Journal of Chemical Ecology, 1Iol. 19, No. 4, 1993
C18 DIENES AS ATTRACTANTS FOR EIGHTEEN CLEARWING (SESIIDAE), TINEID (TINEIDAE), AND CHOREUTID (CHOREUTIDAE) MOTH SPECIES
V . B I J D A 1'* U. M A E O R G , 2 V . K A R A L I U S , 1 G . H . L . R O T H S C H I L D , 3 S. K O L O N I S T O V A , 2 P. I V I N S K I S , l a n d R. M O Z O R A I T I S 1
~Institute of Ecology Akademijos 2, Vilnius, Lithuania 2Tartu University Jakobi 2, Tartu, Estonia 3Australian Centre for International Agricultural Research G.P.O. Box 1571, Canberra A.C.T. 2601, Australia (Received April 30, 1991; accepted December 3, 1992) A b s t r a c t - B y screening singly and binary mixed 2,13- and 3,13-octadecadien-yl acetates and alcohols (2,13- and 3,13-18: Ac/OH)in Lithuania, Armenia, Azerhaijan, Turkmenistan, Ukraine, and the far east of Russia, sex attractants were discovered for 12 Sesiidae, four Tineidae, and one Choreutidae moth species. Males of Sesia yezoensis and Bembeciapuella as well as Nemapogonflavifrons were attracted by mixture of Z3,Z13-18:Ac/OH in a ratio of 9:1, Pyropteron sp. n. by the same mixture (ratio 1:9), Bembecia romanovi and B. zuwandica by Z3,Z13-18:AC and E3,Z1318:Ac (9:1), Synanthedon caucasicum by the same mixture in the opposite ratio (1:9), B. scopigera by 23,213-18:Ac and E2,Z13-18:OH in a ratio 9:1, Synasphecia triannuliformis by Z3,Z13-18:OH and E3,Z13-18:OH (9:1), Similipepsis takizawai and Archimeessia sp. n. by E3,Z13-18:OH and E2,Z13-18:Ac (1:1), Prochoreutis sechestediana by a mixture of E3,Z13-18:Ac plus E2,Z13-18:OH (1:), Microsphecia brosiformis by E3,Z13-18:Ac, Synanthedon conopiformis by the analogous alcohol, Synanthedon scoliaeformis and Nemaxera betulinella by E2,Z13-18:Ac, Triaxomera fulvimitrella by Z3,Z13-18:Ac. An analogous alcohol component is essential for the attraction of B. ichneumoniformis males. Inhibitors for B. romanovi, B. scopigera and B. zuwandica attraction were discovered. Preliminary data on attractants for six other species as well as on the diurnal rhythm of sexual activity of three species are presented. A new method for the stereoselective synthesis of 3,13-18:Ac/OH and E2,Z13-18:Ac/OH is described. *To whom correspondence should be addressed. 799 0098-0331/93/0400-0799507.00/09 1993PlenumPublishingCorporation
800
BUDA ET AL. Key Words--Lepidoptera, Sesiidae, Tineidae, Choreutidae, (E2,Z13)-octadecadien-l-ol/acetate, 3,13-octadecadien-l-ol/acetate, isomers, stereoselectire synthesis, sex attractants, inhibitors, field trapping.
INTRODUCTION
At present, sex pheromones and attractants are known for about 1500-2000 moth species (Am et al., 1986). This number is, however, small when compared with the large number of species in the order Lepidoptera and is insufficient to solve chemotaxonomic or evolutionary problems in pheromone communication systems. Hence, the search for attractants aimed at finding new chemical substances as well as the detection of new species with already known compounds remains of great importance. The identification of sex attractants for clearwing species is equally likely to be of use in taxonomic research. Moths in this family are strong fliers, but are not captured by light traps and, consequently, are more difficult to collect than many other moth species. Many sesiids have limited distributions and appear to be relatively scarce. Thus, their attractants could be useful for both taxonomic and biological studies. The main components of sex pheromones that have been identified from the females of sesiids are dienes with a chain length of 18 carbon atoms and unsaturation at the 13 as well as the 2 or 3 position (Tumlinson et al., 1974; Schwarz et al., 1983) and, possibly, a corresponding monoene with unsaturation at the same 13 position (Szrcs et al., 1985). In the present paper we describe a new approach to the synthesis of some C~8 dienes and field screening trials with a number of these compounds carried out in Lithuania, Armenia, Azerbaijan, Turkmenistan, Ukraine, as well as in the far east of Russia.
METHODS AND MATERIALS
Chemicals. Four isomers of 3,13-octadecadienes with functional alcohol and acetate groups, one isomer of 2,13-octadecadien-1-oi and the corresponding acetate, as well as one monoene with equal carbon chain lengths were used for field screening (Table 1). Synthesis and Purity. The steps for the synthesis of 3,13-octadecadien1-yl acetates and alcohols are shown in Figures 1 and 2. The methods for the synthesis of 3,13-octadecadienols are mainly based on the alkylation of metalacetylides with protected 8-bromo-l-octanol (Uchida et al., 1978; Doolittle et al., 1980) or on the shift of the triple bond in 3-decyn-l-ol to the terminal position under superbasic media (Zhang et al., 1985; Teng and Yuan, 1985).
DIENEATTRACTANTS
801
TABLE 1. C18 COMPOUNDSUSEDFOR SCREENING Functional group and isomer
Double Bond position 3,13 3,13 3,13 3,13 2,13 13
Ac
OH
ZZ EZ EE ZE EZ
ZZ EZ EE ZE EZ
Z
Z
"'~'~/"~/'~OH "-~~i~s'~1~f" OH b d
e
~ v ~ / ~ / ~ j ~ / ~ B r
W
=
V
V
-
V
~
:
~
O
~
H
W
~
~
N
-
=
'
~
~
~
V
V
V
~
~
Z3, Z13 - 18 OH
F__.3,Z13 - 18 OH
Z3, Z13 - 18Ac
E3, Z13 - 18Ac
O
H
OH
FIG. 1. Stereoselectivesynthesis of Z,Z- and E , Z - 3 , 1 3 - I S : O H / A c . For a g abbreviations, see Figure 3.
Judging by the yields and complexity of reactions, preference is given to the latter method. The triple-bond shift in ethylenediamine was undertaken with NaNH(CH2)2NHa at 25~ The reagent was obtained from NaNHa and ethylenediamine without specific removal of the ammonia formed. The key stage of the synthesis, namely, the reaction between the 9-tetradecenylhalide and tetrahydropyranyl ether of 3-butyn-l-ol resulted in a relatively low yield of 35%, which agreed with previously published data, but not with the results presented by Hoskovec et al. (1990), where the yield of the product is indicated to be up to 65%. For E reduction of the triple bond in 9-tetradecyn-l-ol and 13-octadecen-3-yn-l-ol, Rossi's method appeared to be appropriate (Rossi and Carpita,
802
Bf3DA ET AL.
1977). Preliminary data on the method of synthesis and the biological activity of (Z,Z) and (E,Z)-3,13-octadecadienols and their acetates were presented briefly by the present authors at the thirty-second IUPAC Conference (Miieorg et al., 1989). (E2,Z13)-octadecadien-l-ol and the corresponding acetate were synthesized according to the scheme given in Figure 3. The reaction control and the purity of products were determined by GLC with the chromatograph Chrom-5 using a glass column 2.5 m x 2 mm packed with 10% PDEAS on Chromosorb W-HP 100-120 mesh. The isomer composition and the reduction control of the enynols were checked on a fused silica
'~
= ~OH
'~-
\
$,
OH
$,
Z3, E13 - 18Ac
F_.3,E13 - 18Ac
FIG. 2. Stereoselective synthesis of Z , E and E , E isomers of 3 , 1 3 - | 8: O H / A c . For c-g a b b r e v i a t i o n s , s e e F i g u r e 3.
"~f~/~AOH'=~ ~ ~ O H ,,,N///~OH
b
d~ ,,~,,,,,,~I/~i~.i.~/~Br h~
/vvv'A.vo. e
f ~
OH ~
E2, Z13 - 18Ae
FI6. 3. Stereoselective synthesis of E2,Z13-18:Ac. a: NaNH(CH2)2NH2/ H2N(CH2)2NH2; b: DHP/H § LiNH2/NH3/BuBr, MeOH/H+; c: NiP-2/H2N(CHz)2NH2/ C2H5OH; d: TsC1/Py, LiBr/(CH3)2CO; e: LiC~(CH2)2-(CH2)2-OTHP/THF/HMPA, MeOH/H+; f: Ac20/Py; g: LiAlHJdiglyme; h: Li-C~CH2OH/THF/HMPA, MeOH/ H+; i: LiA1Ha/THF.
D I E NE A T T R A C T A N T S
803
capillary column 25 m • 0.2 mm coated with a liquid crystal p-methoxycinnamoyloxy-p'-methoxyazobenzol. The structure of products obtained was confirmed by IR and 13C NMR spectroscopy with Spekord 711R and Bruker AC200P instruments, respectively. The isomer purity of E2,Z13-18 :Ac/OH was over 99% and that of E3,E 13-18:Ac/OH was 94.5 %, whereas purity of other isomers was lower. Some of the E2,Z13-18:Ac was obtained from Dr. S. Voerman (Wageningen); its isomeric purity exceeded 99%. The compound was used at the initial stage of the screening program. Z13-18:Ac/OH was synthesized according to Klun et al. (1975), the isomeric purity exceeding 99 %. Field Experiments. Each compound tested was dissolved in hexane and applied to red rubber tube dispensers (18 x 15 mm; a single cap for compound mixtures), 0.5 mg/cap. Each lure was fixed into an opaque white delta trap measuring 10 x 11 x 10 cm, which had an exchangeable bottom (11 x 18 cm) coated with sticky material (Atracon A trap and Pestifix glue, both from Flora Co., Tartu, Estonia). Traps were fixed to bush or tree branches 0.5-1 m above the ground and were inspected every one or two days or once a week depending on total screening duration. The distance between traps was not less than 15 m. Screening was carded out from 1986 to 1991 in Lithuania and various areas of European and Asian territories of the former USSR. The trapping period as well as the number of traps in each locality are described below. For brevity, localities are numbered and indicated in the text and Table 2 as follows: I--Lithuania: Ia, vicinity of Vilnius (eastern Lithuania), May 22 to July 24, 1986, two C18 compounds were tested separately (see Table 2), six replicates; Ib, the same locality, June 23 to July 29, 1987, five replicates of each attractant noted in Table 2; Ie, Puvo~iai, Varena District (southern Lithuania), July 22-27, 1990, two replicates. II--Azerbaijan, vicinity of Lenkoran (southem Azerbaijan), August 4-9, 1987, seven replicates of separate components or mixtures noted in Table 2. III--Armenia, the Khosrov Reserve (southwestern Armenia), July 10-19, 1989, four replicates. IV--Turkmenistan: IVa, Kara-Kala, Parkhai (southwestem Turkmenistan), September 17-21, 1991, three replicates for each compound and each mixture; IVb, Syunt Khoserdag Reserve, Yol-Dere Canyon, May 5-17, 1989, four replicates; IVc, the same locality, September 3-15, 1989, five replicates. V--Far east of Russia: Va, Kedrovaya Pad Reserve, Primorye, July 26 to August 12, 1989, four replicates; Vb, Partizansky District, Barabash-Levada, July 14-30, 1989, four replicates. VI--Ukraine, Crimea, Karadag Reserve: Via, August 10-18, 1990, four replicates; VIb, July 4-14, 1991, three replicates. Statistical Analysis. Data from the field tests were transformed x/x + 0.5
804
BI3DA ET AL.
where x is the number of moths captured per trap. Then they were submitted to analysis by Duncan's multiple-range test and significantly different means were separated. Identification of Moth Species. Captured specimens were identified by analysis of genitalia. Scientific names are given according to Leraut (1980) and Zagulyaev (1964) and descriptions of new species according to Gorbunov (1986, 1987) and Arita and Spatenka (1989). Bembecia ichneumoniformis and B. scopigera were identified after (;patenka and La~tuvka (1990), Microsphecia brosiformis after La~tuvka (1985). Representative specimens are kept in the insect collection at the Institute of Ecology, Vilnius, Lithuania. RESULTS AND DISCUSSION
New sex attractants for 13 clearwing (Sesiidae), four tineid (Tineidae), and one choreutid (Choreutidae) moth species as well as inhibitors for three of the sesiids were found during the field screening trials. Detailed data are presented in Table 2. The following species were attracted:
Sesiidae Microsphecia brosiformis Hubn. ( = Zenodoxus dorsalis Le Cerf after Lastuvka, 1985). A total of 42 males were captured: eight in Armenia (HI) and 34 in Turkanenistan (IVa,c). E3,Z13-18:Ac and two mixtures both containing 90 % of this component showed highest attractivity, which differed statistically significant from all tested compositions (Table 2). The binary mixtures were no more attractive than a single E3,Z 13-18:Ac, even tending to be less attractive; therefore, only E3,Z13-18:Ac should be considered a sex attractant for this species. Traps baited with this chemical are quite effective in trapping males. It should be noted that during screening in Portugal the same substance was determined to be a sex attractant for M. tineiformis Esp., a very closely related species (Nielsen et al., 1979). This species and M. brosiformis were for a time considered synonymous (see Heppner and Duckworth, 1981) and only recently were they classified as two separate species (Lagtuvka, 1985). It is not quite clear if the sex attractants for both species are indeed the same. It is highly possible that moths trapped by Nielsen et al. (1979) could be classified as M. brosiformis following the latest morphological study (La~tuvka, 1985). For that reason, clearwings trapped to E3,Z13-18 :Ac in Portugal should be reinvestigated and their taxonomy verified. In case the representative specimens are no longer available, M. tineiformis sex attractant composition should, in our opinion, be reinvestigated. Similipepsis takizawai Arita et Spatenka. A total catch of 34 males of the
DIENE ATTRACTANTS
805
species was obtained in the Far East (u Vb). Most moths (15) were attracted to a mixture of E 3 , Z 1 3 - 1 8 : O H and E 2 , Z 1 3 - 1 8 : A c in a ratio of 1: 1. This is the first attractant blend for the genus and the first record of the species outside Japan. Sesia yezoensis Hampson. Thirty-six males were trapped in the far east of Russia (Va, Vb). Most of them were attracted to a mixture of Z 3 , Z 1 3 - 1 8 : A c and Z 3 , Z 1 3 - 1 8 : O H in a ratio of 9 : 1 (Table 2). No attractivity was registered, when both components were tested separately or in different ratios. Synanthedon caucasicum Gorbunov. Captures in Azerbaijan (II) totaled 145 males. The most attractive blend for them was that of Z 3 , Z 1 3 - 1 8 : A c and E3,Z13-18 :Ac in a ratio of 1:9. Separately, the components were not attractive, and only one moth was captured at baits with the main component E 3 , Z 1 3 18:Ac. Mixtures in other ratios were ineffective. When alcohol (E3,Z1318 : OH) is used in the mixture with E3,Z13-18 : Ac, the blend is also attractive, although significantly less than the two acetates. Thus, the E3,Z13 alcohol might be a syntergist; however, to determine this, tests with three-component mixtures should be made. It is noteworthy that S. caucasicum is morphologically very closely related to S. vespiformis L. (Gorbunov, 1986), as is the chemical composition of their sex attractants (the present paper and Voerman et al., 1983). Preference of the same attractant indicates that interspecific isolation is unlikely to be determined by differences in pheromone composition. Our previous indication on S. caucasicum single-component attractant (M~ieorg et al., 1989) should be considered as preliminary. A two-component mixture for the attraction of males of this species is necessary. Synanthedon conopiformis Esp. A total of 25 males were trapped in Lithuania (Ib). Five binary mixtures of 13 used were attractive, the common component being E3,Z13-18 : OH. Of the compounds tested singly, only this alcohol was attractive. The total number of moths captured was small, as this species is scarce in the trapping area. Indeed, the species has been recently included in the Red Data Book (of rare species) of Lithuania. The attractant E 3,Z 13-18 : OH is effective enough for monitoring S. conopiformis populations, as was proved by our special program for monitoring this rare species throughout Lithuania, started in 1990. Synanthedon scoliaeformis Borkh. In Lithuania (la) 174 moths were captured in traps baited with a single component, E 2 , Z 1 3 - 1 8 : A c . As it appeared to be an effective attractant, binary mixtures with other C18 dienes were not tested. Pyropteron sp. n. Twenty-eight males were captured in traps baited with a mixture of Z3,Z13-18 :Ac and Z 3 , Z 1 3 - 1 8 : O H in Turkmenistan (IVb). The most attractive blend for them was a ratio of 1 : 9. Three males were attracted to the mixture of the same alcohol and E2,Z13-18 :Ac in a ratio of 1 : 1. Synasphecia triannuliformis Freyer. Seventy-three males were trapped in
TABLE 2. TRAP CAPTURES OF MOTHS WITH DIFFERENT C18 DIENES AND THEIR BINARY MIXTURES*
Sesiidae
Chemical Field testinglocation Z3,ZI3-18 : Ac Z3,Z13-18:Ac + Z3,Z13-18:OH Z3,Z13-18 : Ac + Z3,ZI3-1g : OH Z3,Z]3-18:Ac +Z3,ZI3-I8:OH Z3,ZI3-18:Ac + E3,Z13-18:Ac Z3,ZI3-18:Ac + E3,Z13-|8:Ac Z3,ZI3-18:Ac + E3,Z13-18:Ac Z3,ZI3-18:Ac + E3,ZI3-18 : OH Z3,ZI3-18:Ac + E3,ZI3-18:OH Z3,Z13-18:Ac + E3,ZI3-18 : OH Z3,Z!3-18:Ac + E3,EI3-18:Ac Z3,ZI3-18:Ac +E3,E13-18:OH Z3,ZI3-18:Ac + E2,Z13-18:Ac Z3,ZI3-18tAc + E2,ZI3-t8:Ac Z3,ZI3-18:Ac + E2,Z13-18:Ac Z3,ZI3-18:Ac + E2,Z13-18:OH Z3,Z13-18:Ac + E2,Z13-18:OH Z3,Zt3-18:Ac + E2,ZI3-1g:OH Z3,Z13-18 :OH Z3,Z13-18:OH + E3,Z13-18:Ac Z3,ZI3-18:OH + E3,ZI3-1g:Ac Z3,ZI3-18:OH + E3,Z, i3-18:Ac Z3,ZI3-18:OH + E3,ZI3-18:OH Z3,Z13-18:OH + E3,Zt3-18:OH Z3,ZI3-18:OH + E3,Zt3-18:OH Z3,Z13-18:OH + E3,E13-18:Ac Z3~Z13-lS:OH + E3,E13-18:OH Z3,ZI3-18:OH + E2,ZI3-18:Ac Z3,Z13-18:OH + E2,ZI3-18:OH E3,ZI3-18 : Ac E3,Z13-18:Ac + E3,Z13-18 : OH E3,ZI3-18:Ac + E3,Z13-18:OH E3,ZI3-18:Ac + E3,Zt3-18:OH E3,Zi3-18:Ac + E3,E13-18:Ac E3,Z13-18:Ac + E3,E13-18:OH E3,Z13-18:Ac + E2,ZI3-18:Ac E3,ZI3-18:Ac + E2,ZI3-18:OH E3,Z13-18 :OH E3,Z13-18:OH + E3,EI3-18:Ac E3,ZI3-18:OH + E3,E13-18:OH E3,ZI3-18:OH + E2,ZI3-I8:Ac E3,EI3-18 : Ac E3,EI3-18 :OH E2,ZI3-18 : Ac E2,Z13-18 : OH Total
Ratio I11, IVa, IVc I:l 9:1 1:9 I:1 9:1 1:9 1:1 9:1 1:9 1:1 I:1 1:1 9:1 1:9 1:1 9:1 1:9 l:l 9:1 i:9 1:1 9:1 1:9 l:l 1:1 I:I 1:1
Va, Vb
Va, Vb 4ab 19c
11
m
m
la
m
la 2ab 97b
9b la 7b
la
3ab
5ab la 2ab
i
la lib
6a
3ab
17b 1:1 9:1 1:9 1:1 1:1 1:1 1:1
Ia, Ib
la 21a
la
15a 3ab
5ab 2a la la
5a
llb
1:1 I:I l:l
7ab 2a 15b
--
4b
--
174b 42
34
36
145
25
174
e-
N~
~
.=.a
R~
~,~
N
,::,
g
--~ I
t
I
.
9 J
.
I
~
.
~
~
9
.
9
I
I
~
9
i
. . . . . . . .
.
9
I
I
.
.
. . . . . . . .
. . . . . . . .
.
i
I
J
.
.
.
I
I
9
.
.
~
.
I
I
9
.
i
I
I
.
i
. . . .
I
I
#
9 ~
~
-
o
I
J
~
.
I
I
.
.
.
.
c:r
#
I
I
.
.
.
.
I
I
. . . . .
.
.
9 ~,
I
I
.
I
I
.
l
.
I
I
i
. . . . .
J
I
.
g
I
I
9
I
I
.
~-
I
I
.
.
I
I
.
.
I
I
.
.
.
I
I
.
~ - g
. . . . . . .
. . . . . . . . . . . . . . . . . .
. . . . . .
.
.
~
~
I
~
9
9
-
9
9
.
.
.
.
.
.~ .
g.
~
. . . . .
.
.
I
I
~'-
.
i
e = - ~
.
I
I
.
I
I
.
I
.
d
.
I
d
.
I
d
. . . . . .
i
.
.
I
I
.
I
I
I
d
(
E I
d
~-
.
.
C18 DIENES AS ATTRACTANTS FOR EIGHTEEN CLEARWING (SESIIDAE), TINEID (TINEIDAE), AND CHOREUTID (CHOREUTIDAE) MOTH SPECIES
V . B I J D A 1'* U. M A E O R G , 2 V . K A R A L I U S , 1 G . H . L . R O T H S C H I L D , 3 S. K O L O N I S T O V A , 2 P. I V I N S K I S , l a n d R. M O Z O R A I T I S 1
~Institute of Ecology Akademijos 2, Vilnius, Lithuania 2Tartu University Jakobi 2, Tartu, Estonia 3Australian Centre for International Agricultural Research G.P.O. Box 1571, Canberra A.C.T. 2601, Australia (Received April 30, 1991; accepted December 3, 1992) A b s t r a c t - B y screening singly and binary mixed 2,13- and 3,13-octadecadien-yl acetates and alcohols (2,13- and 3,13-18: Ac/OH)in Lithuania, Armenia, Azerhaijan, Turkmenistan, Ukraine, and the far east of Russia, sex attractants were discovered for 12 Sesiidae, four Tineidae, and one Choreutidae moth species. Males of Sesia yezoensis and Bembeciapuella as well as Nemapogonflavifrons were attracted by mixture of Z3,Z13-18:Ac/OH in a ratio of 9:1, Pyropteron sp. n. by the same mixture (ratio 1:9), Bembecia romanovi and B. zuwandica by Z3,Z13-18:AC and E3,Z1318:Ac (9:1), Synanthedon caucasicum by the same mixture in the opposite ratio (1:9), B. scopigera by 23,213-18:Ac and E2,Z13-18:OH in a ratio 9:1, Synasphecia triannuliformis by Z3,Z13-18:OH and E3,Z13-18:OH (9:1), Similipepsis takizawai and Archimeessia sp. n. by E3,Z13-18:OH and E2,Z13-18:Ac (1:1), Prochoreutis sechestediana by a mixture of E3,Z13-18:Ac plus E2,Z13-18:OH (1:), Microsphecia brosiformis by E3,Z13-18:Ac, Synanthedon conopiformis by the analogous alcohol, Synanthedon scoliaeformis and Nemaxera betulinella by E2,Z13-18:Ac, Triaxomera fulvimitrella by Z3,Z13-18:Ac. An analogous alcohol component is essential for the attraction of B. ichneumoniformis males. Inhibitors for B. romanovi, B. scopigera and B. zuwandica attraction were discovered. Preliminary data on attractants for six other species as well as on the diurnal rhythm of sexual activity of three species are presented. A new method for the stereoselective synthesis of 3,13-18:Ac/OH and E2,Z13-18:Ac/OH is described. *To whom correspondence should be addressed. 799 0098-0331/93/0400-0799507.00/09 1993PlenumPublishingCorporation
800
BUDA ET AL. Key Words--Lepidoptera, Sesiidae, Tineidae, Choreutidae, (E2,Z13)-octadecadien-l-ol/acetate, 3,13-octadecadien-l-ol/acetate, isomers, stereoselectire synthesis, sex attractants, inhibitors, field trapping.
INTRODUCTION
At present, sex pheromones and attractants are known for about 1500-2000 moth species (Am et al., 1986). This number is, however, small when compared with the large number of species in the order Lepidoptera and is insufficient to solve chemotaxonomic or evolutionary problems in pheromone communication systems. Hence, the search for attractants aimed at finding new chemical substances as well as the detection of new species with already known compounds remains of great importance. The identification of sex attractants for clearwing species is equally likely to be of use in taxonomic research. Moths in this family are strong fliers, but are not captured by light traps and, consequently, are more difficult to collect than many other moth species. Many sesiids have limited distributions and appear to be relatively scarce. Thus, their attractants could be useful for both taxonomic and biological studies. The main components of sex pheromones that have been identified from the females of sesiids are dienes with a chain length of 18 carbon atoms and unsaturation at the 13 as well as the 2 or 3 position (Tumlinson et al., 1974; Schwarz et al., 1983) and, possibly, a corresponding monoene with unsaturation at the same 13 position (Szrcs et al., 1985). In the present paper we describe a new approach to the synthesis of some C~8 dienes and field screening trials with a number of these compounds carried out in Lithuania, Armenia, Azerbaijan, Turkmenistan, Ukraine, as well as in the far east of Russia.
METHODS AND MATERIALS
Chemicals. Four isomers of 3,13-octadecadienes with functional alcohol and acetate groups, one isomer of 2,13-octadecadien-1-oi and the corresponding acetate, as well as one monoene with equal carbon chain lengths were used for field screening (Table 1). Synthesis and Purity. The steps for the synthesis of 3,13-octadecadien1-yl acetates and alcohols are shown in Figures 1 and 2. The methods for the synthesis of 3,13-octadecadienols are mainly based on the alkylation of metalacetylides with protected 8-bromo-l-octanol (Uchida et al., 1978; Doolittle et al., 1980) or on the shift of the triple bond in 3-decyn-l-ol to the terminal position under superbasic media (Zhang et al., 1985; Teng and Yuan, 1985).
DIENEATTRACTANTS
801
TABLE 1. C18 COMPOUNDSUSEDFOR SCREENING Functional group and isomer
Double Bond position 3,13 3,13 3,13 3,13 2,13 13
Ac
OH
ZZ EZ EE ZE EZ
ZZ EZ EE ZE EZ
Z
Z
"'~'~/"~/'~OH "-~~i~s'~1~f" OH b d
e
~ v ~ / ~ / ~ j ~ / ~ B r
W
=
V
V
-
V
~
:
~
O
~
H
W
~
~
N
-
=
'
~
~
~
V
V
V
~
~
Z3, Z13 - 18 OH
F__.3,Z13 - 18 OH
Z3, Z13 - 18Ac
E3, Z13 - 18Ac
O
H
OH
FIG. 1. Stereoselectivesynthesis of Z,Z- and E , Z - 3 , 1 3 - I S : O H / A c . For a g abbreviations, see Figure 3.
Judging by the yields and complexity of reactions, preference is given to the latter method. The triple-bond shift in ethylenediamine was undertaken with NaNH(CH2)2NHa at 25~ The reagent was obtained from NaNHa and ethylenediamine without specific removal of the ammonia formed. The key stage of the synthesis, namely, the reaction between the 9-tetradecenylhalide and tetrahydropyranyl ether of 3-butyn-l-ol resulted in a relatively low yield of 35%, which agreed with previously published data, but not with the results presented by Hoskovec et al. (1990), where the yield of the product is indicated to be up to 65%. For E reduction of the triple bond in 9-tetradecyn-l-ol and 13-octadecen-3-yn-l-ol, Rossi's method appeared to be appropriate (Rossi and Carpita,
802
Bf3DA ET AL.
1977). Preliminary data on the method of synthesis and the biological activity of (Z,Z) and (E,Z)-3,13-octadecadienols and their acetates were presented briefly by the present authors at the thirty-second IUPAC Conference (Miieorg et al., 1989). (E2,Z13)-octadecadien-l-ol and the corresponding acetate were synthesized according to the scheme given in Figure 3. The reaction control and the purity of products were determined by GLC with the chromatograph Chrom-5 using a glass column 2.5 m x 2 mm packed with 10% PDEAS on Chromosorb W-HP 100-120 mesh. The isomer composition and the reduction control of the enynols were checked on a fused silica
'~
= ~OH
'~-
\
$,
OH
$,
Z3, E13 - 18Ac
F_.3,E13 - 18Ac
FIG. 2. Stereoselective synthesis of Z , E and E , E isomers of 3 , 1 3 - | 8: O H / A c . For c-g a b b r e v i a t i o n s , s e e F i g u r e 3.
"~f~/~AOH'=~ ~ ~ O H ,,,N///~OH
b
d~ ,,~,,,,,,~I/~i~.i.~/~Br h~
/vvv'A.vo. e
f ~
OH ~
E2, Z13 - 18Ae
FI6. 3. Stereoselective synthesis of E2,Z13-18:Ac. a: NaNH(CH2)2NH2/ H2N(CH2)2NH2; b: DHP/H § LiNH2/NH3/BuBr, MeOH/H+; c: NiP-2/H2N(CHz)2NH2/ C2H5OH; d: TsC1/Py, LiBr/(CH3)2CO; e: LiC~(CH2)2-(CH2)2-OTHP/THF/HMPA, MeOH/H+; f: Ac20/Py; g: LiAlHJdiglyme; h: Li-C~CH2OH/THF/HMPA, MeOH/ H+; i: LiA1Ha/THF.
D I E NE A T T R A C T A N T S
803
capillary column 25 m • 0.2 mm coated with a liquid crystal p-methoxycinnamoyloxy-p'-methoxyazobenzol. The structure of products obtained was confirmed by IR and 13C NMR spectroscopy with Spekord 711R and Bruker AC200P instruments, respectively. The isomer purity of E2,Z13-18 :Ac/OH was over 99% and that of E3,E 13-18:Ac/OH was 94.5 %, whereas purity of other isomers was lower. Some of the E2,Z13-18:Ac was obtained from Dr. S. Voerman (Wageningen); its isomeric purity exceeded 99%. The compound was used at the initial stage of the screening program. Z13-18:Ac/OH was synthesized according to Klun et al. (1975), the isomeric purity exceeding 99 %. Field Experiments. Each compound tested was dissolved in hexane and applied to red rubber tube dispensers (18 x 15 mm; a single cap for compound mixtures), 0.5 mg/cap. Each lure was fixed into an opaque white delta trap measuring 10 x 11 x 10 cm, which had an exchangeable bottom (11 x 18 cm) coated with sticky material (Atracon A trap and Pestifix glue, both from Flora Co., Tartu, Estonia). Traps were fixed to bush or tree branches 0.5-1 m above the ground and were inspected every one or two days or once a week depending on total screening duration. The distance between traps was not less than 15 m. Screening was carded out from 1986 to 1991 in Lithuania and various areas of European and Asian territories of the former USSR. The trapping period as well as the number of traps in each locality are described below. For brevity, localities are numbered and indicated in the text and Table 2 as follows: I--Lithuania: Ia, vicinity of Vilnius (eastern Lithuania), May 22 to July 24, 1986, two C18 compounds were tested separately (see Table 2), six replicates; Ib, the same locality, June 23 to July 29, 1987, five replicates of each attractant noted in Table 2; Ie, Puvo~iai, Varena District (southern Lithuania), July 22-27, 1990, two replicates. II--Azerbaijan, vicinity of Lenkoran (southem Azerbaijan), August 4-9, 1987, seven replicates of separate components or mixtures noted in Table 2. III--Armenia, the Khosrov Reserve (southwestern Armenia), July 10-19, 1989, four replicates. IV--Turkmenistan: IVa, Kara-Kala, Parkhai (southwestem Turkmenistan), September 17-21, 1991, three replicates for each compound and each mixture; IVb, Syunt Khoserdag Reserve, Yol-Dere Canyon, May 5-17, 1989, four replicates; IVc, the same locality, September 3-15, 1989, five replicates. V--Far east of Russia: Va, Kedrovaya Pad Reserve, Primorye, July 26 to August 12, 1989, four replicates; Vb, Partizansky District, Barabash-Levada, July 14-30, 1989, four replicates. VI--Ukraine, Crimea, Karadag Reserve: Via, August 10-18, 1990, four replicates; VIb, July 4-14, 1991, three replicates. Statistical Analysis. Data from the field tests were transformed x/x + 0.5
804
BI3DA ET AL.
where x is the number of moths captured per trap. Then they were submitted to analysis by Duncan's multiple-range test and significantly different means were separated. Identification of Moth Species. Captured specimens were identified by analysis of genitalia. Scientific names are given according to Leraut (1980) and Zagulyaev (1964) and descriptions of new species according to Gorbunov (1986, 1987) and Arita and Spatenka (1989). Bembecia ichneumoniformis and B. scopigera were identified after (;patenka and La~tuvka (1990), Microsphecia brosiformis after La~tuvka (1985). Representative specimens are kept in the insect collection at the Institute of Ecology, Vilnius, Lithuania. RESULTS AND DISCUSSION
New sex attractants for 13 clearwing (Sesiidae), four tineid (Tineidae), and one choreutid (Choreutidae) moth species as well as inhibitors for three of the sesiids were found during the field screening trials. Detailed data are presented in Table 2. The following species were attracted:
Sesiidae Microsphecia brosiformis Hubn. ( = Zenodoxus dorsalis Le Cerf after Lastuvka, 1985). A total of 42 males were captured: eight in Armenia (HI) and 34 in Turkanenistan (IVa,c). E3,Z13-18:Ac and two mixtures both containing 90 % of this component showed highest attractivity, which differed statistically significant from all tested compositions (Table 2). The binary mixtures were no more attractive than a single E3,Z 13-18:Ac, even tending to be less attractive; therefore, only E3,Z13-18:Ac should be considered a sex attractant for this species. Traps baited with this chemical are quite effective in trapping males. It should be noted that during screening in Portugal the same substance was determined to be a sex attractant for M. tineiformis Esp., a very closely related species (Nielsen et al., 1979). This species and M. brosiformis were for a time considered synonymous (see Heppner and Duckworth, 1981) and only recently were they classified as two separate species (Lagtuvka, 1985). It is not quite clear if the sex attractants for both species are indeed the same. It is highly possible that moths trapped by Nielsen et al. (1979) could be classified as M. brosiformis following the latest morphological study (La~tuvka, 1985). For that reason, clearwings trapped to E3,Z13-18 :Ac in Portugal should be reinvestigated and their taxonomy verified. In case the representative specimens are no longer available, M. tineiformis sex attractant composition should, in our opinion, be reinvestigated. Similipepsis takizawai Arita et Spatenka. A total catch of 34 males of the
DIENE ATTRACTANTS
805
species was obtained in the Far East (u Vb). Most moths (15) were attracted to a mixture of E 3 , Z 1 3 - 1 8 : O H and E 2 , Z 1 3 - 1 8 : A c in a ratio of 1: 1. This is the first attractant blend for the genus and the first record of the species outside Japan. Sesia yezoensis Hampson. Thirty-six males were trapped in the far east of Russia (Va, Vb). Most of them were attracted to a mixture of Z 3 , Z 1 3 - 1 8 : A c and Z 3 , Z 1 3 - 1 8 : O H in a ratio of 9 : 1 (Table 2). No attractivity was registered, when both components were tested separately or in different ratios. Synanthedon caucasicum Gorbunov. Captures in Azerbaijan (II) totaled 145 males. The most attractive blend for them was that of Z 3 , Z 1 3 - 1 8 : A c and E3,Z13-18 :Ac in a ratio of 1:9. Separately, the components were not attractive, and only one moth was captured at baits with the main component E 3 , Z 1 3 18:Ac. Mixtures in other ratios were ineffective. When alcohol (E3,Z1318 : OH) is used in the mixture with E3,Z13-18 : Ac, the blend is also attractive, although significantly less than the two acetates. Thus, the E3,Z13 alcohol might be a syntergist; however, to determine this, tests with three-component mixtures should be made. It is noteworthy that S. caucasicum is morphologically very closely related to S. vespiformis L. (Gorbunov, 1986), as is the chemical composition of their sex attractants (the present paper and Voerman et al., 1983). Preference of the same attractant indicates that interspecific isolation is unlikely to be determined by differences in pheromone composition. Our previous indication on S. caucasicum single-component attractant (M~ieorg et al., 1989) should be considered as preliminary. A two-component mixture for the attraction of males of this species is necessary. Synanthedon conopiformis Esp. A total of 25 males were trapped in Lithuania (Ib). Five binary mixtures of 13 used were attractive, the common component being E3,Z13-18 : OH. Of the compounds tested singly, only this alcohol was attractive. The total number of moths captured was small, as this species is scarce in the trapping area. Indeed, the species has been recently included in the Red Data Book (of rare species) of Lithuania. The attractant E 3,Z 13-18 : OH is effective enough for monitoring S. conopiformis populations, as was proved by our special program for monitoring this rare species throughout Lithuania, started in 1990. Synanthedon scoliaeformis Borkh. In Lithuania (la) 174 moths were captured in traps baited with a single component, E 2 , Z 1 3 - 1 8 : A c . As it appeared to be an effective attractant, binary mixtures with other C18 dienes were not tested. Pyropteron sp. n. Twenty-eight males were captured in traps baited with a mixture of Z3,Z13-18 :Ac and Z 3 , Z 1 3 - 1 8 : O H in Turkmenistan (IVb). The most attractive blend for them was a ratio of 1 : 9. Three males were attracted to the mixture of the same alcohol and E2,Z13-18 :Ac in a ratio of 1 : 1. Synasphecia triannuliformis Freyer. Seventy-three males were trapped in
TABLE 2. TRAP CAPTURES OF MOTHS WITH DIFFERENT C18 DIENES AND THEIR BINARY MIXTURES*
Sesiidae
Chemical Field testinglocation Z3,ZI3-18 : Ac Z3,Z13-18:Ac + Z3,Z13-18:OH Z3,Z13-18 : Ac + Z3,ZI3-1g : OH Z3,Z]3-18:Ac +Z3,ZI3-I8:OH Z3,ZI3-18:Ac + E3,Z13-18:Ac Z3,ZI3-18:Ac + E3,Z13-|8:Ac Z3,ZI3-18:Ac + E3,Z13-18:Ac Z3,ZI3-18:Ac + E3,ZI3-18 : OH Z3,ZI3-18:Ac + E3,ZI3-18:OH Z3,Z13-18:Ac + E3,ZI3-18 : OH Z3,Z!3-18:Ac + E3,EI3-18:Ac Z3,ZI3-18:Ac +E3,E13-18:OH Z3,ZI3-18:Ac + E2,Z13-18:Ac Z3,ZI3-18tAc + E2,ZI3-t8:Ac Z3,ZI3-18:Ac + E2,Z13-18:Ac Z3,ZI3-18:Ac + E2,Z13-18:OH Z3,Z13-18:Ac + E2,Z13-18:OH Z3,Zt3-18:Ac + E2,ZI3-1g:OH Z3,Z13-18 :OH Z3,Z13-18:OH + E3,Z13-18:Ac Z3,ZI3-18:OH + E3,ZI3-1g:Ac Z3,ZI3-18:OH + E3,Z, i3-18:Ac Z3,ZI3-18:OH + E3,ZI3-18:OH Z3,Z13-18:OH + E3,Zt3-18:OH Z3,ZI3-18:OH + E3,Zt3-18:OH Z3,Z13-18:OH + E3,E13-18:Ac Z3~Z13-lS:OH + E3,E13-18:OH Z3,ZI3-18:OH + E2,ZI3-18:Ac Z3,Z13-18:OH + E2,ZI3-18:OH E3,ZI3-18 : Ac E3,Z13-18:Ac + E3,Z13-18 : OH E3,ZI3-18:Ac + E3,Z13-18:OH E3,ZI3-18:Ac + E3,Zt3-18:OH E3,Zi3-18:Ac + E3,E13-18:Ac E3,Z13-18:Ac + E3,E13-18:OH E3,Z13-18:Ac + E2,ZI3-18:Ac E3,ZI3-18:Ac + E2,ZI3-18:OH E3,Z13-18 :OH E3,Z13-18:OH + E3,EI3-18:Ac E3,ZI3-18:OH + E3,E13-18:OH E3,ZI3-18:OH + E2,ZI3-I8:Ac E3,EI3-18 : Ac E3,EI3-18 :OH E2,ZI3-18 : Ac E2,Z13-18 : OH Total
Ratio I11, IVa, IVc I:l 9:1 1:9 I:1 9:1 1:9 1:1 9:1 1:9 1:1 I:1 1:1 9:1 1:9 1:1 9:1 1:9 l:l 9:1 i:9 1:1 9:1 1:9 l:l 1:1 I:I 1:1
Va, Vb
Va, Vb 4ab 19c
11
m
m
la
m
la 2ab 97b
9b la 7b
la
3ab
5ab la 2ab
i
la lib
6a
3ab
17b 1:1 9:1 1:9 1:1 1:1 1:1 1:1
Ia, Ib
la 21a
la
15a 3ab
5ab 2a la la
5a
llb
1:1 I:I l:l
7ab 2a 15b
--
4b
--
174b 42
34
36
145
25
174
e-
N~
~
.=.a
R~
~,~
N
,::,
g
--~ I
t
I
.
9 J
.
I
~
.
~
~
9
.
9
I
I
~
9
i
. . . . . . . .
.
9
I
I
.
.
. . . . . . . .
. . . . . . . .
.
i
I
J
.
.
.
I
I
9
.
.
~
.
I
I
9
.
i
I
I
.
i
. . . .
I
I
#
9 ~
~
-
o
I
J
~
.
I
I
.
.
.
.
c:r
#
I
I
.
.
.
.
I
I
. . . . .
.
.
9 ~,
I
I
.
I
I
.
l
.
I
I
i
. . . . .
J
I
.
g
I
I
9
I
I
.
~-
I
I
.
.
I
I
.
.
I
I
.
.
.
I
I
.
~ - g
. . . . . . .
. . . . . . . . . . . . . . . . . .
. . . . . .
.
.
~
~
I
~
9
9
-
9
9
.
.
.
.
.
.~ .
g.
~
. . . . .
.
.
I
I
~'-
.
i
e = - ~
.
I
I
.
I
I
.
I
.
d
.
I
d
.
I
d
. . . . . .
i
.
.
I
I
.
I
I
I
d
(
E I
d
~-
.
.
