53 dispar sex pheromone is a behavioral antagonist to pheromonal attraction of male Lymantria mathura

Regine Gries Department of Biological Sciences, Simon Fraser University. 5888 University Drive, Burnaby. British Columbia, Canada V5A I S6 Paul W. Schaefer United States Department of Agriculture. Agricultural Research Service, Beneficial Introduction Research. Nessark. Delaware 19713, United States of America Katsunori Nakamura Forestry and Forest Products Research Institute, Tohoku Research Center, Naheyashiki 92-25, Sliimo-Kurivagawa. Morioka Iwate 020-0123, Japan Gerhard Gries1 Department of Biological Sciences, Simon Fraser University. 8888 University Drive, Burnaby. British Columbia, Canada V5A I S6

Abstract—In a trapping study conducted in the experimental research forest of the Tohoku Re- search Center, Morioka. Honshu, Japan, we investigated the effect ol heterospecilic pheromone on pheromonal attraction of male Japanese gypsy . Lvmaniria dispar japonica (Motsehuisky), and male pink gypsy moth. L inathora Moore (: Noctuidae: ). Traps baited with synthetic pheromone of L. d. japonica 0 7R.8S)-ci.-7.8-epoxy-2methvloctadecane = +)-disparlure (100 p g)) or L. ,nath,,ra ( ( 9R. I OS .3Z.6Z)-cLs-9. I 0-epoxynonadecadiene = (+)-mathuralLlre (20 ftC) and (95. I OR,3Z.6Z)-cis-9, I 0-epoxynonadecadiene = (-)-niathuralure (80 ftg)) attracted male L. d. japonna or L. inal/olm. respectively. Traps baited with synthetic pheromone of both species captured significantly fewer male L. /oat/niro than traps baited solely with syn- thetic L. ,nathu,a pheromone. Numbers of male L. d. japonica captured in traps baited with (+)-disparlure were unaffected by the addition of L. ,,iathioa pheromone. (+)-Disparlure is it behav- ioral antagonist to pheromonal attraction of male L. ma/hum, whereas male L. d. japan/ca are in- ditlerent to the presence of synthetic L. ,naihu,-a pheromone.

Résumé—Dans uric étude de piégeage réalisée dans la forét expérinientale de recherche du Centre de recherche de Tohoku (Morioka, Honshu. Japon). nous avons examine Les effets de hi pheromone hCtCrospCciOque sur I attraction phéromonale des males de la spongicuse japonaise. L1-mantra th.spar japonica (Motschulsky) et de Ia spongieuse rose, L. maihura Moore (Lepidop- tera: Noctuidae: Lymantriinae). Les piCges garnis de pheromone synthCtique de L. d. japonica (( 7R.8S)-cis-7.8-Cpoxy-2-méthyloctadCcane = (+)-disparlure (100 pg)) on de L. mathuta ((9R.IOS,3Z,6Z)-d.s-9,10CpoxynonadécadiCne = (+)-rnathuralure (20 pg) et (95, /OR,3Z,6Z)-ci.r- 9. lO-époxynonadCeadiène = (-)-mathuralure (80 ftg)) attirent respective ment les males dc L. d. japonica et ceux de L. ,oarhora. Les pièges garnis des pheromones synthCtiques des deux espèces c:ipturent signilicativement moms (IC males de L. mnathura Clue les pièges garnis seulement de la pheromone synthCtique (IC L. mnaihora. Les captures de L. il. japonica dans Ics piCges munis de (+)-disparlure no sont pas atiectCes par laddition de la pheromone de L. ,nathi,ra. Le (+)-dispar- lure est un antagoniste comportemental de lattraction phéromonale des mhles de L. niathura, alors Clue les males de L. d. japonica restent inditlCrents i la presence de Ia pheromone synthC- tiqtie de L. ,nathura. [Traduit par ]a Redaction]

Received 27 September 2008. Accepted 25 November 2008. Corresponding author (e-mail: [email protected]). doi: I 0.4039/n08-069

Can. Entomol. 141: 3-55 (2009) 1) 2009 Entomological Society of Canada

54 Can. Entomol. Vol. 141, 2009 captured between 2 and Fig. 1. Numbers (mean + SE) of male Lvinantrw thsparjciponica and male L. inciihura 4 August 2008 in the experimental forest of the Tohoku Research Center. Morioka. Honshu. Japan. in delta- shaped traps hailed with synthetic pheromone of L. ci. Japonica ((7R.8S)-cis-7,8-epox y 2- methyloctadecane = (+)-disparlure), L. ,nathura ((91?, I 0S.3Z.6L)-cis-9. I 0-epoxynunadecadiene = (+)- (n = 6 replicates). A mathuralure plus (9S. I OR.3464-cis-9 .1 0-expoxynonadecadiene = (-)-mathuralure). or both different letter above the bar indicates a statistical difference (I < 0.05). -o25 a -I .20-I Ct, I 15-I L. mathura

L. d. japonica 4- I be o 5-I C5 Z 0 d 1 d 1 I 100 100 - (+)-Disparlure 20 - 20 - (+)-Mathuralure 80 - 80 - (-)-Mathuralure Test stimulus (Jg)

The Japanese gypsy moth. Lvmantria ciispai expoxynonadecadiene (= (-)-mathuralure) in a japonica (Motschulsky). and the pink gypsy 1:4 ratio (Gries ci al. 1999). moth, L. mat/inca Moore (Lepidoptera: Noctu- The trapping study was conducted in the ex- idae: Lymantriinae), occur in sympatry in Japan. perimental research forest of the Tohoku Re- feed on the same or similar host plants, and search Center. Morioka, Honshu. Japan (3945N. pose comparable risks of inadvertent introduc- 141 005E). At 15-20 m intervals, delta-shaped tion into North America (Pogue and Schaefer traps (Gray ci al. 1984) coated with adhesive 2007). Pheromone-based surveys will need to Tanglefoot (Tanglefoot Company, Grand Rapids. be deployed at ports of entry into Canada and Michigan) were suspended from deciduous trees the United States of America to ensure the ear- 1.7-2.0 m above ground in randomized com- plete blocks. In each of six blocks (replicates), liest possible detection of both species. Time and money would be saved if traps could be traps were baited with a grey sleeve stopper (West Pharmaceutical Services, Lionville. Penn- baited with lures emitting the pheromone of sylvania) impregnated with test chemicals in both species instead of one. Our objective was solution with high-performance liquid chroma- to determine whether such a two-species phero- tography-grade hexane. In each block, four test mone lure would have adverse effects on attrac- stimuli were randomly allocated among traps: L. mcithura. tion of male L. d. japonica or (1)(+ )-disparlure (IOU pg): (2) (+)-mathuralure The single-component sex pheromone of (20 .ig) plus (-)-mathuralure (80 pg): (3) 1 plus the European gypsy moth. L. ci. dispar (L.). and (4) a solvent control. (+)-Disparlure was is (7R.8S)_cis_7.8_epoxy_2_methylOctadeCafle purchased from Dr. Roger Hahn (Department of (= (+)-disparlure) (Bierl ci al. 1970; Klirnetzek Chemistry. Syracuse University, Syracuse, New ci 0/. 1976: Cardé ci cii. 1977: Miller ci al. York) and both (+)- and (-)-mathuralure were 1977; Plimmer ci al. 1977). Pheromone analy- available from previous research (Gries et cii. ses of the Asian subspecies L. ci. a,siatica 1999). The enantiomers of mathuralure were pu- Vnukovskij and L. ci. japonica revealed addi- rified by high-performance liquid chromatogra- tional candidate pheromone components. but none phy using equipment and procedures described appeared to enhance the attractiveness of (+)- in Gries ci cil. (2005). All test chemicals were disparlure (Ciries ci al. 2005; G.G. and P.W.S., >96% chemically pure. and >99% ((+)-disparlure) unpublished data). Male L. mat/inca are attracted and >88% ((+)- and (-)-mathuralurn) enantionieically to the female-pRxluced pheromone blend consisting pure. Enantiomeric purity was taken into ac- of (91?. 1 OS.3Z.6Z)-cis-9, 1 0-expoxynonadecadiene count in the preparation of the mathuralure blend. The experiment started on 2 August 2008 (= (+)-mathuralure) and (9S. I OR.3Z.6Z)-cis-9. I 0-

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Gries et al 55

and terminated 3 days later. Trap-catch data G.G., with Pherotech International Inc.. S.C. were subjected to transformation (log(x + I)) to Johnson Canada. and Global Forest Science as ensure normal distribution and homogeneity of industrial sponsors. variance and analyzed by two-way ANOVA ( F 0530 = 6.23) followed by a least significant Bier], BA.. Bcroza. M.. and Collier. C.W. 1970. Po- means Students I test (Zar 1984, SAS Institute tent sex attractant of the gypsy moth: its isolation, Inc. 1988). The zero trap-catch data obtained identification. and synthesis. Science (Washing- from nonbaited control traps were not included (on. D.C.). 170: 87-89. in the analyses because of a lack of variance as- Card, R.T., Duane, CC.. Baker, T.C., Iwaki, S., and Marumo. S. 1977. Attractancy of optically active sociated with them. pheromone for male gypsy moth. Environmental Traps baited with (+)-disparlurc or the Entomology, 6: 768-772. mathuralure blend captured 77 male L. ci. ja- Gray. T.G .. Slessor, K.N.. Shepherd, R.F., Grant. G.G.. ponica and 13 1 male L. niathum, respectively and Manville I.F. 1984. European pine shoot moth. (Fig. I ). (+)-Disparlure was a significant behav- Rhvacionia him/lana (Lepidoptera: Tortricidae): ioral antagonist to pheromonal attraction of identification of additional pheromone components male L. mathura (F 30 = 34.5770), whereas at- resulting in an improved lure. The Canadian Ento- mologist, 116: 1525-1532. traction of male L. d. japonica to (+)-disparlure Gries, CL, Goes, R.. Schaefer, P.W., Gotoh. T.. and was not affected by the presence of the rnathui- Higashiura. Y. 1999. Sex pheromone components alure blend (Fig. I). Unhaited control traps did Of pink gypsy moth, Lvmanuria ,uathura. Natur- not capture a single male moth. wisscnschaftcn. 86: 235-238. The antagonistic effect of (+)-disparlure on Grics, R., Khaskin. G., Schaefer, P.W., IIahn, R., pheromonal attraction of male L. inathura is Gotoh, T.. and Gries. G. 2005. (7R,85)-cis7,8- surprising because the sexual-communication Epoxy-2-rnethyloctadcc- I 7-enc: a novel trace component from the sex pheromone gland of periods of the two species are well separated. gypsy moth. Lv,iiwitria dispar. Journal of Chcnii- Lvman!ria d. japonica engages in pheronion al n cal Ecology. 31: 49-62. commu ication throughout the day, whereas Klimetzek, D.. Loskant, G., Viti.L J.P., and Mon. K. L. maihura "communicates around midnight 1976. Disparlure: differences in pheromone per- (unpublished data). The data suggest that the ception between gypsy and nun moth. Natur- sexual-com niunication periods of L. d. japonica wissenschaften, 65.581-582. and L. inathura might have overlapped for some Miller, JR., Mon, K.. Roelofs. W.L. 1977. Gypsy time during their evolution. Alternatively, there moth held trapping and clectroantennogram stud- is another, as yet unknown sympatric moth spe- ies with phermune enantiomers. Journal of Physiology, 23: 1447-1454. cies that uses (+)-disparlure as a communica- Plimmer, J.R.. Schwalhe, C.R. Paszek. E.C.. Bierl, tion signal and signals at the same time BA., Webb, RE., Marumo, S.. and twaki, S. as L. mat/inca. 1977. Contrasting effects of (+)- and (—)-enantio- Based on the results of our investigation, our mers of disparlure for trapping native populations recommendation is that single-species phero- of the g ypsy moth in Massachusetts. Environmen- nione lures in separate traps be deployed in tal Entomology, 6: 518-522. trapping programs for detecting L. d. japonica Pogue. MG.. and Schaefer. P.W. 2007. A review of selected species of Lvinantria Hiibner II 8 191 and L. niathura at ports of entry. (Lepidoptera: Noeluidue: Lymantriinac) from sub- tropical and temperate regions of Asia including We thank Tadao Gotoh for logistic support. the description of three new species, some poten- Mike Cheng for word processing. Bob Birtch tially invasive to North America. Publication for the graphic illustration, Stephen TakiIcs for FHTET-2006-2007, United States Department of statistical analysis. Eberhard Kiehlmann for proof- Agriculture Forest Service. Forest Health Tech- nology Enterprise Tcarn, Fort Collins. Colorado. reading the manuscript, and two anonymous re- SAS Instiute Inc. 1988. SAS/STAIIM users guide. viewers for constructive comments. The research Release 6.03 ed. SAS institute Inc.. Cary, North was financially supported by a contract from Carolina. the Canadian Food Inspection Agency and by a Zar. J.H. 1984. Biostatistical analysis. Prentice—Hall. Natural Sciences and Engineering Research Englewood Cliffs, New Jersey. Council of Canada Industrial Research Chair to

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