Hydrocarbons with a Homoconjugated Polyene System and Their Monoepoxy Derivatives: Sex Attractants of Geometrid and Noctuid Moths Distributed in Japan

Journal of Chemical Ecology, Vol. 19, No. 4, 1993

HYDROCARBONS WITH A HOMOCONJUGATED POLYENE SYSTEM AND THEIR MONOEPOXY DERIVATIVES: SEX ATTRACTANTS OF GEOMETRID AND NOCTUID MOTHS DISTRIBUTED IN JAPAN

TETSU ANDO, l'* HIROKAZU OHSAWA, 1'3 TADAHIRO UENO, 1 HIDEKI KISHI, 1 YUTAKA OKAMURA, l and SATOSHI HASHIMOTO 2

1Department of Applied Biological Science, Faculty of Agriculture Tokyo University of Agriculture and Technology Fuchu, Tokyo 183, Japan 2Department of Zoology Natural History Museum and Institute, Chiba 955-2 Aoba-cho, Chuo-ku, Chiba 260, Japan

(Received August 31, 1992; accepted December 1, 1992)

Abstract--Although several sex pheromones of the family Geometridae have been characterized, investigations on Japanese species are limited. In order to obtain more information, screening using known sex pheromones and their analogs was carried out. The (Z,Z,Z)-3,6,9-triunsaturated and (Z,Z)-6,9-diun- saturated hydrocarbons with straight C~9-C~ chains were synthesized by the Grignard coupling reaction as a key step starting from linolenic and linoleic acids, respectively. Oxidation of the homoconjugated trienes with m-chloroperoxybenzoic acid yielded a 1 : 1 : 1 mixture of three monoepoxy derivatives that could be separated by silica gel chromatography. The chemical structure of each positional isomer was confirmed using two-dimensional NMR techniques and MS measurements, which enabled characteristic fragment ions from the isomers to be identified. Field tests using lures incorporating only one of the above six hydrocarbons or nine epoxides were carried out in a forest in Tokyo. Consequently, attraction of male moths of 14 geometrid species in addition to four species in another family, the Noctuidae, was observed. It was concluded that hydrocarbons with a homoconjugated polyene system and the monoepoxy derivatives are important components of sex pheromones produced by Japanese lepidopterous insects, particularly the geometrid moths.

*To whom correspondence should be addressed. 3present address: Tsukuba Research Institute, Banyu Pharmaceutical Co., Ltd., Okubo 3, Tsukuba 300-33, Japan

787

0098-0331/93/0400-0787507.00/0 • 1993 Plenum Publishing Corporation 788 ANDO ET AL.

Key Words--Sex pheromone, lepidopterous attractant, field test, unsaturated hydrocarbon, epoxydiene, Geometridae, Noctuidae.

INTRODUCTION

Lepidopterous sex pheromones have been identified from females of more than 300 species since the first investigation on bombykol was carried out. Analyses of their chemical structures (Am et al., 1986; Ando, personal data base), showed that ca. 85 % of them contained linear aliphatic alcohols, aldehydes, or acetates, with a Cm-C~ 8 straight chain and one or two olefinic linkages. These types of compounds with a terminal functional group, which have been identified from 16 families of Lepidoptera, are among the most ubiquitous pheromone components. Another type of pheromone component lacking in a functional group at the terminal position was identified in the last decade, as shown in Table 1 (Ando, personal data base). To date, hydrocarbons with a homoconjugated triene or diene system and their monoepoxy derivatives have been identified from 38 species mainly in the families of Geometridae and Arctiidae. Pheromone components with the above terminal functional group, however, have not been identified from female moths in these two families. Furthermore, screening tests using these types of synthetic pheromones and the analogs revealed the attraction

TABLE 1. NUMBER OF LEPIDOPTEROUS SPECIES, SEX PHEROMONES OR ATTRACTANTS REPORTED TO BE HYDROCARBONS WITH A POLYENE SYSTEM, AND/OR THEIR MONOEPOXY DERIVATIVES

Number of species Superfamily Family Subfamily Sex pheromone Attractant

Geometroidea Geometridae Ennominae 16 20 Larentiinae 4 10 Oenochrominae 1 1 Noctuoidea Arctiidae 9 0 Ctenuchidae 2 0 Lymantriidae 1 0 Noctuidae Catocalinae 4 3 Herminiinae 0 3 Hypeninae 0 3 Ophiderinae 1 1 Total 38 41 POLYENE HYDROCARBONS 789 of 41 lepidopterous species in Canada and Europe (Millar et al., 1990b; and references therein). Although some of them are distributed in Japan, information about sexual communication utilizing the unsaturated hydrocarbons and epoxides is very limited. Therefore attempts were made to synthesize these types of compounds and to examine their field attractancy against Japanese Lepidoptera.

METHODS AND MATERIALS

Synthesis of 3,6,9-Trienes and 6,9-Dienes. Three homoconjugated trienes (Z3,Z6,Z9-19 : H, Z3,Z6,Z9-20: H, and Z3,Z6,Z9-21 : H) 4 and three dienes (Z6,Z9-19:H, Z6,Z9-20: H, and Z6,Z9-21 :H) were synthesized by a modi- fication of the method described by Conner et al. (1980) and Underhill et al. (1983). A mixture of linolenic and linoleic acids in a ratio of ca. 3:1 (25 g, 90 mmol), purchased from Tokyo Kasei Kogyo Co., Ltd. (Tokyo, Japan), was esterified with ethanol and reduced to alcohols with LiA1H4 in dry ether. After silica gel column chromatography, this alcohol mixture was treated with p-toluenesulfonyl chloride in pyridine to yield the tosylates (27 g, 72% yield from the acids). A portion of the tosylates (5.0 g, 12 mmol) was supplied for Grignard coupling with methylmagnesium bromide (16.7 ml, 15.9 mmol) [0.95 M tetrahydrofuran (THF) solution, Kanto Chemical Co., Inc., Tokyo, Japan] in dry THF (100 ml) under the mediation of Li2CuC14 (Fouquet et al., 1974) (1 ml) (0.1 M THF solution, Aldrich Chemical Co., Inc., Milwaukee, Wis- consin) to prepare a mixture of Z3,Z6,Z9-19:H, and Z6,Z9-19:H. The C~9 triene (1.5 g, 48% yield from the tosylates) and diene (0.6 g, 19% yield) were separated by a silica gel column impregnated with 20% AgNO3 using a n-hexane- benzene solvent system. The C20 and C~ analogs were synthesized in the same manner using ethylmagnesium bromide (0.93 M THF solution, Kanto Chemical Co.) and n-propylmagnesium bromide prepared from n-propyl bromide, respectively. Synthesis of Monoepoxy Derivatives. m-Chloroperoxybenzoic acid (Tokyo Kasei Kogyo Co., 70% pure, 490 mg, 2.0 mmol) was added to a solution of Z3,Z6,Z9-19 : H (470 mg, 1.8 mmol) in dry CH2C12 (30 ml) and stirred at 0~ for 1 hr. After further stirring at room temperature for 2 hr, the reaction mixture was washed with a saturated aqueous solution of NaHCO 3 (20 ml • 2) and dried with Na2SO4. Silica gel column chromatography gave a mixed product of the three cis-monoepoxy derivatives (240 mg, 48% yield) in a ratio of ca. 1 : 1 : 1. Since the epoxydiene mixture showed three separate spots on a silica gel TLC plate (60 F254, Merck, Darmstadt, Germany) [solvent system: n-hexane-benzene (1:1), Rf values: epo3,Z6,Z9-19:H, 0.34; Z3,epo6,Z9-19:H, 0.43; and

4Compounds were abbreviated as follows: Z3,Z6,Z9-19:H is (Z,Z,Z)-3,6,9-nonadecatriene, epo3,Z6,Z9-19 : H is (Z,Z)-6,9-cis-3,4-epoxynonadecadiene. 790 ANDO ET AL.

Z3,Z6,epo9-19:H, 0.40], 4 each pure sample (a racemic mixture) was obtained by preparative TLC purification. This chromatographic behavior corresponded to that of C~7 epoxydienes in a normal phase HPLC (Millar et al., 1987). In the same manner, the C20 and C2~ epoxydienes were prepared and purified. Spectroscopy. NMR spectra of each compound in CDC13 were analyzed with a JEOL GX 270 Fourier transform spectrometer (270.2 MHz for ~H and 67.9 MHz for J3C) using TMS as an internal standard. Signal assignments were made by two-dimensional techniques (1H-~H and IH-~3C COSY spectra) using ordinal pulse sequences (Ando et al., 1988). Electron-impact (EI) GC-MS was achieved using a JEOL JNM DX-300 mass spectrometer with a OV-1 capillary column (0.25 mm ID x 25 m, Gasukuro Kogyo Inc., Tokyo, Japan). Ionization voltage of every measurement was 70 eV and ion source temperature was 240~ Field Evaluation. Each chemical at 1 mg with purity >95% by GC (i.e., TIC trace from GC-MS) was applied to a white rubber septum (8 mm OD, Aldrich Chemical Co., Ltd.), which was placed in a sticky-type trap (30 • 27 cm bottom plate with a roof, Takeda Chemical Ind., Ltd., Osaka, Japan). A parallel experiment was conducted for each chemical, and the traps were set at a 1.5-m height from the ground. The screening test was carried out from August 1991 to July 1992 in a mixed forest area in the suburbs of Tokyo (Rolling Land Laboratory, Tokyo University of Agriculture and Technology, Hachiohji-shi, Tokyo), and species, sex, and number of moths caught were recorded every two weeks. Each lure was renewed every two months.

RESULTS AND DISCUSSION

Characterization of Synthetic Compounds. The ~H NMR and MS spectro- scopic data of the synthetic hydrocarbons with a homoconjugated triene and diene system corresponded well to those previously reported (Comer et al., 1980; Underhill et al., 1983). The number of C=C double bonds and the carbon length were determined by integration of their olefinic 1H signals and molecular ions, respectively. The 13C NMR spectra of the unsaturated hydrocarbons were very similar to those of the parent fatty acids. Signals from C-1 to C-15 of the hydrocarbons corresponded to the signals from C-18 to C-4 of the acids. These 13C signals were assigned based on 1H-~3C COSY experiments (see Table 2 for C2~ diene and triene). Allylic carbon signals (C-5, C-8 and C-11 of dienes, and C-2 of trienes additionally) resonated in a higher field than that calculated for the geometrical isomers (Rossi and Veracini, 1982), indicating all Z configu- rations in the hydrocarbons and the parent fatty acids. Tables 2 and 3 indicate the 1H and 13C NMR assignments for three C2j cis-epoxydienes in addition to Cz~ diene and triene. The NMR spectra of the C19 and C2o analogs were almost the same as those of the C2~ compounds except z

> Oz TABLE 2. ~H NMR ASSIGNMENTS FOR HENEICOSADIENE (Z6,Z9-21 :H), HENEICOSATRIENE (Z3,Z6,Z9-21 :H), AND EPOXYHENEICOSADIENE (epo3,Z6,Z9-21 :H, Z3,epo6,Z9-21 :H, and Z3,Z6,epo9-21 :H)

Chemical shift (ppm)

Compound H-1 H-2 H-3 H-4 H-5 H-6 H-7 H-8 H-9 H-10 H-11 H-12-H-20 H-21

Z6,Z9-21 :H 0.89 1.2-1.4 2.05 -5.35 -5.35 2.78 -5.35 -5.35 2.05 1.2-1.4 0.88 Z3,Z6,Z9-21 :H 0.98 2.08 -5.35 -5.35 2.81 -5.35 -5.35 2.81 -5.35 -5.35 2.08 1.2-1.4 0.88 epo3,Z6,Z9-21 :H 1.06 - 1.6 2.90 2.96 2.22, 2.41 -5.45 -5.5 2.81 -5.3 -5.35 2.04 1.2-1.4 0.88 Z3,epo6,Z9-21 :H 0.99 2.07 -5.5 -5.4 2.22, 2.42 2.95 2.95 2.22, 2.42 -5.4 -5.5 2.07 1.2-1.4 0.88 Z3,Z6,epo9-21 :H 0.97 2.08 -5.4 -5.35 2.81 -5.5 -5.45 2.22, 2.40 2.95 2.93 - 1.5 1.2-1.5 0.88 bo

TABLE 3. ~3C NMR ASSIGNMENTS FOR HENEICOSADIENE (Z6,Z9-21 :H), HENEICOSATRIENE (Z3,Z6,Z9-21 :H), AND EPOXYHENEICOSADIENE (epo3,Z6,Z9-21 : H, Z3,epo6,Z9-21 : H, AND Z3,Z6,epo9-21 : H)

Chemical shift (ppm)

Compound C-1 C-2 C-3 C-4 C-5 C-6 C-7 C-8 C-9 C-10 C-11 C-12-C-18 C-19 C-20 C-21

Z6,Z9-21 :H 14.08 22.6 31.6 -29.5 27.3 130.20" 127.97b 25.7 127.99I' 130.21~ 27.3 -29.5 32.0 22.7 14.13 Z3,Z6,Z9-21 :H 14.3 20.6 132.0 127.2 25.6 128.29" 128.34" 25.6 127.7 130.4 27.3 -29.5 32.0 22.7 14.1 epo3,Z6,Z9-21 :H 10.6 21.1 58.3 56.6 26.2 124.2 130.8 25.8 127.2 130.7 27.3 -29.5 31.9 22.7 14. l Z3,epo6,Z9-21 : H 14.2 20.8 134.4 123.2 26.1 56.5 56.5 26.1 123.7 132.9 27.5 -29.5 31.9 22.7 14.1 Z3,Z6,epo9-21 !H 14.3 20.6 132.2 126.7 25.7 130.8 124.2 26.3 56.4 57.2 27.8 -29.5 J 31.9 22.7 14.1 o-CChemical shift values may be reversed. dC-12 26.6 ppm.

> POLYENE HYDROCARBONS 793 for the intensity of the overlapping signals in the methylene proton region (1.2- 1.4 ppm) and methylene carbon region (ca. 29.5 ppm). Chemical structure of each epoxydiene was confirmed based on a IH-1H COSY experiment. Starting from the methyl protons at the 1 position (H-l), correlation peaks of 3,4-epoxides were traced in the following order: protons adjacent to an epoxy ring (H-2), protons at an epoxy ring (H-3 and H-4), allylic protons (H-5), olefinic protons (H-6 and H-7), doubly allylic protons (H-8), olefinic protons (H-9 and H-10) and allylic protons (H-11). The cis-configuration of the epoxy ring, pre- dicted based on the reaction property of olefinic epoxidation with a peracid, was indicated by the coupling constant (4.5 Hz) between H-3 and H-4. Homonuclear COSY spectra of 6,7-epoxides and 9,10-epoxides also clearly revealed the position of the epoxy ring. Our 1H signal assignments corresponded to the published data for C17 compounds (Millar et al., 1987). Signal separation of the epoxides was adequate in the 13C NMR spectra, which were very useful to estimate the purity and the structure of contaminating components. A usual 1H-13C COSY spectrum and a spectrum emphasizing long-range couplings were analyzed in order to assign the ~3C signals. EI-MS data of all nine epoxydienes synthesized for the field screening tests are listed in Table 4. Fragment ions were observed that were characteristic of the position of the epoxy ring. These ions are as follows: 3,4-epoxides: m/z M-29, M-72, and 79; 6,7-epoxides: m/z M-29, M-69, and 111; 9,10-epoxides: m/z M-29, M-69, M-109, 108, and 79. The origin of these ions is indicated in Figure 1. The values of the relative intensity, affixed with a in Table 4, were larger than those of the other epoxy ring positional isomers. These fragmentation properties, which were also observed in the mass spectra measured under the other instrumental conditions (Millar et al., 1987; Hansson et al., 1990), are useful in the identification of epoxydienes from a natural pheromone extract. Detection of Sex Attractants by Field Tests. Field screening tests using the compounds consisting of one major group of sex pheromones, monoenyl (Ando et al., 1977, 1981) and dienyl compounds (Ando et al., 1987) with a terminal functional group, resulted in the specific attraction of more than 200 lepidopterous species in Japan. In the current experiment, a one-year survey was conducted from the summer of 1991 using analogs of another type of pheromonal structure, hydrocarbons with a homoconjugated polyene system and their monoepoxy derivatives, resulted in the attraction of 18 species. Table 5 lists the scientific names of the attracted male moths, abbreviated names of the attractants, time of flight, and total number of moths captured by dual traps. These data suggest that some Japanese lepidopterous insects also utilize hydrocarbons and epoxides as sex pheromones. Chemicals of these types have been identified from the female moths in the following five families: Geometridae, Arctiidae, Ctenuchidae, Lymantriidae, and Noctuidae (see Table 1). Although species in Arctiidae, Ctenuchidae, and Lymantriidae were not captured in these tests, 14 4~

TABLE 4. RELATIVE ]INTENSITY ( % ) OF MOLECULAR [ON (~[+) AND SOME FRAGMENT IONS IN MASS SPECTRA OF EPOXYD1ENES

Fragment ions

m/z 79 m/z 108 m/z 111 M-109 M-72 M-69 M-29 M-18 M--

3,4-Epoxide epo3,Z6,Z9-19 : H 100 " 9 14 0 23" 0 6 a 11 4 h epo3,Z6,Z9-20 : H 100" 2 19 0 22" 0 7" 9 7' epo3,Z6,Z9-21 : H 100" 14 14 0 21" 0 5" 10 3 '1 6,7-Epoxide Z3,epo6,Z9-19 : H e 36 6 41" 0 1 10" 3" 10 6 h Z3,epo6,Z9-20 : H j 41 6 47 ~ 0 1 12" 4" 10 8" Z3,epo6,Z9-21 : t-If 34 7 39" 1 0 6" 2" 7 4 a 9,10-Epoxide Z3 ,Z6,epo9-19 : H 100" 66" 3 5" 2 2" 2" 9 4/' Z3,Z6,epo9-20: H 100" 77" 4 3" 1 3" 2" 9 4" Z3,Z6,epo9-21 : H 100" 81" 5 4" 1 2" 2 ~ 8 3 d

"Characteristic ions assumed to be produced by the fragmentation as shown in Figure 1. b m/z 278. " m/z 292. d m/z 306. eBase ion peak was observed at m/z 53. fBase ion peak was observed at m/z 83. > z o

> t- POLYENE HYDROCARBONS 795

(A) (B) (C) 108

. I '

I M-29 , ] - I M-29 Fro. 1. Characteristic fragment ions of 3,4-epoxides (A), 6,7-epoxides (B), and 9,10- epoxides (C). Molecular ions of epoxynonadecadienes (Ci9 compounds, R = n-CsHt7), epoxyeicosadienes (C20 compounds, R = n-C9H~9), and epoxyheneicosadiene (C21 compounds, R = n-C~0Hz0 were observed at m/z 278, 292, and 306, respectively.

species in Geometridae and four species in Noctuidae were attracted by these synthetic pheromone analogs, indicating that the broad screening test is appli- cable for research on the pheromones of Japanese geometrid and noctuid moths. The Geometridae, one of the two families attracted in this experiment, is a large family in the Lepidoptera, which consists of six subfamilies and ca. 800 species in Japan. The pheromones hitherto identified from the geometrid females, which are classified into the subfamilies Ennominae, Larentiinae or Oenochrom- inae, consisted of hydrocarbons and epoxy derivatives. Furthermore sex attraction by the synthetic hydrocarbons and epoxides has been observed for some other European and Canadian species of the above subfamilies. In addition to some species in these subfamilies, two species of Geometrinae, one of the other three subfamilies, were attracted for the first time by the unsaturated hydrocarbons in this field survey. Males in the subfamilies Sterrhinae and Arcchiearinae had not been attracted by these types of compounds, whereas attraction by the compounds with a terminal functional group had been reported for some species of Sterrhinae (Am et al., 1986). The Noctuidae, another large family, is composed of 18 subfamilies and ca. 1200 species in Japan. A large number of pheromone components with a terminal functionality were identified from the species of the following subfamilies: Amphipyrinae, Eustrotiinae, Hadeniinae, Heliothidae, Noctuidae, Plusi- inae, and Westermanniinae. Unsaturated hydrocarbons and monoepoxy derivatives have been recently identified as sex pheromones of some species of the subfamilies Catocalinae and Ophiderinae and sex attractants of some species of the subfamilies Herminiinae and Hypeninae. In the current field test, the attraction of males was recorded in Herminiinae and Ophiderinae. It is inter- esting to note that noctuid moths are classified into two groups according to the chemical structure of their sex pheromones. One group utilizes the compounds 796 ANDO ET AL.

TABLE 5. SEX ATTRACTANTS DETECTED IN FIELD SCREENING TESTS IN A FOREST IN TOKYO FROM 1991 tO 1992

Family Number of Subfamily attracted Species Attractant Time of flight moths

Geometridae Ennominae Aclis angulifera (Butler) epo3,Z6,Z9-19: H Sep.-Oct. 18 Ascotis selenaria epo3,Z6,Z9-19: H Aug.-Sep. 36 cretacea (Butler) Colotois pennaria epo3 ,Z6 ,Z9-20: H Nov. -Dec. 16 ussuriensis Bang-Haas Z3,epo6,Z9-19 : H 3 Pachyligia dolosa Butler Z3,Z6,epo9-21 : H Mar. 9 Z3,epo6,Z9-21 : H 8 Geometrinae Agathia carissima Butler Z3,Z6,Z9-20: H Aug. 6 Agathia visenda visenda Z6,Z9-20: H Apr., Aug. 14 Prout Z6,Z9-21 :H 3 Pachyodes superans Butler Z3,Z6,Z9-20: H Aug.-Sep. 17 Larentiinae Epirrita viridipurpurescens Z3,Z6,Z9-21 : H Nov.-Dec. 111 (Prout) Esakiopteryx volitans Z3,Z6,X9-19 : H Mar.-Apr. 384 (Butler) Operophtera relegata Prout Z 3 ,epo6 ,Z9-19 : H Nov.-Dec. 188 Z3,Z6,epo9-19 : H 149 epo3,Z6,Z9-19 : H 41 Sibatania mactata (Felder & Z3,Z6,Z9-21 : H Sep.-Oct. 11 Rogenhofer) Oenochrominae Alsophila japonensis Warren Z3,Z6,Z9-19 : H Jan. 88 lnurois fumosa Inoue epo3,Z6,Z9-21 : H Jan.-Feb. 10 lnurois membranaria epo3,Z6,Z9-21 : H Jan.-Feb. 36 (Christoph) Noctuidae Herminiinae Paracolax pryeri (Butler) Z3,epo6,Z9-20: H May-Aug. 407 Ophiderinae Paragabara flavornacula Z3,epo6,Z9-21 : H June-Aug. 65 (Oberthfir) Rivula sericealis (Scopoli) Z3,epo6,Z9-19 : H Sep.-Oct. 48 Rivula sasaphila Sugi Z3,epo6,Z9-19 : H Oct. 6 POLYENE HYDROCARBONS 797 with the most common chemical structure of lepidopterous sex pheromones, and another group utilizes the key components of taxonomically unrelated geometrid moths. Recently, sex pheromone components of Ascotis selenaria (Boarmia selenaria: Hampson) and Colotois pennaria have been analyzed. From the former species distributed in Israel, the epo3,Z6,Z9-19 :H with 3S,4R configuration has been identified as a major active component (Becket et al., 1990; Coss6 et al., 1992). One of the Japanese subspecies A. s. cretacea, a pest insect in a tea garden, was captured by the racemic mixture in this test. ZS,epo6,Z9-19 :H has been identified from C. pennaria in Hungary, and a 10:3 mixture with the parent triene exhibited field attractancy (Hansson et al., 1990). Although we have not examined the two-component lure, the Japanese subspecies C. p. ussuriensis was more strongly attracted to epo3,Z6,Zg-20: H than to ZS,epo6,Z9- 19:H. Sex pheromones and attractants of 16 other species captured in this experiment had not been detected so far, although those of some related species had been reported. A C~9 tetraene (1 ,Z3,Z6,Z9-19:H) was isolated from Oper- ophtera accidentalis, and this compound also attracted O. occidentalis (Roelofs et al., 1982). Meanwhile O. relegata, one of the Japanese representatives, was attracted by C t9 epoxydienes in this test. The sex pheromone of Alsopfiila pometaria was found to consist of a mixture of Z3,Z6,Z9-19:H and another C19 tetraene (Z3,Z6,Z9,Z11-19:H or ZS,Z6,Z9,E11-19:H) (Wong et al., 1984), while attraction of A. quadripunctata to a mixture of Z3,Z6,Z9-19:H and Z6,Z9-19:H was reported in Hungary (Sz6cs et al., 1984). In the current screening test, A. japonensis was attracted to a lure baited with only the C19 triene. Rivula propinqualis was attracted to a mixture of Z3,Z6,Z9-19:H and Z3,epo6,Z9-19:H in Canada (Millar et al., 1990a), and R. sericealis and R. sasaphila were attracted to a single component lure of the epoxydiene in Japan. Considering the large number of species of Lepidoptera, information about sex pheromones and attractants is still limited. In order to further understand the structure-activity relationship of sex attractants, it is necessary to system- aticaUy test other compounds with different chain lengths as well as multicom- ponent lures.

Acknowledgments--The authors are grateful to Prof. Y. Ambe of Rolling Land Laboratory (Tokyo University of Agriculture and Technology) for his assistance in the field screening tests.

REFERENCES

ANDO, T., YOSHIDA, S., TATSUKI, S., and TAKAHASHI, N. 1977. Sex attractants for male Lepidop- tera. Agric. Biol. Chem. 41:1485-1492. ANDO, T., KUROKO, H., NAKAGAKI, S., SAITO, O., OKU, T., and TAKAHASrtl, N. 1981. Multi- component sex attractants in systematic field tests of male Lepidoptera. Agric. Biol. Chem. 46:487-495. 798 ANDO ET AL.

ANDO, T., KOIKE, M., UCHIYAMA, M., and KUROKO, H. 1987. Lepidopterous sex attractants with a conjugated diene system. Agric. Biol. Chem. 51:2691-2694. ANDO, T., OGURA, Y., KOYAMA,M., KURANE,M., UCHIYAMA,M., and SEOL, K.Y. 1988. Syntheses and NMR analyses of eight geometrical isomers of 10,12,14-hexadecatrienyl acetate, sex pheromone candidates of the mulberry pyralid. Agric. Biol. Chem. 52:2459-2468. ARN~ H., TOTH, M, and PR]ESNER, E. 1986. List of Sex Pheromones of Lepidoptrea and Related Attractants. OILB-SROP/IOBC-WPRS, Secr6tariat General, 149 Rue de Bemy, F-75595 Paris cedex 12; ISBN 92-9067~002-9, 123 pp. BECKER, D., CYJON, R., COSSE, A., MOORE, 1., KIMMEL, T., and WYSOKI,M. 1990. Identification and enantioselective synthesis of (Z,Z)-6,9-cis-3S,4R)-epoxynonadecadiene, the major sex pheromone component of Boarmia selenaria. Tetrahedron Lett. 31:4923-4926. CONNER, W.E., EISNER, T, MEER, R.K.V., GUERRERO, A., GH1RINGELLI, D., and MEINWALD, J. 1980. Sex attmctant of an arctiid moth (Utetheisa ornatrix): A pulsed chemical signal. Behav. Ecol. Sociobiol. 7:55-63. Coss~, A.A., CYJON, R., MOORE, I., WYSOKI, M., and BECKER, D. 1992. Sex pheromone components of the giant looper, Boarmia selenaria Schiff. (Lepidoptera: Geometridae): Identifi- cation, synthesis, electrophysiological evaluation, and behavioral activity. J. Chem. Ecol. 18:165-181. FOUQUET, G., and SCHLOSSER, M. 1974. Improved carbon-carbon linking by controlled copper catalysis. Angew. Chem. (Int. Ed. Engl.) 13:82-83. HANSSON, B.S., Sz6cs, G., SCHMEDT, F., FRANCKE, W., LOFSTEDT, C., and TOTH, M. 1990. Electrophysiological and chemical analysis of sex pheromone communication system of the mottled umber, Erannis defoliaria (Lepidoptera: Geometridae). J. Chem. Ecol. 16:1887-1897. M1LLAR, J.G., UNDERHILL, E.W., G1BLIN, M., and BARTON, D. 1987. Sex pheromone components of three species of Semiothisa (Geometridae), (Z,Z,Z)-3,6,9-heptadecatriene and two mono- epoxydiene analogs. J. Chem. Ecol. 13:1371-1383. MILLAR, J.G., GIBLIN, M., BARTON, D., and UNDERHILL, E.W. 1990a. (3Z,6Z,9Z)-Nonadecatriene and enantiomers of (3Z,9Z)-cis-6,7-epoxynonadecadiene as sex attractants for two geometrid and one noctuid moth species. J. Chem. Ecol. 16:2153-2166. MILLAR, J.G., GIBLIN, M., BARTON, D., MORRISON, A., and UNDERHILL, E.W., 1990b. Synthesis and field testing of enantiomers of 6Z,9Z-cis-3,4-epoxydienes as sex attractants for geometrid moths. Interactions of enantiomers and regioisomers. J. Chem. Ecol. 16:2317-2339. ROELOFS, W.L., HILL, A.S., LINN, C.E., MEINWALD, J., JA1N, S.C., HERBERT, H.J., SMITH, R.F. 1982. Sex pheromone of the winter moth, a geometrid with unusually low temperature pre- copulatory responses. Science 217:657-659. Ross!, R, and VERACINI, C.A. 1982. Insect pheromone components. Use of ~3C NMR spectroscopy for assigning the configuration of C=C double bonds of monoenic or dienic pheromone components and for quantitative determination of Z/E mixtures. Tetrahedron 36:639-644. SzOcs, G., Tt3TH, M., BESTMANN, H.J., and VOSTROWSKY, O. 1984. A two-component sex attractant for males of the geometrid moth Alsophila quadripunctata. Entomol. Exp. Appl. 38:287- 291. UNDERHILL, E.W., PALANISWAMY,P., ABRAMS,S.R., BAILEY, B.K., STECK, W.F., and CHISHOLM, M.D. 1983. Triunsaturated hydrocarbons, sex pheromone components of Caenurgina erechtea. J. Chem. Ecol. 9:1413-1423. WONG, J.W., PALANISWAMY,P., UNDERHILL, E.W., STECK, W.F., and CHISHOLM, M.D. 1984. Sex pheromone components of fall cankerworm moth, Alsophila pometaria. Synthesis and field trapping. J. Chem. Ecol. 10:1579-1596.