University of Nebraska - Lincoln DigitalCommons@University of Nebraska - Lincoln

USDA Systematic Entomology Laboratory Entomology Collections, Miscellaneous

2010

History, Distribution, and Identification Of dodecella (L.) (: ) in North America, With Insights into the Systematics of Exoteleia Wallengren using Characters of the Adult, Immatures, Bionomics, and DNA Barcodes

David Adamski Department of Entomology, National Museum of Natural History, Smithsonian Institution, P.O. Box 37012, MRC 168, Washington, D.C., 20013- 7012, U.S.A., [email protected]

Jean-Francois Landry Agriculture and Agri-Food Canada, Neatby Building, C.E.F., 960 Carling Avenue, Ottawa, Ontario, K1A 0C6, Canada, [email protected]

Steven C. Passoa USDA, APHIS, PPQ, USDA Forest Service Northern Research Station and The Ohio State University, Museum of , 1315 Kinnear Road, Columbus, OH 43212, U.S.A., [email protected]

Robert Tracy USDA, APHIS, PPQ, 2500 Brunswick Avenue, Building G, Linden, NJ 07036, U.S.A., [email protected] Follow this and additional works at: https://digitalcommons.unl.edu/systentomologyusda

Part of the Entomology Commons

Adamski, David; Landry, Jean-Francois; Passoa, Steven C.; and Tracy, Robert, "History, Distribution, and Identification Of (L.) (Lepidoptera: Gelechiidae) in North America, With Insights into the Systematics of Exoteleia Wallengren using Characters of the Adult, Immatures, Bionomics, and DNA Barcodes" (2010). USDA Systematic Entomology Laboratory. 54. https://digitalcommons.unl.edu/systentomologyusda/54

This Article is brought to you for free and open access by the Entomology Collections, Miscellaneous at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in USDA Systematic Entomology Laboratory by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. PROC. ENTOMOL. SOC. WASH. 112(2), 2010, pp. 183–206

HISTORY, DISTRIBUTION, AND IDENTIFICATION OF EXOTELEIA DODECELLA (L.) (LEPIDOPTERA: GELECHIIDAE) IN NORTH AMERICA, WITH INSIGHTS INTO THE SYSTEMATICS OF EXOTELEIA WALLENGREN USING CHARACTERS OF THE ADULT, IMMATURES, BIONOMICS, AND DNA BARCODES

D. ADAMSKI, J.-F. LANDRY,S.PASSOA, AND R. A. TRACY

(DA) Department of Entomology, National Museum of Natural History, P.O. Box 37012, NHB-E523, Smithsonian Institution, Washington, D.C. 20013-7012, U.S.A. (e-mail: [email protected]); (J-FL) Agriculture and Agri-Food Canada, Neatby Building, C.E.F., 960 Carling Avenue, Ottawa, Ontario, K1A 0C6 Canada (e-mail: [email protected]; (SP) USDA, APHIS, PPQ, USDA Forest Service Northern Research Station and The Ohio State University, Museum of Biodiversity, 1315 Kinnear Road, Columbus, OH 43212, U.S.A. (e-mail: steven. [email protected]); (RAT) USDA, APHIS, PPQ, 2500 Brunswick Avenue, Building G, Linden, NJ 07036, U.S.A. (e-mail: [email protected])

Abstract.—Exoteleia dodecella (L.) (Lepidoptera: Gelechiidae), a native of Europe, was first documented from North America at several locations in eastern Canada. Additional records indicate this has now spread throughout New England and west to northern Pennsylvania, New York, and possibly into Michigan in the United States. A second introduction of E. dodecella has occurred near the Vancouver area of British Columbia in Canada. To help with the identification of E. dodecella, morphological, biological, and molecular evidence are presented. Key features of the adult, larval, and pupal morphology are compared to other of Exoteleia and illustrated with line drawings or scanning electron micrographs. The high sequence divergence (.7%)ofE. dodecella compared to samples of related native North American species demonstrates that DNA barcodes are a useful identification tool for this . A summary of the biology of E. dodecella, including 12 species of larval and pupal parasitoids (most representing new host records), is also included. Key Words: bud-miner, chaetotaxy, DNA barcode, Europe, , invasive species, life cycle, morphology, needle-miner, North America, parasitoids, pine DOI: 10.4289/0013-8797-112.2.183

The pine bud moth, Exoteleia dode- gren, was first recorded in North cella (L.) (Lepidoptera: Gelechiidae), America as an established population the type species of Exoteleia Wallen- on the Canadian side of the Niagara Peninsula (Ontario) in 1928 (Sheppard * Accepted by Robert R. Kula 1930, Martin 1959). Attempts to erad- 184 PROCEEDINGS OF THE ENTOMOLOGICAL SOCIETY OF WASHINGTON icate this pest failed, and it spread to at conditions (Martin 1959). Both P. least Quebec (Handfield 1992) in east- sylvestris and P. mugo have been ern Canada. Exoteleia dodecella is also introduced into the United States; the known from the Vancouver area of former species during colonial times for British Columbia (specimens in the lumber (Elias 1989) and both species CNC), which likely represents a second more recently for ornamental use or in introduction to North America. tree plantations on state forests (Braun New World records for E. dodecella 1961, White and Hosie 1980). usually cite only Ontario (USDA 1986, In Europe, Lemarie (1958a, b, 1959a, Mattson et al. 1994) or North America b, 1961) reared 50 species of parasitoids (Hodges 1983, 1986). The history of that have been associated with the life this species in the United States is cycle of E. dodecella, while Martin poorly documented. Exoteleia dodecel- (1959) reared seven species of parasit- la was first collected from New York oids of E. dodecella in Canada. State in 1934 (see Material Examined). In addition to E. dodecella, there are These records apparently were never currently seven nominal species of published, or perhaps the population native Exoteleia recorded from North never established. Tracy collected E. America (if not described in Exoteleia, dodecella from Maine in 1978 and sent original combinations are in parenthe- voucher specimens to R. W. Hodges ses): E. anomala Hodges, 1985; E. (formerly of the Systematic Entomolo- burkei Keifer, 1932; E. californica gy Laboratory, Washington, DC) for (Busck, 1907) (Paralechia); E. chillcotti confirmation. His discovery became the Freeman, 1963; E. graphicella (Busck, first published United States record 1903) (); E. nepheos (Tracy 1980). The New York or Maine Freeman, 1967; and E. pinifoliella records were never entered into any of (Chambers, 1880) (). the older databases documenting the Nearctic immigrant fauna Exoteleia chillcotti has been errone- (USDA/ARS/BBII 1986, Knutson et ously listed under al. 1990, Kim and Wheeler 1991). (Hodges 1983, Lee and Brown 2008). Additionally, USDA-APHIS-PPQ has Examination of its holotype (CNC) no record of E. dodecella in the United shows that it is properly placed in States (J. Cavey pers. comm.), this pest Exoteleia. Hodges (1985) correctly is absent from their most current indicated that E. californica and E. database (NAPIS pest tracker), and no graphicella are misplaced in Exoteleia New Pest Advisory Group Report ever without giving a proper generic place- documented this introduction. ment. The genitalia of E. graphicella Exoteleia dodecella is a serious pest show that it is not a Gnorimoschema of pine in Europe (Martin 1959, Bland and suggests that it may belong in et al. 2002) and more recently in , although we are not certain. Canada, where up to 60% of the buds The genitalia of E. californica confirm of Scots (or Scotch) pine, Pinus syl- its misplacement, but we cannot make a vestris L. (Pinaceae), were destroyed in statement as to its positive generic Ontario (Martin 1959). Although the placement although the wing pattern is preferred hosts are Scots and Mugo similar to Coleotechnites. After remov- pines ( Turra), E. dodecella ing two possible misplaced species (see occasionally lays eggs on larch (Larix) details in Results section), five native (Pinaceae) (Bland et al. 2002) and feeds nominal species of North American on other pine species under certain Exoteleia remain: E. anomala, E. bur- VOLUME 112, NUMBER 2 185 kei, E. chillcotti, E. nepheos, and E. tella with the caveat that too few female pinifoliella. specimens were available to assess Thereisnosingleworkforthe variability and reliability of the charac- identification of North American Exo- ters (Huemer and Karsholt 1999). teleia. All existing publications present Interestingly, upon describing E. neph- individual species descriptions. Only eos, Freeman (1967) indicated that the two show a black-and-white photo of initial discovery and distribution near an adult (Hain and Wallner 1973, Lake Erie suggested that it could be an Hodges 1985). Genitalia have been introduced species. Specimens of E. illustrated only for the following three nepheos are similar in pattern to the species: E. chillcotti Freeman, 1963; E. smaller and dark brown ‘‘variants’’ of E. nepheos Freeman, 1967; and E. anom- dodecella from central Europe. ala Hodges, 1985, but they show no Taxonomic problems concerning reliable differences. Exoteleia raise the following two ques- In discussing the of Exo- tions. Is weak morphological differen- teleia, Hodges (1985) expressed frus- tiation evidence of distinct species, or is tration when attempting to delineate it simply intraspecific variation? Can species. Four taxa were recognized (E. the existing nominal species concepts in anomala, E. pinifoliella, E. dodecella, Exoteleia, based on morphological and one undescribed entity from the characters, be applied with any degree eastern U.S.), but he was unable to of confidence to any or all of the define three nominal species (burkei, clusters delineated by DNA barcodes? chillcotti, and nepheos). He also felt The purpose of this study is to (1) that there were possibly two additional document that E. dodecella is estab- undescribed ‘‘entities based on adult lished in the United States and Canada, characters,’’ and that pupal characters (2) provide new records of its present might be useful to define species if a distribution in North America, (3) make larger series of immatures could be available a modern description and associated with known adults. His diagnosis of E. dodecella, including conclusion was that it was not possible the adult and immature stages, for the ‘‘to resolve the question of separation of accurate identification of this species species in Nearctic Exoteleia.’’ He where it is discovered, and (4) assess provided a discussion of how characters the usefulness of classical morphology vary within the groups as he perceived and DNA barcodes as identification them but did not formally attach names tools for E. dodecella and other North to those groups or illustrate any of the American species of Exoteleia. variation he observed. Two species of Exoteleia, E. dode- MATERIALS AND METHODS cella and E. succinctella (Zeller), are Late-instar larvae were field collected recognized in Europe (Huemer and by removing twigs about a centimeter Karsholt 1999). Exoteleia dodecella is from the infested buds. One-half of the variable with respect to wing pattern infested buds were dissected to obtain and size; smaller and darker specimens larvae, and the remaining buds were occur in central Europe. The smallest placed in empty plastic artificial diet specimens of E. dodecella are quite cups closed with paper lids. Moth pupae similar in size and pattern compared to were dissected from infested buds at a some of the North American ‘‘species.’’ later time. Larvae were preserved by Minor genital differences are given to placing live specimens in boiling H2O separate E. dodecella and E. succinc- for less than a minute before transfer- 186 PROCEEDINGS OF THE ENTOMOLOGICAL SOCIETY OF WASHINGTON ring them to 70% ethyl alcohol. Pupae immature stages were not available, were preserved by placing live speci- we have placed quotations around mens directly into 70% ethyl alcohol. species identifications of larval and For SEM study, larvae and pupae pupal Exoteleia taken from the litera- were cleaned in a full-strength solution ture and collections to acknowledge that of Formula 409 TM detergent, rinsed in these names are tentative. The only distilled water, and subsequently dehy- exception is of E. dodecella because our drated in increasing concentrations of field-collected immature specimens are alcohol, ending with absolute alcohol. associated with reared adults. After dehydration, specimens were For molecular analysis specimens critical point dried using a Tousimis were collected live and killed using critical point dryer, mounted on SEM ammonium hydroxide or cyanide prior stubs using carbon paste, and coated to mounting and spreading. Specimens with gold-palladium (40/60%) using a were labeled with individual voucher Cressington sputter coater. Forewings codes (Specimen IDs), databased, and were detached from pinned specimens photographed. All collateral data, im- and mounted on stubs using carbon ages, sequences, and trace files were adhesive tabs. The finestructure of the uploaded to the project entitled, ‘‘Ge- , pupa, and the male sex scales on lechiidae of North America 02 (GO- the undersurface of the forewing was NAB)’’ and ‘‘Lepidoptera of Eastern studied with an Amray 1810 scanning North America’’ in the Barcode of Life electron microscope at an accelerating Database (BOLD) (www.barcodingli- voltage of 10 kV. fe.org, Ratnasingham and Hebert Morphological observations and 2007). Barcode sequences (658 bp measurements of the adults, larvae, segment from the 50 end of mitochon- and pupae were made using a Leitz drial cytochrome c oxidase I—or RS dissecting microscope with a cali- COI—gene) have been submitted to brated micrometer. Genitalia were dis- GenBank and have the following acces- sected as described by Clarke (1941) sion numbers: FJ412321 - FJ412323, except mercurochrome and chlorazol FJ412931 - FJ412939, GU088331 - black were used as stains. The Methuen GU088335, GU089773 - GU089774, Handbook of Colour (Kornerup and GU092285, GU095766 - GU095772, Wanscher 1978) was used as a color and GU358079 - GU358181. standard. Voucher specimens of adults One or two legs were removed from and immature stages of E. dodecella each specimen and stored in individual and its parasitoids from this study are tubes within a 96-tube sample box deposited in Smithsonian Institution obtained from Matrix Technologies. National Museum of Natural History, Analysis of DNA sequences followed Washington, D.C. (USNM), the DNA barcoding methods of Hajibabaei Collection, Department of Entomology, et al. (2005) and deWaard et al. (2008). University of Maine at Orono, Orono, Barcode sequences were obtained at Maine (UMDE), and the S. Passoa the Biodiversity Institute of Ontario at collection, Columbus, Ohio. Larval the University of Guelph. Tissue was nomenclature follows Stehr (1987), placed in a 96-well plate of proteinase and pupal nomenclature follows Mosh- K lysis buffer and incubated for about er (1916). taxonomy, including 18 hours. The lysate was then processed nomenclature and authorship, follows following the glass fibre-protocol of GRIN (2008). Because voucher speci- Ivanova et al. (2006) on a Beckman mens to associate adults with the Coulter BiomekFxp liquid handler. For VOLUME 112, NUMBER 2 187

PCR amplification, 2 ll of DNA extract containing gaps and missing data were was added to each well of a premade omitted from the analysis. Maximum PCR plate stored at 208C and contain- parsimony analysis was performed with ing 2 llofH20, 6.25 llof10% PAUP* 4.0b10 (Swofford 2003) set to trehalose, 1.25 llof103 buffer, 0.625 heuristic search, stepwise addition with llof50mMMgCl2, 0.0625 llof10 100 random addition sequence repli- mM dNTPs, 0.06 ll of Platinum Taq cates (max ¼ 500 trees). All other polymerase (Invitrogen), and 0.125 ll options were set to default. Prior to of each of the 10 lM primers LepF1 performing parsimony analysis, redun- and LepR1 (Hebert et al. 2004). Ther- dant sequences were merged with mocycling conditions consisted of an MacClade (Maddison and Maddison initial denaturation at 948C for 1 min, 2002). Bootstrap values were estimated five cycles of 948C for 30 sec, annealing using the stepwise algorithm and 1000 at 458C for 40 sec, and extension at repetitions. For comparison, barcode 728C for 1 min, followed by 35 cycles sequences for three species of Coleo- of 948C for 30 s, 518C for 40 s, and technites and one species of Recurvaria 728C for 10 min. The PCR reactions were included as outgroups in the were visualized with an Introgen E-Gel analysis based upon Lee and Brown 96 agarose electrophoresis system be- (2008) who suggested that these two fore performing the sequencing reac- genera are closely related to Exoteleia. tions, again in premade and frozen plates. Both the forward and reverse RESULTS AND DISCUSSION direction plates contained 0.25 llof Exoteleia dodecella (Linnaeus, 1758) Applied Biosystems Dye terminator (Figs. 1–32) mix v3.1, 1.875 llof53 sequencing buffer, 5 llof10% trehalose, and 1 ll Adult diagnosis and remarks.—In of 10 lM PCR primer. Sequencing addition to the grayish ground color reactions were run at an initial denatur- with darkly pigmented scale tufts on the ation at 968C for 2 min, followed by 30 forewings, E. dodecella can be distin- cycles of 968C for 30 s, annealing at guished from native North American 558C for 15 s, and extension at 608C for Exoteleia by having the male genitalia 4 min. The reactions were purified with the costal part of the valva using an Agencourt Bioscience CleanS- extending at least 1/5 its length beyond EQ system on a Biomek Fxp liquid the saccular part and a reduced base of handler before being run on an Applied the saccular part of the valva. In Biosystems 3730XL DNA Analyzer. females, the inception of the ductus Electropherograms were edited and seminalis is anterior to the seventh manually aligned in an Applied Biosys- segment, and an invaginated bulla is tems Seqscape v.2.5, and the resultant near to the posterior end of seventh sequences were uploaded into BOLD. segment. Genetic distances were analyzed with The male sex scales located on the the BOLD analysis tool (Ratnasingham undersurface of the forewing are pre- and Hebert 2007) and with MEGA4 sent in E. dodecella, E. anomala, E. (Tamura et al. 2007) using the Kimura burkei, E. chillcotti, E. nepheos, and E. 2-parameter model of base substitution pinifoliella but are absent in ‘‘californ- and to produce neighbor-joining (NJ) ica’’ or ‘‘graphicella.’’ We found that similarity trees. For the MEGA analy- there are some differences in the lengths sis, the default ‘complete deletion’ of the scale cluster and that these option was used whereby positions differences may help to discriminate 188 PROCEEDINGS OF THE ENTOMOLOGICAL SOCIETY OF WASHINGTON

Fig. 1. Adult of Exoteleia dodecella. species. Furthermore, the finestructure They noted that males have black scent of these sex scales has not been scales on the underside of the forewing examined, and it too shows promise between R1 and R5 but lack hair pencils for species recognition within Exote- at the base of the hind wing. Female leia. The absence of the male sex scales specimens of Exoteleia lack a signum in on the undersurface of the forewing the corpus bursae, a very unusual may indicate monophyly within Exote- character for the tribe. Species of leia and adds to the evidence of the Coleotechnites feed on the same hosts misplacement of ‘‘californica’’ or as Exoteleia but differ in having ‘‘graphicella.’’ asymmetrical male genitalia and a The adult of E. dodecella was rhomboid signum in females. described and illustrated by Freeman Redescription.—Adult. Head: Scales (1960), Piskunov (1989), Huemer and of vertex agouti patterned (transversely Karsholt (1999), Bland et al. (2002), tri-banded from base to apex); basal 2/3 Go´mez de Aizpu´rua (2003), and Lee pale gray, distal 1/3 gray or dark gray and Brown (2008). Freeman (1960), with narrow pale-gray margin; fronto- Piskunov (1989), and Bland et al. clypeus pale gray to white; outer (2002) included Exoteleia in their keys surface of labial palpus with scales emphasizing forewing pattern and the patterned as above; basal article dark male genitalia. With regard to the gray; 2nd article dark gray with paler Holarctic Teleiodini, Lee and Brown scales along distal margin, or scales on (2008) diagnosed Exoteleia by having distal half paler with white band of an ocellus and forewing scale tufts in scales along distal margin; distal article addition to several other characters with three irregularly-shaped alternat- depending on the sex of the individual. ing white bands and two dark-gray VOLUME 112, NUMBER 2 189

Figs. 2–7. Wing venation and scanning electron micrographs of male sex scales on undersurface of forewing of E. dodecella. 2, Venation of forewing with shaded area indicating location of male sex scales; 3, Venation of hind wing; 4, Undersurface of forewing of male, arrows ¼ positional area of sex scales, scale bar ¼ 1 mm; 5, Male sex scales, scale bar ¼ 100 lm; 6, Comparison of male sex scales and other wing scales, scale bar ¼ 10 lm; 7, Finestructure of male sex scales, scale bar ¼ 1 lm. 190 PROCEEDINGS OF THE ENTOMOLOGICAL SOCIETY OF WASHINGTON

Figs. 8–11. Male genitalia and 8th sternal and tergal plates of Exoteleia dodecella. 8, Eighth sternum; 9, Eighth tergum; 10, Genital capsule (sternal elements detached on one side and folded to opposite side); 11, Aedeagus. bands; inner surface as above except 2nd Thorax: Scales of tegula and meso- article with more white; scape of notum agouti patterned; basal 1/2 pale antenna dark gray with narrow band of gray, distal 1/2 gray with narrow pale- white along distal margin, each flagel- gray margin; mesonotum with dark gray lomere of antenna dark gray basally, to black scale tuft (one or two scales pale gray distally. Ocellus absent. wide) on posterolateral surface near Proboscis with pale-gray scales. margin. Scales of legs as above; foreleg VOLUME 112, NUMBER 2 191 and midleg dark gray with narrow white bands on distal margin of tibia and tarsomeres; hind leg similarly patterned with more white scales. Forewing (Fig. 1): Length 5.9–7.8 mm (n ¼ 30) scales agouti patterned; basal, median, and postmedian fasciae distally demarcated with paler scales; one black scale tuft near middle of base (three scales wide); two on opposite sides of CuP near end of basal fascia (anterior scale tuft two scales wide, posterior scale tuft five scales wide); two on opposite sides of CuP near end of median fascia (anterior scale tuft two scales wide, posterior scale tuft four to five scales wide); and two scale tufts on proximal margin of post median fascia above CuP (anterior scale tuft two scales wide, posterior scale tuft five scales wide); four or five marginal black spots present or absent; apex pale gray or white. Venation (Fig. 2) with all veins not reaching margin; R4 and R5 stalked near basal 1/4; M2 absent. Undersurface gray, contrasted by darkly pigmented male sex scales within shaded area (Fig. 2); scanning electron micrographs (Figs. 4–7) of male sex scales indicate palmate struc- ture with 9–12 digitate processes, in contrast to other wing scales (Figs. 5–6) having only irregularly-serrated distal margins; finestructure of palmate sex scales with latticelike pattern of cross- braces between scutes (Fig. 7). Hind wing pale gray. Venation (Fig. 3) with M1 weak; cubitus 4-branched with M2 originating closer to M3 than to M1; frenulum with one acanthus in male, three acanthae in female. Abdomen (Figs. 8–9): Dorsal surface pale gray; ventral surface pale gray, gradually darkening to distal end. Eighth sternum (Fig. 8) elongate, broadly rounded distally, with 2 short Fig. 12. Female genitalia of Exoteleia dode- basal arms; 8th tergum (Fig. 9) crescent- cella. shaped, wider than long, with broad- ened posterolateral margins. 192 PROCEEDINGS OF THE ENTOMOLOGICAL SOCIETY OF WASHINGTON

Figs. 13–18. Scanning electron micrographs of Exoteleia dodecella. 13, Head, frontal view, scale bar¼ 100 lm; 14, Head, ventral view, scale bar¼ 100 lm; 15, Apical sensilla of left antenna, 1¼ sensilla basiconica, 2 ¼ sensilla chaetica, 3 ¼ sensillum styloconicum, 4 ¼ sensillum trichodeum, scale bar ¼ 10 lm; 16, Apical sensilla of left maxillary palpus, A2 ¼ sensillum styloconicum, A1, A3, M1, M2, L1, L2, L3 ¼ sensilla basiconica, SD ¼ sensillum digitiform, scale bar ¼ 10 lm; 17, Left proleg on A5, dorsolateral view, scale bar ¼ 100 lm; 18, Setal arrangement of anal plate on A10, scale bar ¼ 100 lm. VOLUME 112, NUMBER 2 193

Figs. 19–22. Setal maps and mandible of Exoteleia dodecella. 19, T1–T3, lateral view; 20, A1– A2, lateral view; 21, Right mandible, view of inner surface; 22, A6 – 10, lateral view.

Male genitalia (Figs. 10–11): Uncus extending slightly beyond anterior end; hoodlike, setose; gnathos with short- an elongate lateral lobe fused with ened lateral arms extending inwards, vinculum near 1/3 length, extending acutely curved anteriorly and dilated posteriorly, forming slightly curved distally, each arm fusing with a median digitate lobe; lateral lobe fused at base conical process; tegumen gradually with anterolateral arm of tegumen widened basally forming two broad, ventral to an elongate, inwardly curved anteriorly divergent arms; vinculum an spinelike valva and a slightly shorter elongate median process widened pos- digitate lobe; aedeagus acutely bent teriorly and narrowed anteriorly; vincu- near 1/3 length from base, without lum with two short, convergent, digitate cornuti. lobes on posterior end and one elongate Female genitalia (Fig. 12): Oviposi- process originating from 2/3 length, tor telescopic, with three membranous 194 PROCEEDINGS OF THE ENTOMOLOGICAL SOCIETY OF WASHINGTON

Figs. 23–26. Pupa of Exoteleia dodecella. 23, A8 – 10, ventral view; 24, Pupa, ventral view; 25, Pupa, lateral view; 26, A8 – 10, lateral view. subdivisions posterior to 8th segment; imens for Tracy (1980). In addition, papillae anales lobate and setose; these specimens also serve as voucher apophyses posteriores about 2.53 lon- specimens for the first documentation of ger than apophyses anteriores; part of the introduction of this species into the ductus bursae from ostium to anterior United States. Two specimens collected margin of 7th sternum about 1.53 longer from Croton Dam, New York, June 9, than part of ductus bursae from poste- 1934, and one specimen from Valhalla, rior margin of 7th sternum to inception New York, June 12, 1934, reared from of ductus seminalis; corpus bursae Pinus resinosa Aiton are in the USNM, longer than wide; medioposterior mar- suggesting that E. dodecella may have gin of 7th sternum broadly emarginate; been established in the United States an opening within pleural membrane prior to its documented introduction demarcates an invaginated bulla near into Canada in 1928 (Sheppard 1930). posterior end of 7th segment. Other specimens examined: United Specimens examined: Four speci- States: Howland, Maine, 24 ex; Royal- mens from Howland, Penobscot Coun- ton, Vermont, 5 ex; Meriden, Connect- ty, Maine and three specimens from icut, 5 ex; and Great Bend, Pennsylva- Medford, Piscataquis County, Maine, nia, 1 ex. All adult specimens were reared from P. sylvestris by R. Tracy, reared from P. sylvestris [USNM]; (USNM Coll.), serve as voucher spec- Europe: 22 ex [USNM]. Canada: Ontar- VOLUME 112, NUMBER 2 195 io, Grimsby, reared from P. sylvestris, hole in the last instar mine instead of 1938, 1 ex; Ontario, Lorraine, 1932, 11 two holes that is characteristic of their ex; Ottawa, reared from Pinus mugo biology in Europe. Another complicat- and P. sylvestris, various dates 1934– ing factor is that E. dodecella may expel 1971, 7 ex; Ridgeville, reared from P. frass from the last mine of the season, sylvestris, 1932, 1 ex; Simcoe, reared whichisusedasanoverwintering from P. sylvestris, 1962, 8 ex; Vernon, shelter. Thus, biological characteristics reared from P. sylvestris, 1955, 30 ex; alone cannot be used to identify this Port Franks, collected at light, 4 ex; species. British Columbia: Port Coquitlam, col- Host plant data can aid in identifica- lected at blacklight, 2006, 15 ex [all tion. Scots and Mugo pine are readily Canada specimens in CNC]. colonized by E. dodecella. Exoteleia Larval diagnosis and remarks.—Exo- ‘‘pinifoliella’’ from New York preferred teleia dodecella can be distinguished (Pinus banksiana Lamb.) and from native species of North American pitch pine ( Mill.) but did Exoteleia by having a bisetose SV- not survive well on Scots pine (Bennett group on A1 and a single row of 8–14 1954a). Thus, miners on Scots pine are crochets on the anal prologs. Biological more likely to be E. dodecella than E. characters also provide important clues; ‘‘pinifoliella,’’ at least in New York the needle-mine of E. dodecella usually State. Unfortunately, Mugo pine was contains frass, and any specimen of not tested by Bennett (1954a). Exoteleia on Scots or Mugo pine is Because E. dodecella will opportu- most likely E. dodecella because spe- nistically feed on many species of pines cies of Exoteleia that occur in the U.S. if they are growing near their preferred tend to avoid those trees. hosts (Martin 1959), species of Exote- The larva of E. dodecella was leia on native pines are unlikely to be E. partially described or illustrated by dodecella unless they are near Scots or Martin (1959), Lindquist and Trinnell Mugo pine. Exoteleia nepheos is found (1967), and Go´mez de Aizpu´rua (2003). mostly on red pine, P. resinosa, less A combination of biology, host , frequently on Scots pine, and rarely on and morphology are all useful for Mugo pine (Lindquist and Trinnell identifying E. dodecella. 1967) again suggesting that E. dode- According to Freeman (1960), Exo- cella is the most likely species on Scots teleia ‘‘pinifoliella,’’ E. dodecella, and or Mugo pine. Coleotechnites ardas (Freeman 1960) Morphological characters used to (Gelechiidae) are the only North Amer- separate pine-needle miners in Ontario ican pine leaf-miners to retain frass in were given by Lindquist (1963) and their mines. Coleotechnites ardas, Lindquist and Trinnell (1967). Both ‘‘E. known only from Montana, has a silk nepheos’’ and ‘‘E. pinifoliella’’ have the ramp near a large exit hole on the anal crochets reduced in number and middle of the mine. This differs from divided into two groups. The anal the above two species of Exoteleia, proleg of E. dodecella has a single which have an exit hole at the base of row of 8–14 crochets (Lindquist and the mine without a silken ramp (Free- Trinnell 1967). In addition, Lindquist man 1960: fig. 64). Exoteleia ‘‘pinifo- (1963) found that E. ‘‘pinifoliella’’ has a liella’’ pupates in the mine, whereas E. unisetose SV group on A1. This is dodecella pupates in the bud. Interest- unusual because E. dodecella and E. ingly, the North American population of ‘‘nepheos’’ (Lindquist and Trinnell E. dodecella produces only a single 1967), E. ‘‘burkei’’ from the western 196 PROCEEDINGS OF THE ENTOMOLOGICAL SOCIETY OF WASHINGTON

U.S. (Burdick and Powell 1960), and plate, and large pinaculum on anal most Gelechiidae (Hodges 1998) have proleg of A10 dark brown. the SV group of A1 bisetose. Head (Figs. 13–16, 21): Hypogna- Exoteleia dodecella can sometimes thous; adfrontal area extends to epicra- be confused with two unrelated species, nial notch, frontal area extends about 3/ Rhyacionia bouliana () and 4 that distance; integument shallowly Recourvaria resinosae (Freeman 1960) wrinkled; AF2 slightly above apex of (Gelechiidae). Exoteleia dodecella can frons, about equal in length to F1; AF1 be separated from R. bouliana by shorter than and closer to F1 than to examination of the cuticular texture. AF2; C2 about 1/4–1/3 longer than C1; Exoteleia dodecella has a smooth tex- P1 in line with A1, about twice as long ture, whereas R. bouliana is covered as P2; P2 slightly posterodorsal of P1; with course microspinules (Martin L1 dorsal to stemma 2 and dorsolateral 1959). In addition, larvae of Tortricidae to A3; A3 and A1 about equal in length, and Gelechiidae can be separated by the about twice the length of A2; A2 chaetotaxy of A9 (Stehr 1987). As a slightly lateral to line connecting P1 leaf-miner, E. dodecella and R. resin- and A1; six stemmata in an irregular C- osae can be confused because both lack shaped pattern, with stemma 3 and 4 an anal comb. Exoteleia dodecella has approximate, and stemma 6 slightly 8–14 crochets on the anal proleg, and below stemma 4; S-group setae in an the SV setae of A3 – A6 are on separate arclike pattern below stemmata; S3 posteroventral to stemma 6; S2 posteri- pinacula (Lindquist 1963). These con- or to stemma 1; S1ventroposterior to trast with R. resinosae, in which there stemma 3; SS1 beneath and between are 16 crochets on the anal proleg, and base of antenna and condyle of mandi- the SV setae of A3 – A6 are on a single ble; SS2 ventral to and between stemma pinaculum. In addition, R. resinosae 5 and 6; SS3 in near vertical line with feeds on P. resinosa and occasionally on antenna and ventral to SS2; SS4 P. banksiana (Lindquist 1963), two ventroposterior to SS3; labrum with hosts rarely colonized by E. dodecella. six pairs of setae, two equal median As pointed out by Lindquist (1963), a pairs, two subequal frontomarginal large number of taxa are leaf-miners pairs, and two subequal lateromarginal only in the early instars. Our diagnosis pairs; hypopharyngeal complex with is for the most common associates of E. broad spinneret, gradually widening dodecella that are leaf miners through- from base, longer than labial palpus; out their entire life. Identification of proximomedial region spinulate; sensil- early instar pine leaf miners is more la of antenna as figured (Fig. 15); complicated. sensilla of maxillary palpus as figured Redescription.—Larva (Figs. 13–22). (Fig. 16); mandible with four teeth, Length 5.5–9.6 mm (n ¼ 10 preserved middle two larger than outer two, two larvae). Body pale gray, with dense subequal mandibular setae present covering of microspinnules; most pina- above condyle (Fig. 21). cula less than twice diameter of setal Thorax (Fig. 19): T1 with L1 2.0– socket of A1 or absent, pinacula on A8 2.53 longer than L2 and L3; L1 in – 10 slightly darker than pinacula on horizontal line or ventral to L3, both other segments of body, and pinacula on setae beneath spiracle, L2 equal in A8 – 10 greater than twice diameter of length or slightly shorter than L3, in setal sockets on A1; head capsule, horizontal line to L1, or slightly ante- prothoracic shield, thoracic legs, anal rodorsal to L1 at level of center of VOLUME 112, NUMBER 2 197 spiracle; SV-group bisetose, SV1 about as in A1 – 3 except A7 with SV group 2.53.03 longer than SV2; prothoracic bisetose, approximate, near parallel shield with SD1 slightly posterior to with median longitudinal axis; crochets and about 1 1/2 longer than XD1 and of A3 – 6 in uniordinal mesal pennel- XD2; XD2 about 2.03 farther from lipse with small gap or circle closed by XD1 than SD1; SD2 slightly anterior to a few small crochets. A8 with SV group D2 and D1, about same length as D1, unisetose; D1 and SD1 each on slightly both setae slightly shorter than XD1 and widened pinaculum, L1 and L2 usually XD2; D1 in line with XD1; D2 about on same, slightly widened pinaculum; same length as SD1, equidistant to XD1 L3, SV1, and V1 almost aligned and XD2, slightly posterior to D1. T2 - vertically. A9 with all setae approxi- 3 (Fig. 19): D2 about twice length of mating a straight vertical line; D2, D1, D1, both slightly posterior to SD-group the hairlike SD1, L1, and SV1 about setae; SD1 about twice length of SD2; equal in length, each on slightly wid- L1 about same length as D2 and SD1, ened pinaculum, about twice length of about 3.0–4.03 longer than L2, both L2, L3, and V1. A10 (Figs. 18, 22): anal setae slightly anterior to SD setae; L3 plate with SD1 slightly longer than SD2 slightly longer than L2, posterodorsal to and D2, D1 slightly shorter than SD1 L1; SV1 in vertical line with or slightly and D2; anal comb absent; anal prolegs posterior to L3; MD, MSD, and MV bearing 8–14 crochets in uninterrupted setae along anterior margin of seg- arc. ments; MD1 in line with or slightly Specimens examined: 28 Specimens ventral to D2; MSD1, in line with or from Howland, Penobscot County, slightly dorsal to SD2, MSD2 antero- Maine from P. sylvestris by R. Tracy ventral to MSD1; MV1 beneath L setae; (USNM Coll.). V1 setae on T3 slightly farther apart Pupal diagnosis and remarks.—The than V1 setae on T2, each pair at least pupa of E. dodecella was described or twice as far apart as V1 setae on T1; illustrated by Martin (1959), Patocˇka MV3 (not shown) farther apart than (1987), Go´mez de Aizprua (2003), and V1setae; MV2 absent. Patocˇka and Turcˇa´ni (2005). It can be Abdomen: A1 – A2 (Figs. 17–18, 20, distinguished from native species of 22): D1 about 2.0–2.53 length of D2; Exoteleia by having a vertex that lacks a SD1 about as long as D1, in vertical line cutting plate, a body that is widest with or slightly posterior to D2 on A1, medially and not parallel-sided, a single directly above or posterodorsal to pair of proleg scars on A6, and a spiracle on A2; SD2 minute, anterodor- notched terminal abdominal segment. sal to spiracle (need high magnification The pupation site, in the bud of the host, to see); L1 about 2.0–2.53 length of L2, is unusual, however, E. nepheos is both setae approximate, L2 in vertical rarely known to pupate within buds of line with or slightly posterior to spira- its hosts. cle; L3 slightly anterior of D1 and Compared to the few North Ameri- posterior of SV group; SV group can genera studied by Mosher (1916), bisetose on A1 (in transverse line), E. dodecella is most similar to Coleo- trisetose (in triangular pattern) on A2; technites (listed in her key as Recurva- MD1 and MV1 as above; V1 setae ria group B). Both genera lack modified equidistant to or slightly closer than abdominal setose knobs, dense body V1setae on T2 – 3; MV3 (not shown) setae, a fringe of setae around the farther apart than V1 setae, MV2 posterior margin of A7, and stout spines absent. A3 – 10 (Figs. 17–18, 22): setae on the end of the abdomen. Exoteleia 198 PROCEEDINGS OF THE ENTOMOLOGICAL SOCIETY OF WASHINGTON and Coleotechnites are similar in that scattered and apically hooked setae the antennae reach the caudal margin of (Figs. 24, 26) on segments A8–A10, the wings, and apically hooked setae are whereas in E. ‘‘pinifoliella or near’’ the present on the terminal segments. apically hooked setae are restricted to a Exoteleia differs from other known small clump on A10 (similar to Bennett Holarctic gelechiids in the relative 1954b). length of the maxillae and prothoracic Bennett (1954b, 1966) described the legs. In particular, the maxillae are pupa of E. ‘‘pinifoliella’’ and E. ‘‘chill- equal to or shorter than the prothoracic cotti.’’ He illustrated E. ‘‘pinifoliella’’ legs in E. dodecella and other species of with a spined vertex called the ‘‘cutting Exoteleia (Bennett 1954b, 1966; Lind- plate.’’ The vertex of E. dodecella is quist and Trinnell 1967). This diagnosis rounded without a spine. The cylindri- of E. dodecella was used by Patocˇka cal body shape of E. ‘‘chillcotti’’ is more and Turcˇa´ni (2005) and formed the similar to E. ‘‘pinifoliella’’ than to E. basics for Lee and Brown’s (2008) dodecella. Hodges (1985) noted that it placement of E. dodecella as the sole was unclear which species of Exoteleia member of their group I in the tribe. was illustrated by Bennett (1954b, Undoubtedly, other species of Exoteleia 1966), but they cannot be of E. will be added to this group when their dodecella. pupal morphology is confirmed. Lindquist and Trinnell (1967) men- Hodges (1985) noted that pupal tioned characters to separate three morphology may help define species species of Exoteleia found on pine in of Exoteleia. We found substantial Ontario. They noted that E. dodecella differences between our specimens of has a dark reddish brown pupa with the E. dodecella and those of an Exoteleia anal end tending to be broadly notched labeled as ‘‘pinifoliella’’ from North inwardly (see Fig. 23). The pupa of E. Carolina. In agreement with the warn- ‘‘nepheos’’ is yellow brown, and the ings by Hodges (1985) that pinifoliella anal end is not notched. Larval exuvium may be a complex of three sibling will show the characteristic crochet species, our unassociated pupal speci- arrangement that readily separates these mens of E. pinifoliella are listed as two species. Details and references on Exoteleia ‘‘pinifoliella or near’’ in the mounting microlepidopteran larvae on following. microscope slides are listed in Passoa Characters that help separate E. (2008: 305–306). dodecella from related species include These examples show that pupal the shape of the body and vertex, morphology is useful in separating E. number of proleg scars, and the termi- dodecella from related species. Future nal abdominal segments. In particular, problems will remain in assigning the the body of E. dodecella is widest at the correct species name to the various middle (Fig. 24) compared to E. phenotypes of Exoteleia pupae that can ‘‘pinifoliella or near,’’ which is cylin- be recognized in collections and in the drical and more parallel-sided (similar literature, especially for the rarer spe- to Bennett 1954b). There is only one cies. pair of proleg scars in E. dodecella Redescription.—Pupa (Figs. 23–26). visible on A6 (Fig. 24); the scars on A5 Length 5.1–5.7 (n ¼ 5): Brownish are hidden by the overlying wings. The orange, smooth, slightly flattened dor- pupa of E. ‘‘pinifoliella or near’’ has two soventrally; vertex rounded, with fine exposed proleg scars on A5 and A6. In punctures almost forming longitudinal addition, A8–A10 of E. dodecella has rows; frontoclypeus bilobed, suture not VOLUME 112, NUMBER 2 199

Figs. 27–30. Larval damage of Exoteleia dodecella to Scots pine in Maine. 27, ‘‘Bunched growth’’ resulting from repeated infestations; 28, Needle mine of third instar. Arrow pointing to entrance; 29, Bud attacked by fourth instar. Arrow indicates white, pitch-impregnated silken tube at base; 30, Shoots attacked after beginning of spring growth. Arrow indicates silken tube at base. straight; labrum U-shaped, labial palpi present on A6; thoracic spiracle a small hidden or minutely exposed; maxillary tubercle; abdominal spiracles small and palpi exposed, not extending beyond circular; dorsum of abdomen anterior margin of eye; maxillae slightly shagreened; A8–10 with apically shorter than or equal to the length of hooked setae on dorsal and ventral prothoracic legs; mesothoracic legs surfaces (Figs. 23, 26). shorter than antennae; antennae meet Specimens examined: Four speci- mesially; apical parts of metathoracic mens of E. dodecella from Howland, legs exposed; pair of proleg scars Penobscot County, Maine from P. 200 PROCEEDINGS OF THE ENTOMOLOGICAL SOCIETY OF WASHINGTON

Table 1. Percent mitochondrial cytochrome c oxidase I (COI) sequence divergence among species of Exoteleia, Coleotechnites, and Recurvaria. Uncorrected average pairwise distances are shown. Cells below diagonal ¼ mean between-species distances in %. Cells above diagonal ¼ mean number of base pair changes between species. Diagonal (shaded) cells ¼ mean within-species distances in %, with mean number of base pair changes in parentheses. Species abbreviations are as follows: dod ¼ Exoteleia dodecella, ex1 ¼ ‘‘E. pinifoliella complex’’ group1, ex2 ¼ ‘‘E. pinifoliella complex’’ group2, ex3 ¼ ‘‘E. pinifoliella complex’’ group3, ex4 ¼ ‘‘E. pinifoliella complex’’ group4, atr ¼ Coleotechnites atrupictella,flo¼ Coleotechnites florae, que ¼ Coleotechnites quercivorella, nan ¼ .

sylvestris, (USNM); 12 specimens of to the fourth instar and enters a bud ‘‘Exoteleia pinifoliella’’ from Lunen- before spring growth begins (Fig. 29). burg from P. rigida needles, Massachu- The larva constructs a silken tube at the setts, (USNM); six specimens from base of the bud, which becomes im- Bent Creek Experimental Forest, North pregnated with pitch. If the bud is not Carolina, from P. resinosa needles large enough for the larva to complete (USNM). its development, it will leave the bud Biology (Figs. 27–30).—The life and enter into an elongated shoot (Fig. history of E. dodecella was studied in 30). The presence of dead buds and detail by Lemarie (1958a) in Slovakia, shoots with a tube of silk and pitch at Martin (1959) and Freeman (1960) in the base is unique for E. dodecella. Ontario (Canada), and Tracy (1980) in Other species that feed on Scots and Maine (United States). Larvae of E. Mugo pine, such as E. nepheos and R. dodecella cause extensive damage to bouliana, enter the stem after growth buds of Scots pine. Infested trees show has begun and the buds have elongated a bunched growth (Fig. 27). Severe into shoots. infestations can cause a witches’ broom Pupation occurs in the bud or shoot effect as these trees suffer repeated from late May to early June. Adults attacks. emerge from mid June to early July. The elliptical eggs are laid on the Distribution.—In North America, E. current year’s growth or sometimes on dodecella is known from the Niagara one-year-old stems. The first instar Peninsula of Ontario, Canada south into enters a needle tip in July. Second and the northeastern United States from third instar larvae continue to feed on Maine, Vermont, Connecticut, and west needles (Fig. 28) until later in the into New York and northeastern Penn- summer when the caterpillar seals the sylvania. We also have unconfirmed mine after ejecting most of the frass. reports that E. dodecella occurs in This shelter becomes the overwintering Michigan (R. Tracy pers. obs.), but the site. Feeding continues the following records could not be confirmed by spring, and in early May the larva molts voucher specimens. In the West, it is VOLUME 112, NUMBER 2 201

Fig. 31. Neighbor-joining tree based on Kimura-2-Parameter distances for 658 bp of cytochrome c oxidase I (COI) in species of Exoteleia, Coleotechnites, and Recurvaria (total ¼ 130 specimens). Numbers below branches indicate % of sequence divergence. Numbers in parentheses after species names indicate the number of specimens analyzed and the range of sequence lengths obtained. Individual species branch clusters were collapsed. also present in the Vancouver area of (Cresson) (); Telenomus British Columbia, Canada. sp. (Scelionidae). The citation for E. dodecella in China Results from DNA barcodes.—There may represent another introduction of were 523 positions in the final dataset; this species to an Old World locality 135 positions were omitted in the (Zhang 1994). China is outside the analysis due to gaps or missing data. normal range of this species (Bland et This was due primarily to four speci- al. 2002). mens in the ‘‘Exoteleia pinifoliella complex’’ with shorter barcode se- Parasitoids.—Tracy (1980) reared the quences (ca. 560–600 bp). following species of parasitoids from E. Samples of E. dodecella from the dodecella in Maine: Diptera. Phyto- United States exhibited more than 7% myptera usitata (Coquillett) (Tachini- sequence divergence from the closest dae). . Goniozus sp. (Be- samples that represented native North thylidae); Chelonus recurvariae American species of Exoteleia, indicat- McComb, Orgilus sp. (Braconidae); ing strong evidence for a distinct Elachertus sp., Sympiesis stigmatipen- species. While these findings are con- nis Girault, Peckekachertus sp. (Eulo- gruent with morphological differences phidae); Copidosoma geniculatum in the adult and immature stages, it is (Dalman) (Encyrtidae); Scambus sp., notable that this high amount of Phaeogenes sp., Exeristes comstockii divergence translates into a relatively 202 PROCEEDINGS OF THE ENTOMOLOGICAL SOCIETY OF WASHINGTON

Fig. 32. Strict consensus tree of 42 most parsimonious trees (length ¼ 224, CI ¼ 0.763, RI ¼ 0.918) based on 658 bp of cytochrome c oxidase I (COI) in species of Exoteleia, Coleotechnites, and Recurvaria (total ¼ 130 specimens). Bremer support indices are shown above branches; bootstrap support values are below branches. Branch terminal labels indicate unique specimen identifiers (Specimen IDs in BOLD) and taxonomic assignment. small amount of morphological differ- ly low level of sequence divergence ence. (,3%). Geographic representation of In contrast, samples of native North the samples was uneven with only American taxa revealed a comparative- eastern specimens and a stronger rep- VOLUME 112, NUMBER 2 203 resentation from the southeastern region geographic distribution) are either in Tennessee and Florida. Four weak insufficient or not congruent with each clusters can be detected (Table 1, Fig. other and do not support species 31) with distances ranging from 0.04– separation. Hodges’ (1985) ‘‘entities’’ 3.03%. The clusters are moderately to based on color pattern and genitalia strongly corroborated by parsimony appear incongruent with our sequence analysis (Fig. 32), which lends to data. For example, specimen support the contention based on mor- CNCLEP00026043 from Florida is a phology that there are several weakly very pale moth whose barcode clusters differentiated species. An alternative tightly with very dark specimens from interpretation would suggest that native Tennessee and Florida. All ‘‘E. pini- populations of Exoteleia in North foliella complex’’ sensu lato barcodes America represent a single variable obtained in the present study were species with different haplotypes that obtained from specimens collected at reflect its phylogeographic history light; consequently, there is no host through ancestral polymorphism, host information for these samples. All races, or some other biological pattern. reared specimens of E. pinifoliella, Available evidence is insufficient to as well as the other nominal species, make a convincing argument either had degraded DNA that is not suitable way. However, we prefer to retain the for sequencing. Consequently, addi- hypothesis of multiple species at this tional data will be necessary to time because it agrees with the tradi- achieve better taxonomic resolution tional classification (although it lacks of Exoteleia, notably preservation of clarity) and is consistent with interspe- the immature stages associated with cific barcode divergences observed in reared adults, careful host and biolog- other genera of Lepidoptera with well ical observations, and preservation of defined species (Hebert et al. 2009). material for barcode data from sam- Intra-cluster divergences are consistent- ples with known hosts and detailed ly lower than inter-cluster divergences biological data. (Table 1). DNA barcodes can reliably distin- ACKNOWLEDGMENTS guish E. dodecella from native species of Exoteleia sampled from eastern We thank the following staff, for- North America. However, the inade- merly of the Systematic Entomology quate state of current taxonomy pre- Laboratory, Smithsonian Institution cludes the unambiguous naming of the National Museum of Natural History, barcode clusters of native species, so Washington, DC, for identifications of they have been informally labeled parasitoids: R. W. Carlson (Ichneumo- ‘‘group 1’’ and ‘‘group 2.’’ For conve- nidae), E. E. Grissell (), P. nience, and to distinguish them from the M. Marsh (Braconidae), A. S. Menke E. dodecella samples, we use the ‘‘E. (Bethylidae), and C. W. Sabrosky pinifoliella complex’’ based upon the (Diptera). We are grateful to Scott oldest name. This is the name that Whittaker, Lab Manager, Scanning would apply if additional data led to the Electron Microscopy Laboratory, conclusion that they represent but a Smithsonian Institution, Washington, single and variable species. DC, for the suggestions on larval All other attributes as reported in fixation prior to SEM analysis and the literature (i.e., adult coloration, scheduling time on the SEM. Diana genitalia, larval hosts, immatures, Marques, Lisboa, Portugal, constructed 204 PROCEEDINGS OF THE ENTOMOLOGICAL SOCIETY OF WASHINGTON the fine computer generated illustra- Elias, T. S. 1989. Field Guide to North American tions of the male and female genitalia Trees. Grolier Book Clubs, Danbury. 51 pp. and produced the plates. We also thank Freeman, T. N. 1960. Needle-mining Lepidoptera of pine in North America. The Canadian J. R. deWaard, D. Handfield, P. D. N. Entomologist, Supplement 16, 51 pp. Hebert, and D. Holden for supplying ———. 1963. Two new species of coniferous some barcoded specimens. Support for needle miners from Louisiana and the DNA barcoding was provided by the description of a new (Lepidoptera: Canadian Barcode of Life Network Gelechiidae). The Canadian Entomologist from Genome Canada through the 95: 727–730. Ontario Genomics Institute, NSERC ———. 1967. A new Nearctic species of Exoteleia Wallengren (Gelechiidae) on Pine. (to JFL), and other sponsors listed at Journal of the Lepidopterists’ Society 21: www.BOLDNET.ca. 9–11. Germplasm Resources Information Network LITERATURE CITED (GRIN). 2008. United States Department of Agriculture, Agricultural Research Service, Bennett, W. H. 1954a. The effect of needle Beltsville Area. Website: http://www. structure upon the susceptibility of hosts to ars-grin.gov/cgi-bin/npgs/html/taxon. the pine need miner Exoteleia pinifoliella pl?401686. Accessed 2 February 2008. (Chamb.) (Lepidoptera: Gelechiidae). The Go´mez de Aizpu´rua, C. 2003. Orugas y Mar- Canadian Entomologist 86: 49–54. iposas de Europa. I. Ministerio de Medio ———. 1954b. The pupal morphology of the Ambiente. 352 pp. pine needle miner. Proceedings of the Hain, F. P. and W. E. Wallner. 1973. The life Entomological Society of Washington history, biology, and parasites of the pine 56(1): 41–42. candle moth, Exoteleia nepheos (Lepidop- ———. 1966. Pupal morphology of Exoteleia tera: Gelechiidae), on Scotch pine in Mich- chillcotti Freeman. Proceedings of the Ento- igan. The Canadian Entomologist 105: mological Society of Washington 68(3): 157–164. 181–183. Hajibabaei, M., J. R. deWaard, N. V. Ivanova, S. Bland, K. P., M. F. V. Corley, A. M. Emmet, R. J. Ratnasingham, R. Dooh, S. L. Kirk, P. M. Heckford, P. Huemer, J. R. Langmaid, S. M. Mackie, and P. D. N. Hebert. 2005. Critical Palmer, M. S. Parsons, L. M. Pitkin, T. factors for the high volume assembly of Rutten, K. Sattler, A. N. B. Simpson, and P. DNA barcodes. Philosophical Transactions: H. Sterling. 2002. Gelechiidae. The Biological Sciences 360: 1959–1967. and Butterflies of Great Britain and Ireland. Handfield, L. 1992. Liste des Le´pidopte`res du Vol. 4 (part 2). Harley Books, England. 277 Que´bec et du Labrador. Fabreries. Supple- pp. ment 7, 155 pp. Braun, E. L. 1961. The Woody Plants of Ohio. Hebert, P. D. N., E. H. Penton, J. M. Burns, D. H. Ohio State University Press, Columbus. 362 Janzen, and W. Hallwachs. 2004. Ten species pp. in one: DNA barcoding reveals cryptic Burdick, D. J. and J. A. Powell. 1960. Studies on species in the Neotropical skipper butterfly the early stages of two California moths Astraptes fulgerator. Proceedings of the which feed in the staminate cones of digger National Academy of Sciences of USA pine (Lepidoptera: Gelechiidae). The Cana- 101: 14812–14817. dian Entomologist 92: 310–320. Hebert, P. D. N., J. R. deWaard, and J.-F. Landry. Clarke, J. F. G. 1941. The preparation of slides of 2009. DNA barcodes for 1/1000 of the the genitalia of Lepidoptera. Bulletin of the kingdom. Biology Letters doi: 10. Brooklyn Entomological Society 36: 1098/rsbl.2009.0848. 149–161. Hodges, R. W. 1983. Gelechiidae, pp. 19–25. In deWaard, J. R., N. V. Ivanova, M. Hajibabaei, R.W. Hodges, ed. Check List of the Lepi- and P. D. N. Hebert. 2008. Assembling DNA doptera of America North of Mexico. E. W. Barcodes: Analytical Protocols, pp. 275–293. Classey Ltd., Faringdon, Oxfordshire, Eng- In C. Martin, ed. Methods in Molecular land, and The Wedge Entomological Re- Biology: Environmental Genetics. Humana search Foundation, Washington, DC, USA, Press Inc., Totowa. xxiv þ 284 pp. VOLUME 112, NUMBER 2 205

———. 1985. A new species of Exoteleia knowledge of the parasites of Exoteleia (Gelechiidae) reared from Ponderosa Pine. dodecella L.]. Beitra¨ge Zur Entomologie 9: Journal of the Lepidopterists’ Society 39: 805–809. 139–144. ———. 1959b. Prˇ´ıspeˇvek k pozna´nı´ cizopasnı´ku˚ ———. 1986. , Gelechiidae (part). makadlovky borove´ Exoteleia (Heringia) Moths of America North of Mexico. E.W. dodecella L. Cˇ a´st II. Celedi Ichneumonidae, Classey Ltd., and the Wedge Entomological Braconidae, Larvaevoridae. [Contribution to Research Foundation. Fascicle 7.1, 195 pp. the knowledge of parasites of the pine-bud ———. 1998. Gelechioidea, pp. 131–158. In N. wandering moth Exoteleia (Heringia) dode- P. Kristensen, ed. Handbuch der Zoologie, cella L. 2. Ichneumonidae, Braconidae, and Lepidoptera, Part 1, Volume 35, Walter de Larvaevoridae.]. Zoologicke Listy 22: Gruyter & Co, Berlin, New York. 495 pp. 309–314. Huemer, P. and O. Karsholt. 1999. Microlepi- ———. 1961. Prˇ´ıspeˇvek k pozna´nı´ cizopasnı´ku˚ doptera of Europe. Vol. 3, Gelechiidae. makadlovky borove´ Exoteleia (Heringia) Apollo Books, Stenstrup. 356 pp. dodecella L. Cˇ a´st IV. Ichneumonidae, Bra- Ivanova, N. V., J. R. de Waard, and P. D. N. conidae, Chalcidoidea, Bethyloidea. [Contri- Hebert. 2006. An inexpensive, automation- bution to knowledge of the parasites of the friendly protocol for recovering high quality pine bud-moth Exoteleia (Heringia) dode- DNA. Molecular Ecology Notes. doi: 10.111/ cella L. Part 4. Ichneumonidae, Braconidae, j.1471–8286.2006.01471.x. Chalcidoidea, Bethyloidea.]. Zoologicke Kim, K. C. and A. G. Wheeler, Jr. 1991. Listy 24: 119–126. Pathways and Consequences of the Introduc- Lindquist, O. H. 1963. Larvae of pine needle tion of Non-indigenous and Arach- miners in Ontario. The Canadian Entomolo- nids in the United States. Report to the U.S. gist 95(5): 517–521. Congress Office of Technology Assessment. Lindquist, O. H. and J. R. Trinnell. 1967. Biology 74 pp. and description of immature stages of Knutson, L., R. I. Sailer, W. L. Murphy, R. W. Exoteleia nepheos (Gelechiidae) on pine in Carlson, and J. R. Dogger. 1990. Computer- Ontario. Journal of the Lepidopterists’ Soci- ized data base on immigrant . ety 21(1): 15–21. Annals of the Entomological Society of Maddison, W. P. and D. R. Maddison. 2002. America 83(1): 1–8. MacClade 4.05 OSX [computer program]. Kornerup, A. and J. H. Wanscher. 1978. Methuen Sinauer Associates, Sunderland, Massachu- Handbook of Colour. 2nd ed., Methuen and setts. Co., Ltd., London. 243 pp. Martin, J. L. 1959. The bionomics of the pine bud Lee, S. and R. L. Brown. 2008. Revision of moth, Exoteleia dodecella L. (Lepidoptera: Holarctic Teleiodini (Lepidoptera: Gelechii- Gelechiidae), in Ontario. The Canadian dae). Zootaxa 1818: 1–55. Entomologist 91: 5–14. Lemarie, J. 1958a. Druhy´ prˇ´ıspeˇvek k pozna´nı´ Mattson, W. J., P. Niemela, I. Millers, and Y. bionomie makadlovky borove´ Exoteleia Inguanzo. 1994. Immigrant phytophagous (Heringia) dodecella L. [Second investiga- insects on woody plants in the United States tion on the bionomics of the pine bud moth, and Canada: an annotated list. United States Exoteleia dodecella L.] Zoologicke Listy 7: Department of Agriculture, Forest Service, 208–220. North Central Forest Experiment Station ———. 1958b. Prˇ´ıspeˇvek k pozna´nı´ cizopasnı´ku˚ General Technical Report NC-169. 27 pp. makadlovky borove´ Exoteleia (Heringia) Mosher, E. 1916. A classification of the Lepi- dodecella L. Cˇ a´st I. Nadcˇeled’ Chalcidky, doptera based on characters of the pupa. Chalcidoidea; Vejrˇitky, Proctotrupoidea; Bulletin of the Illinois State Laboratory of Hbiteˇnky, Bethyloidea. [Contribution to Natural History 12: 17–159, pls. 19–27. knowledge of the parasites of the pine bud- National Agricultural Pest Information System moth Exoteleia (Heringia) dodecella L. Part (NAPIS) Pest Tracker. http://pest.ceris. I. Chalcidoidea, Proctotrupoidea, Bethyloi- perdue.edu/searchnews.php. Accessed Janu- dea.] Zoologicke Listy 21: 221–230. ary 2009. ———. 1959a. Erster Beitrag zur Kenntnis der Passoa, S. 2008. Part III. Immature Stages, pp. Parasiten von Exoteleia dodecella L. (Hy- 295–314. In T. M. Gilligan, D. J. Wright, and menoptera: Chalcidoidea; Proctotrupoidea; L. D. Gibson, eds. Olethreutine Moths of the Bethyloidea). [First contribution to the Midwestern United States, An Identification 206 PROCEEDINGS OF THE ENTOMOLOGICAL SOCIETY OF WASHINGTON

Guide. Ohio Biological Survey Bulletin, New Analysis Using Parsimony (*and other meth- Series, Vol. XVI, No. 2, The Ohio Biological ods). Version 4.0b.10. Sinauer Associates, Survey, Columbus. vii þ 334 pp. Sunderland, Massachusetts. Patocˇka, J. 1987. Uber die Puppen der Mitteleur- Tamura, K., J. Dudley, M. Nei, and S. Kumar. opaischen Gelechiide (Lepidoptera). 3. Teil 2007. MEGA4: Molecular Evolutionary Ge- Tribus Teleiodini und benachbarte Gattun- netics Analysis (MEGA) software version gen. Vestnik-Ceskoslovenske-Spolecnosti- 4.0. Molecular Biology and Evolution Zoologicke 51(4): 286–299. 10.1093/molbev/msm092. Patocˇka, J. and M. Turcˇa´ni. 2005. Lepidoptera Tracy, R. A. 1980. Distribution, abundance, Pupae. Central European Species. Apollo damage and parasites of Exoteleia dodecella Books, Stentrup. 863 pp. L., Dioryctria sp. and Petrova albicapitana Piskunov, V. I. 1989. Gelechiidae, pp. 889–1,024. (Busck) on pines in Maine. Masters Thesis. In G. S. Medvedev, ed. Keys to the Insects of Department of Entomology, University of the European Part of the USSR. Lepidoptera. Maine at Orono, Orono, Maine. vii þ 67 pp. Vol. 4, part 2. Oxonian Press Ltd., New United States Department of Agriculture Re- Delhi. 1,092 pp. [English translation]. search Service, Biosystematics and Benefi- Ratnasingham, S. and P. D. N. Hebert. 2007. BOLD: The Barcode of Life Data System cial Insects Institute. 1986. Western Hemi- (http://www.barcodinglife.org). Molecular sphere Immigrant Arthropod Database Ecology Notes 7: 355–364. (WHIAD). Unpublished printout. Doc Sheppard, R. W. 1930. Notes on the occurrence 0402M. 95 pp. of the pine bud moth, Exoteleia dodecella L. White, J. H. and R. C. Hosie. 1980. The Forest in Welland County, Ontario. Annual Report Trees of Ontario. Ontario Ministry of Natural th of the Entomological Society of Ontario 61: Resources, Toronto, Canada 7 edition. 114 79–81. pp. Stehr, F. W. 1987. Immature Insects. Kendall/ Zhang, B. C. 1994. Index of Economically Hunt Pub. Co., Dubuque. xiv þ 754 pp. Important Lepidoptera. CAB International, Swofford, D. L. 2003. PAUP*: Phylogenetic United Kingdom. 599 pp.