(2S,12Z)-2-acetoxy-12-heptadecene: Major sex pheromone component of pistachio twig borer, Kermania pistaciella

Article abstract of DOI:10.1007/s10886-006-9190-0

The sex pheromone of the pistachio twig borer, Kermania pistaciella (Lepidoptera: Oinophilidae), one of the most important insect pests of pistachio, Pistacia vera, in Turkey and Iran, was identified. In gas chromatographic-electroantennographic detection (GC-EAD) and GC-mass spectrometric analyses of pheromone gland extracts of female K. pistaciella from Turkey, (2S,12Z)-2-acetoxy-12-heptadecene was identified as the major candidate pheromone component. In field experiments in Turkey, lures containing synthetic (2S,12Z)-2-acetoxy-12-heptadecene attracted large numbers of male moths. Its attractiveness was significantly reduced by the presence of the R-enantiomer or of either enantiomer of the corresponding alcohol. (2S,12Z)-2-Acetoxy-12- heptadecene is the first pheromone component identified in the Oinophilidae and the first secondary acetate pheromone component identified in the Lepidoptera.

Full text of DOI:10.1007/s10886-006-9190-0

J Chem Ecol (2006) 32:2667–2677
DOI 10.1007/s10886-006-9190-0
(2S,12Z)-2-Acetoxy-12-heptadecene: Major Sex
Pheromone Component of Pistachio Twig
Borer, Kermania pistaciella
Regine Gries & Grigori Khaskin & Hassan Daroogheh &
Cafer Mart & Serpil Karadag & M. Kubilay Er &
Robert Britton & Gerhard Gries
Received: 17 February 2006 /Revised: 24 July 2006 /Accepted: 5 August 2006 /
Published online: 23 November 2006
#
Springer Science + Business Media, Inc. 2006
Abstract The sex pheromone of the pistachio twig borer, Kermania pistaciella
(Lepidoptera: Oinophilidae), one of the most important insect pests of pistachio, Pistacia
vera, in Turkey and Iran, was identified. In gas chromatographic-electroantennographic
detection (GC-EAD) and GC-mass spectrometric analyses of pheromone gland extracts of
female K. pistaciella from Turkey, (2S,12Z)-2-acetoxy-12-heptadecene was identified as the
major candidate pheromone component. In field experiments in Turkey, lures containing
synthetic (2S,12Z)-2-acetoxy-12-heptadecene attracted large numbers of male moths. Its
attractiveness was significantly reduced by the presence of the R-enantiomer or of either
enantiomer of the corresponding alcohol. (2S,12Z)-2-Acetoxy-12-heptadecene is the first
pheromone component identified in the Oinophilidae and the first secondary acetate
pheromone component identified in the Lepidoptera.
.
.
.
Keywords Pistachio twig borer Kermania pistaciella (2S,12Z)-12-Heptadecen-2-ol
.
.
(2R,12Z)-12-Heptadecen-2-ol (2S,12Z)-2-Acetoxy-12-heptadecene
(2R,12Z)-2-Acetoxy-12-heptadecene Sex pheromone Secondary acetate
.
.
:
:
R. Gries G. Khaskin G. Gries (*)
Department of Biological Sciences, Simon Fraser University, Burnaby, British Columbia,
Canada, V5A 1S6
e-mail: gries@sfu.ca
:
C. Mart M. K. Er
Department of Crop Protection, Faculty of Agriculture, Kahramanmaras Sutcu Imam University,
Kahramanmaras, Turkey
S. Karadag
Pistachio Research Institute, Gaziantep, Turkey
R. Britton
Department of Chemistry, Simon Fraser University, Burnaby, British Columbia, Canada, V5A 1S6
Present Address:
H. Daroogheh
Nelson Ave, West Vancouver, British Columbia, Canada, V7V 2R6

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Introduction
The pistachio twig borer, Kermania pistaciella (Lepidoptera: Oinophilidae), is one of the
most important insect pests in plantations of pistachio, Pistacia vera, in Turkey and Iran.
Females lay eggs close to shoot tips and fruit clusters. Larvae bore into and feed on terminal
buds and in twigs and shoots formed the previous year. Larval feeding causes abscission of
fruit buds and die-back of twigs (Küçükarslan 1966) and may directly damage fruit clusters
(Mart et al., 1995). Pupation takes place in cocoons attached to the branch surface near
larval exit holes. After 20–25 days, adults eclose, and females lay up to 60 eggs. The
1-month flight period of first-generation adults occurs in April–May (Küçükarslan 1966).
Many pistachio plantations in Turkey and Iran are treated annually with insecticides to
reduce damage caused by K. pistaciella larvae. For example, in southeast Turkey in 2005,
548,600 trees were sprayed with insecticides to suppress K. pistaciella populations
(Anonymous 2005). Identification of the K. pistaciella sex pheromone might be a first step
toward pheromone-based control tactics (e.g., attract and kill) of K. pistaciella populations.
We report the identification, synthesis, and field testing of (2S,12Z)-2-acetoxy-12-
heptadecene as the major component of the sex pheromone of K. pistaciella.
Methods and Materials
Experimental Insects In early March 2001, in several locations of the Nizip province of
Gaziantep (Turkey), sections of pistachio tree branches bearing pupal cocoons were cut off
and sent under import permit from Agriculture and Agri-Food Canada (Food Production
and Inspection Branch) to Simon Fraser University. Insects were kept at 24°C in mesh
cages under a photoperiod of 13-hr light/11-hr dark. Eclosed males were transferred to and
kept individually in filter paper-lined Petri dishes. The abdominal tip with pheromone gland
of 1- to 2-d-old females was removed and extracted with hexane for 15 min.
Analyses of Extracts Aliquots of 1 female equivalent (FE) of such extracts were analyzed
by coupled gas chromatographic-electroantennographic detection (GC-EAD; Arn et al.,
1975; Gries et al., 2002), employing a Hewlett-Packard (HP) 5890 gas chromatograph fitted
with a column (30 m×0.25 or 0.32 mm ID) coated with DB-5, DB-23, or DB-210 (J&W
Scientific, Folsom, CA, USA). Helium was used as carrier gas (35 cm/sec), with
temperature programs described in the captions of Figs. 2 and 4. GC-mass spectrometry
(MS) of compounds eliciting responses from antennae and of synthetic standards employed
a Saturn 2000 Ion trap GC-MS (Varian Instruments) fitted with the DB-5 column referred to
above.
Instrumentation Nuclear magnetic resonance (NMR) spectra of synthetic compounds were
1
taken on a Varian AS500 spectrometer at 499.77 MHz for H and 125.68 MHz for 13C
spectra with chemical shifts reported in parts per million relative to tetramethylsilane (1H, δ
0.00) and CDCl3 (13C, δ 77.00). Elemental analyses were performed using a Carlo-Erba
model 1106 elemental analyzer. Optical rotations were measured with a Perkin-Elmer 341
polarimeter.
The major candidate pheromone component in gland extracts was isolated by high-
performance liquid chromatography (HPLC), employing a Waters LC 625 HPLC equipped
with a Waters 486 variable wavelength UV–visible detector set at 210 nm, HP Chemstation

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software (Rev.A.07.01), and a reverse phase Nova Pak® C18 (3.9×300 mm) column
(Waters) eluted with acetonitrile (1 ml/min).
The absolute configuration of the HPLC-isolated natural pheromone component B and
the enantiomeric excess of synthetic standards were determined by analyses of samples on a
custom-made chiral GC column coated with a 1:1 mixture of heptakis-(2,6-di-O-methyl-3-
O-pentyl)-β-cyclodextrin and OV-1701 (König et al., 1992; Pietruszka et al., 1992).
Syntheses
(Z)-12-Heptadecen-2-ol (compound 2 in scheme 1, Fig. 1). Racemic (Z)-12-heptadecen-2-ol
was synthesized starting with (Z)-11-hexadecenal (1, scheme 1, Fig. 1; Bedoukian
Research, Danbury, CT, USA). Compound 1 (225 g, 0.915 mol) in 400 ml of ether was
added dropwise under argon at 0–10°C in 3 hr to a stirred solution of methylmagnesium
chloride (3 M in ether; 333 ml, 1 mol). The mixture was allowed to warm to room
temperature, quenched with 200 ml of saturated aqueous NH4Cl, and extracted with ether
(3×200 ml). Extracts were combined, washed with brine, dried (anhydrous MgSO4),
filtered, and evaporated in vacuo, affording 233 g of (Z)-12-heptadecen-2-ol (2, 99% pure
by GC, 0.906 mol, 99% yield) as a slightly yellowish oil. 1H NMR (CDCl3) δ: 0.89 (t, 3H,
J=7.1 Hz), 1.18 (d, 3H, J=6.3 Hz), 1.21–1.49 (m, 24H), 2.05 (m, 1H), 3.78 (m, 1H), 5.34
(m, 2H). 13C NMR (CDCl3) δ: 129.84, 129.82, 68.15, 39.34, 31.94, 29.74, 29.63, 29.59,
29.55, 29.51, 29.27, 27.16, 26.88, 25.76, 23.45, 22.32, 13.98. Anal. calcd. for C17H34O
(%): C, 80.24; H, 13.47; found C, 79.96; H, 13.24.
(Z)-2-Acetoxy-12-heptadecene (compound 3 in scheme 1, Fig. 1). Racemic (Z)-2-
acetoxy-12-heptadecene was synthesized by adding 94 ml (0.99 mol) of acetic anhydride
dropwise at room temperature to a stirred solution of 233 g (0.906 mol) of alcohol 2 in
100 ml (1.23 mol) of pyridine. After stirring for 6 hr, the reaction was quenched with water,
and the product was extracted with ether/hexane (1:1) (3×250 ml). Combined extracts were
washed successively with saturated aqueous NaHCO3, HCl (10%), water, and brine, then
dried (anhydrous MgSO4). Filtration and removal of solvents afforded 270 g of (Z)-2-
1
acetoxy-12-heptadecene (3, 99.5% pure by GC, quantitative yield). H NMR (CDCl3) δ:
0.87 (t, 3 H, J=7.0 Hz), 1.18 (d, 3 H, J=6.3 Hz), 1.21–1.35 (m, 22 H), 1.50 (m, 2 H), 2.00
(s, 3 H), 4.86 (m, 1 H), 5.33 (m, 2 H). 13C NMR (CDCl3) δ: 170.68, 129.79, 129.77, 70.99,
35.88, 31.92, 29.72, 29.50, 29.49, 29.47, 29.42, 29.24, 27.14, 26.87, 25.37, 22.30, 21.31,
19.90, 13.94. Anal. calcd. for C19H36O2 (%): C, 80.24; H, 13.47; found C, 79.96; H, 13.24.
(2S,12Z)-12-Heptadecen-2-ol, (2R,12Z)- and (2S,12Z)-2-acetoxy-12-heptadecene (com-
pounds 4, 5, and 6 in scheme 2, Fig. 1). Enantioselective syntheses of these compounds
were initiated by adding 70 mg of immobilized lipase Novozym 435 (10,000 units per
gram, Sigma Chemical, St. Louis, MO, USA; Xiao and Kitazume, 1997) to a stirred
solution of 0.600 g of racemic alcohol 2 (2.36 mmol; scheme 2, Fig. 1) and 0.53 ml of vinyl
acetate (5.75 mmol, Aldrich Chemical, Milwaukee, WI, USA) in 3 ml of hexane. After
stirring for 4 hr at 40°C, the Novozym-containing resin was filtered off, solvents were
evaporated in vacuo, and alcohol 4 (0.280 g) and acetate 5 (0.340 g) were separated by
flash chromatography [10 g of SiO2, ether/hexane (1:20) as eluent]. Subsequent acetylation
of alcohol 4 with acetic anhydride in pyridine and usual work-up (see above) afforded
0.320 g (1.08 mmol) of (2S,12Z)-2-acetoxy-12-heptadecene [6, 99.5% pure, 45.8% yield
20
(theory: 50%)], ½aꢀD ¼ þ0:83ꢁ (c 4.5; CHCl3). The NMR spectrum of acetate 6 was
consistent with that of racemic acetate 3. The enantiomeric excesses (ee) of 4, 5, and 6 were

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Fig. 1 Synthesis of racemic (Z)-12-heptadecen-2-ol (2) and (Z)-2-acetoxy-12-heptadecene (3) (scheme 1);
enantioselective syntheses of (2S,12Z)-heptadecen-2-ol (4), (2R,12Z)-2-acetoxy-12-heptadecene (5) and
(2S,12Z)-2-acetoxy-12-heptadecene (6) (scheme 2); and synthesis of (2S,12Z)-2-acetoxy-12-heptadecene (6)
from (2R,12Z)-2-acetoxy-12-heptadecene (5) (scheme 3)

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99, 95, and 99%, respectively, as determined by gas chromatography that separated
enantiomers with baseline resolution (Fig. 2).
Additional quantities of 6 were obtained by a three-step synthesis (scheme 3, Fig. 1),
based on inversion of the absolute configuration of 5. A mixture of 5 (0.280 g) and K2CO3
(1.0 g) in methanol (10 ml) was stirred for 18 hr at room temperature. After removal of 9 ml
of methanol in vacuo, ether (20 ml) and water (5 ml) were added to the reaction mixture.
The organic layer was washed with water and brine, dried (anhydrous MgSO4) and solvents
were removed in vacuo to afford (2R,12Z)-12-heptadecen-2-ol (7, 95% pure by GC). Without
further purification, alcohol 7 was mesylated at 0°C with methanesulfonyl chloride (1.5 ml)
in the presence of triethylamine (2 ml) in dichloromethane (10 ml). After 1 hr, the reaction
was quenched with aq. NaHCO3. Extraction with ether (50 ml), washing of the extract with
10% HCl, water, and brine, and drying and evaporation of solvent afforded the (R)-mesylate
8. Without further purification, mesylate 8 was stirred for 96 hr at 60–70°C in dry
dimethylformamide (10 ml) with 3 g of sodium acetate. Usual work-up of the reaction
mixture and purification of the product by flash chromatography yielded 0.180 g of pure 6
with 90% ee. Overall yield based on 5 was 64%.
Assignment of the absolute configuration of 6 was confirmed by an alternative synthesis,
which comprised converting (Z)-1-bromo-9-tetradecene to the Grignard reagent and then to
the monocuprate, adding (S)-propylene oxide and acetylating the resulting alcohol 4. This
Fig. 2 Representative recording of flame ionization detector (FID) and electroantennographic detector
(EAD: male Kermania pistaciella antenna) responses to one female equivalent of female K. pistaciella
pheromone extract. Chromatography: DB-5 column; splitless injection, temperature of injection port and
FID: 240°C; temperature program: 1 min at 50°C, then 20°C/min to 280°C

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approach also proved conducive to large-scale synthesis of 6 (Britton and Khaskin,
unpublished data).
Acetoxyheptadecanes (Table 1) were produced from the corresponding alcohols
synthesized by Grignard reactions of suitable aldehydes with n-alkylmagnesium bromides.
2-Acetoxyheptadecenes with various (E)- and (Z)-double bond positions (Table 1) were
synthesized from the corresponding hexadecenals according to scheme 1.
Field Experiments Field experiments were conducted in a pistachio orchard of the
Gaziantep Pistachio Research Institute in Gaziantep, Turkey. Experiments employed a
randomized complete block design with 8–10 replicates each. Delta-like traps were made
from 2-L milk carton (Gray et al., 1984), coated with Tanglefoot (The Tanglefoot, Grand
Rapids, MI, USA) and suspended from trees ∼1.5 m above ground with ∼16-m spacing.
Traps were baited with a gray sleeve stopper (West Pharmaceutical Services, Lionville, PA,
USA) impregnated with candidate pheromone components in HPLC-grade hexane.
Experiment 1 tested whether racemic (Z)-2-acetoxy-12-heptadecene was effective in
attracting male K. pistaciella. Experiment 2 tested whether the corresponding alcohol, (Z)-
12-heptadecen-2-ol, enhanced attractiveness of the acetate or was attractive by itself.
Experiment 3 explored which enantiomer of (Z)-2-acetoxy-12-heptadecene was attractive
and whether there were synergistic or inhibitory interactions between enantiomers.
Considering the inhibitory nature of the R-enantiomer in experiment 3, experiment 4
retested the S-enantiomer by itself and in combination with either one or both enantiomer(s)
of (Z)-12-heptadecen-2-ol. Doses used in field experiments are shown in Fig. 5.
Trap catch data were subjected to nonparametric analysis of variance by ranks
(Friedman’s test) followed by comparison of means (Scheffé test; Zar 1984; SAS/STAT
1988).
Table 1 Retention indices (RI) of synthetic standards and of components A and B in pheromone gland
extracts of female Kermania pistaciella
Compounds
RI on GC column
DB-23
DB-5
DB-210
A in Fig. 2
B in Fig. 2
2,376
2,355
2,309
2,376
2,355
2,342
2,348
2,342
2,363
2,450
2309
2284
2254
2240
2231
2227
1,890
2,011
2,021
1,890
2,011
2,002
2,007
2,014
2,019
2,105
2021
2001
1981
1970
1965
1962
2,093
2,290
2,286
2,093
2,290
2,281
2,284
2,276
2,297
2,386
2286
2262
2237
2226
2217
2215
B1 (= hydrogenated B)
(Z)-12-Heptadecen-2-ol
(Z)-2-Acetoxy-12-heptadecene
(Z)-2-Acetoxy-10-heptadecene
(Z)-2-Acetoxy-11-heptadecene
(E)-2-Acetoxy-13-heptadecene
(Z)-2-Acetoxy-13-heptadecene
Heptadec-1-yl acetate
2-Acetoxyheptadecane (= B1)
3-Acetoxyheptadecanea
4-Acetoxyheptadecanea
5-Acetoxyheptadecanea
6-Acetoxyheptadecanea
7-Acetoxyheptadecanea
a Mass spectra of 2-, 3-, 4-, 5-, 6-, and 7-acetoxyheptadecane did not have fragment ions diagnostic of the
acetoxy position.

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Results and Discussion
GC-EAD analyses of pheromone gland extracts from female K. pistaciella revealed two
components (A and B) that elicited strong responses from male K. pistaciella antennae
(Fig. 2). The mass spectrum of compound B (Fig. 3) with diagnostic fragment ion m/z 61
indicated an acetate functionality. Moreover, fragment ion m/z 236 [MW of C19H36O2 (296)−
60], instead of m/z 238 [MW (298)−60] expected for a saturated compound (Table 1),
suggested that compound B had one double bond.
Hydrogenation (Millar and Haynes, 1998 and references cited therein) of pheromone
gland extract, followed by repeated GC-EAD and GC-MS analyses, revealed a new EAD-
active compound (B1) with different retention characteristics (Table 1). B1 did not co-
chromatograph with synthetic heptadec-1-yl acetate, suggesting that it might be a saturated
secondary or tertiary acetate. It was determined to be 2-acetoxyheptadecane by GC-MS
analyses and comparison of its GC retention indices (Table 1) with those of seven synthetic
acetoxyheptadecanes (acetoxy group at C1, C2, C3, C4, C5, C6, and C7, respectively).
To determine the position of the double bond in 2-acetoxyheptadecene (B), HPLC-
isolated B was treated with dimethyl disulfide (DMDS; Dunkelblum et al., 1985). The mass
spectrum of the DMDS-derivative (MW=390) had adduct fragment ions [m/z 117 (26%)
(CH3–S=CH–(CH2)3–CH3)+; m/z 213 (100%) (273-HOAc)+; m/z 273 (15%) (CH3–
CHOAc–(CH2)9–CH=S–CH3)+; and m/z 390 (M+, 16%)] that revealed a double bond at
C12. Synthetic (Z)-2-acetoxy-12-heptadecene, but not the (E)-isomer, and none of (Z)-2-
acetoxy-10-heptadecene, (Z)-2-acetoxy-11-heptadecene, and (Z)- or (E)-2-acetoxy-13-
heptadecene co-chromatographed with female-produced B (Table 1).
Comparative GC analyses of synthetic racemic (Z)-2-acetoxy-12-heptadecene, the
synthetic S-enantiomer, and insect-produced B revealed that female K. pistaciella produces
the S-enantiomer of B (Fig. 4). Component A (Fig. 2) had retention indices (Table 1)
Fig. 3 Ion trap mass spectrum of compound B in Fig. 2 (compound 6 in Fig. 1)

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Fig. 4 Flame ionization detector (FID) chromatograms of racemic (Z)-2-acetoxy-12-heptadecene (3 in
scheme 1, Fig. 1), (2S,12Z)-2-acetoxy-12-heptadecene (6 in schemes 2, 3; Fig. 1); and HPLC-isolated, insect-
produced B (Fig. 2); chromatography: custom-made GC column coated with a 1:1 mixture of heptakis-(2,6-
di-O-methyl-3-O-pentyl)-β-cyclodextrin and OV-1701 (König et al., 1992; Pietruszka et al., 1992); splitless
injection; temperature of injection port and FID, respectively: 240 and 250°C; temperature program: 1 min at
100°C, then 10°C/min to 180°C (20 min)
indicative of an alcohol functionality. It was shown to be (Z)-12-heptadecen-2-ol by
comparison of its GC and GC-MS data with those of authentic 2.
In field experiment 1, traps baited with racemic (Z)-2-acetoxy-12-heptadecene (3)
captured large numbers of male K. pistaciella (Fig. 5). The corresponding alcohol, (Z)-12-
heptadecen-2-ol (2), was not attractive, and a 1:1 (w/w) mixture of 2 and 3 (as found in
pheromone gland extracts) was significantly less attractive than 3 alone (Fig. 5, experiment
2). However, the addition of different ratios of alcohol 2 to the acetate 3 may have
enhanced, instead of reduced, the attractiveness of the lure. This remains to be determined
by further field tests. (2S,12Z)-2-Acetoxy-12-heptadecene attracted many males, whereas
the R-enantiomer was not attractive and, when added to the S-enantiomer, significantly
reduced its attractiveness (Fig. 5, experiment 3). Attractiveness of the S-acetate was reduced
by either or both enantiomers of the corresponding alcohol (Fig. 5, experiment 4).
(2S,12Z)-2-Acetoxy-12-heptadecene is the first secondary acetate reported as a sex
pheromone component in the Lepidoptera. In contrast, secondary alcohols have been
identified in several species of the Lepidoptera, including Stigmella malella [(S)-(E)-6,8-
nonadien-2-ol and (S)-(Z)-6,8-nonadien-2-ol (Tóth et al., 1995)], Eriocrania cicatricella
[(2R)-heptan-2-ol (Zhu et al., 1995)], Eriocrania sangii [(2S,6Z)-nonen-2-ol (Kozlov et al.,
1996)], Eriocrania semipurpurella [(2S,6Z)-nonen-2-ol and (2R,6Z)-nonen-2-ol (Kozlov
et al., 1996)], and Orgyia detrita [(11S,6Z,9Z)- and (11R,6Z,9Z,)-6,9-heneicosadien-11-ol
(Gries et al., 2003)].
Straight-chain secondary acetate sex pheromones also commonly occur in cecidomyiid
midges, including the Hessian fly Mayetiola destructor [(2S,10E)-2-acetoxy-10-tridecene;
Foster et al., 1991; Harris and Foster, 1991; Millar et al., 1991], pea midge Contarinia pisi
[(S,S)-2,11-diacetoxytridecane, (S,S)-2,12-diacetoxytridecane and 2-acetoxytridecane;

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Fig. 5 Mean numbers (+SE) of male Kermania pistaciella captured per experimental period in experiments
1–4 in sticky traps baited with various candidate pheromone components, placed in pistachio orchards of the
Gaziantep Pistachio Research Institute in Gaziantep, Turkey. In each experiment, bars with different letter
superscripts are significantly different; nonparametric analysis of variance by ranks (Friedman’s test)
followed by comparison of means (Scheffé test), P<0.05 (Zar 1984; SAS/STAT 1988)

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Hillbur et al., 1999, 2000, 2001], Douglas fir cone gall midge Contarinia oregonensis
[(2S,4Z,7Z)-2-acetoxy-4,7-tridecadiene; Gries et al., 2002], Aphidoletes aphidimyza [(R,S)-
2,7-diacetoxytridecane; Choi et al., 2004], swede midge Contarinia nasturdii [(S,S)-2,9-
diacetoxyundecane, (S,S)-2,10-diacetoxyundecane, (S)-2-acetoxyundecane; Hilbur et al.,
2005], and western red cedar cone midge Mayetiola thujae [(S,S)-2,12-diacetoxyheptade-
cane, (S,S)-2,13-diacetoxyheptadecane, (S,S)-2,14-diacetoxyheptadecane; Gries et al.,
2005]. Cecidomyiid midges and K. pistaciella belong to discrete taxonomic orders (Diptera
and Lepidoptera, respectively) of the Insecta. However, these two orders are related, which
might explain the similarity of some of their sex pheromone structures.
With the K. pistaciella pheromone identified and shown to attract large numbers of male
moths, it may be possible to develop pheromone-based tactics for control of K. pistaciella.
In a 75-ha experiment in commercial pistachio orchards in Iran in 2005, plots treated with a
proprietary attract-and-kill formulation containing chiral pheromone had significantly fewer
larvae-infested fruit bunches (>120,000 assessed) than insecticide-treated plots or control
plots (unpublished data; two PCT patent applications filed in 2006).
Acknowledgements We thank Wittko Francke for lending us a custom-made chiral column and for review
of the manuscript, Eberhard Kiehlmann and one anonymous reviewer for constructive comments, Micky
Yang for elemental analyses, Sharon Oliver and Renée Picard for word processing, and Bob Birtch for
graphical illustrations. The research was supported by a Discovery grant from the Natural Sciences and
Engineering Research Council of Canada (NSERC) and by an NSERC-Industrial Research Chair to G.G.
with Phero Tech Int., SC Johnson Canada, and Global Forest Science as industrial sponsors.
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