Ceratitis Capitata, to a SPECTRUM of PLANT VOLATILES
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Journal of Chemical Ecology, Vol. 14, No. i, 1988 ELECTROANTENNOGRAM RESPONSES OF THE MEDITERRANEAN FRUIT FLY, Ceratitis capitata, TO A SPECTRUM OF PLANT VOLATILES DOUGLAS M. LIGHT ~ ERIC B. JANG, 2 and JOSEPH C. DICKENS 3 USDA-ARS Western Regional Research Center 800 Buchanan Street Albany, California 94710 2 USDA-ARS Tropical Fruit and Vegetable Laboratory P.O. Box 4459 Hilo, Hawaii 96720 3 USDA-ARS Boll Weevil Research Laboratory P.O. Box 5367 Mississippi State, Mississippi 39762 (Received September 30, 1986; accepted January 15, 1987) Abstract--Electroantennograms (EAGs) were recorded from unmated, lab- oratory-reared, male and female Ceratitis capitata (medfly) in response to a range of Cl and C2 to C~z carbon chain-length aliphatic alcohols, aldehydes, acetates, and acids, and lactones, some of which are known volatiles from leaves and fruits. A large degree of EAG response uniformity between the sexes was observed, with only eight of the 70 compounds tested eliciting significantly larger amplitude EAG responses from female than male anten- nae. In general, for the five functional-group series tested, aldehydes and alcohols elicited greater responses than acetates, lactones, and acids. The unsaturated alcohols, aldehydes, acetates, and acids elicited equal or larger amplitude EAG responses than their comparable saturated compounds. For four of the functional-group series tested, the EAG response amplitude was significantly greater for a particular carbon chain length, with responsiveness to primary alcohols and aldehydes peaking at C6, acids peaking at C 5 6, and acetates peaking at both C5 and Cs. The EAG responses to both the 2- and 3-position monoenic alcohols peaked at C6 and Cs, while the secondary al- cohols peaked at C7. The greatest EAG responses of all compounds tested were elicited by monoenic C6 alcohols and aldehydes that are constituents of the "general green-leaf odor" that emanates from most plants. The potential adaptive benefit of selective sensitivity to green-leaf volatiles is discussed in regard to foraging behavior of medflie8. 159 0098-0331/88/0100-0159506.00/0 1988 Plenum Publishing Corporation 160 LIGHT ET AL. Key Words--Diptera, Tephritidae, Mediterranean fruit fly, Ceratitis capi- rata, plant volatiles, fruit volatiles, green-leafvolatiles, olfaction, electro- physiology,electroantennogram. INTRODUCTION The Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (medfly), is a polyphagous pest worldwide, attacking over 250 varieties of fruits, nuts, and vegetables (Hagen et al., 1981). It is widely assumed that medflies and other tephritid fruit flies use olfactory cues, in addition to visual cues (Cytrynowicz et al., 1984; Nakagawa et al., 1978; Prokopy and Roitberg, 1984; Prokopy and Economopoulos, 1976), to seek and assess habitat, food, and ovipositional re- sources (Prokopy and Roitberg, 1984). Such a phenomenon has been observed in studies of the behavior of the apple maggot, Rhagoletis pomonella (Walsh), in response to fruit model traps emanating apple volatiles (Prokopy et al., 1973; Reissig et al., 1982). However, there is little behavioral evidence (Bateman, 1972; Fdron, 1962; Keiser et al., 1975) to substantiate this assumption of ol- factory orientation to plant volatiles by C. capitata. The olfactory fragrance of a plant and, in particular, a ripening fruit, is a complex blend of usually over a hundred(s) detectable volatile compounds, pos- sessing various functional groups and ranging in structure from simple, short, straight carbon chains to complex multiring sesquiterpenes (e.g., Buttery, 1981; Van Straten and Maarse, 1983) (Table 1). Although the host-plant range of medflies is extremely broad and diverse, most of the host-plant and fruit odors will share some constituents from various classes of volatiles, e.g., aliphatic alcohols, aldehydes, acetates and acids, and monoterpenes, sesquiterpenes, and lactones (Buttery, 1981; Van Straten and Maarse, 1983; Visser et al., 1979) (Table 1). Little is known about the neurophysiological reception of plant volatiles by Diptera in general, let alone C. capitata and other tephritid species. To date, only four electrophysiological studies have been reported on tephritid fruit flies. Recently, Van Der Pets et al. (1984) utilized the electroantennogram (EAG) technique to study the responsiveness and sensitivity of male and female Dacus oleae (Gmelin) (olive fruit fly) to six components of its putative pheromones and five chemical analogs, some of which are commonly occurring plant vol- atiles. An EAG study on tephritids that tested "generally occurring fruit vola- tiles" was briefly reported by Guerin et al. (1983a). They recorded EAGs from female antennae of three species, D. oleae, Rhagoletis cerasi (L.) (cherry fruit fly), and C. capitata. Although 30 volatiles were tested (series of aliphatic C5 to C12 terminal position alcohols, aldehydes, and esters), only heptanal, oc- tanal, nonanal, and (E)-2-nonenal were reported to elicit the highest amplitude TABLE 1. SOURCE AND PURITY OF CHEMICALS USED IN ELECTROPHYSIOLOGICAL STUDIES AND THEIR PRESENCE IN A VARIETY OF HOST FRUITS OF Ceratitis capitata Presence in fruit of Chemical Compound purity (%)a Source~, Citrus,.,~ Papaya e Passion fruitd Peach a Atiphatic alcohols Ethanol 100.0 A + § § § Propan-l-ol 95.0 B + § § 2-Propen- 1-ol 99.0 C Butan-l-ol 100.0 D + § § § (E)-2-Buten-l-ol 92.3 B 3-Buten-l-ol 99.1 D Pentan-l-ol 99.5 D + § § + (E)-3-Penten-l-ol 97.0 C Hexan-l-ol 100.0 D + § § q- (E)-2-Hexen-l-ol 95.9 D + § § (Z)-2-Hexen-l-ol 99.5 D (E)-3-Hexen-l-ol 100.0 D + (Z)-3-Hexen- 1-ol 99.9 D + + § Heptan-l-ol 98.0 D + + + (+)-Heptan-2-ol 99.2 D § Octan-l-ol 100.0 D + § § + (E)-2-Octen-l-ol 92.0 C (E)-3-Octen-l-ol 84.7 C (+)-l-Octen-3-ol 99.5 D + Nonan-l-ol 100.0 D + (• 98.8 D + + Decan-l-ol 99.0 D + Undecan-l-ol 96.5 B + (• 98.0 E Dodecan-l-ol 96.9 F + + TABLE 1. (Continued) Presence in fruit of Chemical Compound purity (%)" Source b Citrus c, d Papaya e Passion fruit a Peach d Aliphatic aldehydes Propanal 54.0 B Butanal 67.0 B + + (E)-2-Butenal 93.9 F Pentanal 99.0 D + Hexanal 98.3 D + + (E)-2-Hexenal 98.6 D + + Heptanal 98.1 D + + Octanal 95.5 D + Nonal 91.1 G + + Decanal 97.9 D + Undecanal 91.0 D + Dodecanal 89.5 O + Aliphatic acids Formic 88.0 H § Acetic 99.9 I + + + Propanoic 99.8 J + 2-Propenoic 95.0 B Butanoic 99.0 J + + + Pentanoic 99 + K + + + Hexanoic 98.2 D + § § (E)-2-Hexenoic 99.0 D § + Heptanoic 98.2 D + § Octanoic 100.0 D + + + Nonanoic 94.1 D + § Decanoic 99 + D + + + Dodecanoic 99 + F + + Aliphatic esters Ethyl acetate 99 + H + + + + Propyl acetate 98.0 F + Butyl acetate 98,0 B + + + + Pentyl acetate 37.0 O + + + Hexyl acetate 92.0 O + + + (E)-2-Hexenyl acetate 99.7 L + Heptyl acetate 97.0 B Octyl acetate 98.4 O Nonyl acetate 97.0 C Decyl acetate 95.0 C Lactones gamma-Butyrolactone 99 + M gamma-Pentalactone 96.6 D + gamma-Hexalactone 98.0 C + + + gamma-Heptalactone 88.2 M + + + gamma-Octalactone 95.0 D + q- + gamma-Nonatactone 88.6 N + + + gamma-Decalactone 97.0 D + + + gamma-Undecalactone 97.1 N -k + mixture of C8-C12 delta-lactones 90 + P + + + aCapillary GLC analysis (12.5-m x 0.2-mm methyl silicone cross-linked column) at USDA-ARS-WRRC, Albany, California. h A, U.S. Industrial Chemicals; B, Eastman Kodak Co. ; C, synthesized at USDA-ARS-WRRC, Albany, California; D, Aldrich Chemical Co. ; E, Fluka Chemical Co.; F, Chem Service, Inc.; G, Fritzsche, Dodge and Olcott, Inc.; H, Fisher Scientific Co.; I, J.T. Baker Chemical Co.; J, Mallinckrodt Chemical Co.; K, Sigma Chemical Co.; L, CTS Organics; M, K&K Laboratories; N, Norda Chemical Co.; O, source presently unknown, tu file at USDA-ARS-WRRC, Albany, California; P, Oril Chemical Co. ~Keflbrd and Chandler (1970). dVan Straten and Maarse (1983). eFlath and Forrey (1977). 164 LIGHT ET AL. EAGs. In addition, Fein et al. (1982) recorded the EAGs ofR. pomenella to a group of seven esters they identified from apple odor. The purpose of this initial study was to investigate, by means of EAGs, peripheral olfactory selectivity of adult C. capitata to commonly occurring classes of host-plant and fruit volatiles, including the ubiquitous "general green- leaf volatiles" (Visser et al., 1979). The volatiles tested ranged from C1 or C2 to C12 saturated and unsaturated aliphatic alcohols, aldehydes, acids, acetates, and a series of lactones (Table 1). This research was further intended to provide a basis for studies of single-cell responses by surveying and assessing the degree that the antennal olfactory system of medflies is receptive to classes and/or particular plant/fruit volatiles. METHODS AND MATERIALS Insects. Pupae of C. capitata were obtained from a laboratory colony, periodically infused with feral flies, maintained at the USDA, Tropical Fruit and Vegetable Research Laboratory, Honolulu, Hawaii. Upon their arrival, pu- pae were segregated by sex and placed in separate cages. After eclosion, adult flies were provided sucrose cubes, hydrolyzed protein, and water until they were tested two to five days after emergence. Olfactory Stimuli. Table 1 lists the compounds tested, their supply sources, and purities. The compounds were dissolved in spectrometric grade hexane (that was additionally distilled and treated with an antioxidant) at a rate of one part test compound and nine parts hexane solvent, forming 10% volume per volume solutions. From these solutions, test cartridges were produced for each com- pound by pipetting 1-/zl aliquots onto separate 1 • 2-cm pieces of fluted, glass- fiber filter paper, which were then inserted into individual Pasteur pipets.