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A gas chromatographic survey of the volatile fractions of twenty species of insects from eight orders
J. Insect Physiol., 1972, Vol. 18,pp. 555 to 564. Pergamon Press. Printed in Great Britain
A GAS CHROMATOGRAPHIC SURVEY OF THE VOLATILE FRACTIONS OF TWENTY SPECIES OF INSECTS FROM EIGHT ORDERS* P. A. HEDIN,
Entomology
Research
C. S. NIEMEYER, R. C. GUELDNER, and A. C. THOMPSON
Division, Agricultural Research Service, U.S. Agriculture, State College, Mississippi 39762
Department
of
(Received 26 July 1971)
Abstract-The profiles of the essential oil of twenty species of insects from eight orders were obtained by steam distillation and analysis by gas chromatography. For six of the seven species analysed separately by sex, the profiles showed prominent differences. In Coleoptera, phylogenetic similarities were apparent. Sex attractant activity has been reported in only seven of the twenty species investigated.
INTRODUCTION TUMLINSON et al. (1969, 1970) recently isolated and identified four monoterpene compounds from the adult male boll weevil, Anthonomw grandis Boh. When these compounds were combined and assayed in the laboratory, the mixture was as attractive to females as live males. In subsequent tests in the field, HARDEE et al. (1971) found that all four elicited both an attractant and aggregant response ; however, the response of insects in nature to the pheromone complex (grandlure mixture) was somewhat less, 50 to 80 per cent of that elicited by live males. Thus, though the problem may simply be one of formulation, the possibility exists that still other components are required. A preliminary effort was therefore made to identify any other potential pheromones produced by the boll weevil, by analysing the total volatile fraction of both sexes with an integrated gas chromatography-mass spectrometry system. Evidence was obtained of the presence of a number of mono- and sesquiterpene hydrocarbons, some substituted anilines including o-toluidine, some C, and C, alcohols and monoterpene alcohols, and at least one sesquiterpene alcohol. The major components were found to be a series of alkanes, alkenes, and alkyl alcohols of high molecular weight (Hedin et al., 1972). Subsequently, when the volatile fractions from males and females were analysed separately, so we could determine whether any differences could be attributed to sex, we found that the profile of males possess small maxima at the expected retention volumes of the known
* Mention of a proprietary product in this paper does not constitute an endorsement of this product by the U.S. Department of Agriculture. 555
556
P. A. HEDIN, C. S. NIEMEYER,R. C. GUELDNER, AND
A. C. THOMPSON
sex attractants; they appeared to be absent in that of females. Also, several other differences in the profiles were apparent. The logical extension of this approach was to conduct a gas chromatographic profile survey of a number of species of insects, sexed when feasible, encompassing several orders. Of course, some would not necessarily be producing any attractant when they were analysed, the pheromones might not survive steam distillation and solvent extraction, and any pheromones present might represent less than 1 per cent of the total volatiles and so escape observation. Nevertheless, the approach appeared to have some promise, and a phylogenetic application also appeared feasible. The only known exhaustive investigation of the entire volatile profile of an insect previous to the study of the boll weevil was that of KULLENBERG et al. (1970) who investigated the volatile components of the cephalic marking secretion of male bumblebees, Bombus sp. They reported acyclic mono-, sesqui-, and diterpenes as well as aliphatic normal chain compounds with 10 to 27 carbon atoms.
MATERIALS
AND METHODS
Collections of insects Insects in 5 to 50 g quantities were collected during the summer of 1970 by research entomologists stationed at the Boll Weevil Research Laboratory and at the Department of Entomology, Mississippi State University, State College, Miss. The female pink bollworms, Pectinophora gossypiella (Saund.), were provided by the Western Cotton Research Laboratory, Phoenix, Arizona. The insects were classified by the co-operating entomologists, and sexing was performed either by the entomologist or at his direction.
Preparation of insect essential oils On receipt, the insects were weighed, hilled in redistilled methylene chloride or ethyl ether, and stored at -20°C until steam distilled. The aqueous distillate was extracted three times with the organic solvent, the solvent was partially removed under vacuum, the sample was dried with anhydrous sodium sulphate, and the remaining material was reduced in volume to 5 ml under vacuum. The distillate was then stored at -20°C until analysis.
Gas chromatographic analysik For GLC, the sample was reduced in volume with a stream of N, and slight warming to about O-2 ml. Then, successive GLC analyses (2 to 5) were performed until reproducible profiles were obtained. As little as O-4 g of insects (e.g. 25 boll weevils) will provide sufficient distillate for two determinations if a microdistillation system is used, but in this study, we had 5 g samples or larger. Analyses were performed on an F.I.D. instrument with the following column and conditions:
VOLATILE FRACTIONS OF INSECTS
557
a 0.0032 x 6.145 m stainless steel column packed with 10% SE-30 on 60/80 mesh Chromosorb W treated with HMDS. Carrier gas flow N, at 50 ml/mm, column temperature 175”C, injector 185”C, detector 190°C. The profiles obtained from the SE-30 column were used in preparing the figures because with this column the retention volumes were lower than those obtained with the Carbowax-4000 columns. KOVATS (1961) indices (Ik) were calculated from the retention volumes, e.g. pentadecane = 1500, hexadecane = 1600, and used as a guide in the prediction of structures.
RESULTS
AND DISCUSSION
The insects in Table 1 are listed by chromatogram number and classified by order, family, genus, and species. The common names, weight per insect, and contributing entomologist are also included. Sex pheromones and/or identification work have been reported for seven of the insects surveyed as follows: the male boll weevil (TUMLINSON et al., 1969) ; the female tobacco budworm (GENTRY et aE., 1964); the female bollworm (GASTONand SHOREY,1964); the female pink bollworm (JACOBSON,1969) ; the female American cockroach (JACOBSON et al., 1963) ; a male fruit fly (MAYR, 1950); and a female fruit fly (STANDFUSS,1896). Figs. 1 to 7 contain the 28 gas chromatographic profiles obtained with the SE-30 column; they are grouped phylogenetically. Since the data were accumulated over 4 months, retention times may be subject to variances of perhaps 3 to 5 per cent. Table 2 gives the retention times in terms of Kovats’ indices and their structural equivalency so the chromatographic maxima may be better understood. Several phylogenetic similarities seem apparent. The Curculionidae (1 to 4), the Chrysomelidae (9 to 12), and the Coccinellidae (5 to 7) all possess similar profiles within families. Differences that can be attributed to sex seem obvious in all instances except for the American cockroach (19, 20). All the Coleoptera (1 to 12) possess a series of maxima which may be attributed in part to the monoterpenes which are known to be biosynthesized by this order. The components of the boll weevil sex attractant components (TUMLINSONet al., 1969) are included among maxima at 5 to 8 min (1); they are essentially absent in the female (2). Among the Lepidoptera, the large maxima in the male tobacco budworm (13) (the female is presumed to be attractive) and the female bollworm (16) are probably C,, to C,, alcohols and/or esters. The pheromones of this order are usually represented by these classes of compounds (and have this or a slightly higher molecular weight). Little is known about pheromones in Diptera, but both sexes of D. melanogaster (21 to 22) appear to produce unique compounds. The defence secretions of the Say stink bug (26) may be in part attributed to maxima 8 to 11, but 2-hexenal, a common repellent known to be produced by this insect, should have been eluted in the first 2 min. In summary, GLC profile analyses appear promising for surveys of insect pheromones. Th ey also appear promising for phylogenetic classification and for monitoring isolation work.
Curculio Curculio Hippodamia Coleomegilla Coleomegilla Chauliognathus Chrysomela Chrysomela Galerucella Diabrotica
Curculionidae
Curculionidae
Coccinellidae
Coccinellidae
Coccinellidae
Cantharidae
Chrysomelidae
Chrysomelidae
Chrysomelidae
Chrysomelidae
Noctuidae Noctuidae Noctuidae Noctuidae Gelechiidae
3
4
5
6
7
8
9
10
11
12
13 14 15 16 17
Heliothis Heliothis Heliothis Heliothis F’ectinophora
Anthomomus
Curculionidae
__-
2
Genus
Anthonomus
Family
Curculionidae
__-____ COLEOPTERA
(Saunders)
gossypiella
virescens
LEPIDOPTERA (F.) virescens (F.) zea (Boddie) zea (Boddie)
undecimpunctata howardi Barber
(Schrank)
xanthomelaean
scripta
scripta
DeG.
(Muls.)
(Muls.)
G&r.
pennsylvanicus
fuscilabrus
fuscilabrus
convergens
caryae (Horn)
caryae (Horn)
grandis Boh.
grandis Boh.
Species
beetle
Tobacco budworm Tobacco budworm Bollworm Bollworm Pink bollworm
Spotted cumcumber beetle
Cottonwood leaf beetle Cottonwood leaf beetle Elm leaf beetle
Soldier
Convergent lady beetle Spotted ladybird beetle Larvae
Q
d ? d ?
M
306.0
292.0
F. F. F. F. J.
G. G. G. G. C.
Maxwell Maxwell Maxwell Maxwell Keller
J. L. Hamer and L. N. Latson
R. B. Head
M
Hamer and N. Latson Hamer and N. Latson Head R. B. Head
J. L. L. J. L. L. R. B.
E. P. Lloyd
D. D. Hardee and 0. H. Lindig D. D. Hardee and 0. H. Lindig W. W. Neel and 0. M. Baughman W. W. Neel and 0. M. Baughman E. P. Lloyd
Entomologist
?
d
75.8
9.8
M
M
27.2
M*
weevil
Pecan
57.5 68.0
c3
weevil
Pecan
12.0
13.1
(mg)
Weight of insect
Q
Q
Boll weevil
Sex
6
name ~._
Boll weevil
Common
~--CLASSIFICATION OF INSECTS, THEIR SIZE,AND SOURCE _ ..__....__~----
1
Chromatogram No.
TABLE
Drosophilidae
Drosophilidae
Tabanidae Tabanidae
Membracidae
Pentatomidae
Chrysopidae
A mixture of spiders from cotton
21
22
23 24
25
26
27
28
* M = mixed.
Periplaneta Periplaneta
Blattidae Blattidae
19 20
Chrysopa
Chlorochroa
Stictocephala
Tabanus sp. Tabanus sp.
Drosophila
Drosophila
Microcentrum
Tettigoniidae
18
Genus
Family
Chromatogram No. Common name
Meig.
DIPTERA Meig.
oculata
Say
sayi Stll
ARACHNIDA
NEUROPTERA Goldeneye lacewing
Say stink bug
HEMIPTERA(HOMOPTERA) Buffalo treehopper
bubalus (F.)
melanogaster
melanogaster
rhombifolium
ORTHOPTERA (Sauss.) Broadwinged katydid American cockroach americana (L.) American cockroach americana (L.)
Species
TABLE 1 (cont.)
M
M
M
M
M M
?
8
6 e
M
Sex
13.2
300.0 100.0
1180.0
Weight of insect (mg)
E. P. Lloyd
J. L. Hamer
J. L. Hamer and L. N. Latson E. P. Lloyd
D. M. Elliot and C. H. Thomas D. M. Elliot and C. H. Thomas C. S. Niemeyer C. S. Niemeyer
H. B. Green H. B. Green
E. P. Lloyd
Entomologist
P. A. HEDIN, C. S. NIEMEYER,R. C. GUELDNER,ANDA. C. THOMPSON
560
.2 46 10 I 0
!
1
4
‘1
1”
8
12
1 ” I6 2G ” 24 ” 25 ” 32 ” 36
12 13 11 0
11 4
II
4
0
’
1 8
1
”
6
”
12
”
12
”
13
12
II
16
”
16
”
20
” 20
”
24
”
” 24
28
” 2s
”
32
I
,
I
40
I
44
14 ”
36
”
”
” 32
40
”
’
36
40
44
” 44
FIG. 1. Insect volatile profiles of (1) male boll weevils, (2) female boll weevils, (3) male pecan weevils, (4) female pecan weevils. A 0.0032 x 6.145 m SE-30 column; retention time in min.
TABLE 2--KOV.~TS
Rt
*
o-2 2-9
AND STRUCTURAL
EQUIVALENCY
O-900
GLC
RETENTION
TIMES
Compound
Ikt
900-l 300
OF
Solvent, C&C, hydrocarbons, Ci-C,
alcohols, carbonyls
Cl0 (Terpene) hydrocarbons, carbonyls, alcohols, C&i,, esters, boll weevil pheromones, bark beetle pheromones, verbenol, brevicomin Hydrocarbons,
C&C,,
254
1500-1700
Hydrocarbons, Ci,-Ci, alcohols, C&-esters, C1&1, pheromones of cabbage looper and fall armyworm
44-100
1700-2000
Propylure, bombykol, gyptol
* Retention time in min. t Kovats indices for 6.145 m SE-30 column.
C,,-C,,
alcohols
1300-1500
9-25
VOLATILE FRACTIONS OF INSECTS
561
-::
-Z -4 LD -:
-,” - hg,
-w
N -
-s
-::
-x
-2
:
-x
P
LD
-z
-n”
-4 IO.. -
-x -2
N
562
P. A. HEDIN, C. S. NIEMEYER, R. C. GUELDNER, AND A. C. THOMPSON
VOLATILE FRACTIONS OF INSECTS
564
P. A. HEDIN, C. S. NIEMEYER,R. C. GUELDNER,ANDA. C. THOMPSON
Acknowledgements-The authors thank FRANK BENCI and his staff at the Boll Weevil Research Laboratory for photographic and art work. REFERENCES GASTONL. K. and SHOREY H. H. (1964) Sex pheromones of noctuid moths-IV. An apparatus for bioassaying the pheromones of six species. Ann. ent. Sot. Am. 57,779-780. GENTRY C. R., LAWSONF. R., and HOFFMANJ. D. (1964) A sex attractant in the tobacco budworm. J. econ. Ent. 57, 819-821. HARDEED. D., MCKIBBEN G. H., GUELDNERR. C., MITCHELL E. B., TUMLINSONJ. H., and CROSSW. H. (1971) Boll weevils in nature respond to grandlure, a synthetic pheromone. J. econ. Ent. In press. HEDIN P. A., THOMPSONA. C., GUELDNERR. C., and MINYARDJ. P. (1972) Volatile constituents of the boll weevil. r. Insect Physiol. 18, 79-86. JACOBSONJ., BEROZAM., and YAMAMOTO R. T. (1963) Isolation and identification of the sex attractant of the American cockroach. Science, Wush. 139, 4849. JACOBSONM. (1969) Sex pheromone of the pink bollworm: biological masking by its geometric isomer. Science, Wash. 163, 190-191. Kov.4~~ E. az (1961) Zusammenhange zwischen Struktur und Gas chromatographischen Daten organischer Verbindungen. 2. anal. Chem. 181, 351-366. KIJLLENBERCB., BERGSTROM G., and STALLBERG-STENHAGEN S. (1970) Volatile components of the cephalic marking secretion of male bumblebees. Acta them. Stand. 24, 149-154. MAYR E. (1950) The role of the antennae in the mating behavior of female Drosophila. Evolution 4, 149-l 54. STANDFUSSM. (1896) Handbuch der Pakihrktischen Gross-Schmetterlinge fiir Forscher und Sammler, 2nd ed. Fischer, Jena. TUMLINSONJ. H., HARDEED. D., GUELDNERR. C., THOMPSONA. C., HEDIN P. A., and MINYARDJ. P. (1969) Sex pheromones produced by the boll weevil: isolation, identification and synthesis. Science, Wash. 166, 1010-1012. TUMLINSONJ. H., HARDEED. D., GUELDNERR. C., THOMPSONA. C., HEDIN P. A., and MINYARDJ. P. (1970) The boll weevil sex attractant. In Chemicals Controlling Insect Behavior (Ed. by BEROZAM.), pp. l-59. Academic Press, New York.
A GAS CHROMATOGRAPHIC SURVEY OF THE VOLATILE FRACTIONS OF TWENTY SPECIES OF INSECTS FROM EIGHT ORDERS* P. A. HEDIN,
Entomology
Research
C. S. NIEMEYER, R. C. GUELDNER, and A. C. THOMPSON
Division, Agricultural Research Service, U.S. Agriculture, State College, Mississippi 39762
Department
of
(Received 26 July 1971)
Abstract-The profiles of the essential oil of twenty species of insects from eight orders were obtained by steam distillation and analysis by gas chromatography. For six of the seven species analysed separately by sex, the profiles showed prominent differences. In Coleoptera, phylogenetic similarities were apparent. Sex attractant activity has been reported in only seven of the twenty species investigated.
INTRODUCTION TUMLINSON et al. (1969, 1970) recently isolated and identified four monoterpene compounds from the adult male boll weevil, Anthonomw grandis Boh. When these compounds were combined and assayed in the laboratory, the mixture was as attractive to females as live males. In subsequent tests in the field, HARDEE et al. (1971) found that all four elicited both an attractant and aggregant response ; however, the response of insects in nature to the pheromone complex (grandlure mixture) was somewhat less, 50 to 80 per cent of that elicited by live males. Thus, though the problem may simply be one of formulation, the possibility exists that still other components are required. A preliminary effort was therefore made to identify any other potential pheromones produced by the boll weevil, by analysing the total volatile fraction of both sexes with an integrated gas chromatography-mass spectrometry system. Evidence was obtained of the presence of a number of mono- and sesquiterpene hydrocarbons, some substituted anilines including o-toluidine, some C, and C, alcohols and monoterpene alcohols, and at least one sesquiterpene alcohol. The major components were found to be a series of alkanes, alkenes, and alkyl alcohols of high molecular weight (Hedin et al., 1972). Subsequently, when the volatile fractions from males and females were analysed separately, so we could determine whether any differences could be attributed to sex, we found that the profile of males possess small maxima at the expected retention volumes of the known
* Mention of a proprietary product in this paper does not constitute an endorsement of this product by the U.S. Department of Agriculture. 555
556
P. A. HEDIN, C. S. NIEMEYER,R. C. GUELDNER, AND
A. C. THOMPSON
sex attractants; they appeared to be absent in that of females. Also, several other differences in the profiles were apparent. The logical extension of this approach was to conduct a gas chromatographic profile survey of a number of species of insects, sexed when feasible, encompassing several orders. Of course, some would not necessarily be producing any attractant when they were analysed, the pheromones might not survive steam distillation and solvent extraction, and any pheromones present might represent less than 1 per cent of the total volatiles and so escape observation. Nevertheless, the approach appeared to have some promise, and a phylogenetic application also appeared feasible. The only known exhaustive investigation of the entire volatile profile of an insect previous to the study of the boll weevil was that of KULLENBERG et al. (1970) who investigated the volatile components of the cephalic marking secretion of male bumblebees, Bombus sp. They reported acyclic mono-, sesqui-, and diterpenes as well as aliphatic normal chain compounds with 10 to 27 carbon atoms.
MATERIALS
AND METHODS
Collections of insects Insects in 5 to 50 g quantities were collected during the summer of 1970 by research entomologists stationed at the Boll Weevil Research Laboratory and at the Department of Entomology, Mississippi State University, State College, Miss. The female pink bollworms, Pectinophora gossypiella (Saund.), were provided by the Western Cotton Research Laboratory, Phoenix, Arizona. The insects were classified by the co-operating entomologists, and sexing was performed either by the entomologist or at his direction.
Preparation of insect essential oils On receipt, the insects were weighed, hilled in redistilled methylene chloride or ethyl ether, and stored at -20°C until steam distilled. The aqueous distillate was extracted three times with the organic solvent, the solvent was partially removed under vacuum, the sample was dried with anhydrous sodium sulphate, and the remaining material was reduced in volume to 5 ml under vacuum. The distillate was then stored at -20°C until analysis.
Gas chromatographic analysik For GLC, the sample was reduced in volume with a stream of N, and slight warming to about O-2 ml. Then, successive GLC analyses (2 to 5) were performed until reproducible profiles were obtained. As little as O-4 g of insects (e.g. 25 boll weevils) will provide sufficient distillate for two determinations if a microdistillation system is used, but in this study, we had 5 g samples or larger. Analyses were performed on an F.I.D. instrument with the following column and conditions:
VOLATILE FRACTIONS OF INSECTS
557
a 0.0032 x 6.145 m stainless steel column packed with 10% SE-30 on 60/80 mesh Chromosorb W treated with HMDS. Carrier gas flow N, at 50 ml/mm, column temperature 175”C, injector 185”C, detector 190°C. The profiles obtained from the SE-30 column were used in preparing the figures because with this column the retention volumes were lower than those obtained with the Carbowax-4000 columns. KOVATS (1961) indices (Ik) were calculated from the retention volumes, e.g. pentadecane = 1500, hexadecane = 1600, and used as a guide in the prediction of structures.
RESULTS
AND DISCUSSION
The insects in Table 1 are listed by chromatogram number and classified by order, family, genus, and species. The common names, weight per insect, and contributing entomologist are also included. Sex pheromones and/or identification work have been reported for seven of the insects surveyed as follows: the male boll weevil (TUMLINSON et al., 1969) ; the female tobacco budworm (GENTRY et aE., 1964); the female bollworm (GASTONand SHOREY,1964); the female pink bollworm (JACOBSON,1969) ; the female American cockroach (JACOBSON et al., 1963) ; a male fruit fly (MAYR, 1950); and a female fruit fly (STANDFUSS,1896). Figs. 1 to 7 contain the 28 gas chromatographic profiles obtained with the SE-30 column; they are grouped phylogenetically. Since the data were accumulated over 4 months, retention times may be subject to variances of perhaps 3 to 5 per cent. Table 2 gives the retention times in terms of Kovats’ indices and their structural equivalency so the chromatographic maxima may be better understood. Several phylogenetic similarities seem apparent. The Curculionidae (1 to 4), the Chrysomelidae (9 to 12), and the Coccinellidae (5 to 7) all possess similar profiles within families. Differences that can be attributed to sex seem obvious in all instances except for the American cockroach (19, 20). All the Coleoptera (1 to 12) possess a series of maxima which may be attributed in part to the monoterpenes which are known to be biosynthesized by this order. The components of the boll weevil sex attractant components (TUMLINSONet al., 1969) are included among maxima at 5 to 8 min (1); they are essentially absent in the female (2). Among the Lepidoptera, the large maxima in the male tobacco budworm (13) (the female is presumed to be attractive) and the female bollworm (16) are probably C,, to C,, alcohols and/or esters. The pheromones of this order are usually represented by these classes of compounds (and have this or a slightly higher molecular weight). Little is known about pheromones in Diptera, but both sexes of D. melanogaster (21 to 22) appear to produce unique compounds. The defence secretions of the Say stink bug (26) may be in part attributed to maxima 8 to 11, but 2-hexenal, a common repellent known to be produced by this insect, should have been eluted in the first 2 min. In summary, GLC profile analyses appear promising for surveys of insect pheromones. Th ey also appear promising for phylogenetic classification and for monitoring isolation work.
Curculio Curculio Hippodamia Coleomegilla Coleomegilla Chauliognathus Chrysomela Chrysomela Galerucella Diabrotica
Curculionidae
Curculionidae
Coccinellidae
Coccinellidae
Coccinellidae
Cantharidae
Chrysomelidae
Chrysomelidae
Chrysomelidae
Chrysomelidae
Noctuidae Noctuidae Noctuidae Noctuidae Gelechiidae
3
4
5
6
7
8
9
10
11
12
13 14 15 16 17
Heliothis Heliothis Heliothis Heliothis F’ectinophora
Anthomomus
Curculionidae
__-
2
Genus
Anthonomus
Family
Curculionidae
__-____ COLEOPTERA
(Saunders)
gossypiella
virescens
LEPIDOPTERA (F.) virescens (F.) zea (Boddie) zea (Boddie)
undecimpunctata howardi Barber
(Schrank)
xanthomelaean
scripta
scripta
DeG.
(Muls.)
(Muls.)
G&r.
pennsylvanicus
fuscilabrus
fuscilabrus
convergens
caryae (Horn)
caryae (Horn)
grandis Boh.
grandis Boh.
Species
beetle
Tobacco budworm Tobacco budworm Bollworm Bollworm Pink bollworm
Spotted cumcumber beetle
Cottonwood leaf beetle Cottonwood leaf beetle Elm leaf beetle
Soldier
Convergent lady beetle Spotted ladybird beetle Larvae
Q
d ? d ?
M
306.0
292.0
F. F. F. F. J.
G. G. G. G. C.
Maxwell Maxwell Maxwell Maxwell Keller
J. L. Hamer and L. N. Latson
R. B. Head
M
Hamer and N. Latson Hamer and N. Latson Head R. B. Head
J. L. L. J. L. L. R. B.
E. P. Lloyd
D. D. Hardee and 0. H. Lindig D. D. Hardee and 0. H. Lindig W. W. Neel and 0. M. Baughman W. W. Neel and 0. M. Baughman E. P. Lloyd
Entomologist
?
d
75.8
9.8
M
M
27.2
M*
weevil
Pecan
57.5 68.0
c3
weevil
Pecan
12.0
13.1
(mg)
Weight of insect
Q
Q
Boll weevil
Sex
6
name ~._
Boll weevil
Common
~--CLASSIFICATION OF INSECTS, THEIR SIZE,AND SOURCE _ ..__....__~----
1
Chromatogram No.
TABLE
Drosophilidae
Drosophilidae
Tabanidae Tabanidae
Membracidae
Pentatomidae
Chrysopidae
A mixture of spiders from cotton
21
22
23 24
25
26
27
28
* M = mixed.
Periplaneta Periplaneta
Blattidae Blattidae
19 20
Chrysopa
Chlorochroa
Stictocephala
Tabanus sp. Tabanus sp.
Drosophila
Drosophila
Microcentrum
Tettigoniidae
18
Genus
Family
Chromatogram No. Common name
Meig.
DIPTERA Meig.
oculata
Say
sayi Stll
ARACHNIDA
NEUROPTERA Goldeneye lacewing
Say stink bug
HEMIPTERA(HOMOPTERA) Buffalo treehopper
bubalus (F.)
melanogaster
melanogaster
rhombifolium
ORTHOPTERA (Sauss.) Broadwinged katydid American cockroach americana (L.) American cockroach americana (L.)
Species
TABLE 1 (cont.)
M
M
M
M
M M
?
8
6 e
M
Sex
13.2
300.0 100.0
1180.0
Weight of insect (mg)
E. P. Lloyd
J. L. Hamer
J. L. Hamer and L. N. Latson E. P. Lloyd
D. M. Elliot and C. H. Thomas D. M. Elliot and C. H. Thomas C. S. Niemeyer C. S. Niemeyer
H. B. Green H. B. Green
E. P. Lloyd
Entomologist
P. A. HEDIN, C. S. NIEMEYER,R. C. GUELDNER,ANDA. C. THOMPSON
560
.2 46 10 I 0
!
1
4
‘1
1”
8
12
1 ” I6 2G ” 24 ” 25 ” 32 ” 36
12 13 11 0
11 4
II
4
0
’
1 8
1
”
6
”
12
”
12
”
13
12
II
16
”
16
”
20
” 20
”
24
”
” 24
28
” 2s
”
32
I
,
I
40
I
44
14 ”
36
”
”
” 32
40
”
’
36
40
44
” 44
FIG. 1. Insect volatile profiles of (1) male boll weevils, (2) female boll weevils, (3) male pecan weevils, (4) female pecan weevils. A 0.0032 x 6.145 m SE-30 column; retention time in min.
TABLE 2--KOV.~TS
Rt
*
o-2 2-9
AND STRUCTURAL
EQUIVALENCY
O-900
GLC
RETENTION
TIMES
Compound
Ikt
900-l 300
OF
Solvent, C&C, hydrocarbons, Ci-C,
alcohols, carbonyls
Cl0 (Terpene) hydrocarbons, carbonyls, alcohols, C&i,, esters, boll weevil pheromones, bark beetle pheromones, verbenol, brevicomin Hydrocarbons,
C&C,,
254
1500-1700
Hydrocarbons, Ci,-Ci, alcohols, C&-esters, C1&1, pheromones of cabbage looper and fall armyworm
44-100
1700-2000
Propylure, bombykol, gyptol
* Retention time in min. t Kovats indices for 6.145 m SE-30 column.
C,,-C,,
alcohols
1300-1500
9-25
VOLATILE FRACTIONS OF INSECTS
561
-::
-Z -4 LD -:
-,” - hg,
-w
N -
-s
-::
-x
-2
:
-x
P
LD
-z
-n”
-4 IO.. -
-x -2
N
562
P. A. HEDIN, C. S. NIEMEYER, R. C. GUELDNER, AND A. C. THOMPSON
VOLATILE FRACTIONS OF INSECTS
564
P. A. HEDIN, C. S. NIEMEYER,R. C. GUELDNER,ANDA. C. THOMPSON
Acknowledgements-The authors thank FRANK BENCI and his staff at the Boll Weevil Research Laboratory for photographic and art work. REFERENCES GASTONL. K. and SHOREY H. H. (1964) Sex pheromones of noctuid moths-IV. An apparatus for bioassaying the pheromones of six species. Ann. ent. Sot. Am. 57,779-780. GENTRY C. R., LAWSONF. R., and HOFFMANJ. D. (1964) A sex attractant in the tobacco budworm. J. econ. Ent. 57, 819-821. HARDEED. D., MCKIBBEN G. H., GUELDNERR. C., MITCHELL E. B., TUMLINSONJ. H., and CROSSW. H. (1971) Boll weevils in nature respond to grandlure, a synthetic pheromone. J. econ. Ent. In press. HEDIN P. A., THOMPSONA. C., GUELDNERR. C., and MINYARDJ. P. (1972) Volatile constituents of the boll weevil. r. Insect Physiol. 18, 79-86. JACOBSONJ., BEROZAM., and YAMAMOTO R. T. (1963) Isolation and identification of the sex attractant of the American cockroach. Science, Wush. 139, 4849. JACOBSONM. (1969) Sex pheromone of the pink bollworm: biological masking by its geometric isomer. Science, Wash. 163, 190-191. Kov.4~~ E. az (1961) Zusammenhange zwischen Struktur und Gas chromatographischen Daten organischer Verbindungen. 2. anal. Chem. 181, 351-366. KIJLLENBERCB., BERGSTROM G., and STALLBERG-STENHAGEN S. (1970) Volatile components of the cephalic marking secretion of male bumblebees. Acta them. Stand. 24, 149-154. MAYR E. (1950) The role of the antennae in the mating behavior of female Drosophila. Evolution 4, 149-l 54. STANDFUSSM. (1896) Handbuch der Pakihrktischen Gross-Schmetterlinge fiir Forscher und Sammler, 2nd ed. Fischer, Jena. TUMLINSONJ. H., HARDEED. D., GUELDNERR. C., THOMPSONA. C., HEDIN P. A., and MINYARDJ. P. (1969) Sex pheromones produced by the boll weevil: isolation, identification and synthesis. Science, Wash. 166, 1010-1012. TUMLINSONJ. H., HARDEED. D., GUELDNERR. C., THOMPSONA. C., HEDIN P. A., and MINYARDJ. P. (1970) The boll weevil sex attractant. In Chemicals Controlling Insect Behavior (Ed. by BEROZAM.), pp. l-59. Academic Press, New York.