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Pheromone components of the female elephant hawk-moth,Deilephila elpenor, and the silver-striped hawk-moth,Hippotion celerio

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Abstract

By means of gas chromatographic and mass spectroscopic methods, and combined GC-electroantennogram and electrosensillogram techniques, (E)-11-hexadecenal and (10E, 12E)-10,12-hexadecadienal [(E,E)-bombykal is also the main constituent of the pheromone of the silver-striped hawk-mothHippotion celerio. The biological activity of the substances was demonstrated with electroantennogram and single cell recording, and the physiological efficacy of the different hexadecadienal isomers compared.
610 Experientia 48 (1992), Birkh/iuser Verlag, CH-4010 Basel/Switzerland
Research Articles
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0014-4754/92/060606-0551.50 + 0.20/0
9 Birkh~iuser Verlag Basel, 1992
Pheromone components of the female elephant hawk-moth, Deilephila elpenor, and the silver-striped
hawk-moth, Hippotion celerio 1
H. J. Bestmann, J. Erler, W. Garbe, F. Kern, V. Martischonok, D. Sch/ifer, O. Vostrowsky and * L. T. Wasserthal
Department of Organic Chemistry, University Erlangen-Niirnberg, Henkestr. 42, D-8520 Erlangen, and * Institute of
Zoology, University Erlangen-Niirnberg, Staudtstr. 5, D-8520 Erlangen (Germany)
Received 27 March 1991; accepted 29 November 1991
Abstract.
By means of gas chromatographic and mass spectroscopic methods, and combined GC-electroantennogram
and electrosensillogram techniques, (E)-I 1-hexadecenal and (10 E, 12 E)-I 0,12-hexadecadienal [(E,E)-bombykal]
were identified as components of the sex pheromone of the female sphingid moth
DeilephiIa elpenor.
The (E,E)-bom-
bykal is also the main constituent of the pheromone of the silver-striped hawk-moth
Hippotion celerio.
The biological
activity of the substances was demonstrated with electroantennogram and single cell recording, and the physiological
efficacy of the different hexadecadienal isomers compared.
Key words. DeiIephila elpenor ; Hippotion Celerio ;
Sphingidae; pheromone components; (E,E)-bombykal; (E)-I 1-hex-
adecenal.
The elephant hawk-moth (German: Mittlerer Wein-
schw/irmer),
Deilephila elpenor
L. (Lepidoptera, Sphingi-
dae), is one of the most common sphingid species of
Central Europe, with a distribution throughout the
palearctic region. The silver-striped hawk-moth (Ger-
man: Grosser Weinschw/irmer),
Hippotion celerio L.
(Sphingidae), is a sphingid species of the tropics and
subtropics of the old world, occasionally migrating to
Central Europe.
Already in 1979 Starrat et al. described the identification
of (10E, 12Z)-10,12-hexadecadienal [bombykal] as an
active component of the sex attractant of the female
tobacco hornworm moth,
Manduca sexta
L. (Sphingi-
dae), using an electroantennogram biosasay z. Bombykal
was originally found as a sex pheromone component
in the silkworm moth
Bombyx mori
L. (Bombycidae)3.
A more detailed analysis of solvent rinses of pheromone
glands of
M. sexta
revealed the presence of a series
of twelve saturated, mono-, di- and triunsaturated
C16-
and Cls-aldehydes4; (10E, 12Z)-10,12-hexadecadienal
[bombykal] and (10E, 12E, 14Z)-10,12,14-hexadeca-
trienal represented the active principle required to stimu-
late a 4
complete behavioral sequence . This is, to the best
of our knowledge, the only pheromone from any member
of the sphingid family of Lepidoptera of which the com-
position is known.
Materials and methods
Insect rearing. D. elpenor
L. were caught at Erlangen
(FRG), and subsequent generations reared on wil-
lowherb
EpiIobium
(Onagraceae) in the laboratory. H.
celerio
L. were collected on the Canary Island "la
Gomera" and also reared on
Epilobium
species in sum-
mer and on vine
Parthenocissus
(Vitaceae) in winter, un-
der a 14 : 10 h light: dark regime. Pupae were sexed and
separated a few days before hatching.
Research Articles
HO ~ OH
2
+
HO PPh3 1
4
Experientia 48 (1992), Birkh/iuser Verlag, CH-4010 Basel/Switzerland
HBr HO ~ Br PPh3
3
1. PhLi, LiBr; 2. O ~ 5 ;
Br- ,~
--. -x CHzOH PCC
6
Synthesis of (10 E, 12 E)-10,12-hexadecadiena1 1 [E,E-bombykal].
3. PhLi, LiBr; 4. HC1, ether;
5. KO-tert-Bu
~, "~ CH=O
1
611
Isolation and identification. From newly-hatched calling
virgin females the eight and ninth abdominal segments
with the corresponding intersegmental membrane, the
pheromone gland, were excised under a stereo micro-
scope, and each preparation extracted with 15 ~tl hexane
or CS2 for GC analysis. In addition separated glands
were sealed in glass capillaries and analyzed gas-chro-
matographically using the solid sampling technique 5.
The GC analyses were performed on a HP model 5890A
gas chromatograph equipped with a splitless injector, a
flame ionization detector [FID] and a solid sampler 5.
The volatiles were chromatographed on a 25-m FSCC
SE 54 capillary column (0.25 mmID). Injection port and
detector temperatures were 200~ and 240~ respec-
tively, and the column was programmed from 80 ~ ini-
tial temperature (3 min) to 230~ final temperature at
4~ Carrier gas (N2) flow rate was 1 ml/min.
GCMS analyses were conducted with a Finnigan
MAT90 GC-mass spectrometer in electron impact [EI]
mode, 70 eV, with splitless injection, 25 m FSCC SP2340
(0.25 mmID), injector 200 ~ carrier gas He, 2 ml/min.
Electrophysiologieal bioassays. Electroantennograms
[EAG] were recorded with separated male antennae, us-
ing filter paper loaded with test chemicals as stimulus
source. For the electrosensillograms [ESG], single-cell
recording was performed according to Kaissling's
method 6 by cutting off the tip of a sensillum trichodeum
and inserting it into the "different" electrode, the "indif-
ferent" one being inserted into the base of the antenna.
GLC-coupled EAG and ESG recordings were made us-
ing a Packard United Technologies A49 model with a
1 :
1 effluent splitter which partitioned the column efflu-
ent to FID and electroantennogram detector. Injector
240 ~ FID detector 260 ~ column 25 m FSCC SP2340
(0.25 mmID), temperature program 5 min at 70 ~ 70-
195~ at 4~ carrier gas N z.
Synthesis of E, E-bombykal. ( I O E, 12E)-10,12-Hexadeca-
dienal 1 [E,E-bombykal] was synthesized according to
Schlosser's method of (E)-stereoselective carbonyl olefi-
nation 7, starting from 1,10-decandiol 2. Reaction with
HBr and subsequent with triphenyl phosphane yielded
the 10-hydroxydecyl triphenylphosphonium bromide 4,
which was converted into the corresponding ylide with
PhLi and olefinated with (E)-2-hexenal 5. Treatment of
the reaction mixture with additional PhLi and LiBr, HC1
and KO-tert-Bu gave (10E, 12E)-10,12-hexadecadien-1-
ol 6 [36%, Kp 115~ Torr (bath temp.), IR (film):
3350, 1670, 980cm-1; 1H-NMR (CDC13): 0.90-2.40
(23aliph. H), 3.15 (s, OH, H-D), 3.48 (t, J = 6Hz,
CH/O) and 4.89-6.44 (mc, 4 CH=) ppm; 13C-NMR
(CDC13): 13.95 (CH3), 22.45, 25.60, 27.54, 29.08, 29.20,
29.25, 29.31, 31.32, 32.60, 32.75 (10 CH/), 62.67 (CH20),
125.57, 128.47, 129.98 and 134.36 (4CH) ppm; MS
(70 eV): 238 (M+), 220 ( - H20), 67 (100)], from which
E,E-bombykal 1 [33 %, Kp 115~ (bath temp.); IR
(film): 1740, 1670, 980 cm-1; 1H-NMR (CDC13): 0.90-
2.40 (25 aliph. H), 4.90-6.35 (mc, 4 CH=) 9.93 (t, 2 Hz,
CHO); 13C-NMR (CDC13): 13.81 (CH3), 22.63, 23.95,
26.91, 28.80, 29.02, 29.31, 29.44, 37.82, 43.66 (all CH2),
126.03, 128.51, 129.11,132.3 (4 CH) and 204.22 (CHO)
ppm; MS (70 eV): 236 (6, M+), 192(1), 67 (100)] was
obtained by pyridinium chlorochromate [PCC] oxidation
(formula).
lO,12-Hexadecadienal isomers as GC standards. (10E,
12 Z)-10,12-Hexadecadienal [bombykal] and (10 Z,
12E)-10,12-hexadecadienal [Z,E-bombykal] were avail-
able in our laboratory from previous work s. (10 Z, 12 Z)-
10,12-Hexadecadienal [Z,Z-bombykal] was obtained as a
minor isomeric reaction product only, resulting from an
attempt to synthesize it starting from commercially avail-
able (Z)-2-hexenol. The mixture, mainly comprising
(Z,E)-bombykal, still contained enough of the (Z,Z)-iso-
mer for determination of GC retention and comparison.
Results and discussion
About 8 h after onset of the scotophase female D. elpenor
exhibited calling behavior, that is, they visibly protruded
their ovipositors. From groups of two or three female
insects, the 8th and 9th abdominal segments together
with the common intersegmental membrane were dissect-
ed and extracted with solvent. In initial attempts to iden-
tify pheromonally active compounds in the gland extract,
the extract equivalent to two females was analyzed by
gas chromatography with synchronous EAG-detection
[EAD] 9, using a male antenna. These are most senSitive
towards the conspecific sex pheromone. Two physiologi-
cally active compounds (a and c in fig. 1 A) were found to
612
A
B
Experientia 48 (1992), Birkh~iuser Verlag, CH-4010 Basel/Switzerland
Deilephila elpenor 2 fe L0-
EAD chromatogram
a I[ [ 0
II
0.8.
RGC chromatogram
3re
Research Articles
C
RGC chromatogram
9
of four isomeric
t I
I0,12 -hexadecadienals
d f
I , I ~ I
900 1000 -- scan no. 1, 1100
--ret. time ~ rain
Figure 1. Structure elucidation of female D. elpenor sex pheromone com-
ponents. A Gas chromatogram with electroantennogram detection
[EAD] of volatiles of two female gland equivalents [re]; B GCMS recon-
structed gas chromatogram [RGC] of three insects' glands [re]; a) (E)-I 1-
hexadecenal, b) hexadecanal, c) (lOE, 12E)-lO,12-hexadecadienal;
C RGC chromatogram of the four synthetic isomers of 10,12-hexadecadi-
enal: d) (10Z, 12E)-10,12-hexadecadienal, e) (10E, 12Z)-10,12-hex-
adecadienal, f) (10Z, 12Z)-10,12-hexadecadienal and c) (10E, 12E)-
10,12-hexadecadienal.
have the retention time: of oxygenated C16-compounds.
Recording the GC mass spectra, in the corresponding
elution range the chromatogram of the gland extract of
three female insects revealed three peaks (a, b and c in
fig. 1 B). The two which eluted first were identified as
(E)-ll-hexadecenal (a) and hexadecanal (b) by compar-
ing their mass spectrum and retention time with those of
authentic compounds. The spectrum of c was almost
identical with that of bombykal, but c had a slightly
longer retention time. Because of this retention differ-
ence, substance c was thought to be an isomer of bom-
bykal. Since originally only (10E, 12Z)- [bombykal] and
(10Z, 12E)-10,12-hexadecadienal [Z,E-bombykal] were
available in our laboratory, and the retention time of the
(10Z, 12E)-isomer also differed from that of c,
(10E, 12E)-10,12-hexadecadienal 1 [E,E-bombykal] had
to be prepared according to the formula scheme.
The mixture of the four geometrical isomers of 10,12-
hexadecadienal was separated gas-chromatographically
under the same GC conditions, and the retention se-
quence determined with (10Z, 12E) (peak d in the fig-
ure), (10E, 12Z) (e), (10Z, lZZ) (f) and (10E, 12E) (c),
as shown in figure 1 C. The mass spectrum and the reten-
tion time of compound c were in full agreement with
those of 1, defining the natural hexadecadienal of D.
elpenor as (10E, 12E)-10,12-hexadecadienal 1.
a
DeilephilaEAG elpenor /
(n = s)
/i/ i ~ 6 []
0.4-
0.2- / | [EIO,ZI2"I6:AI 1 e
0.001 O.O1 0.1 1 10 100 10~
gg
stimulus source loading
Figure 2. Normalized electroantennogram [EAG] dose response curves
of (E)-ll-hexadecenal [Ell-16:AI a ( = 1.0)], (10E, 12E)-10,12-hex-
adecadienal [El0 E12-16:A1 c] and the 10,12-hexadecadienal isomers
ZIOE12-16:AI d and EIOZ12-16:A1 e obtained from male D.
elpenor antennae (mean n of 5 recordings). Stimulus loading is given in
~tg.
Both synthetic substances, (E)-ll-hexadecenal a and
(10E, 12E)-10,12-hexadecadienal c (1), were cochro-
matographed with an extract equivalent on a polar
SP 2340 column with EAD detection for final identifica-
tion. Additionally, comparative EAGs were recorded
with (E)-ll-hexadecenal a together with the hexadecadi-
enal isomers c-e. The (E,E)-bombykal c was about ten
times less active than the monounsaturated aldehyde a,
both giving typically sigmoid dose response curves
(fig. 2). The other bombykal isomers d and e revealed no
significant electrophysiological activity at all. The recep-
tor potential responses to a and c recorded by single cell
measurements (ESG) reflected similar dose response
types like the EAG experiments.
Since for the analysis of the female H. celerio pheromone
an even smaller number of moths was available, the iden-
tification had to be based on more specific electrophysio-
logical techniques. An extract of pheromone glands was
produced by similar methods to those used before, gas-
chromatographed, and monitored with a male antenna
as a species-specific detector. The chromatogram of the
extract revealed the presence of one physiologically ac-
tive compound only. Its retention time was the same as
that of (10E, 12E)-10,12-hexadecadienal c (1), as it had
already been determined for the sex pheromone compo-
nent of D. elpenor. To ascertain this identification, (E,E)-
bombykal (1, c) was monitored with a male moth anten-
na, and gave a high response signal (fig. 3 A). A sensillum
trichodeum, the pheromone-specific olfactory hair of a
male antenna, was then prepared for an electrosensillo-
gram (ESG, single cell recording) 1~ and this most
specific detector used in the GC analysis of the
pheromone extract. With this ESG detector, only one
type of spike was observed, reflecting the occurrence
of only one physiologically active component in the
Research Articles
Experientia 48 (1992), Birkhfiuser Verlag, CH-4010 Basel/Switzerland 613
A
B
- ~ celerio
chromatogram of
(E,E)-bombykal c
GC coupled ESG
I~ gland solid sampling
C~sensillum
trichodeum ]
T I r
23 25
mm
! i
27
--.ret. time .~ rain
Figure 3. Pheromone identification of female
H. celerio
moths. A Gas
chromatogram of (E,E)-bombykal e monitored with a male insect anten-
na; B chromatogram of the pheromone extract monitored by single cell
recording (ESG) (receptor potential) with a male olfactory hair.
1.0-
0.8.
0.6.
0.4.
0.2-
Hippotion celerio
EAG
(~ = 7)
/~
Se~.e~ e e
JZ10,E12-16:AI]
r )
........---O v~ LEI0,E12-16:AIJ e
D~
/
d
I l I I I I
0.001 0.01 0.1 1 10 100 1000 /.tg
stimulus source loading
9
Figure 4. Normalized dose response curves of EAG tests obtained from
male
H. celerio
antenna and the bombykal isomers E 10 E 12-16
:A1 (c),
Z10
El2
16:At (d) and El0 Z12-16:A1 (e) (mean n of 7 recordings),
stimulus source loading in gg.
pheromone extract. The spike signal could be correlated
with one significant receptor response in the extract
(fig. 3B), again at the retention time of (10E, 12E)-
10,12-hexadecadienal 1. Finally, the mass spectrum was
recorded and the exact retention time determined, and
the spectrum as well as the retention time of the physio-
logically active compound were found to be identical
with those of authentic (E,E)-bombykal 1.
In additional electrophysiological experiments, the same
spike type of response was obtained from a male H.
celerio antenna with (10E, 12E)-10,12-hexadecadienal 1
by separate ESG recordings. The spike frequency reflec-
ted a typical dose-response dependence. Comparing the
electrophysiological activities of the isomeric bombykals
using the electroantennogram test (EAG), the highest
antennal responses were found (fig. 4) for (E,E)-bom-
bykal 1.
Acknowledgment. We want to thank Martin Grund, Claudia Obermeier
and Regina Walther for their help in rearing the caterpillars. We thank the
Deutsche Forschungsgemeinschaft and the Volkswagen-Stiftung for fi-
nancial assistance. V.M. is indebted to the Deutscher Akademischer
Austauschdienst for awarding a fellowship, D. S. for one from the Stu-
dienstiftung des Deutschen Volkes.
1 Pheromones, 79; as Pheromones, 78 is taken: Wu, Cai-Hong, and
Bestmann, H.J., Chinese Science Bulletin
34
(1989) 1475;
Pheromones, 77: Attygalle, A. B., Steghaus-Kovac, S., Ahmed, V. U.,
Maschwitz, U., Vostrowsky, O., and Bestmann, H. J., Naturwissen-
schaften
78
(1991) 90.
2 Starrat, A.N., Dahm, K.H., Allen, N., Hildebrand, J. G., Payne,
T. L., and Roeller, H., Z. Naturforsch.
34
(1979) 9.
3 Kasang, G., Kaissling, K. E., Vostrowsky, O., and Bestmann, H. J.,
Ang. Chem.
90
(1978) 74; int. edn, Engl.
17
(1978) 60.
4 Tumlinson, J. H., Brennan, M. M., Doolittle, R. E., Mitchell, E. R.,
Brabham, A., Mazomenos, B.E., Baumhover, A.H., and Jack-
son, D. M., Arch. Insect Biochem. Physiol.
10
(1989) 255.
5 Attygalle, A. B., Herrig, M., Vosrowsky, O., and Bestmann, H. J., J.
chem. Ecol.
13
(1987) 1299.
6 Kaissling, K. E., in: Biochemistry of Sensory Functions, p. 243. Ed.
L. Jaenicke. Springer Verlag, Berlin/Heidelberg/New York 1974.
7 Schlosser, M., Tuong, H. B., and Schaub, B., Tetrahedron Lett.
26
(1985) 311.
8 Bestmann, H. J., Siiss, J., and Vostrowsky, O., Liebigs Ann. Chem.
1981
2117.
9 Struble, D., and Arn, H., in: Techniques in Pheromone Research,
p. 161. Eds H. E. Hummel and T. A. Miller. Springer Verlag, New
York 1984.
10 Wadhams, L. J., in: Techniques in Pheromone Research, p. 179. Eds
H. E. Hummel and T. A. Miller. Springer Verlag, New York 1984.
0014-4754/92/060610-0451.50 + 0.20/0
9 Birkh~iuser Verlag Basel, 1992
... However, sex pheromones or sexual attractants of the Sphingidae remain largely unexplored as compared to the Tortricidae, Noctuidae, Pyralidae, Gelechiidae, and Sessiidae, which include many important agricultural pests (El-Sayed 2011). Hexadecadienyl compounds such as (10E, 12Z )-10,12-hexadecadienal (bombykal, E10,Z12-16:Ald) have been reported as sex pheromones or sexual attractants for a few sphingid species (Bestmann et al. 1992;Landolt et al. 1989;Reed et al. 1987;Starratt et al. 1979;Tumlinson et al. 1994), and (11E,13Z )-11, 13-hexadecadienal (E11,Z13-16:Ald) attracted Agrius convolvuli (Wakamura et al. 1996). Most of these studies involved attraction of male moths in field screening tests, with no detailed analyses of pheromone gland extracts. ...
... (Starratt et al. 1979;Tumlinson et al. 1994). Although no field trap tests were conducted, E10,Z12-16:Ald and E11-16:Ald were reported as sex pheromone candidates in Deilephila elpenor (Bestmann et al. 1992). By field screening with synthetic lures, Reed et al. (1987) demonstrated that Z10,E12-16:Ald was attractive to male Smerinthus jamaicensis and Hemaris diffinis, and E10,E12-16:Ald was attractive to Proserpinus flavofasiata. ...
Article
Full-text available
Homologs of bombykal, (10E,12Z)-10,12-hexadecadienal, have been reported to be sex pheromones or sexual attractants of several species of sphingid moths. In this study, we identified novel bombykal analogs as sex pheromone components from a Japanese sphingid moth, Dolbina tancrei. Staudinger (Sphingidae: Lepidoptera). Sex pheromone gland extracts from calling female moths were subjected to gas chromatography/electroantennograhic detection (GC/EAD), gas chromatography/mass spectrometry (GC/MS), and gas chromatography (GC) analyses. GC/EAD analyses showed two active components in the crude pheromone extracts. GC/MS analysis determined these two components to be pentadecadienals. GC/MS of their MTAD derivatives showed conjugated double bonds at the 9- and 11-positions, indicating 9,11-pentadecadienals. The isomeric configurations of these candidates were determined by comparison of their Kováts retention indices with those of synthetic compounds. Field bioassays with the four isomers of 9,11-pentadecadienal and their mixtures confirmed that the two sex pheromone components of D. tancrei are (9E,11Z)-9,11-pentadecadienal and (9Z,11Z)-9,11-pentadecadienal, with the highest male catches observed for a 90:10 blend. This is the first report of 9,11-pentadecadienals as sex pheromone components in lepidopteran species.
... Unsaturated aliphatic acetates are common chemicals in the sex pheromone of moths. The sex pheromones of hawk moths are mostly mono-or di-unsaturated aldehydes with C 16 or C 15 (Bestmann et al. 1992;Landolt et al. 1989;Reed et al. 1987;Tumlinson et al. 1994;Uehara et al. 2012Uehara et al. , 2013Uehara et al. , 2015Uehara et al. , 2016Wakamura et al. 1996). Reed et al. (1987) reported that two species of Smerinthinae were attracted to C 16 -conjugated diene acetate. ...
Article
Hawk moths are classified into the subfamilies Sphinginae, Macroglossinae and Smerinthinae. The sex pheromones of hawk moths have been intensively investigated recently. However, these reports were mainly on Sphinginae and Macroglossinae and there are only a few reports on Smerinthinae. Here, we identified sex pheromone components from the Smerinthinae, Smerinthus tokyonis Matsumura (Lepidoptera: Sphingidae). Observation of female calling behavior showed that the behavior started immediately after the photo-phase started. Gas chromatography (GC) coupled with electroantennography detection analysis indicated that male antenna responded to three components in the pheromone gland extract. GC–MS and GC analyses demonstrated that the three components were (10Z,12E)–, (10E,12Z)–, and (10Z,12Z)–hexadecadienyl acetates in a 6:7:87 ratio. We subsequently performed behavioral assays in cages. We observed the orientation and contact behavior of males in response to different odor sources, including a solvent control, calling female, pheromone gland extract, and synthetic blend. Males did not respond to the solvent control, but did respond to the other sources. Since males responded more to the calling female than to the synthetic blend, additional cues seem to be required for complete mating behavior. Nevertheless, the pheromone components determined in this first study of a Smerinthinae species are important chemicals in mating communication.
... E11-16:Ald was first identified as a sex attractant for male moths in several lepidopteran families by systematic field screening tests (Ando et al., 1981). This compound has also been reported as a pheromone component of crambid moths, Diaphania nitidalis , and D. hyalinata (L.) , the sphingid moth Deilephila elpenor (Bestmann et al., 1992), and the pyralid moths, Cryptoblabes gnidiella (Anshelevich et al., 1993) and Neoleucinodes elegantalis (Cabrera et al., 2001). ...
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Coupled gas chromatographic-electroantennographic detection (GC-EAD) of both gland extracts and effluvial collections from female blueberry leafminer, Caloptilia porphyretica Braun (Lepidoptera: Gracillariidae), showed that females produced a single EAD-active compound. The amount of the compound collected from virgin female C. porphyretica was below GC and mass spectrometry (MS) detection thresholds, even with highly concentrated gland extracts (∼150 female equivalent). (E)-11-Hexadecenal (E11-16:Ald) was determined to be a sex pheromone component mainly by comparison of retention times with authentic standards on both polar and nonpolar capillary columns, microreaction-GC-EAD analyses, and field trapping tests. GC-EAD experiments showed that synthetic E11-16:Ald exhibited extraordinarily high electrophysiological activity, stimulating significant male antennal responses at as low as 10 fg. Traps baited with E11-16:Ald alone were attractive to males. Addition of 1 or 3% of its geometric isomer, Z11-16:Ald, to E11-16:Ald did not significantly increase trap captures, but an inhibitory effect was observed at the 10% level. The influence of two kinds of rubber septa on attraction was also evaluated. Male moth captures were higher in traps baited with red rubber septa than with gray rubber septa at 30-300-µg doses. Monitoring of adult flight activity with 3-µg doses of E11-16:Ald indicated at least three distinct flight periods throughout the 2003 season.
... Based on our study, E10E2-16:Ald was the main sex pheromone component of D. angustalis female chemical communication, whereas to date, diunsaturated 16 C aldehydes have been identified as the attractant or sex pheromone in many lepidopteran species. For example, E10Z2-16:Ald was found in the gland extracts of Manduca sexta, Deilephila elpenor, and Theretra oldenlandiae oldenlandiae (Starratt et al. 1979;Bestmann et al. 1992;Uehara et al. 2012). Z10E2-16:Ald was attractive to Smerinthus jamaicensis and Hemaris diffinis, and Z10Z2-16:Ald was the only sex pheromone component of Diaphania hyalinata (Reed et al. 1987;Cabezas and Oehlschlager 1999). ...
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Insect sex pheromones play a crucial role in the mate finding and calling behavior of Lepidoptera pests. Currently, little is known about the chemical ecology of Diaphania angustalis Snellen (Lepidoptera: Crambidae), a severe and important defoliator attacking the medicinal plant, Alstonia scholaris. In the present study, the pheromone components of D. angustalis females were investigated using electrophysiological and behavioral methods. Distilled hexane extracts of female pheromone glands were analyzed through electroantennogram (EAG) and gas chromatography-electroantennogram detector (GC-EAD), and the active compounds were identified through gas chromatography-mass spectrometry (GC-MS). Production peak of female sex pheromone occurred on the third day of age at 5 h into the scotophase with the EAG test, and the hexane extracts were attractive to males in the wind tunnel test. GC-EAD analysis of virgin males to gland extracts that were subsequently evaluated showed two active compounds, (E,E)-10,12-hexadecadienal (E10E12-16:Ald) and (E,E)-10,12-hexadecadien-1-ol (E10E12-16:OH), based on comparison of retention time and mass spectrum, with suitable synthetic compounds. Under laboratory conditions, the blend of E10E12-16:Ald and E10E12-16:OH in a ratio of 9:1 elicited a stronger EAG response than other treatments or a single component. In the field, more male moths were captured by traps baited with the mixture of E10E2-16:Ald and E10E2-16:OH in a ratio of 9:1, whereas a mixture of 8:1 and 10:1 also caught males. Accordingly, E10E2-16:Ald and E10E2-16:OH were regarded as the major sex pheromone components in D. angustalis females.
... It is classified into three subfamilies, Sphinginae, Smerinthinae, and Macroglossinae (Kawahara et al. 2009). To date, female sex pheromones have been described for six species in the family (Tumlinson et al. 1994;Uehara et al. 2012Uehara et al. , 2013Uehara et al. , 2015Wakamura et al. 1996), and a few sex attractants have been reported (Bestmann et al. 1992;Landolt et al. 1989;Reed et al. 1987). Sex pheromones of most of these species are composed of unsaturated aliphatic aldehydes, such as 11-hexadecenal and 10,12-hexadecadienal. ...
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Recent work has suggested that hawk moths share pheromone components but are sexually separated by qualitative and quantitative differences in their pheromone blends. During field assays on the sex pheromones of other species, a diurnal hawk moth, Neogurelca himachala sangaica (Lepidoptera: Sphingidae), was frequently captured, but the composition of the sex pheromone of this species was not known. Analysis of hexane extracts of the pheromone glands of calling female by gas chromatography (GC) using an electroantennographic detector (EAD) revealed two components that elicited EAD responses from male moth antennae. These components were identified by their mass spectra and retention indices on two GC columns as (10E,12Z)-10,12-hexadecadienal (E10,Z12–16:Ald) and a trace of its (10E,12E)-isomer (E10,E12–16:Ald) in 98:2 ratio. In field experiments, E10,Z12–16:Ald alone attracted male moths, and addition of E10,E12–16:Ald significantly reduced the attractiveness, even at the naturally-occurring ratio. Analysis of the data using a generalized linear mixed model showed that E10,Z12–16:Ald positively contributed to attractiveness, whereas E10,E12–16:Ald did so negatively, and it was concluded that the sex pheromone of N. himachala sangaica consists solely of E10,Z12–16:Ald, bombykal. The negative effect of E10,E12–16:Ald on attractiveness could promote the species-specificity of this single-component pheromone system.
... 우리나라에는 대암산 용늪, 선유담을 비 롯하여 그 분포 범위가 매우 제한되어 있는 북방계 식물이 다 (Kim et al., 2013). 이와 같은 조름나물의 생육 특징으로 인해 지구온난화에 따라 점점 서식처의 범위가 축소될 것 으로 예상되며, 이를 보존하기 위하여 조름나물의 서식처 특성 (Han and Kim, 2006;Kim et al., 2013), 서식기질에 따른 타 식물과의 경쟁관계 (Lee and Kim, 2011), 최대 영 양 번식을 위한 지하경 삽목 조건 (Lee and Kim, 2012 (Bestmann, 1992). 주홍박각시 애벌레는 주 로 습지에 서식하는 것으로 알려졌으며, 터키에서는 Vitis (포도속), Parthenocissus(담쟁이덩굴속), Epilobium(바늘꽃 속), Chamaenerion angustifolium(분홍바늘꽃), Clarkia(클 라키아속), Galium(갈퀴덩굴속), Calla palustris(산부채), Impatiens(봉선화속), Fucsia(후크시아속), Menyanthes(조 름나물속)를 섭식하는 것으로 알려졌다 (Akkuzu et al. 2007). ...
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Even though many researches are conducted for the conservation and restoration of endangered species Menyanthes trifoliata, recently, there is no study on the threatening factors to this plant. This is the first time in Korea to study growth and feeding characteristics of Deilephila elpenor as a threatening factor to Menyanthes trifoliata through an experiment. Experiment was done with 6 Eephant hawk-moth larvae and change of body weight, food preference, and ingestion amount of Bog-bean were investigated. It took 27 days from larva to pupa and maximum body weight of lavae was in the range of 4-7.5g. The food preference sequence of the lavae was Menyanthes trifoliata, Impatiens balsamina, Ampelopsis brevipedunculata var. heterophylla, Parthenocissus tricuspidata. Ingestion model shows the total amount of ingestion by a larva is 11-30g and this amount can be acquired at 0.03-0.08m^2 of Menyanthes trifoliata pure stand. This study showed Deilephila elpenor as a potential threatening factor and suggests that the conservation and restoration plan of endangered species Menyanthes trifoliata include the control plan of Deilephila elpenor, also.
... 1992), Notarcha basipunctalis Bremer (Honda et al. 1994), Maruca vitrata Fabricius (Downham et al. 2003), and Conogethes pluto Butler (El-Sayed et al. 2013). These components are also found in pheromones of other moth families, including Noctuidae (Cork et al. 1988), Saturniidae (McElfresh et al. 2001, Schreckensteiniidae , and Sphingidae (Bestmann et al. 1992;Uehara et al. 2012). However, only in Diaphania were two pheromone components found at the same time. ...
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The aim of this study is to identify the pheromone active component of female moths, Diaphania glauculalis, an important pest of Anthocephalus chinensis in China. The sex pheromone was extracted from sex pheromone gland extracts of virgin female moth of D. glauculalis using n-hexane, and the pheromone gland extracts of females were analyzed using coupled gas chromatography-electroantennogram detection (GC-EAD) and gas chromatography-mass spectrometry (GC-MS). The sex pheromone active components were based on the comparison the retention time and mass spectrum, with suitable synthetic compounds. (E)-11-hexadecenal (E11-16:Ald) and (E,E)-10,12-hexadecadienal (E10E12-16:Ald) were identified as the major sex pheromone components in the females. Their biological activities were evaluated in a series of electroantennogram (EAG) experiments and four-arm olfactometer assays using synthetic compounds. D. glauculalis males could be attracted by any single component, but a mixture of the E11-16:Ald and E10E12-16:Ald in a ratio of 5:5 elicited a substantial response, demonstrating that the binary blend is essential in male attraction. We therefore conclude that the aldehyde compounds, a mixture of E11-16:Ald and E10E12-16:Ald, comprise the sex pheromone components of D. glauculalis, which might be applied for insect field trapping.
Chapter
Sphingidae (hawk moths) is one of the largest families in Bombycoidea and includes approximately 1450 species worldwide. Hawk moths have varying life cycles, interesting flight behavior (hovering at flowers), include both nocturnal and diurnal species, and have a large body size and long proboscis, making them valuable subjects for studies across a broad range of biological fields. Hawk moths also provide useful subjects for studies on sex pheromone communication systems; however, they remain to be further explored. We have determined the chemical structures of sphingid sex pheromone components, and evaluated their biological activity in both the field and the laboratory. The compound (10E,12Z)-hexadecadienal, known as bombykal, and its analogues are sex pheromone components in many hawk moth species. In this chapter, we discuss the sex pheromone communication system of hawk moths with reference to the similarity and dissimilarity of pheromone components.
Article
Extracts from the abdominal tips of female moths of the cotton caterpillar moth, Diaphania indica (ca. 15.4 ng/female and 7.6 ng/female, respectively) yielded major EAG-active components that were identified as (E)-11-hexadecenal (E11-16:Ald) and (E,E)-10,12-hexadecadienal (E10E12-16:Ald). Hexadecanal (16:Ald) was also found and determined to be at a ca. 2% level compared with E11-16:Ald. Mixture of synthetic E10E12-16:Ald and E11-16:Ald showed attractiveness to males in the field, although it was inferior to three 2-d-old virgin females. The addition of 16:Ald at 0.5% to 10% amounts of E11-16:Ald did not effect the trap catches.
Chapter
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Sensory transduction means the conversion of a physical or chemical stimulus into the excitation of a sensory (or receptor) cell. This process involves a number of steps which can be fully understood only by a combined effort of biophysicists, biochemists, morphologists and physiologists. To study sensory transduction it is also worthwhile to compare sensory systems adapted to different stimulus modalities, since the receptor organs differ considerably depending on their adequate form of stimulus energy.
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Analyses of solvent rinses of the external surfaces of pheromone glands excised from calling female tobacco hornworm moths, Manduca sexta (L.), revealed the presence of the following compounds: (Z)-9-hexadecenal, (Z)-11-hexadecenal, (E)-11-hexadecenal, hexadecanal, (E,Z)-10,12-hexadecadienal, (E,E)-10,12-hexadecadienal, (E,E,Z)-10,12,14-hexadecatrienal, (E,E,E,)-10,12,14-hexadecatrienal, (Z)-11-octadecenal, (Z)-13-octadecenal, octadecanal, and (Z,Z)-11,13-octadecadienal. The two trienals were identified by mass and PMR spectral analyses and by ozonolyses, and their structures were confirmed by synthesis. In a wind tunnel male tobacco hornworm moths exhibit the same behaviors in response to a synthetic blend of all of the components, the gland rinse, or a calling female. Both (E,Z)-10,12-hexadecadienal and (E,E,Z)-10,12,14-hexadecatrienal are required to stimulate males to complete the characteristic behavioral sequence: anemotaxis, approaching and touching the pheromone source, and bending their abdomens in apparent copulatory attempts. The other components of the blend may play more subtle roles.
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The first sex pheromone of a Sphinx moth has been isolated from Manduca sexta (Lepidoptera: Sphingidae) with the aid of an electroantennogram assay. It is attractive to males in a field test where its activity might be augmented by a second component in female extracts. Chemical tests and chromatographic properties identify the pheromone as a C16-aldehyde with a pair of conjugated double bonds. (E , Z)-0,12-hexadecadienal, (“bombykal”) was identical with the natural product. The comparison includes chromatography on three different capillary columns and determination of specific activities in the electroantennogram test.
Book
This volume contains the Proceedings of the 25th Mosbach Colloquium, the general theme of which is the Biochemistry of Sensory Functions. It was intended, continuing the silver-tradition of these Colloquia, to provide the uninitiated biochemist with an insight into the current status of a line of research in Molecular Biology which, more than many other fields in Biochemistry, has maintained its contacts with and respect for Physiology. The speakers were asked to attempt to outline their topic sufficiently to define the fundamentals and to build up upon this basis the more sophisticated details of their own studies. It is for the reader to evaluate how well both organizer and participants 1 have attained this end • These Proceedings not only mirror the hubs around which several groups of scientists wheel but may also serve as a source of literature references and for the advanced student as an introduction to this highly up-to-date branch of Biochemistry, although no index is provided as the table of con­ tents is considered sufficient to locate most of the s
Chapter
Electroantennogram (EAG)3 recording of insect olfactory responses has been a great asset as a method of bioassay for chemical identifications of pheromones and for the development of synthetic attractants (Roelofs, Chapter 5). When EAG is used separately from the purification method, e.g., gas chromatography (GC), it is necessary to isolate and transfer small amounts (ng) of purified components to the EAG preparation. This does not take full advantage of the GC resolution capabilities and it may result in a loss of essential minor pheromone components. It is also time consuming.
Chapter
The first identification of a pheromone in 1961 by Butenandt et al. (1961) was achieved without the aid of a gas chromatograph-a tool which subsequent workers have found essential. Although gas chromatography (GC) is a powerful technique for the separation and isolation of pheromones, additional techniques are required to establish which components in the mixture are involved in the behavior of the insect.
Article
The structure elucidation of sex pheromones of Lepidoptera by a solid-sample injection technique in conjunction with capillary gas chromatography is described. The applicability of this method in GC and GC-MS modes was demonstrated by reanalyzing the sex attractants of females ofOstrinia nubilalis andBombyx mori. The pheromone complex ofMamestra brassicae was reinvestigated and (Z)-9-hexadecenyl acetate and (Z)-11-hexadecenol were found in addition to already known pheromone components of this species. By using the solid-sample injection, the exact site of pheromone release could be determined inM. brassicae.
Article
Nach Art eines “Baukastensystems” werden konjugiert-ungesättigte Alkadienylacetate, Alkadienole und Alkadienale, wie sie als Sexualpheromone weiblicher Schmetterlinge bekannt sind, mit unterschiedlichen Positionen und Geometrien der Doppelbindungen synthetisiert.Pheromones XXXIV. — Synthesis of Conjugated-Unsaturated Lepidoptera Pheromones and AnaloguesConjugated-unsaturated alkadienyl acetates, alkadienols, and alkadienals — known as sex pheromones of female butterflies and moths — with varying positions and configurations of double bonds were synthesized using a “unitized construction principle”.
Endocrinology I, p. 363
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