm pouch is highly membranous, spherical metrical, in that only the left paramere is present (the (Fig. 1), and more-or-less ßattened in virgin bugs, and right paramere having been lost). The paramere is expands noticeably as it Þlls with seminal products variable in size and shape among species, and is used from the copulating male (Horton et al. 2002). A extensively in identifying species of Anthocoris (Hill specialized bridge of conductive tissues connects the 1957, Pe´ricart 1972, Carayon 1972, Kelton 1978, Bu and sperm pouch to the ovarian wall (Carayon 1953). Zheng 2001). The male seems to use the paramere as These conductive tissues assemble to form a trunk a tool to gain entry into the opening of the femaleÕs (“tronc du tissu conducteur,” Carayon 1953, p. 1207) copulatory tube. A longitudinal groove (“gouttie`re du 1262 ANNALS OF THE ENTOMOLOGICAL SOCIETY OF AMERICA Vol. 104, no. 6

basal primary foramen ejaculatory gonopore bulb

phallus A

B phallus t V C D

t VI ductus seminis paramere t VII

basal foramen t VIII genital capsule

articulatory paramere apparatus

E ductus seminis

Fig. 2. Male genitalia. (A and B) Location of genital capsule (ninth abdominal segment) showing paramere, ejaculatory bulb, and phallus. (C) Cutaway model of genital capsule showing location of ejaculatory bulb, phallus, and primary gonopore. (D) Phallus showing location of basal foramen with primary gonopore (A. antevolens). (E) Photograph of phallus (with articulatory apparatus) of A. confusus, showing articulatory apparatus, basal foramen, and ductus seminis. Line drawing of phallus by Debra Broers. parame`re,” Carayon 1972, p. 322; “rainure,” Pe´ricart is everted through the femaleÕs copulatory tube (Fig. 1972, p. 22) is present on the paramere of some species, 3C). Knobs, bumps, or spiniform projections may line and apparently functions to guide the membranous the surface of the fully everted endosoma (Fig. 3C and intromittent organ (described below) as it enters the D). As the endosoma is retracted into the phallosoma for femaleÕs copulatory tube. storage, these processes are carried with the endosoma, The genital capsule holds the phallus, which com- eventually ending on the interior of the membranous prises the sclerotized articulatory apparatus and in- organ when it is in repose (Fig. 3A and B). The retracted tromittent organ (Fig. 2CÐE). The articulatory appa- endosoma is a narrow, ribbon-shaped structure, showing ratus is composed of a system of plates and apodemes extensive looping while in storage within the phallosoma that secure the phallus to the reßexed internal wall (Fig. 3A and E). Length and shape of the everted organ, (diaphragm) of the genital capsule, and to which are as well as presence of projections, lobes, or looping, are attached the muscles that move the phallus. The two variable among species. primary plates of the articulatory apparatus (the basal Anterior of the phallus is the ejaculatory bulb (Fig. plates) join together ventrally to partially encircle the 2A and C), which varies noticeably in size and shape entrance (basal foramen) to the lumen of the intro- among species. The ejaculatory bulb narrows poste- mittent organ (Fig. 2D and E). The intromittent organ riorly into the ejaculatory duct, which enters the phal- includes two structures, a proximal phallosoma and a lus through the basal foramen at the primary gonopore distal endosoma (Fig. 3D). The phallosoma is a mem- (Fig. 2C and D). branous or partially sclerotized structure that houses the endosoma when that organ is in repose (Fig. 3A Materials and Methods and E). The endosoma is a membranous structure surrounding the ductus seminis distalis, through Source of Insects. We examined male and female which the male seminal products are transported, and genitalia of specimens from each of 10 species of An- acts to carry the opening of the duct (secondary go- thocoris (Table 1). The 10 species include all but three nopore) to the femaleÕs sperm pouch as the endosoma of the species of Anthocoris listed in the catalog of November 2011 HORTON AND LEWIS:GENITALIA OF NORTH AMERICAN Anthocoris 1263

Fig. 3. Phallosoma and endosoma. (A and E) Endosoma in storage within the phallosoma. (B) Partially everted endosoma. (C and D) Fully everted endosoma showing ductus seminis and secondary gonopore. Line drawings by Debra Broers.

Henry (1988) for North America north of Mexico. We been synonymized with A. whitei (Lewis et al. 2003). were unable to obtain specimens of Anthocoris fulvi- Specimens include two species (A. nemoralis, A. con- pennis Reuter, 1884 (reported from California and fusus) of Old World origin and known to be estab- New Mexico in Henry 1988) and Anthocoris nigripes lished in North America (Horton et al. 2004, Lewis et Reuter, 1884 (New Mexico). Anthocoris tristis Van al. 2005, Horton and Lewis 2009), and a third species Duzee, 1921 was listed by Henry (1988) but has since (Anthocoris sp. L) similar in appearance to A. tomen-

Table 1. Geographic distribution of North American Anthocoris and sources of specimens examined in this study

Species Known distribution in North America Geographic and plant sources of insects Anthocoris albiger Reuter, 1884 Southwestern United States; Mexico Tucson, AZ (Cercocarpus) Anthocoris antevolens White, 1879 Transcontinental in Canada; western United Yakima, WA (Populus) States, Alaska, Mexico Anthocoris bakeri Poppius, 1913 California, Oregon, Arizona Bandon, Oregon (Arctostaphylos) Anthocoris confusus Reuter, 1884 British Columbia, Canada (?)a; western Seattle, WA (Tilia) Washington state; eastern Canada and eastern United States; Great Lakes region (?) Anthocoris dimorphicus Anderson and Eastern Canada and westward to Alberta; Oswego County, NY; Rutland and Bennington Kelton, 1963 Montana, Michigan, northeastern United counties, VT ( Salix) States, Colorado Anthocoris musculus (Say, 1832) Apparently transcontinental in Canada; Oswego County, NY (Salix, Alnus) northeastern United States westward to Montana Anthocoris nemoralis (F., 1794) Western North America; Ontario, Canada (?) San Francisco, CA (Acacia) Anthocoris tomentosus Pe´ricart, 1971 Western North America into Alaska Yakima, WA (Pyrus, Salix) Anthocoris whitei Reuter, 1884 Western North America Tieton, WA (Purshia) Anthocoris sp. L Western north America into Alaska Silverthorne and Crested Butte, CO; Lookout Pass, Montana; Chinook Pass and White Pass, WA (Salix)

a (?), decades-old records only; not known whether species is currently established in the region. 1264 ANNALS OF THE ENTOMOLOGICAL SOCIETY OF AMERICA Vol. 104, no. 6 tosus but not yet described (T.M.L., unpublished the copulatory tube of Anthocoris, consult Ke and Bu data). Two species (A. antevolens and A. musculus) are (2007). part of a species complex containing an unknown Specimens were killed either by crushing the fe- number of undescribed and reproductively isolated maleÕs head or by freezing the insect at Ϫ80ЊC. Spec- cryptic species, whose genitalia have the same general imens were dissected within 20 min after killing or appearance among species, albeit with subtle but mea- after removal from the freezer. Virgin females were sureable differences in size or shape (Horton and used almost exclusively in the dissections; specimens Lewis 2005; Horton et al. 2007, 2008). The specimens of A. confusus and Anthocoris sp. L that had been of A. antevolens examined here were of the genitalic collected from the Þeld probably included previously phenotype and genetic haplotype designated as “GC” mated females. The abdomen of each specimen was in a previous publication (Horton et al. 2007). detached from the insect between segments 4 and 5. Specimens were collected from the Þeld either as Sclerites were removed to expose the sperm pouch. adults or nymphs. Nymphs of most species were Each sperm pouch was then measured for length (base reared to the adult stage by feeding them aphids and of pouch to apical edge) and maximum width. We psyllids from laboratory cultures. Nymphs of the spe- found that the sperm pouch from previously frozen cialized A. bakeri were collected from Arctostaphylos females expanded noticeably in saline. Thus, measure- and were fed a gall-forming aphid that associates with ments of the sperm pouch taken from frozen speci- this host plant. For all but three species, dissections mens were obtained quickly after dissection. were made using Þrst generation adult offspring from After measuring the sperm pouch, additional scle- Þeld-collected specimens. Specimens of A. bakeri rites were removed, leaving intact the ventral portion were obtained by rearing Þeld-collected nymphs to of the abdomen distal to segment 7. An insect pin was the adult stage, supplemented with a few Þeld-col- used to remove fat, ovaries, and alimentary tract, ex- lected adult females. Dissected specimens of A. con- posing the copulatory tube. Conductive tissue adher- fusus comprised primarily Þeld-collected adults but ing to the base of the sperm pouch was left in place, included a few females reared from Þeld-collected but the branches of conductive tissue leading to the nymphs. Dissected specimens of Anthocoris sp. L were ovaries were severed. Strands of connective tissue obtained from the Þeld as adults. were removed until the copulatory tube was free Methods of Dissection. Dissections were done in in the body cavity except at the point of attachment to saline beneath a dissection microscope (Stemi 2000, the intersegmental membrane. We then straightened Carl Zeiss, Thornwood, NY) by using microdissection the copulatory tube for measurement and illustration tools. Reproductive structures and genitalia were ex- by pulling the apical portion of the dissected abdomen amined under a dissection microscope or compound across the glass slide in a thin layer of saline. The microscope (DMLS, Leica, Buffalo Grove, IL). Pho- dissected material was manipulated only enough to tographs were taken using a Spot Insight color digital straighten (but not stretch) the copulatory tube. The camera (Diagnostic Instruments, Sterling Heights, length of the copulatory tube (basal attachment to MI) or MPS 60 analog Þlm camera (Leica). Measure- point at which it meets the sperm pouch) and its width ments were made using an ocular micrometer. Sample were then measured. Sketches were made of the cop- sizes are provided in Results. ulatory tube as viewed through a dissecting micro- Female: Copulatory Tube and Sperm Pouch. Ke and scope, with a few details enhanced by use of a com- Bu (2007) provided a geographically broad compari- pound microscope. son of the copulatory tube among 40 species of An- Male: Reproductive Tract. The majority of speci- thocoris, including all but two of the North American mens were killed by crushing the head. A few speci- species included here (A. dimorphicus and Anthocoris mens were examined after killing and storage in an sp. L). Our approach differs from that of Ke and Bu ultrafreezer; however, we found that freezing altered (2007) in that we were able to dissect fresh specimens, the structural integrity of the more fragile organs, thus whereas Ke and Bu (2007) necessarily dissected these sclerotized organs only were examined in previously organs from dried specimens, and then treated the frozen specimens. Structures associated with the re- structures in KOH to dissolve muscle and other tissues. productive tract were sketched and measured at 50ϫ We found that dissection of fresh specimens in saline beneath a dissecting microscope, with additional de- allowed us to view structures which dissolve in KOH tail obtained as needed at 100Ð400ϫ beneath a com- but are seen in freshly killed specimens. For example, pound microscope. Some chitinous structures associ- the trunk of conductive tissue at the base of the sperm ated with the genital capsule were examined at 100Ð pouch is easily seen in fresh specimens but largely 400ϫ after soaking the capsule for 1Ð3 d in 10% KOH disappears if immersed in KOH. Conversely, our dis- at room temperature. We observed fairly substantial section methods made it more difÞcult to observe levels of intraspeciÞc variation in size of organs within certain details near the apical portion of the copula- the maleÕs reproductive tract. Therefore, structures tory tube where the tube meets the sperm pouch, in were measured for 9Ð12 specimens of each species, that it was difÞcult to determine for some species except Anthocoris sp.L(n ϭ 5Ð9 specimens for each whether the apical end of the tube actually entered organ). Illustrations show structures from the average the sperm pouch or was attached externally to the of the multiple measurements. membrane of the pouch. Our illustrations show this Male: Paramere. Dissection methods are described apical region as it appears in saline. For details about in Horton and Lewis (2005). Glycerine jelly was November 2011 HORTON AND LEWIS:GENITALIA OF NORTH AMERICAN Anthocoris 1265

Fig. 4. Internal reproductive organs of males of A. antevolens, A. dimorphicus, A. musculus, and A. confusus. As viewed through dissecting microscope (50ϫ). eb, ejaculatory bulb; m, mesadenia; md ϩ vd, paired mesadene duct and vas deferens; mr, mesadene reservoir; p, phallus; sv, seminal vesicle; t, testicular follicle; vd, vas deferens; ve, vas efferens. Illustrations based upon examination and measurement of structures for 9Ð12 specimens per species. Illustrations by Janet Lane. melted onto a microscope slide. A shallow well was paramere had a longitudinal groove, the hair could then created by placing a drop of glycerine on the be seen moving in and out of the groove, and the cooling jelly. The paramere of each male was removed depth, length, and positioning of the hollowed area from his genital capsule, placed in the well in a drop was more easily viewed. of glycerine, and covered with a glass slip. We exam- Male: Endosoma. For most species, inßated endo- ined parameres in multiple orientations and with the somata were obtained by freezing copulating pairs light source from various directions, at magniÞcations in liquid nitrogen and then gently pulling the male of 80Ð400ϫ under dissecting and compound micro- and female apart. In those pairings in which this scopes. method failed to produce a fully inßated endosoma, The paramere of several species was found to gentle pressure applied to the phallosoma often have a longitudinal groove (Carayon 1972). It was resulted in full extension of the organ. We were not often difÞcult to determine the presence and exact able to free the inßated endosoma of male A. whitei conÞguration of this structure. To conÞrm presence and A. albiger from the femaleÕs copulatory tube, of a groove, the paramere was rinsed in water and because a dilated region present at the apical end of allowed to dry. Then, a very Þne hair glued to the the fully inßated endosoma (see Results) anchored end of a pin was moved along the face of the the maleÕs organ within her copulatory tube. How- paramere while observing at 80ϫ under a dissecting ever, we found that by rapidly freezing males (in microscope. For those species in which the liquid nitrogen), we occasionally obtained speci- 1266 ANNALS OF THE ENTOMOLOGICAL SOCIETY OF AMERICA Vol. 104, no. 6

Fig. 5. Internal reproductive organs of males of A. tomentosus, Anthocoris sp. L, A. bakeri, and A. nemoralis. As viewed through dissecting microscope (50ϫ). eb, ejaculatory bulb; m, mesadenia; mr, mesadene reservoir; p, phallus; sv, seminal vesicle; t, testicular follicle; vd, vas deferens. Illustrations for A. tomentosus, A. bakeri, and A. nemoralis based upon examination and measurement of structures from 9 to 12 specimens per species; sample sizes for Anthocoris sp. L were Þve to nine specimens per structure measured. Illustrations by Janet Lane. mens in which the endosoma was completely in- saline solution to conÞrm that the organ had indeed ßated. The genital capsule with inßated endosoma been fully everted. The phallus was examined under was removed from the insect and placed in saline a compound microscope to conÞrm complete ex- solution. The endosoma coiled more or less strongly tension of the endosoma. in saline, depending on the species. To measure length of the organ, the structure was pulled along Results a thin Þlm of saline until it had been straightened on the microscope slide. The organ was then measured Reproductive System and Genitalia. Reproductive (at 50ϫ) from its apex to the point at which it Tract of Male. Paired testes each composed of two or entered the genital capsule. Once the endosoma had Þve to seven follicles; testicular follicles (t) connected been measured, the genital capsule was dissected in by short vasa efferentia (ve; illustrated here only for November 2011 HORTON AND LEWIS:GENITALIA OF NORTH AMERICAN Anthocoris 1267

gresses to ejaculatory bulb in parallel and in close contact with vas deferens (Fig. 7; see also illustration in Carayon 1972, p. 327); the paired mesadene duct and vas deferens illustrated as single structure (md ϩ vd) in Figs. 4Ð6. Ejaculatory bulb generally round or oval, occasionally more elongate; short ejaculatory duct at posterior end of ejaculatory bulb leading to phallus (p). Genitalia of Male. Genital capsule (ninth abdominal segment) houses the phallus and support structures (Figs. 2Ð3, 8Ð12). Phallus composed of sclerotized articulatory apparatus and oval-shaped phallosoma, the latter housing the membranous endosoma when in repose. Fully inßated endosoma of some spe- Fig. 6. Internal reproductive organs of males of A. whitei cies with relatively pronounced ßeshy lobe at basal and A. albiger. As viewed through dissecting microscope end, and in other species with conspicuous looping at (50ϫ). eb, ejaculatory bulb; m, mesadenia; mr, mesadene basal end; surface of fully inßated endosoma often reservoir; p, phallus; sv, seminal vesicle; t, testicular follicle; with spinulate, dentate, or tuberculate projections. vd, vas deferens. Illustrations based upon examination and Paramere arising near distal end of genital capsule; measurement of structures for 10 specimens per species. with longitudinal groove or depression in some spe- Illustrations by Janet Lane. cies; surface of paramere invariably with several setae. Copulatory Tube and Sperm Pouch of Female. Cop- A. antevolens; Fig. 4) to vasa deferentia (vd); vasa ulatory tube membranous, elastic, and tubular in deferentia near testes enlarged to form seminal vesi- cross-section, opening on ventral surface of female cles (sv), then narrowing below seminal vesicles be- and terminating at sperm pouch; receives inßated en- fore entering ejaculatory bulb (eb). Mesadene reser- dosoma of male (Figs. 1 and 13-16). Sperm pouch a voirs (mr) arising near posterior end of seminal spherical or ellipsoid structure arising at termination vesicles, each reservoir with Þnger-like mesadenia of copulatory tube; highly membranous, more-or-less (m) attaching near posterior end. Each mesadene ßattened in virgin females, expands during copulation reservoir with duct emerging from posterior end (me- as it receives seminal products from male. Trunk of sadene duct; not illustrated in Figs. 4Ð6); duct pro- conductive tissue generally visible at base of pouch.

vas deferens

mesadene duct

Fig. 7. Paired mesadene duct and vas deferens below seminal vesicle (A. whitei). 1268 ANNALS OF THE ENTOMOLOGICAL SOCIETY OF AMERICA Vol. 104, no. 6

Fig. 8. Fully everted and straightened endosomata of Anthocoris, showing interspeciÞc differences in length and appearance of the organ. Illustrations by Debra Broers.

Characteristics of Each Species [width ϫ length]; n ϭ 9) and readily apparent (Figs. 13, and 14A,D). A. antevolens (Figs. 4, 8Ð9, 10A, 13, 14A and D) A. dimorphicus Male Reproductive Tract. Intermediate in size to (Figs. 4, 8, 9, 10B, 13) reproductive tract of A. tomentosus/Anthocoris sp. L and A. whitei/A. albiger species groups. Seminal ves- Male Reproductive Tract. Very similar in appear- icles somewhat parallel-sided, ejaculatory bulb round ance to reproductive tract of A. antevolens but some- or oval and approximately same size as phallus; two what smaller and with wider seminal vesicles (Fig. 4). testicular follicles per testis (Fig. 4). Paramere. Similar in appearance to paramere of A. Paramere. Lamellate, broadest at basal end, nar- antevolens but slightly longer and with more pro- rowing apically with hook at apex; pronounced lon- nounced hook at apex; pronounced longitudinal gitudinal groove formed by inßection of anterior edge groove formed by inßection of anterior edge of of paramere (Fig. 9). paramere (Fig. 9). Endosoma. Everted organ 0.96 mm in length (n ϭ Endosoma. Everted organ 0.85 mm in length (n ϭ 5); basal Ϸ70% densely covered with dentate projec- 7); similar in appearance to endosoma of A. antevolens tions, distal 30% smooth; short, ßeshy lobe near base, but with two basal lobes (Fig. 10B, inset 1), the second covered with spiniform projections (Figs. 8 and 10A, lobe smaller than the most basal lobe, located distal inset). and ventral to basal lobe, may be partially hidden by Female Copulatory Tube and Sperm Pouch. Length basal lobe in dorsal view; short spiniform projections 0.71 mm (n ϭ 10); diameter largest at basal end, be- on both lobes (Fig. 10B, inset 2); surface of endosoma coming gradually more narrow toward apex; apical with densely scattered dentate projections adjacent portion of copulatory tube s-shaped (not shown in to and posterior to lobes, distal Ϸ70% of organ Fig. 13), the curved section hidden within trunk of smooth (Figs. 8 and 10B). conductive tissue (Figs. 1 and 14D). Sperm pouch 0.38 Female Copulatory Tube and Sperm Pouch. Cop- by 0.25 mm (width ϫ length; n ϭ 10); trunk of con- ulatory tube similar in appearance to that of A. ante- ductive tissue spherical in shape (0.23 by 0.17 mm volens but somewhat shorter in length (0.67 mm; n ϭ November 2011 HORTON AND LEWIS:GENITALIA OF NORTH AMERICAN Anthocoris 1269

volens, but somewhat shorter (0.62 mm; n ϭ 7). Sperm pouch similar in appearance to that of A. antevolens (0.35 by 0.25 mm [width ϫ length]; n ϭ 7); trunk of conductive tissue spherical in shape and readily apparent (Fig. 13).

A. confusus (Figs. 4, 8, 9, 10D, 13) Male Reproductive Tract. Intermediate in size to reproductive tract of A. tomentosus/Anthocoris sp. L and A. whitei/A. albiger species groups (Fig. 4). Sem- inal vesicles somewhat oval shaped; ejaculatory bulb large and parallel-sided; two testicular follicles per testis. Paramere. Lamellate, slender with sharply deÞned preapical tooth, pronounced curve in organ at base; without longitudinal groove (Fig. 9). Endosoma. Everted organ long (2.21 mm; n ϭ 5; photograph in Fig. 10D of partially everted organ) with basal loop, thin and of uniform diameter distal to loop; low rounded protuberances sparsely distributed in loop area, Ϸ0.2 mm of tube immediately distal to loop noticeably shagreened (Figs. 8 and 10D). Female Copulatory Tube and Sperm Pouch. Cop- ulatory tube 1.69 mm in length (n ϭ 8), of relatively uniform diameter. Sperm pouch 0.35 by 0.25 mm (width ϫ length; n ϭ 3); trunk of conductive tissue not pronounced; thin-walled, sac-like structure of unknown function visible within sperm pouch at junction of pouch and copulatory tube (Fig. 13). Fig. 9. Parameres of Anthocoris; posterolateral view. g, longitudinal groove. Illustrations based upon two to Þve specimens examined per species. Illustrations by Debra Broers. A. tomentosus (Figs. 5, 8, 9, 11A, 13, 15C) Male Reproductive Tract. Overall size largest 8). Sperm pouch 0.50 by 0.33 mm (width ϫ length; n ϭ among the 10 species (Fig. 5). Large seminal vesicles 10), noticeably larger than that of A. antevolens; trunk and mesadene reservoirs; mesadenia attached to me- of conductive tissue spherical in shape and readily sadene reservoir well away from junction of reservoir apparent (Fig. 13). and seminal vesicle; small testes, two testicular follicles per testis; very large ejaculatory bulb but small phallus. A. musculus Paramere. Slender and somewhat curved, distinct (Figs. 4, 8, 9, 10C, 13) lateral bend in dorsal view with pronounced twist at Male Reproductive Tract. Similar in appearance to apex; groove in form of a shallow depression (Fig. 9). reproductive tract of A. antevolens but somewhat Endosoma. Everted organ very short, 0.4 mm in smaller (Fig. 4). length (n ϭ 8), thin in cross section, of relatively Paramere. Similar in shape to paramere of A. ante- uniform diameter, abruptly tapering to point at apex; volens but slightly smaller in length; pronounced lon- distal one third densely covered with long spiniform gitudinal groove formed by inßection of anterior edge projections oriented basad, basal 2⁄3 sparsely covered of paramere (Fig. 9). with minute dentate projections (Figs. 8 and 11A). Endosoma. Everted organ 0.85 mm in length (n ϭ Female Copulatory Tube and Sperm Pouch. Cop- 10), similar in appearance to endosoma of A. antevo- ulatory tube very thin and fragile, 0.44 mm in length lens, spiniform projections on lobe shorter than those (n ϭ 11); largest diameter at basal end, then rapidly of A. antevolens; surface of endosoma with sparsely becoming very narrow; funnel-shaped at apex. Sperm scattered and minute dentate projections adjacent pouch 0.48 by 0.37 mm (width ϫ length; n ϭ 11); trunk and posterior to the lobe, distal Ϸ60% of surface of conductive tissue hemispherical in shape; thin- smooth (Figs. 8 and 10C). walled, sac-like structure of unknown function visible Female Copulatory Tube and Sperm Pouch. Cop- within sperm pouch at junction of pouch and copu- ulatory tube similar in appearance to that of A. ante- latory tube (Figs. 13, and 15C). 1270 ANNALS OF THE ENTOMOLOGICAL SOCIETY OF AMERICA Vol. 104, no. 6

A B 0.2 mm 0.2 mm

1. lobes

A. antevolens A. dimorphicus

C D 0.2 mm 0.2 mm

A. musculus A. confusus

Fig. 10. Everted endosomata of Anthocoris. (A) A. antevolens; inset shows spiniform projections on lobe of endosoma near base of organ. (B) A. dimorphicus; inset one shows paired ßeshy lobes near base of organ; inset two shows short spiniform projections on most basal lobe. (C) A. musculus. (D) A. confusus (organ not fully inßated).

Anthocoris sp. L lateral bend in dorsal view with pronounced twist at (Figs. 5, 8, 9, 11B, 13, 15A and B) apex; groove in form of a shallow depression (Fig. 9). Male Reproductive Tract. Large in overall size (Fig. Endosoma. Everted organ similar in appearance to 5). Small testes, as in A. tomentosus, with two testicular endosoma of A. tomentosus except longer (0.85 mm in ϭ follicles per testis; seminal vesicles prominent, as in A. length; n 6); distal one half densely covered with tomentosus, but more parallel-sided; large and heavily spiniform projections oriented basad, basal one half lobed mesadene glands; mesadenia attached to me- sparsely covered with minute dentate projections sadene reservoir well away from junction of reservoir (Figs. 8 and 11B). and seminal vesicle, as in A. tomentosus; ejaculatory Female Copulatory Tube and Sperm Pouch. Cop- bulb oval, slightly larger than phallus. ulatory tube 0.54 in length (n ϭ 4); largest diameter Paramere. Similar in appearance to paramere of A. along basal Ϸ60% of tube and then rapidly becoming tomentosus; slender and somewhat curved; distinct narrow apically; funnel-shaped at apex. Sperm pouch November 2011 HORTON AND LEWIS:GENITALIA OF NORTH AMERICAN Anthocoris 1271

A B 0.1 mm 0.1 mm

A. tomentosus Anthocoris sp. L

Fig. 11. Everted endosomata of Anthocoris. (A) A. tomentosus; inset shows positioning of endosoma along paramere. (B) Anthocoris sp. L.

0.52 by 0.52 mm (width ϫ length; n ϭ 4); trunk of Paramere. Slender and curved; without longitudinal conductive tissue hemispherical in shape; thin-walled, groove (Fig. 9). sac-like structure of unknown function visible within Endosoma. Everted organ 1.60 mm in length (n ϭ sperm pouch at junction of pouch and copulatory tube 5), conspicuously larger diameter at base than at apex; (Figs. 13 and 15A,B). with distinct bend or loop at basal end, followed dis- tally by noticeable increase in diameter before becoming more narrow apically; surface of looped region A. bakeri with noticeable striations (Fig. 12B, inset), bulbous (Figs. 5, 8, 9, 12A, 13, 14B) portion distal to loop Þnely shagreened, remaining Male Reproductive Tract. Intermediate in size to surface smooth (Figs. 8 and 12B). reproductive tract of A. tomentosus/Anthocoris sp. L Female Copulatory Tube and Sperm Pouch. Cop- and A. whitei/A. albiger species groups (Fig. 5). Testes ulatory tube 1.13 mm in length (n ϭ 11); diameter each with Þve to seven testicular lobes; ejaculatory largest at base, with gradual narrowing toward apical bulb somewhat pear-shaped; seminal vesicles parallel- end. Sperm pouch 0.35 by 0.29 mm (width ϫ length; sided. n ϭ 10); trunk of conductive tissue not pronounced Paramere. Lamellate, slender and curved; without (Figs. 13 and 14C,E). longitudinal groove (Fig. 9). Endosoma. Everted organ 1.81 mm in length (n ϭ A. whitei 9); noticeable loop present at basal end; surface of (Figs. 6, 8, 9, 12C, 13, 16) organ mostly smooth, looped portion with moderate density of small rounded protuberances (Figs. 8 and Male Reproductive Tract. Overall size small (Fig. 12A). 6). Ejaculatory bulb parallel-sided and relatively large; Female Copulatory Tube and Sperm Pouch. Cop- two testicular follicles per testis; phallus roundish and ulatory tube 1.42 mm in length (n ϭ 11), of uniform relatively large. diameter. Sperm pouch large (0.50 by 0.44 mm, Paramere. Slender with sharp apical tip, pro- width ϫ length; n ϭ 9); trunk of conductive tissue not nounced curve in organ at basal end; anterior edge pronounced (Figs. 13 and 14B). rolled inwards to create pronounced groove, the groove wide and shallow at apex of organ, becoming very narrow in basal half to form almost fully enclosed A. nemoralis tube (Fig. 9). (Figs. 5, 8, 9, 12B, 13, 14C and E) Endosoma. Fully everted organ very long (3.52 mm; Male Reproductive Tract. Intermediate in size to n ϭ 6), exceeding length of body; of uniform diameter reproductive tract of A. tomentosus/Anthocoris sp. L until distal 10% of organ, where it widens substantially and A. whitei/A. albiger species groups (Fig. 5). Ejac- but then narrows rapidly at very end; line of minute ulatory bulb spherical; seminal vesicles parallel-sided; teeth present over entire length except at distal end two testicular follicles per testis. (Fig. 12C, inset 1); surface of organ at the enlarged 1272 ANNALS OF THE ENTOMOLOGICAL SOCIETY OF AMERICA Vol. 104, no. 6

AB0.2 mm 0.2 mm

A. bakeri A. nemoralis

C teeth

0.2 mm 2.

A. whitei A. albiger

Fig. 12. Everted endosomata of Anthocoris. (A) A. bakeri. (B) A. nemoralis; inset shows striations on surface of looped region. (C) A. whitei; inset one shows row of teeth on surface of endosoma; inset two shows raised bumps on enlarged distal region for A. albiger (thin terminal process not everted in photograph), very similar in appearance to same structure of A. whitei. distal end covered with raised bumps (Fig. 12C, inset ulatory bulb and seminal vesicles are less parallel- 2 [shown for closely related species A. albiger]) (Figs. sided; two testicular follicles per testis (Fig. 6). 8 and 12C). Paramere. Similar in size and shape to paramere of Female Copulatory Tube and Sperm Pouch. Cop- A. whitei; anterior edge rolled inwards to create pro- ulatory tube long (2.48 mm; n ϭ 9); distal end in- nounced groove, the groove wide and shallow at apex creases in diameter, followed by abrupt decrease in of organ, becoming very narrow in basal half to form diameter at very end of tube. Sperm pouch spherical, almost fully enclosed tube (Fig. 9). relatively small (0.21 by 0.23 mm, width ϫ length; n ϭ Endosoma. Fully everted organ 3.31 mm in length 2); trunk of conductive tissue not pronounced; thin- (n ϭ 6), similar in appearance to endosoma of A. walled, sac-like structure of unknown function visible whitei, swollen apical area somewhat shorter than that within sperm pouch at junction of pouch and copu- of A. whitei, densely covered with raised bumps (Figs. latory tube (Figs. 13 and 16). 8 and 12C [insert 2]). Female Copulatory Tube and Sperm Pouch. Cop- ulatory tube similar in appearance to that of A. whitei, A. albiger but somewhat shorter (2.13 mm; n ϭ 7). Sperm pouch (Figs. 6, 8, 9, 12C [inset 2], 13) spherical, relatively small (0.21 by 0.21 mm; n ϭ 6); Male Reproductive Tract. Similar in size and ap- trunk of conductive tissue not pronounced; thin- pearance to reproductive tract of A. whitei, but ejac- walled, sac-like structure of unknown function visible November 2011 HORTON AND LEWIS:GENITALIA OF NORTH AMERICAN Anthocoris 1273

Fig. 13. Copulatory tubes and sperm pouches of Anthocoris. As seen when dissected in saline and viewed through dissecting microscope (50ϫ); some details at apical end of tube examined under compound microscope. Illustrations of sperm pouches from examination of virgin females with the exception of Anthocoris sp. L. Copulatory tubes straightened for illustration. Illustrations by Debra Broers. within sperm pouch at junction of pouch and copu- of the female fail to allow identiÞcation (Yasunaga latory tube (Fig. 13). 1997, Postle et al. 2001). Size and shape of the copulatory tube and sperm pouch of females of Anthocoris have been used to explore phylogenetic groupings of Discussion species within this genus (Ke and Bu 2007). Divergence among insect species in morphology of We found extensive variation in length of the cop- the genitalia, especially genitalia in male insects, has ulatory tube among North American species of An- made these structures useful for distinguishing among thocoris. The differences in morphology among spe- even closely related species. For the Anthocoridae, cies are often strikingly large. For example, there is a the paramere is routinely used in keys or descriptions Þve-fold difference in length of the copulatory tube to identify species (Hill 1957, Herring 1966, Carayon between the species having the shortest structure (A. 1972, Pe´ricart 1972, Kelton 1978, Bu and Zheng, 2001). tomentosus) and the species having the longest struc- Indeed, in some difÞcult complexes, identiÞcations ture (A. whitei). Our results are consistent with the cannot be made without examining the paramere taxonomically broader (40 species) examination of the (Pe´ricart 1972, Yasunaga 1997). Characteristics of the copulatory tube of Anthocoris provided by Ke and Bu female genitalia are used less frequently, but are of (2007), with minor exceptions. By dissecting fresh importance in complexes of Anthocoridae in which specimens, we were able to view a spherical structure males may be difÞcult to Þnd or in which other traits at the base of the sperm pouch where the copulatory 1274 ANNALS OF THE ENTOMOLOGICAL SOCIETY OF AMERICA Vol. 104, no. 6

A. antevolens A. bakeri A. nemoralis 0.2 mm 0.2 mm 0.2 mm

A B C A. antevolens A. nemoralis sperm pouch 0.1 mm

D trunk E 0.2 mm Fig. 14. Copulatory tubes and sperm pouches of Anthocoris. (A) A. antevolens, showing pronounced “trunk” of connective tissue immediately below sperm pouch. (B) A. bakeri. (C) A. nemoralis; sperm pouch damaged during dissection. (D) A. antevolens, showing curve of copulatory tube within trunk of connective tissue. (E) A. nemoralis. tube meets the pouch (Figs. 1, 13, and 14A). Carayon identiÞcations within this genus (Hill 1957, Pe´ricart (1953) referred to this structure as the “trunk” of the 1972, Kelton 1978). In seven of the 10 species, a groove conductive tissues which bridge the sperm pouch and or shallow depression runs longitudinally down the ovaries. This trunk of conductive tissues was not men- length of the organ (Fig. 9; as also discussed in Car- tioned by Ke and Bu (2007), presumably because it ayon 1972 and Pe´ricart 1972). This groove apparently had disappeared during preparation of specimens in functions as a guide for the endosoma as it is enters the KOH. femaleÕs copulatory tube (Fig. 11A, inset). The groove Internal reproductive organs of males show sub- was not easily seen, particularly for species in which stantial variation in size and shape among species it is shallow (A. tomentosus and Anthocoris sp. L). We (Figs. 4Ð6). There are noticeable differences among found that moving a Þne hair back-and-forth across species in size and shape of testicular follicles, seminal the surface of the paramere was useful for conÞrming vesicles, ejaculatory bulb, phallus, and mesadene res- presence of the groove (see Materials and Methods). ervoirs. Variation among species in internal reproduc- Setae are present on the paramere of all species (Fig. tive organs includes differences in the number of tes- 9; also illustrated for several European Anthocoris in ticular follicles. Anthocoris bakeri has Þve to seven Pe´ricart 1972). Function of the setae is unknown. follicles per testis, whereas the remaining species each Apparent length and orientation of setae was found to have two follicles per testis. The typical number in depend upon orientation of the paramere while view- Anthocoris is two follicles per testis, although one Old ing it beneath the microscope. World species, Anthocoris nemorum (L., 1761), has Our descriptions of the endosomata are the Þrst for four or more follicles per testis (Pendergrast 1957, any species of Anthocoris. We observed substantial Carayon 1972). Within the Anthocoridae, the ten- variation among species in length of the endosoma, in dency is to have two follicles per testis, except in the whether the structure exhibits looping, and in the Xylocorini (with Þve follicles; Carayon 1972) and presence or absence of spiniform projections, bumps, some Almeidini (seven follicles; Woodward 1977). roughening, and lobes on the surface of the inßated The paramere varies among species in size, shape, organ. Presumably, looping of the endosoma or the and presence of a longitudinal groove. Size and shape presence of projections, lobes, and swellings (Figs. 8 of the paramere are regularly used to conÞrm speciesÕ and 10Ð12) assist in anchoring the fully inßated organ November 2011 HORTON AND LEWIS:GENITALIA OF NORTH AMERICAN Anthocoris 1275

A B 0.1 mm 0.2 mm

Anthocoris sp. L Anthocoris sp. L

C A. tomentosus 0.2 mm

Fig. 15. Copulatory tubes and sperm pouches of Anthocoris. (A) Anthocoris sp. L. (B) Anthocoris sp. L, showing extensive pleating of copulatory tube; sperm pouch destroyed during dissection. (C) A. tomentosus (base of copulatory tube obscured by fatty tissue). within the femaleÕs copulatory tube during copulation 2004, Eberhard 2010). In some models of sexual se- (Edvardsson and Tregenza 2005). Indeed, we found it lection, morphology of genitalia is expected to co- impossible to separate freshly killed male and female evolve in males and females (Hosken and Stockley partners of copulating A. whitei or A. albiger without 2004, Eberhard 2010), leading to correlation in size or damaging the genitalia, apparently because the swol- shape of the genitalia between males and females. len portion of the endosoma near its apex (Figs. 8 and Genitalic traits that lead to a sexual advantage for one 12C) was too large to pull through the femaleÕs cop- sex and a cost to the other sex could lead to coevo- ulatory tube. lutionary change in genitalic morphology between Extensive divergence among closely related species males and females (Arnqvist and Rowe 2002, Hosken in morphology of genitalia, especially genitalia of and Stockley 2004, Brennan et al. 2007). Correlation in males, is one of the most general patterns of evolution size or shape of male and female genitalia across spe- (Eberhard 1985, 2010; Hosken and Stockley 2004). cies have been described in several insect taxa, in- Evidence supports the idea that sexual selection is cluding Diptera (Iwaizumi et al. 1997, Ilango and Lane largely responsible for rapid evolution in genitalic 2000), Lepidoptera (Mikkola 1992), and Coleoptera morphology (Arnqvist 1997, Hosken and Stockley (Sota and Kubota 1998). 1276 ANNALS OF THE ENTOMOLOGICAL SOCIETY OF AMERICA Vol. 104, no. 6

A. tomentosus have a short, thin copulatory tube (Figs. A 13 and 15C). Correlation in length of endosoma and copulatory tube is not unexpected. Males seem to deposit sperm directly into the femaleÕs sperm pouch, thus length of his endosoma relative to length of the femaleÕs copulatory tube probably is important in de- termining whether insemination is successful. Horton and Lewis (2005) showed that mismatched endosoma and copulatory tubes among geographic populations of A. antevolens led to reduced success of the male in everting his endosoma during copulation, accompanied by reduced success in inseminating the female. Size and shape of different structures within sexes also may also covary. For example, size of the phallosoma seems to vary across species in parallel with length of the endosoma. This particular relationship is

0.2 mm not unexpected, as the phallosoma houses the endosoma when the latter is in repose (Fig. 3A and E). Thus, the endosoma in A. whitei and A. albiger is quite long (Fig. 8), and the phallosoma in both species is correspondingly large (Fig. 6). Conversely, the endosoma of A. tomentosus is quite short (Fig. 8), and the phallosoma is correspondingly small (Fig. 5). Varia- B tion in other structures such as the testes, seminal vesicles, ejaculatory bulb, and mesadene reservoirs across species is also noticeable, but cannot at this time be explained. Size of these structures has been shown for other insects to depend upon traits such as sperm length, volume of sperm inseminated by the male, intensity of sperm competition, and copulation duration (Dybas and Dybas 1981, Gage 1994, Pitnick 1996, Minder et al. 2005). Too little is known about sexual selection in Anthocoris to speculate about the potential role of these factors affecting size of the various reproductive organs in members of this genus. endosoma Finally, characteristics of the endosoma, paramere, and copulatory tube of these 10 North American spe- of male cies produce some distinct groupings of species: (1) A. whitei and A. albiger; (2) A. antevolens, A. musculus, and A. dimorphicus; and (3) A. tomentosus and the as yet undescribed species. These clusters are consistent with groupings suggested by external traits. Anthocoris antevolens, A. musculus, and the macropterous form of A. dimorphicus are quite similar in external appear- Fig. 16. Sperm pouch and distal end of copulatory tube ance. Indeed, traits used to separate species within this of A. whitei. (B) Shows distal end of maleÕs endosoma within group vary enough geographically that there can be copulatory tube where copulatory tube meets sperm pouch. difÞculties in identifying insects from certain geographic regions (Hill 1957, Harper 1959, Horton et al. We observed correlation between the sexes in 2008). Moreover, we now know that A. musculus and structures associated with the genitalia. One espe- A. antevolens are actually part of a complex of cryptic cially noticeable correlation seems to be that between species that can be difÞcult to separate morphologi- the length of the endosoma of the male, and the length cally, suggesting that these species are evolutionarily of the femaleÕs copulatory tube. Thus, the very long close to one another (Horton et al. 2008). Thus, sim- endosoma of male A. whitei and A. albiger is mirrored ilarity in genitalia among species within this group is by the extreme length of the femaleÕs copulatory tube not unexpected. A. albiger and A. whitei are similar to of these two species (Figs. 8 and 13). The distal portion one another in external traits, and group together here of the inßated endosoma of A. whitei and A. albiger in characteristics of the genitalia. We have found that narrows sharply, in parallel with the rapid narrowing heterotypic crosses in the laboratory lead to insemi- of the femaleÕs copulatory tube where it enters the nation, albeit without production of offspring (T.M.L., sperm pouch (Fig. 16B shows apex of endosoma unpublished data). Finally, A. tomentosus and the un- within femaleÕs copulatory tube). Males of A. tomen- described species seem to group together based upon tosus have a short, thin endosoma (Fig. 8); females of characteristics of the genitalia. These two species are November 2011 HORTON AND LEWIS:GENITALIA OF NORTH AMERICAN Anthocoris 1277 also quite similar in external appearance (T.M.L., un- of Cimicidae. The Thomas Say Foundation, volume VII, published data). Entomological Society of America, Lanham, MD. We have an incomplete understanding of the phy- Carayon, J. 1972. Caracte`res syste´matiques et classiÞcation logenetic relationships among species of Anthocoris des Anthocoridae (Hemipt.). Ann. Soc. Entomol. France (Ke and Bu 2007). Similarities among some species in (N.S.). 8: 309Ð349. characteristics of genitalia suggest that these struc- Carayon, J. 1977. Inse´mination extra-geńitale traumatique, tures may be useful in developing superspeciÞc group- pp. 351Ð390. In P. P. Grasse´ (ed.), Traite´ de Zoologie. Anatomie, Systematique, Biologie. Tome VIII: Insectes. ings. Ke and Bu (2007) used morphology of the cop- ´ Game´togene`ses, Fećondation, Me´tamorphoses. Masson, ulatory tube of female Anthocoris to subdivide the Paris, France. genus. They examined the copulatory tubes of 40 spe- Dupuis, C. 1970. Heteroptera, pp. 190Ð209. In S. L. Tuxen cies from several geographic regions, and identiÞed 13 (ed.), TaxonomistÕs glossary of genitalia in insects. speciesÕ groupings based upon similarity in size and Munksgaard, Copenhagen, Denmark. shape of the copulatory tube. Ke and Bu (2007) com- Dybas, L. K., and H. S. Dybas. 1981. Coadaptation and tax- pared their results with similar efforts made by Hill onomic differentiation of sperm and spermathecae in (1957), Pe´ricart (1972), and Bu and Zheng (2001), featherwing beetles. Evolution 35: 168Ð174. who based groupings on external traits. Our groupings Eberhard, W. G. 1985. Sexual Selection and animal genita- with 10 North American species of Anthocoris based lia. Harvard University Press, Cambridge, MA. upon male genitalia are entirely consistent with Eberhard, W. G. 2010. Evolution of genitalia: theories, ev- groupings made by Ke and Bu (2007) from their ex- idence, and new directions. Genetica 138: 5Ð18. Edvardsson, M., and T. Tregenza. 2005. Why do male Cal- amination of the female genitalia. In sum, character- losobruchus maculatus harm their mates? Behav. Ecol. 16: istics of both male and female genitalia in the Antho- 788Ð793. coridae may allow inferences to be made about Gage, M.J.G. 1994. Associations between body size, mating superspeciÞc groupings (Yasunaga 1997; Ke and Bu pattern, testis size and sperm lengths across butterßies. 2005, 2007). Proc. R. Soc. Lond. B 258: 247Ð254. Correction of Terminology Used in Previous Pub- Harper, A. M. 1959. Gall aphids on poplar in Alberta. II. lications. In other accounts of Anthocoris, Horton in- Periods of emergence from galls, reproductive capacities, correctly used the terms “phallus” and “genital cap- and predators of aphids in galls. Can. Entomol 91: 680Ð sule” in referring to “endosoma” and “phallosoma,” 685. respectively (Horton and Lewis 2005, Horton et al. Henry, T. J. 1988. Family Anthocoridae Fieber, 1837. The 2007, 2008). minute pirate bugs, pp. 12Ð28. In T. J. Henry and R. C. Froeschner (eds.), Catalog of the Heteroptera, or true bugs, of Canada and the continental United States. E. J. Acknowledgments Brill, Leiden, The Netherlands. Herard, F. 1986. Annotated list of the entomophagous com- Illustrations and layout for Figs. 4Ð6 are by Janet Lane. We plex associated with pear psylla, Psylla pyri (L.) (Hom.: are grateful to Debra Broers for providing the line drawings Psyllidae) in France. Agronomie 6: 1Ð34. in Figs. 2, 3, 8, 9, and 13.We thank Laura Corley Lavine, Tom Herring, J. L. 1966. The genus Orius of the Western Hemi- Henry, and Kazutaka Yamada for reviewing an earlier version sphere (Hemiptera: Anthocoridae). Ann. Entomol. Soc. of this manuscript. Am. 59: 1093Ð1109. Hill, A. R. 1957. A key to the North American members of the genus Anthocoris Fallen. Pan-Pac. Entomol 33: 171Ð References Cited 174. Horton, D. R., and T. M. Lewis. 2000. Seasonal distribution Anthocoris Anderson, N. H. 1962. Bionomics of six species of of Anthocoris spp. and Deraeocoris brevis (Heteroptera: (Heteroptera: Anthocoridae) in England. Trans. R. En- Anthocoridae, Miridae) in orchard and non-orchard hab- tomol. Soc. Lond. 114: 67Ð95. itats of central Washington. Ann. Entomol. Soc. Am. 93: Arnqvist, G. 1997. The evolution of animal genitalia: distin- 476Ð485. guishing between hypotheses by single species studies. Horton, D. R., and T. M. Lewis. 2005. Size and shape dif- Biol. J. Linn. Soc. 60: 365Ð379. Arnqvist, G., and L. Rowe. 2002. Correlated evolution of ferences in genitalia of males from sympatric and repro- male and female morphologies in water striders. Evolu- ductively isolated populations of Anthocoris antevolens tion 56: 936Ð947. White (Heteroptera: Anthocoridae) in the Yakima Val- Brennan, P.L.R., R. O. Prum, K. G. McCracken, M. D. So- ley, Washington. Ann. Entomol. Soc. Am. 98: 527Ð535. renson, R. E. Wilson, and T. R. Birkhead. 2007. Coevo- Horton, D. R., and T. M. Lewis. 2009. Anthocoris confusus lution of male and female genital morphology in water- collected from western Washington State, with a sum- fowl. PLoS ONE 2: e418. mary of North American records (Hemiptera: Heterop- Bu, W.-J., and L.-Y. Zheng. 2001. Fauna Sinica. Insecta vol. tera: Anthocoridae). Proc. Entomol. Soc. Wash. 111: 609Ð 24. Hemiptera. Lasiochilidae Lyctocoridae Anthocori- 616. dae. Chinese Academy of Sciences, Science Press, Bei- Horton, D. R., T. M. Lewis, and T. Hinojosa. 2002. Copu- jing, China. lation duration in three species of Anthocoris (Heterop- Carayon, J. 1953. Existence dÕun double oriÞce geńital et tera: Anthocoridae) at different temperatures and effects dÕun tissue conducteur des spermatozoõ¨des chez les An- on insemination and ovarian development. Pan-Pac. En- thocorinae (Hemipt. Anthocoridae). C.R. Acad. Sci. Paris tomol. 78: 43Ð55. 236: 1206Ð1208. Horton, D. R., T. M. Lewis, and D. A. Broers. 2004. Eco- Carayon, J. 1966. Traumatic insemination and the parageni- logical and geographic range expansion of the introduced tal system, pp. 81Ð166. In R. L. Usinger (ed.), Monograph predator Anthocoris nemoralis (Heteroptera: Anthocori- 1278 ANNALS OF THE ENTOMOLOGICAL SOCIETY OF AMERICA Vol. 104, no. 6

dae) in North America: potential for non-target effects? Maw, H.E.L., R.G. Foottit, K.G.A. Hamilton, and G.G.E. Am. Entomol. 50: 18Ð30. Scudder. 2000. Checklist of the Hemiptera of Canada Horton, D. R., T. R. Unruh, T. M. Lewis, and K. Thomsen- and Alaska. National Research Council of Canada, NRC Archer. 2007. Morphological and genetic divergence in Research Press, Ottawa, ON, Canada. three populations of Anthocoris antevolens (Heteroptera: Mikkola, K. 1992. Evidence for lock-and-key mechanisms in Anthocoridae). Ann. Entomol. Soc. Am. 100: 403Ð412. the internal genitalia of the Apamea moths (Lepidoptera, Horton, D. R., T. M. Lewis, K. Thomsen-Archer, and T. R. Noctuidae). Syst. Entomol. 17: 145Ð153. Unruh. 2008. Morphology, genetics, and male mating Minder, A. M., D. J. Hosken, and P. I. Ward. 2005. Co-evolu- success compared between Anthocoris musculus and A. tion of male and female reproductive characters across the antevolens (Hemiptera: Heteroptera: Anthocoridae). Scathophagidae (Diptera). J. Evol. Biol. 18: 60Ð69. Proc. Entomol. Soc. Wash. 110: 960Ð977. Pendergrast, J. G. 1957. Studies on the reproductive organs Hosken, D. J., and P. Stockley. 2004. Sexual selection and of the Heteroptera with a consideration of their bearing genital evolution. Trends Ecol. Evol. 19: 87Ð93. on classiÞcation. Trans. R. Entomol. Soc. Lond. 109: 1Ð63. Ilango, K., and R. P. Lane. 2000. Coadaptation of male ae- Pe´ricart, J. 1972. He´mipte`res. Anthocoridae, Cimicidae et deagal Þlaments and female spermathecal ducts of the old Microphysidae de LÕOuest-Pale´arctique. In Faune de world Phlebotomine sand ßies (Diptera: Psychodidae). LÕEurope et du Bassin Me´diterrane´en. Masson et Cie J. Med. Entomol. 37: 653Ð659. Paris 7: 1Ð402. Iwaizumi, R., M. Kaneda, and O. Iwahashi. 1997. Correla- Pe´ricart, J. 1996. Family Anthocoridae Fieber, 1836-ßower tion of length of terminalia of males and females among bugs, minute pirate bugs, pp. 108Ð140. In B. Aukema and nine species of Bactrocera (Diptera: Tephritidae) and C. Rieger (eds.), Catalogue of the Heteroptera of the differences among sympatric species of B. dorsalis com- Palaearctic Region, vol. 2. The Netherlands Entomolog- plex. Ann. Entomol. Soc. Am. 90: 664Ð666. ical Society, Wageningen, The Netherlands. Ke, Y.-L., and W.-J. Bu. 2005. A morphological study on Pitnick, S. 1996. Investment in testes and the cost of making female copulatory tubes of the genus Tetraphleps Fieber long sperm in Drosophila. Am. Nat. 148: 57Ð80. (Hemiptera: Heteroptera: Anthocoridae) from China. Postle, A. C., M. Y. Steiner, and S. Goodwin. 2001. Oriini Acta Entomol. Sin. 48: 391Ð395. (Hemiptera: Anthocoridae) new to Australia. Aust. J. Ke, Y.-L., and W.-J. Bu. 2007. Female copulatory tubes and Entomol. 40: 231Ð244. the subdivision of the genus Anthocoris (Heteroptera: Schuh, R. T., and J. A. Slater. 1995. True bugs of the world Anthocoridae: Anthocorini). Eur. J. Entomol. 104: 89Ð98. (Hemiptera: Heteroptera). Cornell University Press, Kelton, L. A. 1978. The Anthocoridae of Canada and Alaska. Ithaca, NY. Heteroptera Anthocoridae. The Insects and Arachnids of Sota, T., and K. Kubota. 1998. Genital lock-and-key as a Canada. Part 4. Agriculture Canada Research Publication selective agent against hybridization. Evolution 52: 1507Ð 1639. 1513. Lattin, J. D. 1999. Bionomics of the Anthocoridae. Annu. Woodward, T. E. 1977. The occurrence of Almeidini Rev. Entomol. 44: 207Ð231. (Hemiptera: Anthocoridae) in Australia, with the de- Lewis, T. M., D. R. Horton, and J. D. Lattin. 2003. Antho- scription of a new genus. Syst. Entomol. 2: 255Ð260. coris tristis Van Duzee, 1921, a synonym of Anthocoris Yasunaga, T. 1997. The ßower bug genus Orius Wolff whitei Reuter, 1884 (Hemiptera: Heteroptera: Anthocori- (Heteroptera: Anthocoridae) from Japan and Taiwan, dae). Pan-Pac. Entomol. 79: 155. Part I-III. Appl. Entomol. Zool. 32: 355Ð364, 379Ð386: Lewis, T. M., D. R. Horton, and D. A. Broers. 2005. New 387Ð394. state and United States records for Anthocoridae (Hemiptera: Heteroptera). Pan-Pac. Entomol 81: 59Ð67. Received 16 May 2011; accepted 26 August 2011.