Journal of Chemical Ecology, Vol. 25, No.

12, 1999

TERPENE ATTRACTANT CANDIDATES OF Dioryctria

sylvestrella IN MARITIME PINE (Pinus pinaster)
OLEORESIN, NEEDLES, LIBER, AND HEADSPACE
SAMPLES

M. KLEINHENTZ,* H. JACTEL, and P. MENASSIEU

Laboratoire d'Entomologie Forestiere, INRA

BP45, 33611 Gazinet Cedex, France

(Received March 4, 1998; accepted August 12, 1999)

Abstract—Capillary GC analysis was used to determine the proportion and

quantity of terpenes in wood resin, pentane extracts from needles and liber,
and headspace samples of needles and pruning wounds in 24 thirteen-year-old
maritime pine. Fifteen different terpenes were identified in the samples.
Germacrene D and B-pinene were the dominant terpenic compounds in the
needles, while A-pinene and B-pinene were dominant in the liber and wood
resin. Headspace samples of both needles and pruning wounds contained
essentially monoterpenes. Only trace amounts of sesquiterpenes were found in
pruning wound emissions. The presence of an oxygenated compound, linalool,
in the pruning wound emissions is discussed, although this compound is not
found in Maritime pine essential oil. Twelve of the 24 trees studied were
infested by Dioryctria sylvestrella. Maritime pine susceptibility to this insect
was related to the terpene composition of the different samples.

Key Words—Pinus pinaster, wood resin, needle, liber, headspace, terpenes,

insect-plant interaction, Dioryctria sylvestrella.

INTRODUCTION

Terpenes are a major volatile component of coniferous resin and have often
been studied because of their multiple ecological roles (Harborne, 1990; Langen-
heim, 1994). They affect the behavior of a variety of insects (reviewed by
Speight and Wainhouse, 1989; Honda, 1995) and may act as attractants, ovipo-
sition stimulants, or defenses. Monoterpenes and sesquiterpenes have also been

*To whom correspondence should be addressed.

2741

0098-0331/99/1200-2741$16.00/0 © 1999 Plenum Publishing Corporation

2742 KLEINHENTZ, JACTEL, AND MENASSIEU

used as biochemical markers in forest genetics because many of these com-

pounds have been shown to be under strong genetic control and are little influ-
enced by environmental factors (Baradat et al., 1991; Hanover, 1992).
Terpenes have been shown to affect the behavior of Dioryctria species such
as D. amatella (Hulst) (Fatzinger and Merkel, 1985; Hanula et al., 1985), D.
horneana (Dyar) (Valterova et al., 1995), and D. abietivorella (Grote) (Shu et
al., 1997). The European stem borer, D. sylvestrella, is a major pest of maritime
pine plantations in Aquitaine, a region of France that represents the largest unit of
artificial forest in Europe (1.2 million ha). This oligophagous insect is restricted
to the genus Pinus in Eurasia (Menassieu et al., 1989) with occasional damage
to Picea spp. (Menassieu, unpublished observations). Unlike other Dioryctria
spp., D. sylvestrella does not damage cones or terminal shoots; the moth attacks
primarily the main stem and occasionally the branches. In southwest France,
adult moths emerge from mid-June to July and, after mating, eggs are deposited
approximately 10 days later (Menassieu et al., 1989). Eggs are laid individually
on the trunk under bark scales and hatch in 15-20 days (Zocchi, 1961). Larvae
tunnel through the bark into the phloem where they feed until October. The
larvae overwinter in their gallery and recommence boring activity in March until
the pupal stage (May-June). Adult moths emerge approximately 21-30 days after
pupation and have a life-span of 15-21 days (Menassieu et al., 1989). If larvae
succeed in girdling the stem, the tree may suffer from interruption of sap flow,
resulting in a decrease of the annual increment or in a malformation of the trunk
and in increasing the risk of windbreak (Baronio and Butturini, 1988).
Previous studies (Jactel et al., 1994, 1996a; Jactel and Kleinhentz, 1997)
have demonstrated that volatiles exuding from pruning wounds or bark cracks
may attract D. sylvestrella to host trees. Jactel et al. (1996b) related maritime
pine susceptibility to D. sylvestrella to the proportion of some monoterpenes
in the wood resin. Infested trees should exhibit either cracked bark or a high
percentage of terpinolene in the wood resin to release the requisite amount of
attractant into the atmosphere (Kleinhentz et al., 1998).
Wood resin composition appears to play an important role in host selec-
tion by D. sylvestrella. However, resin ducts are found throughout the maritime
pine tree and are located within the parenchyme of buds, leaves, and cortex
and in phloem and xylem (Pauly, 1962; Baradat and Marpeau-Bezard, 1988).
Within the tree, the nature and relative amount of these components varies within
organs or tissues (Bernard-Dagan, 1968; Pauly et al., 1973). Variability in ter-
pene composition also exists between trees (Bernard-Dagan et al., 1971; Baradat
et al., 1978) due to some physiological and genetic properties that control terpene
biosynthesis (reviewed by Baradat et al., 1991). Moreover, terpene emission rates
do not necessarily reflect abundance within the tissue. Environmental factors
(light, temperature, humidity) also influence the emission rate (Rasmussen, 1972;
Yokouchi and Ambe, 1984; Evans et al., 1985).
ATTRACTANTS OF Dioryctria 2743

The high cost of the intensive management of this maritime pine forest
makes it necessary to prevent stem borers from reducing the volume and the
quality of the forest products grown. The impossibility of chemical control,
because the insect is hidden under the bark for more than 11 months each year
and the uncertainty of biological controls, led us to investigate other pest man-
agement strategies, including the use of host chemicals as insect attractants.
Thus, the goal of this study was to identify candidate terpenes that could be
attractants to D. sylvestrella. For this, the terpene compositions of resin from
different tissues (wood, needle and liber) and the terpene emission generating
by pruning wounds and needles were determined.

METHODS AND MATERIALS

The study was conducted in maritime pine from a local provenance grown
on a mesophilous humid heath at the Station de Recherches Forestieres, Gazinet,
France. Trees were 13 years old; 12 infested trees (mean diameter at breast height
= 16.6 ± 2.6 cm) and 12 uninfested trees (mean diameter at breast height = 15.9
± 1.4 cm) were randomly selected from, respectively, the 44 infested and 183
uninfested trees initially identified for the study. All samples were taken during
the period of D. sylvestrella flight (from June to August).
Resin Samples. Wood resin was collected 2 m above ground level using
methods described by Jactel et al. (1996b) and oleoresin samples were stored
at -30°C under nitrogen until analyses were performed (Delorme and Lieutier,
1990). In addition, 5 g of needles (1 and 2 years old) and 5 g of liber were
collected at the 5-year-old internode of each tree. Resins were extracted in pen-
tane for 24 hr. The extracts were filtered through a 70-230 mesh (10 ml) col-
umn of SDS silica gel. Mono- and sesquiterpenes were eluted with 30 ml of
pentane-ether (98:2). The purified extracts were concentrated by evaporation
under moderate vacuum before FID-GC analyses.
Headspace Samples. Two different headspace techniques were developed
to collect terpene emissions from needles and pruning wounds. The first trap
had been designed for needle emission collection in the laboratory and was
adapted by Roques (personal communication). Needles (50 g) were enclosed
in a Tedlar bag perforated with air inlet holes and connected by a Teflon tube to
a glass cartridge filled with 100 mg XAD2. The trap was connected by Teflon
tubes to a vacuum pump (90 ml/min). Eight samples of needles of different
trees (infested or not), including a blank control, were analyzed simultaneously.
The effluvial sampling was run for 15 hr. The adsorbent was eluted using 400
U1 dichloromethane (CH2Cl2). The estimated trapping efficiency for XAD2 and
CH2Cl2 use is between 47 and 78% for low-molecular-weight compounds such
as terpenes (Figure 1), using the method described by Mathieu (1995).
2744 KLEINHENTZ, JACTEL, AND MENASSIEU
ATTRACTANTS OF Dioryctria 2745

Since the infestation rate for D. sylvestrella increases with pruning sever-
ity (Jactel et al., 1994, 1996a; Jactel and Kleinhentz, 1997), a second trap was
designed to collect volatiles emitted by pruning wounds from a living tree. The
sampling apparatus consisted of two polytetrafluoroethylene (PTFE) chambers
(500 ml) with an open rectangular shaped side (4 x 10 cm). The first chamber
was attached by wire to the trunk around a pruning wound. The second cham-
ber was attached to a glass plate (control chamber). The edge of the chamber,
which touched the tree or the glass plate was wrapped with PTFE to prevent gas
exchange between the chamber and the atmosphere. Incoming air was filtered
using an XAD16 cartridge. The chamber was also connected to a glass cartridge
filled with 100 mg XAD2, by a Teflon tube. The cartridge was finally connected
to a vacuum pump (180 ml/min) by Teflon tubes. The effluvial sampling was
run for 48 hours. The adsorbent was eluted using 400 U1 CH2C12.
FID-GC Analysis. The purified extract was analyzed by FID-GC (Hewlett
Packard 5890 series II; temperature program: 60°C to 90°C at a rate of 6°C/min,
90°C to 102°C at a rate of 3°C/min, 5 min at 102°C, and 102°C to 280°C at a rate
of 6°C/min; splitless injector 270°C; detector 290°C) on a 30 x 0.25 mm ID HP-1
column (He 15 psi, 2.5-Ul sample). The relative percentage of mono- and sesquiter-
penes was calculated by adding up all recorded terpene peaks. Components were
also quantified by comparing their peak area with that of an n-dodecane standard.
Statistical Analyses. All statistical analyses were performed using SAS
software (SAS Institute, 1996). Statistical analyses of percentage variables were
computed using the arcsin \/x transformation (Dagnelie, 1973). Nonparamet-
ric analyses of variance (Wilcoxon test) were used for oleoresin composition
data that were proportions of p-terpenes with the constraint Epi = 1. Correlation
between terpenes was estimated using the Pearson correlation coefficient. Mean
differences were tested using Scheffe's multiple range test.

RESULTS AND DISCUSSION

Fifteen terpenes or oxygenated compounds exhibited relative proportions

higher than 0.1% in the different tree tissues and emissions. In order of
increasing retention times, there were seven monoterpenes (A-pinene, camphene,
B-pinene, myrcene, D-3-carene, limonene, terpinolene), an oxygenated monoter-
pene (linalool) and seven sesquiterpenes (longipinene, A-copaene, A-cubebene,
longifolene, B-caryophyllene, A-humulene and germacrene D).

Within-Tree Variation in Terpene Composition of Maritime Pine Resin

The terpene compositions of resin from the different tissues (wood, nee-
dle, and liber) showed some qualitative differences. For example, longipinene
and longifolene were absent from the needle resin. In addition, D-3-carene and
A-copaene were absent from the liber resin (Table 1).
2746 KLEINHENTZ, JACTEL, AND MENASSIEU
ATTRACTANTS OF Dioryctria 2747

Few changes in the relative proportion of terpenes were recorded between

the wood and liber resins (Table 1). A-Pinene and B-pinene were the dominant
terpenic compounds in these two tissues, averaging 80% and 90% of the total
amounts of terpenes in the liber and wood resin, respectively. The terpene com-
position of these two resins was consistent with that found in the literature for
maritime pine liber resin (Bernard-Dagan, 1968) and wood resin (Jactel et al.,
1996b).
Needle resin composition was very different from the terpene profile of the
wood and liber resin (Table 1). A-Pinene and B-pinene averaged only 45% of
the total terpene amounts. Sesquiterpenes, which represented less than 5% in
the wood resin, averaged 45% in the needle resin. Germacrene D and B-pinene
were the dominant terpenic compounds in the needles. The terpene composition
of the needle resin was very similar to that demonstrated by Pauly et al. (1973).
This study has confirmed the existence of two resins that have different
terpene proportions. Baradat and Marpeau-Bezard (1988) also ranked maritime
pine resins into two categories. The primary resin, synthesized in primary tis-
sues such as the needles, is characterized by an equal proportion of mono- and
sesquiterpenes. The secondary resin, synthesized in secondary tissues such as the
liber and wood, is characterized by a high proportion of A-pinene and B-pinene
(between 85% and 95% of total terpenes).
Tree diameter was not correlated with terpene proportion in the three resin
profiles. Kleinhentz et al. (1998) have also shown a lack of any correlation
between tree diameter and terpene proportion in the wood resin. Pinaceae ole-
oresins generally consist of 50-90% nonvolatile compounds (essentially diter-
penes) and 10-50% volatile mono- and sesquiterpenes (Kramer and Kozlowski,
1979). In maritime pine, the terpene quantity in the wood resin averaged 160
mg/g resin (Table 2). Large differences existed between the different tissues.
These differences in terpene quantities resulted from differences in terpene
biosynthesis as well as from anatomical differences such as the number of resin
ducts and duct diameter (Baradat and Marpeau-Bezard, 1988). The quantity of
mono- and sesquiterpenes averaged 13 mg/g fresh needles, while it averaged
only 3 mg/g of fresh liber (Table 2). Baradat and Marpeau-Bezard (1988) have
also shown that primary tissues such as needles have a higher capacity for ter-
pene synthesis than secondary tissues such as liber and wood. Between the sec-
ondary tissues, wood had five to seven times higher terpene production, than did
liber (Baradat and Marpeau-Bezard, 1988).

Maritime Pine Terpene Emission

Foliage Emission. Only monoterpenes were detected in the needle emis-
sions. A-Pinene, B-pinene, D-3-carene, limonene, and myrcene, with low pro-
portions of camphene and terpinolene were present (Table 1). Buchbauer et
2748 KLEINHENTZ, JACTEL, AND MENASSIEU
ATTRACTANTS OF Dioryctria 2749

al. (1994) also identified only monoterpenes in Douglas needle headspace (in
decreasing proportions: sabinene, B-pinene, terpinolene, A-pinene, D-3-carene,
terpinene, and camphene). Larix decidua Mill, foliage emitted A-pinene, cam-
phene, sabinene, limonene and B-phellandrene (Rappaport et al., 1995). Mono-
terpene emissions from maritime pine foliage averaged 14.4 Ug/g/day on a fresh
weight basis (Table 2), which is approximately equivalent to 28 Ug/g/day on a
dry weight basis (Porte, personal communication). For species that emit mainly
monoterpenes, the emission rates usually range between 2.4 and 240 Ug/day
per leaf tissue dry weight (Lamb et al., 1985). Monoterpene emissions from five
Pinus species ranged from 60 to 260 Ug/g/day on a dry weight basis (Tingey
and Burns, 1980).
In maritime pine, significant correlations were found between the same
monoterpene proportions from needle emission and needle resin (Table 3),
except for limonene. The greatest difference between the two profiles was
the absence of sesquiterpenes from needle emissions. This absence could be
explained by the fact that only a few monoterpenes, particularly a- and B-pinene,
dominate terpene emissions during emission measurements (Zimmerman, 1979;
Tingey et al., 1980; Evans et al., 1985; Juuti et al., 1990). The relative abundance
of some monoterpenes in the gas phase has been attributed to their low boiling
points (Hanover, 1972). When there is no difference in the diffusion coefficient
between the two compounds, water solubility could also explain some of the
variations in emissions rates found for the monoterpenes (Tucker and Nelken,
1982).
Pruning Wound Emission. Like needle emissions, maritime pine pruning
wound emissions contained essentially A- and B-pinene and low proportions

TABLE 3. MATRIX OF CORRELATIONS OF PROPORTION OF SAME TERPENE IN Two

DIFFERENT PROFILES"

Terpene

Profile API CAM BPI MYR D3C LIM TPL LGF BCA

Needle resin 0.66 0.62 0.52 0.95 0.83 0.34 0.47

and needle ***b
emission
Wood resin 0.55 0.25 0.68 -0.42 0.34 -0.33 0.20 -0.14 0.32
and pruning ** ***
wound
emission
a
API: A-pinene, CAM: camphene, BPI: B-pinene, MYR: myrcene, D3C: D-carene, LIM: limonene,
TPL: terpinolene, LGF: longifolene, BCA: B-caryophyllene.
b
Two-tailed significance: *0.05, **0.01, ***0.001.
2750 KLEINHENTZ, JACTEL, AND MENASSIEU

of camphene, myrcene, limonene, and linalool. Traces of D-3-carene, terpino-

lene and two sesquiterpenes (longifolene and B-caryophyllene) were also iden-
tified (Table 1). Riba (1991) has reported that the atmosphere above a maritime
pine canopy contains A-pinene, B-pinene, D-3-carene and limonene. Undamaged
boles of Pinus contorta murrayana Engelm. emit B-phellandrene, R-cymene,
B-pinene, an oxygenated monoterpene, and A-pinene (Rhoades, 1990). Terpene
emissions from maritime pine pruning wounds averaged 16 Ug/cm2/day (Table
2). Rhoades (1990) estimated the total volatile flux emitted by the boles of intact
lodgepole pine at 6 ng/cm2/day. It appears that pruning significantly increases
terpene emissions. This increase could result from the elimination of bark as a
natural barrier. Emissions could also have been enhanced by the neosynthesis
of a traumatic resin (Cheniclet, 1987; Baradat and Marpeau-Bezard, 1988). In
pruning wounds, the wood resin exuding from several resin ducts might mix
with the resin of the induced reaction, although it has been demonstrated that
terpene composition of the neosynthesized resin is very similar to that of wood
resin (Cheniclet, 1987).
When terpene composition from pruning wound emissions and wood resin
are compared, only a few correlations were found: for A-pinene, B-pinene, and
myrcene proportions (Table 3). We also observed the almost complete absence
of sesquiterpenes from wound emissions, which could be explained by the lower
volatility and water solubility of these compounds compared to monoterpenes.
However, the most important difference consisted in the presence of linalool,
an oxygenated monoterpene, in the wound emissions, although this compound
is not found in maritime pine resin. The presence of linalool might be an arti-
fact. Pauly (1973) analyzed the volatile leaf oil of maritime pine and showed
that some alcohols, especially linalool, seemed to be formed during the steam
distillation process. In the method used, trapping was performed in situ for 48 hr
in a region (southwestern France) where humidity is very high at night, even in
summer. Terpenes are highly sensitive to oxidation and acid rearrangements on
the adsorbent (Stromvall and Petersson, 1992). Terpenic compounds may also
be transformed into oxygenated varieties through ozonolysis and interaction with
°O and °OH radicals (Lilian, 1972; Graedel, 1979). In our trap, oxydants such
as O3, NO3, etc., might have not been filtered. Added to the long air residence
times in the enclosure, it is possible that they may have reacted on the trap. The
proportion of wood resin terpinolene, which is often absent from surrounding
air due to its high gas-phase reactivity (Stromvall and Petersson, 1992), was the
only one that was both positively and significantly correlated with the proportion
of linalool from the wound emissions (r = 0.58; P = 0.003). Moreover, measure-
ments of needle emissions did not show a disappearance of terpinolene or an
appearance of linalool under controlled conditions. A chemical transformation
from terpinolene to linalool may occur in the trap.
ATTRACTANTS OF Dioryctria 2751

Host-tree Susceptibility to Dioryctria sylvestrella

The terpene composition of the resins and pine emissions showed no sig-
nificant quantitative or qualitative difference between D. sylvestrella trees that
were attacked and not attacked. However, some differences in terpene propor-
tions (semiquantitative) were significant (Figure 2). Attacked trees exhibited
significantly more terpinolene in the wood resin (df= 1; Prob > Z = 0.004).
Needle resin in attacked pines contained a significantly higher proportion of
B-caryophyllene (df= 1; Prob > Z = 0.001) and a lower proportion of B-pinene
( d f = 1 ; Prob > Z = 0.040). Linalool (df= 1; Prob > Z = 0.017) and camphene
(df = 1; Prob > Z- 0.035) were higher in the wound emissions of attacked trees
while myrcene (df = 1; Prob > Z = 0.026) and B-caryophyllene (df= 1; Prob > Z
= 0.003) were found in lower proportions. The terpene profile of the liber resin
and needle emissions did not differ between attacked and unattacked maritime
pines.
Because chemical control is not a realistic option (the pest is hidden under
the bark almost all year) and biological control is uncertain, selection of resistant
maritime pines represents a promising alternative method for pest management.
This study pointed out seven candidates that could be used as biochemical mark-
ers in indirect selection programs: terpinolene, myrcene, camphene, linalool,
B-pinene, and B-caryophyllene (twice). Because of the cumbersome sampling
method, using terpene proportions from pruning wound emissions as biochemi-
cal markers is not feasible. Needle resin composition varies with age and season
(Baradat and Marpeau-Bezard, 1988), and measurements would involve using
the terpene proportion from needle resin as a biochemical marker. Among the
seven candidates, terpinolene proportion in the wood resin is the easiest to mea-
sure and is genetically controlled (Kleinhentz et al., 1998). Its proportion is also
stable at 7 or 8 years of age (Baradat et al., 1972).
Another promising method of pest management is to identify attractants
that could be used in mass trapping. Short-range orientation towards and arrest-
ment on host plants by insects may involve responses to volatile compounds
emitted by the plant (Metcalf, 1987). Therefore, myrcene, B-caryophyllene, and
B-pinene, which are absent or in higher proportions in the pine emissions from
unattacked trees, cannot act as attractants. On the other hand, camphene, ter-
pinolene, and linalool were good candidates for attractants. Linalool and terpino-
lene were the tested terpenes that induced the best electrophysiological response
(the most stimulating) from D. sylvestrella females by EAG, while camphene
was the monoterpene that induced the worst response (not significantly different
from those elicited by the blank) (Jactel et al., 1996b). Because of the major
role played by pruning wounds in host selection (Jactel et al., 1994, 1996a), the
almost total absence of terpinolene from pruning wound emissions might appear
to be inconsistent with its role as an attractant. However, this concurs with the
2752 KLEINHENTZ, JACTEL, AND MENASSIEU

FIG. 2. Comparison of the mean terpene composition in wood resin, pentane extract (nee-
dles and liber), and headspace samples (needles and pruning wounds) from maritime pine
trees attacked or unattacked by D. sylvestrella. Standard errors are indicated by vertical
bars. One star above a pair of bars indicates a significant difference (P > 0.05). api:
A-pinene, bpi: B-pinene, cam: camphene, myr: myrcene, D3c: D-3-carene; lim: limonene,
tpl: terpinolene, Inl: linalool, Igp: longipinene, cop: copaene, cub: cubebene, Igf: longi-
folene, bca: B-caryophyllene, ahu: A-humulene and ger: germacrene D.
ATTRACTANTS OF Dioryctria 2753

FIG. 2. Continued.

hypothesis of the high gas-phase reactivity of terpinolene and may be an artifact

caused by technical problems. These compounds cannot be isolated as poten-
tial attractants. Moreover, terpinolene was the only terpene that was found in
higher proportions and quantities in attacked trees than in all of the analyzed
profiles (Figure 2). Kleinhentz et al. (1998) have also demonstrated the exis-
tence of a threshold for both tree diameter and terpinolene proportion in the
wood resin under which trees are not subject to D. sylvestrella attack. Wright
et al. (1975) have related the resistance of southern Scots pine varieties to D.
zimmermani (Grote) to the low levels of D-3-carene and terpinolene, and many
studies (Fatzinger and Merkel, 1985; Hanula et al., 1985; Shu et al., 1997; Sadof
and Grant, 1997) support the hypothesis that monoterpenes contribute to attacks
by Dioryctria species. Further behavioral experiments should be developed to
verify the real attractiveness of camphene, terpinolene, and linalool in the mar-
itime pine-D. sylvestrella relationship and evaluate their potential use in pest
management.
2754 KLEINHENTZ, JACTEL, AND MENASSIEU

REFERENCES

BARADAT, P., and MARPEAU-BEZARD, A. 1988. Le pin maritime Pinus pinaster Ait. Biologie et
genetique des terpenes pour la connaissance et I'amelioration de I'espece. Doctoral thesis 953.
Universite de Bordeaux, France, 444 pp.
BARADAT, P., BERNARD-DAOAN, C., FILLON, C., MARPEAU, A., and PAULY, G. 1972. Les terpenes
du pin maritime: Aspects biologiques et genetiques. II. Heredite de la teneur en monoterpenes.
Ann. Sci. For. 29(3):307-334.
BARADAT, P., MARPEAU, A., and BERNARD-DAGAN, C. 1978. Variation of Terpenes Within and
Between Populations of Maritime Pine. Biochem. Gen. Forest Trees, Umea, Sweden.
BARADAT, P., MARPEAU, A., and WALTER, J. 1991. Terpene markers, pp. 40-65, in Genetic Variation
in European Populations of Forest Trees. G. Muller-Starck and M. Ziehe (eds.). Sauerlander's
Verlag, Frankfurt am Main.
BARONIO, P., and BUTTIRINI, A. 1988. Gli insetti novici al bosco, Pinus spp. Pyralidae della corteccia
del pino, Dioryctria sylvestrella (Lepidoptera: Phycitinae). Monti Boschi 6:3-4.
BERNARD-DAOAN, C. 1968. Les essences du Pin maritime: Leur repartition dans les divers organes,
nature et evolution des monoterpenes. Bull. Soc. Bot. Fr., Colloq. Physiol Arbre (Paris, Novem-
bre 18-19, 1966), pp. 1-14.
BERNARD-DAOAN, C., FILLON, C., PAULY, G., BARADAT, P., and ILLY, G. 1971. Les terpenes du Pin
maritime: Aspect biologiques et genetiques. I. Variabilite de la composition monoterpeniques
dans un individu, entre individus et entre provenances. Ann. Sci. For. 28:223-258.
BUCHBAUER, G., JIROVITZ, L., WASICKY, M., and NIKIFOROV, A. 1994. Comparative investigation of
Douglas fir headspace samples, essential oils, and extracts (needles and twigs) using GC-FID
and GC-FTIR-MS. J. Agric. Food Chem. 42:2852-2854.
CHENICLET, C. 1987. Effects of wounding and fungus inoculation on terpene producing systems of
maritime pine. J. Exp. Bot. 38(194):1557-1572.
DAGNELIE, P. 1973. Theorie et Methodes Statistiques. Presses agronomiques de Gembloux Editeur,
Gembloux, Belgique. 463 pp.
DELORME, L., and LIEUTIER, F. 1990. Monoterpene composition of the preformed and induced
resin of Scots pine, and their effect on bark beetles and associated fungi. Eur. J. For. Pathol.
20:304-316.
EVANS, R., TIGEY, D., and GUMPERTZ, M. 1985. Interspecies variation in terpenoid emissions from
Engelmann and Sitka spruce seedlings. For. Sci. 31:132-142.
FATZINGER, C. W., and MERKEL, E. P. 1985. Oviposition and feeding preferences of the southern
pine coneworm (Lepidoptera: Pyralidae) for different host-plant materials and observations on
monoterpenes as an oviposition stimulant. J. Chem. Ecol. 11:689-699.
GRAEDEL, T. E. 1979. Terpenoi'ds in the atmosphere. Rev. Geophys. 17:937-947.
HANOVER, J. W. 1972. Factors affecting the release of volatile chemicals by forest trees. Mitt. Forsst-
kichen Bundes-Versuchanst Wien 97:625-644.
HANOVER, J. W. 1992. Applications of terpenes analysis in forest genetics. New For. 6:159-178.
HANULA, J. L., BERISFORD, C. W., and DEBARR, G. L. 1985. Monoterpene oviposition stimulant of
Dioryctria amatella in volatiles from fusiform rust galls and second-year loblolly pine cones.
J. Chem. Ecol. 11:943-952.
HARBORNE, J. B. 1990. Role of secondary metabolites in chemical defence mechanisms in plants,
pp. 126-134, in Bioactive Compounds from Plants. D. J. Chadwick and J. Marsh (eds.). Wiley-
Interscience, Chichester, UK.
HONDA, K. 1995. Chemical basis of differential oviposition by lepidopterous insects. Arch. Insect
Biochem. Physiol. 30:1-23.
JACTEL, H., and KLEINHENTZ, M. 1997. Intensive sylvicultural practices increase the risk of infes-
ATTRACTANTS OF Dioryctria 2755

tation by Dioryctria sylvestrella Ratz (Lepidoptera: Pyralidae), the maritime pine stem borer.
In Integrating Cultural Tactics into the Management of Bark Beetle and Reforestation Pests.
Proceedings of a symposium held September 1-3, 1996, in Vallombrosa, Italy.
JACTEL, H., MENASSIEU, P., and RAISE, G. 1994. Infestation dynamics of Dioryctria sylvestrella
(Ratz.) (Lepidoptera: Pyralidae) in pruned maritime pine (Pinus pinaster Ait). For. Ecol. Man-
age. 67:11-22.
JACTEL, H., MENASSIEU, P., RAISE, G., and BURBAN, C. 1996a. Sensitivity of pruned Maritime pine
(Pinus pinaster Ait) to Dioryctria sylvestrella (Ratz.) (Lepidoptera: Pyralidae) in relation to
tree vigour and date of pruning. J. Appl. Entomol. 120:153-157.
JACTEL, H., KLEINHENTZ, M., MARPEAU-BEZARD, A., MARION-POLL, F., MENASSIEU, P., and BURBAN,
C. 1996b. Terpene variations in maritime pine constitutive oleoresin related to host tree selection
by Dioryctria sylvestrella Ratz. (Lepidoptera: Pyralidae). J. Chem. Ecol. 22(5): 1037-1050.
JUUTI, S., AREY, J., and ATKINSON, R. 1990. Monoterpene emission rate measurements from a Mon-
terey pine. J. Geophys. Res. 95:7515-7519.
KLEINHENTZ, M., RAFFIN, A., JACTEL, H., BROQUET, A., and MENASSIEU, P. 1998. Terpinolene as a
potential marker in indirect selection for Dioryctria sylvestrella Ratz. (Lepidoptera: Pyralidae)
resistance in Maritime pine. Forest genetics 5(3): 147-155.
KRAMER, P. J., and KOZLOWSKI, T. T. 1979. Physiology of Woody Plants. Academic Press, New
York.
LAMB, B., WESTBERG, H. H., ALLWINE, G., and QUARLES, T. 1985. Biogenic hydrocarbon emissions
from deciduous and coniferous trees in the United States. J. Geophys. Res. 90(D1):2380-2390.
LANOENHEIM, J. H. 1994. Higher plants terpenoids: A phytocentric overview of their ecological roles.
J. Chem. Ecol. 20:1223-1280.
LILIAN, D. 1972. Photochemical smog and ozone reaction. Adv. Chem. Ser. 113:211-218.
MATHIEU, F. 1995. Mecanismes de la colonisation de 1'hote chez le scolyte du cafe Hypothenemus
hampei (Ferr.) (Coleoptera: Scolytidae). Doctoral thesis. Universite de Paris VII, 133 pp.
MENASSIEU, P., STOCKEL, J., and LEVIEUX, J. 1989. Donnees actuelles sur la biologie de Dioryctria
sylvestrella Ratz. (Lepidoptera: Pyralidae) ravageur du Pin maritime (Pinus pinaster Ait.) dans
le Sud Ouest de la France. J. Appl. Entomol. 107:238-247.
METCALF, R. L. 1987. Plant volatiles as insect attractants. CRC Crit. Rev. Plant Sci. 5:251-301.
PAULY, G. 1962. Etude de 1'appareil secreteur chez le Pin maritime. These 3eme cycle. Universite
de Bordeaux, 120 pp.
PAULY, G., GLEIZES, M., and BERNARD-DAOAN, C. 1973. Identification des constituants de 1'essence
des aiguilles de Pinus pinaster. Phytochemistry 12:1395-1398.
RAPPAPORT, N. G., JENKINS, M. K., and ROQUES, A. 1995. Cone and foliage volatiles from douglas-
fir and european larch: relationship to attack by cone and seed insects. In G. L. De Barr (ed.).
Proceedings of the Fourth IUFRO Cone and Seed Insects Conference, Beijing, China, 1992.
RASMUSSEN, R. 1972. What do the hydrocarbons from trees contribute to air pollution? J. Air Pollut.
Control Assoc. 22:537-543.
RHOADES, D. F. 1990. Analysis of monoterpenes emitted and absorbed by undamaged boles of lodge-
pole pine. Phytochemistry 29:1463-1465.
RIBA, M. L. 1991. Les isoprenoides dans la plante et dans 1'atmosphere. Etude de leur evolution en
milieu forestier. These de doctoral d'etat de 1'Institut National Polytechnique de Toulouse, 232
pp.
SADOF, C. S., and GRANT, G. G. 1997. Monoterpene composition of Pinus sylvestris varieties resis-
tant and susceptible to Dioryctria zimmermani. J. Chem. Ecol. 23:1917-1927.
SAS Institute. 1996. SAS User's Guide: Statistics, Version 5. SAS Institute, Cary, North Carolina.
SHU, S., GRANT, G. G., LANGEVIN, D., LOMBARDO, D. A., and MACDONALD, L. 1997. Oviposition
and electroantennogram responses of Dioryctria abietivorella (Lepidoptera: Pyralidae) elicited
by monoterpenes and enantiomers from eastern white pine. J. Chem. Ecol. 23:35-50.
2756 KLEINHENTZ, JACTEL, AND MENASSIEU

SPEIGHT, M. R., and WAINHOUSE, D. 1989. Ecology and Management of Forest Insects. Clarendon
Press, Oxford, 374 pp.
STROMVALL, A. M., and PETERSSON, G. 1992. Protection of terpenes against oxidative and acid
decomposition on adsorbent cartridges. J. Chromatogr. 589:385-389.
TINGEY, D. T., and BURNS, W. F. 1980. Hydrocarbon emissions from vegetation, pp. 24-30, in Effects
of Air Pollutants on Mediterranean and Temperate Forest Ecosystems. P. R. Miller (ed.). Pacific
Southwest Forest and Range Experiment Station, Berkeley, California.
TINOEY, D. T., MANNING, M., GROTHAUS, L. C., and BURNS, W. F. 1980. Influence of light and
temperature on monoterpene emission rates from slash pine. Plant Physiol. 65:4013-4018.
TUCKER, W. A., and NELKEN, L. H. 1982. Diffusion coefficients in air and water, pp. 17-1-17-25,
in W. J. Lyman, W. F. Reehl, and D. H. Rosenblatt (eds.). Handbook of Chemical Properties
and Estimation Methods: Environmental Behavior of Organic Compounds. McGraw-Hill, New
York.
VALTEROVA, I., SJODIN, K., VRKOC, J., and NORDIN, T. 1995. Contents and enantiomeric composition
of monoterpene hydrocarbons in xylem oleoresin from four Pinus species growing in Cuba.
Comparison of trees unattacked and attacked by Dioryctria horneana. Biochem. Syst. Ecol.
23:1-5.
WRIGHT, J. W., WILSON, L. F, and BRIGHT, J. N. 1975. Genetic variation in resistance of Scotch
pine to Zimmerman pine moth. Great Lakes Entomol. 8:231-236.
YOKOUCHI, Y, and AMBE, Y. 1984. Factors affecting the emission of monoterpenes from red pine
(Pinus densifom). Plant Physiol. 75:1009-1012.
ZIMMERMAN, P. R. 1979. Tampa Bay area photochemical oxidant study. Determination of emis-
sion rates of hydrocarbons from indigenous species of vegetation in the Tampa/St. Petersburg,
Florida, area. EPA 904/9-77-028. US Environmental Protection Agency, Atlanta, Georgia.
ZOCCHI, R. 1961. Contributi alla conoscenza degli insetti delle plante forestali V il genere Dioryctria
Zell (Lepi. Pyralidae) in Italia. Redia 46:9-13.