Journal of Chemical Ecology, VoL 18, No. 12, 1992

CHEMISTRY OF FRUIT FLIES: GLANDULAR SECRETION OF Bactrocera (Polistomimetes) visenda (HARDY)

S A B I N E K R O H N , J M A R Y T. F L E T C H E R , I W I L L I A M K I T C H I N G , I'* C H R I S T O P H E R J. M O O R E , 2 R I C H A R D A.1. D R E W , 2 a n d W I T T K O F R A N C K E 3

1Department of Chemistry, The University of Queensland, Queensland, 4072 Australia 2Department of Primary Industries Indooroopilly 4068 Australia 31nstitut fur Organische Chemie der Universitat Hamburg D-2000, Hamburg 13, Germany (Received January7, 1992; accepted June 10, 1992) Abstract--The major component ( > 90 % of volatiles) of the male rectal glandular extract of the nonpest species Bactrocera visenda (Hardy) is 3-methyl2-butenyl acetate, with minor components being the isomeric 3-methyl-3butenyl acetate, the homologous esters, 3-methyl-2-butenyl propanoate and 3-methyl-2-butenyl formate, along with 3-methyl-2-buten-l-ol, 3-methyl-2butenal, and 3-methylbutyl acetate. None of these compounds has been identified previously from a Bactrocera species, supporting the view that Bactrocera visenda is taxonomically distant from other Bactrocera species identified from the Australian mainland. This collection of compounds adds to the known types utilized by dipteran species and emphasizes their extensive biosynthetic capability.

Key Words--Bactrocera visenda, Diptera, Tephritidae, fruit fly, 3-methyl2-butenyl acetate.

INTRODUCTION

Within the identified species of fruit flies, a significant number are destructive pests of many forms of horticulture, and methods of monitoring and control are *To whom correspondence should be addressed. 2169 0098-0331/92/1200-2169506.50/09 1992PlenumPublishingCorporation

2170

KROHNET AL.

being sought that are based on the chemical cues that regulate phases of fruit fly biology (Drew et al., 1982). In this connection, the volatile compounds secreted and released from male rectal glands have been studied extensively and shown to be behavior-mediating in certain cases. On the other hand, many species are not of pest status, but study is still warranted to define the considerable biosynthetic capability exhibited by these insects (Blomquist et al., 1987; Blum, 1981; Bradshaw, 1985), as well as to delineate taxonomic features (Perkins et al., 1990a). In previous reports, we described studies of male rectal glands secretions from species that fall into each of these pests (Kitching et al., 1989) and nonpest categories (Perkins et al., 1990b; Krohn et al., 1991). Bactrocera (Polistomimetes) visenda (Hardy) is a medium-sized species (Drew, 1989) that is taxonomically very different from all other species identified from mainland Australia, but does resemble Bactrocera mesonotochra (Drew, 1989), a species localized in Morobe Province in Papua New Guinea. In view of this taxonomic isolation, it appeared that the chemistry of the male rectal gland secretion would be of interest, as previous studies (Perkins et al., 1990a) had shown that the gland contents can provide significant chemical taxonomic information. Bactrocera visenda is widespread in southern coastal areas of Papua New Guinea, northeastern coastal areas of Queensland, and the Torres Strait islands. It infests only native rain-forest hosts such as the "mangosteen" (Garcinia warrenii and Garcinia gibbsiae). In this report we describe the identification of the volatile chemical constituents of the male rectal gland, which are unlike the constituents of any other fruit fly species examined.

METHODS AND MATERIALS

Bactrocera visenda specimens were bred out in the laboratory at the Department of Primary Industries, Indooroopilly, Queensland, from infested mangosteen fruit (Garcinia warrenii and Garcinia gibbsiae) obtained from the rainforests of North Queensland. Gland Secretion Analysis and Identification. Sexually mature adult male flies were chilled briefly (0~ and their rectal glands were excised and stored in spectrograde pentane, as described previously (Kitching et al., 1989). Combined GC-MS examinations were performed using Hewlett-Packard HP5970 MSD, Finnigan Mat 1020 and double-focusing VG 70-750SE instruments with polar (BP21, FFAP) and nonpolar columns (DB5). Proton magnetic resonance spectra were recorded at 400 MHz (FT mode) on JEOL JNM-GX400 and Bruker WP400 spectrometers. Deuterochloroform was employed as solvent and chemical shifts (6 values) are relative to residual chloroform at 6 7.24. 13C NMR spectra were recorded at 100 MHz utilizing CDC13 as solvent, and chemical shifts are relative to the central component of the CDC13 triplet at 77.00 ppm.

FRUITFLIES

2171

Synthesis of Glandular Components. 3-Methyl-2-buten-l-ol (5), 3-methyl2-butenal (6), methylbutylacetate (7), and 3-methyl-3-buten-l-ol were obtained from the Aldrich Chemical Company and exhibited NMR and mass spectra in agreement with the structures. Compounds 1, 2, 3, and 4 have been characterized previously (Babin et al., 1980; Aljancic-Solaja et al., 1987; Nazarov et al., 1952). 3-Methyl-2-butenyl Acetate (1). A stirred solution of 3-methyl-2-buten- 1-ol (5) (1 g) in dry pyridine (20 ml) at 0~ was treated with acetyl chloride (0.94 g), and after about 12 hr was poured into ice-water and then extracted thoroughly with ether. The ether layer was washed with dilute HC1, NaHCO 3 solution, and water. The dried (MgSO4) ether solution was concentrated under reduced pressure and the ester purified by flash chromatography (hexane) to provide 1 (0.89, 54%). 13C NMR (CDC13): 170.96, 138.92, 118.54, 61.27, 25.64, 20.90, 17.87. IH NMR (CDC13): 1.66 (s, 3H), 1.71 (s, 3H), 1.99 (s, 3H), 4.51 (d, 2H, J = 7 Hz), 5.29 (t, 1H, J = 7 Hz). These IH NMR data are in excellent agreement with that reported (Babin et al., 1980). Mass spectrum: 128 (M +', 0.4), 86 (7), 85 (4), 71 (15), 69 (21), 68 (49), 67 (36), 53 (20), 43 (100), 41 (54), 39 (24). 3-Methyl-3-butenyl Acetate (2). 3-Methyl-3-buten-l-ol was treated with acetyl chloride as described above in the preparation of 1. Flash chromatography followed by preparative gas chromatography afforded a pure sample of 2. ~3C NMR (CDCI3): 171.04, 141.67, 112.16, 62.64, 36.65, 22.44, 20.92. IH NMR (CDC13): 1.73 (s, 3H), 2.01 (s, 3H), 2.31 (t, 2H, J = 7 Hz), 4.15 (t, 2H, J = 7 Hz), 4.71 (br s, 1H), 4.78 (br s, 1H). These ~H NMR data agree well with that reported (Aljancic-Solaja et al., 1987). Mass spectrum: 128 (M +', 0), 73 (2), 69 (4), 68 (52), 67 (31), 53 (10), 43 (i00), 41 (12), 39 (15), such data being in agreement with a reported mass spectrum (Aljancic-Solaja et al., 1987). 3-Methyl-2-butenyl Propanoate (3). Treatment of alcohol 5 with propanoyl chloride, in the manner described for acquiring 1, provided the propanoate 3. [Lit. bp 162-164~ mm (Nazarov et al., 1952)]. J3C NMR (CDC13): 174.33, 138.67, 118.65, 61.09, 27.45, 25.59, 17.81, 8.98. IH NMR (CDC13): 1.07 (t, 3H, J = 7.6 Hz), 1.64 (s, 3H), 1.69 (s, 3H), 2.25 (q, 2H, J = 7.6 Hz), 4.50 (d, 2H, J 7 Hz), 5.27 (br t, 1H). Mass spectrum: 142 (M +', t), 114 (1), 100 (3), 85 (5), 71 (3), 69 (42), 68 (58), 67 (41), 57 (100), 53 (16), 41 (71), 39 (22). 3-Methyl-2-butenyl Formate (4). 13C NMR (CDC13): 161:13, 139.91, 117.89, 60.74, 25.65, 17.93. 1H NMR (CDC13): 1.70 (s, 3H), 1.75 (s, 3H), 4.64 (d, 2H, J = 7 Hz), 5.34 (br t, J = 7 Hz), 8.04 (s, 1H). These IH NMR data are in agreement with literature values (Babin et al., 1980). Mass spectrum: 114 (M +', 1), 85 (9), 83 (1), 71 (38), 69 (31), 68 (59), 67 (50), 53 (36), 43 (28), 41 (100), 39 (56), such data being in agreement with that reported (Babin et al., 1980). =

2172

KROHN

ET

AL.

3-Methyl-2-butenal (6). Oxidation of 3-methyl-3-buten-l-ol with pyridinium chtorochromate provided 6, identical with the commercially available material. Distillation [bp 135 ~ lit. 133~ mm (Pauling et al., 1976)] followed by careful preparative gas chromatography yielded 6. ~3C NMR (CDC13): 190.96, 160.50, 128.03, 27.12, 18.80. IH NMR (CDC13): 1.92 (s, 3H), 2.11 (s, 3H), 5.82 (d, 1H, J = 8 Hz), 9.89 (d, 1H, J = 8 Hz). These ~H NMR data are in agreement with literature values (Pauling et al., 1976). Mass spectrum: 84 (M +', 100), 83 (48), 69 (6), 65 (2), 55 (82), 53 (18), 41 (76), 39 (80).

RESULTS AND DISCUSSION Combined GC-MS examination of a pentane extract of approximately 20 rectal glands from Bactrocera visenda males showed the presence of one dominant component ( > 90 % of volatiies), four minor components (each 1-2 %), and a number of trace components ( < 1%) (Figure 1). A library mass spectra search tentatively identified the major component as 3-methyl-2-butenyl acetate (1), and this was confirmed by synthesis and

100.0

,.-•/J•XOCOCH3 ~) (3) P.:C'OCI~CH~ (4) R=COH

.~L'-.....-"~ 0C0C~3 (~

fZ) I

[_

,

m'oo '

500

.~

i~,

, tI

1~oo

TIME (Sec.)

FIG. 1. Gas chromatograph of rectal gland extract form adult male Bactrocera (Polistomimetes) visenda (Hardy). Column and conditions: 30-m DB5 capillary column (J&W). Temperature programmed, 40~ 2 min then 10~ to 260~

FRUITFLIES

2173

coinjection of the authentic sample with the B. visenda extract. Three of the minor gland components showed significant mass spectral similarities with that of 1, with ions at 53 (C4H5), 67 (C5H8), and 69 (C5H9)being important in this regard. GC-MS comparisons with authentic compounds enabled the identification of these components as the isomeric acetate, 3-methyl-3-butenyl acetate (2), and two other esters of 1, 3-methyl-2-butenyl propanoate (3) and 3-methyl2-butenyl formate (4). The fourth minor component showed a mass spectral library match with 3-methyl-2-buten-1-ol (5), and the assignment was confirmed upon examination of a commercial sample of the alcohol and coinjection with the gland extract. An unsaturated Cs-aldehyde was deduced as the most likely structure of one of the trace components based on mass spectral interpretation. 3-Methyl-2butenal (6) proved to be identical with the natural component (coinjection and GC-MS). An additional trace component was identified as 3-methylbutyl acetate (7) on the basis of library matching and coinjection of an authentic sample. A number of less volatile trace components were tentatively identified and included farnesyl acetate and C~6 saturated and unsaturated acids. Some of these compounds are known as components of the aroma of certain fruits with, for example, 1, 2 and 5 in pepino and other fruits (Shiota et al., 1988) and 1, 2, 5, and 7 as minor headspace constituents of intact tree-ripened nectarines (Takeoka et al., 1988). cr aldehyde 6 is a volatile constituent of commercial proteinaceous baits for fruit flies (Morton and Bateman, 1981). With respect to insect derivation, compounds 1, 2 and 7 have been identified (Jang et al., 1989) as minor or trace headspace components from the odor of calling male Mediterranean fruit flies (Ceratitis capitata). Compounds 2 and 7 have also been identified from the venom of the European hornet (Wheeler et al., 1983), while 7 is an important component of the alarm pheromone of honeybee workers (Morse et al., 1967; Btum et al., 1978). Long-chain fatty acid esters of 5 are present in the Dufour's gland of halictid bees (Duffield et al., 1981, 1982). Interestingly, aza-analogs of 7 are also known from insects (Figure 2); 8 is the alarm pheromone of the wasp, Vespula squamosa (Heath and Landolt, 1988) and was also found in Dacus cucurbitae (Baker et al., 1982; Perkins et al., 1990b). The same compound and related amides, including the corresponding propanamide 9, were found in the rectal glands of Dacus tryoni

(8)

O)

(I0)

FIG. 2. Aza-analogues of 7 found in other insect species.

2174

KROHN ET AL.

and D a c u s n e o h u m e r a l i s (Bellas and Fletcher, 1979). Other volatiles showing a 1-aza-4-methylpentyl substructure, such as 10, are known from ponerine ants (Fates et al., 1984). To the best of our knowledge, and propyl ester (3) and formate (4) have not been identified previously from a natural source. The biosynthetic routes to volatile short-chain compounds present in insect species are unclear, but those with branched structures may be derived from isoprenoid units or amino acids (Blomquist et al., 1987; Bradshaw, 1985; Blum, 1981). In this regard, Attygalle et al. (1991) have demonstrated recently that 2H8-L-valine is incorporated with high efficiency into methacrylic and isobutyric acids in the defensive gland of a carabid beetle, S c a r i t e s s u b t e r r a n o u s and that the isotope distribution pattern is consistent with the saturated acid being the precursor of the unsaturated analog. Male rectal glands from other B a c t e r o c e r a species generally contain amides (Bellas and Fletcher, 1979) and/or C9-C13 spiroacetals, ketoalcohols, or diols (Kitching et al., 1989; Krohn et al., 1991; Perkins et al., 1990a,b). Thus the identification of the C 5 alcohols, esters, and aldehydes (1-7) in B. visenda supports the conclusion that this species is taxonomically quite distinct from other Australian species, and examination of B. m e s o n o t o c h r a (Drew, 1989) would be worthwhile from this point of view. In addition, the characterization of these compounds indicates a very diverse biosynthetic capability for Dipteran species (Tumlinson, 1989). Although no assessment of the biological activity of these components has been undertaken, it would be surprising if they lacked a role in the communication system of the species. Acknowledgments--The authors are grateful to Ms. Marlene Elson-Harris and Mr. Edie Hamacek of the Department of Primary Industries, lndooroopilly, Queensland, for collecting the infested fruit and rearing the flies.

REFERENCES ALJANCIC:SOLAJA,I., REY, M., and DREIDING,A.S. 1987. Short synthesis of (+) grandisol and (+)qineatin via a common intermediate. Helv. Chim. Acta 70:1302-1306.

ATTYGALLE,A.B., MEINWALD,J., and EISNER,T. 1991. Biosynthesis of methacrylic acid and isobutyric acids in a carabid beetle, Scarites subterraneus. Tetrahedron Lett. 32:4849-4852. BABiN, D., FOURNERON,J.D., and JULIA,M. 1980. Condensations biomimetiques: edification de squelettes terpeniques. Bull. Soc. Chim. Fr. I1:588-600. BAKER, R., HERBERT,R.H., and LOMER, R.A. 1982. Chemical components of the rectal gland secretions of male Dacus cucurbitae, the melon fly. Experientia 38:232-233. BELLAS,T.E., and FLETCHER,B.S. 1979. Identification of the major components in the secretion from rectal pheromone glands of the Queensland fruit flies Daeus tryoni and Dacus neohumeralis (Diptera: Tephritidae). J. Chem. Ecol. 5:795-803. BLOMQU1ST,G.J., DILLWITH,J.W., and ADAMS,T.S. 1987. Biosynthesis and endocrine regulation of sex pheromone production in Diptera, pp. 217-250, in G.D. Prestwich and G.J. Blomquist (eds). Pheromone Biochemistry. Academic Press, New York.

FRUIT FLIES

2175

BLUM, M.S. 1981. Chemical Defenses of Arthropods. Academic Press, New York. BLUM, M.S., FALES, H.M., TUCKER, K.W., and COLLINS, A.M. 1978. Chemistry of the sting apparatus in the worker honey bees. J. Apic. Res. 17:218-221. BRADSHAW,J.W.S. 1985. Insect natural products--compounds derived from acetate, shikimate and amino acids, pp. 655-703, in G.A. Kerkut and L.I. Gilbert (eds.). Comprehensive Insect Physiology, Biochemistry and Pharmacology, Vol, 11. Pergamon Press, Oxford. DREW, R.A.I. 1989. The tropical fruit-flies (Diptera Tephritidae: Dacinae) of the Australasian and Oceanian regions. Mem. Qld. Mus. 26:1-521. DREW, R.A,I., HOOPER, G.H.S., and BATEMAN, M.A. 1982. Economic Fruit Flies of the South Pacific Region. Queensland Department of Primary Industries, Brisbane. DUFFIELD, R.M., FERANDS,A., LAMB, C., WHEELER,J.W., and EICKWORTH,G.C. 1981. Macrocyclic lactones and isopentenylesters in the Dufour's gland secretion of halictine bees (Hymenoptera: Halictidae). J. Chem. Ecol. 7:319-331. DUFF1ELD, R.M., LA BERGE, W.E., CANE, J.H. and WHEELER,J.W. 1982. Exocrine secretions of bees IV. Macrocyclic lactones and isopentenylesters in Dnfour's gland secretions of Nomia bees (Hymenoptera: Halictidae). J. Chem. Ecol. 8:535-543. FALES, H.M., BLUM, M.S., BIAN, Z., JONES, T.H., and DON, A.W. 1984. Aliphatic amines from ponerine ants in the genus Mesoponera. J, Chem. Ecol. 10:651-665. HEATH, R.R., and LANDOLT, P.J. 1988. The isolation, identification and synthesis of the alarm pheromone of Vespula squamosa (Drury) (Hymenoptera: Vespidae) and associated behaviour. Experientia 44: 82-83. JANG, E.B., LIGHT, D.M., FLATH,R.A., NAGATA,J.T., and MON, T.R. 1989. Electroantennogram responses of the Mediterranean fruit fly, Ceratitis capitata to identify volatile constituents from calling males. Entomol. Exp. Appl. 50:7-19. KITCHING, W., LEWIS, J.A., PERKINS,M.V., DREW, R.A.I., MOORE, C.J., SCHURIG,V., KONIG, W.A., and FRANCKE, W. 1989. Chemistry of fruit-flies. Composition of the rectal gland secretion of (male) Dacus cucumis (cucumber fly) and Dacus halfordiae. J. Org. Chem. 54:3893-3902. KROHN, S., FLETCHER, M.T., K1TCHING, W., DREW, R.A.I., MOORE, C.J., and FRANCKE, W. 1991. Chemistry of fruit flies. Nature of the glandular secretion and volatile emission of Bactrocea (Bactrocera) cacuminatus (Hering). J. Chem. Ecol. 17:485-495. MORSE, R.A., SHEARER, D.A., BOCH, D.A., and BENTON, A.W. 196'7. Observations on alarm substances in the genus Apis. J. Apic. Res. 6:113-118. MORTON, T.C., and BATEMAN,M.A. 1981. Chemical studies on proteinaceous attractants for fruitflies, including the identification of volatile constituents. Aust. J. Agric. Res. 32:905-916. NAZAROV,I.N., RAKCHEEVA,V.N., and SHMONINA,L.I. 1952. Rearrangement of the allyl system. 5. Exchange reactions of 3,3-dimethylallyl chloride with amines, potassium cyanide, and salts of organic acids. Zhur. Obshchei Khim. 22:611-617. (Chem. Abst. 47:5370b, 1953). PAULING, H., ANDREWS,D.A., and HINDLEY, N.C. 1976. The rearrangement of a-acetylenic alcohols to ot,/3-unsaturated carbonyl compounds by silylvanadate catalysts. Helv. Chim. Acta 59:1233-1243. PERKINS,M.V., FLETCHER,M,T., KITCHING,W. DREW,R.A.I., and MOORE, C.J. 1990a. Chemical studies of the rectal secretions of some species of the Bactrocera dorsalis complex of fruit flies (Diptera: Tephritidae). J. Chem. Ecol. 16:2475-2487.' PERKINS, M.V., KITCHING,W., KONIG, W.A., DREW, R.A.I., and MOORE, C.J. 1990b. Chemistry of fruit-flies. Composition of the rectal gland secretions of (male) South-East Asian fruit flies. Re-examination of Dacus cucurbitae (melon fly). J, Chem. Soc. Perkin I: 11 l 1-1117. SHIOTA,n., YOUNG,H., PATERSON,V.J., and IRIE, M. 1988. Volatile aroma constituents of pepino fruit. J. Sci. Food. Agric, 43i343-354 (this paper summarizes the occurrence of 1, 2, 5, and 7 in other fruits).

2176

KROHN ET AL.

TAKEOKA,G.R., FLATH,R.A., GUNTERT,M., and JENNINGS,W. 1988. Nectarine volatiles: Vacuum steam distillation versus headspace sampling. J. Agric. Food. Chem. 36:553-560. TUMLINSON, J.H. 1989. Insect chemical communication systems. Pure Appl. Chem. 61:559-562. WHEELER, J.W., SHAMIN, M.T., BROWNE, P., and DUFF1ELD, R.M. 1983. Semiterpenoid esters from the venom of the European hornet, Vespa crabro (Hymenoptera: Vespidae). Tetrahedron Lett. 24:5811-5819.

Chemistry of fruit flies: Glandular secretion ofBactrocera (Polistomimetes) visenda (Hardy).

The major component (>90% of volatiles) of the male rectal glandular extract of the nonpest speciesBactrocera visenda (Hardy) is 3-methyl2-butenyl ace...
405KB Sizes 0 Downloads 0 Views