Hindawi Publishing Corporation Journal of Volume 2016, Article ID 6340579, 7 pages http://dx.doi.org/10.1155/2016/6340579

Research Article Infestation of Glycaspis brimblecombei Moore on Thirteen Eucalyptus Species and Their Relationship with the Chemical Composition of Essential Oils

Alejandro Lucia, Cecilia Naspi, Eduardo Zerba, and Héctor Masuh

Centro de Investigaciones de Plagas e Insecticidas, CITEDEF, CONICET, J. B. de La Salle 4397, Villa Martelli, B1603ALO Buenos Aires, Argentina

Correspondence should be addressed to Hector´ Masuh; [email protected]

Received 3 September 2015; Revised 23 December 2015; Accepted 6 January 2016

Academic Editor: Dmitri Y. Boudko

Copyright © 2016 Alejandro Lucia et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Glycaspis brimblecombei isapestinsectthataffectsEucalyptus genusandwasfirstlydetectedinArgentinain2005.Themain objective of this study is to determine the correlation between the level of infestation and chemical composition of essential oils extract. In an experimental plantation of 13 Eucalyptus species, the natural presence of the psyllid in the adaxial and abaxial faces of the leaves was determined and the mean number of individuals per species was calculated. The essential oils were extracted by hydrodistillation and analyzed for their chemical composition by gas chromatography-mass spectrometry. The results showed that 7 out of 13 species of Eucalyptus were not affected by G. brimblecombei: E. dunnii, E. globulus maidenii, E. globulus ssp. globulus, E. viminalis, E. cinerea, E. sideroxylon,andE. gunnii.ThemostaffectedEucalyptus species were E. tereticornis and E. camaldulensis whereas the infestation in E. grandis × E. tereticornis and E. grandis × E. camaldulensis was intermediate. E. saligna and E. grandis were the least affected species. The relative concentrations of the compounds 1,8-cineole and 𝛼-and𝛽-phellandrene in the essential oils are highly correlated to the rate of infestation with G. brimblecombei.

1. Introduction last ones plus other Eucalyptus species were also reported as G. brimblecombei hosts: E. brassiana Blake, E. camphora Glycaspis brimblecombei Moore (: ) is Baker, and E. mannifera Baker [7]. In California, the psyllid apsyllidknownasEucalyptus red gum lerp psyllid. This psyl- was also detected in E. diversicolor F. Muell., E. globulus lid has 5 nymphal instars (immatures). Adults (3 mm long) Labill., and E. sideroxylon Cunn. [7]. Eucalyptus camaldu- are brown or pale green, easily distinguished by their long lensis Dehnh. and E. tereticornis Smith, both known as “red genal cones on the head [1–3]. Both adults and immatures Eucalyptus,” were reported as highly susceptible to the psyllid cause great damage to foliage, characterized by discoloration attack; E. grandis Hill (ex Maiden) and E. saligna Smith in leaves, reduction in photosynthetic area, growth reduction, susceptibility are intermediate [5, 6, 8]. and dried sprouts [3]. This pest was originally detected in Australia by Moore [4], later in Queensland, New South Many Eucalyptus species contain essential oils; at least Wales, Brazil, and Chile [1, 5]. In February 2005, it was first twenty of them are commercially exploited for oil production detected in Argentina (Concordia Department, Entre R´ıos [9]. Essential oils are obtained by hydrodistillation of leaves Province) on E. camaldulensis Dehnh. [6]. and are constituted by a mixture of hydrocarbons, terpenes G. brimblecombei is associated exclusively with trees of andsesquiterpenes,andoxygenatedcompoundssuchasalco- Eucalyptus genus (Myrtaceae) and its natural environment hols, esters, ethers, aldehydes, ketones, lactones, and phenols involves a great number of species. Moore [4] detected this [9]. In Argentina, Eucalyptus has been used for reforestation psyllid in E. camaldulensis Dehnh., E. blakelyi Maiden, E. and as a wood resource since the 1950s [10]. nitens Maiden, E. tereticornis Smith, E. dealbata Cunn. ex Terpenoids can act as kairomones. Some monoter- Schauer, and E. bridgesiana Baker, among others [4]. These penes are responsible for attraction of bark insects such as 2 Journal of Insects

Scolytus ventralis LeConte, Dendroctonus frontalis Zimmer- the hydrodistillation method in a modified Clevenger-type mann, Dendroctonus ponderosae Hopkins, and Dendroctonus apparatus. The extraction procedure was performed during brevicomis LeConte [11]. Terpenes that are emitted by Pinus 70 minutes, time at which the yield remained constant sylvestris L. in response to the oviposition of pine sawfly [16]. In March 2006 (30 days after determination of lerp attract Chrysonotomyia ruforum Krausse, an egg parasitoid abundance), fresh leaves from each Eucalyptus individual [12]. Ulmus americana L.,whenaffectedbythepathogen were collected (800 g each). Five independent extractions, Ceratocystis ulmi Buisman, increases the production of an consisting of a mixture of nine individuals, were performed odor, which is different from healthy ones: 𝛼-cubebene for each Eucalyptus species. After extraction, the essential oil [13]. Some authors demonstrate attractiveness and ovipo- was separated from water and dried over anhydrous sodium ∘ sition preference of Glycaspis brimblecombei in Eucalyptus sulfate. The essential oils were maintained at −4 Cuntiltheir camaldulensis [14], and the role of cuticular waxes was also use. studied, related to genetic resistance of Eucalyptus globulus [15]. But none of these authors related these preferences to 2.4. Chemical Analyses of Essential Oils from Eucalyptus. The the chemical composition of their essential oils. chemical composition of Eucalyptus essential oils was deter- The main objective of this work was to study the differen- mined by gas chromatography coupled to mass spectrometry tial susceptibility of Eucalyptus species to G. brimblecombei using a GCMS-QM 5050A instrument (Shimadzu, Kyoto, and the relationship with the chemical composition of their Japan) and it is reported in previous publications of our essential oils. It is expected that a different chemical profile research group [16, 17]. Gas chromatography conditions were of the essential oils as well as the quantities of oil production as follows: injection of 0.4 𝜇L of a hexane solution of Euca- in Eucalyptus (Myrtaceae) influences the infestation rate by lyptus essential oil (1 mg/mL); capillary column HP-1 (cross- G. brimblecombei. Deep knowledge of this interaction could linked methyl silicone gum) (50 m × 0.32 mm × 0.52 𝜇m); and lead us to find chemical compounds useful for monitoring or helium as carrier gas (1.3 mL/min). The analytical conditions controlling the psyllid G. brimblecombei. were as follows: injector and interface temperatures of 250 ∘ and 280 C, respectively, split ratio of 13 : 1, initial isothermic ∘ temperature of 50 C during 10 min, programed temperature 2. Materials and Methods ∘ ∘ of 50 to 68 C(1C/min), programed temperature of 68 to ∘ ∘ ∘ 2.1. Plant Material. An experimental plot consisting of thir- 75 C(0.5C/min),programedtemperatureof75to250C ∘ ∘ teen species of genus Eucalyptus (one block per species (20 C/min), final isothermic temperature of 280 Cduring consisting of 9 individuals each) was constructed in July 2004. ∘ 󸀠 󸀠󸀠 ∘ 󸀠 󸀠󸀠 10 min, and electron impact 70 eV. Compounds identified The field was located at 34 33 42 S, 58 30 39 W, C I P E I N in the samples were confirmed by comparing their GC (Pest and Pesticides Research Center), Villa Martelli, Buenos retention times with standards through a comparison of the Aires, Argentina. The experimental plot was built by two- mass spectra with available NIST or Wiley mass spectral month-old and 15 cm high seedlings. library resident in the system. Quantification of essential oil The following Eucalyptus specieswereacquiredinPaul ∘ components (expressed in relative percentage on total area of Forestal Forest Nursery (INASE N J/5188, San Isidro, Buenos chromatogram) was carried out by peak area normalization Aires, Argentina): E. saligna, E. dunnii Maiden, E. globulus measurements. Based on these results, it can be observed that ssp. maidenii F. v. Muell., E. globulus ssp. globulus Labill., the main component in E. cinerea, E. globulus ssp. maidenii, E. viminalis Labill., E. tereticornis, E. camaldulensis,andtwo E. globulus ssp. globulus, E. sideroxylon,andE. viminalis is clonal hybrids: E. grandis × E. tereticornis and E. grandis × E. 1,8-cineole (Table 1). The hybrids E. grandis × E. tereticornis camaldulensis. Meanwhile, E. cinerea F. v. Muell. (ex Benth.), and E. grandis × E. camaldulensis are characterized by 1,8- E. sideroxylon Cunn., and E. gunnii Hook were purchased 𝛼 ∘ cineole and -pinene. The essential oils from E. tereticornis from Ferrari Hnos Forest Nursery (N INASE JL/2122, La and E. camaldulensis arecomposedof1,8-cineole,𝛼-and𝛽- Plata, Buenos Aires, Argentina). phellandrene, and p-cymene [16]. The chemical composition of the essential oils obtained from E. dunnii and E. gunnii is 2.2. Susceptibility of Eucalyptus spp. to Glycaspis brimble- a complex mixture; the main components are 1,8-cineole, 𝛾- combei. Between January and February 2006, when Euca- terpinene, p-cymene, and spathulenol [16]. Finally, E. grandis lyptus plants were 18 months old, the natural presence of is characterized by high concentrations of 𝛼-pinene [17]. psyllids was observed. In order to determinate the infestation of G. brimblecombei (i.e., lerp abundance), a single and direct 2.5. Statistical Analysis. The number of lerps for each species observation of plant individuals was performed. All plants was expressed by means of nine independent individuals were counted for their lerp abundance in a period of 1-2 days. replicates (±standard deviation). The data were tested for The lerps were 4 mm in diameter and whitish in appearance. their normality distribution by Kolmogorov and Smirnov The number of lerps found on both surfaces (adaxial and test (GraphPad Instat® Vers. 3.01, Copyright 1992–1998, San abaxial) of every leaf in the nine plants for each Eucalyptus Diego, California, USA). The homoscedasticity of data was species was recorded. The trees used for this study were analyzed by Bartlett method (GraphPad Instat Vers. 3.01). similarinsizeandfoliageabundance. These values were submitted to variance analysis (ANOVA) and then compared by Tukey’s HSD (honest significant dif- 2.3. Extraction of Essential Oils from Eucalyptus. The essential ference) test, using Statgraphics Plus Software Version 4.0 for oils of the thirteen Eucalyptus species were obtained by Windows (Statpoint Technologies, Inc., 1994–1999, Herndon, Journal of Insects 3

Table 1: Lerps abundance of Glycaspis brimblecombei on Eucalyptus 600.0 a species. Mean of number of lerps per individual ± SE. Different letters are significantly different (Tukey’s HSD mean multiple com- 500.0 parison test, 𝑃 < 0.05). Uninfested species are not showed. a 400.0 Eucalyptus species Number of lerps/individual ± standard deviation 300.0 E. grandis 3 ± 1.2c 200.0 E. saligna 4.6 ± 2.4c E. grandis × E. camaldulensis ± b 100.0 21.4 5.5 lerps/individual of Number × b b E. grandis E. tereticornis 25 ± 10.2 c c b 0.0 E. camaldulensis 356 ± 63.5a × E. tereticornis 446 ± 95.3a × E. cinerea 0 E. saligna E. grandis

E. globulus ssp. maidenii E. grandis E. tereticornis 0 E. tereticornis E. grandis E. camaldulensis E. globulus ssp. globulus 0 E. camaldulensis E. sideroxylon 0 Figure 1: Lerps abundance of Glycaspis brimblecombei on infested E. viminalis 0 Eucalyptus species.Eachbarrepresentsthemeanofnumberof lerps per individual ± SE. Bars with different letters are significantly E. gunnii 0 different (Tukey’s HSD mean multiple comparison test, 𝑃 < 0.05). E. dunnii 0 Uninfested species (E. dunnii, E. globulus ssp. maidenii, E. globulus ssp. globulus, E. viminalis, E. cinerea, E. sideroxylon,andE. gunnii) are not showed.

𝑃 < VA, USA). A value 0.05 was considered statistically Table 2: Essential oil yields, expressed as the mean of four replicates significant. The regression analysis was performed using the and standard deviation. Data from Lucia et al., 2008 [16]. same software. The quality of each regression model was 2 evaluated using the coefficient of determination𝑅 ( ), 𝑃 value, Eucalyptus species Yield (% w/w) ± standard deviation and 𝐹 ratio value (𝐹). E. cinerea 2.48 ± 0.09 E. globulus ssp. maidenii 2.25 ± 0.05 3. Results E. globulus ssp. globulus 1.66 ± 0.05 E. sideroxylon 1.65 ± 0.06 3.1. Susceptibility of Eucalyptus spp. to Infestation of Glycaspis E. viminalis 1.46 ± 0.05 brimblecombei. The infestation of G. brimblecombei on dif- E. grandis × E. tereticornis 0.88 ± 0.02 ferent Eucalyptus species is showed and presented (Table 1). E. dunnii 0.62 ± 0.02 FromthethirteenspeciesofEucalyptus that we studied, only E. tereticornis 0.59 ± 0.03 six were infested by the psyllid (Figure 1): E. tereticornis, E. grandis × E. camaldulensis 0.54 ± 0.04 E. camaldulensis, E. saligna, E. grandis,andthehybridsE. ± grandis × E. tereticornis and E. grandis × E. camaldulensis. E. camaldulensis 0.38 0.01 ± The infestation level in E. tereticornis (446 lerps/individual) E. saligna 0.36 0.01 and E. camaldulensis (356 lerps/individual) is significantly E. gunnii 0.21 ± 0.01 higher in comparison to the other species (𝑃 < 0.05). The hybrids E. grandis × E. tereticornis and E. grandis × E. camaldulensis showed lower infestation rates, 25 and 21.4 Six from the uninfested species exhibited a high essential lerps/individual, respectively, significantly different from the oil yield and a very high content of the terpene 1,8-cineole: other species (𝑃 < 0.05). Finally, E. grandis and E. saligna E. globulus ssp. maidenii (77.91%), E. globulus ssp. globulus showed the least infestation rates, 3 and 4.6 lerps/individual, (76.66%), E. sideroxylon (91.27%), E. viminalis (85.03%), E. respectively. Based on the lerps abundance per individual, cinerea (79.84%), and E. dunnii (48.48%) (Tables 2 and 3, the Eucalyptus species were classified into different categories: resp.). nonsusceptible (E. globulus ssp. maidenii, E. globulus ssp. The chemical composition and the respective yield ob- globulus, E. sideroxylon, E. viminalis, E. cinerea, E. dunnii, tained for each essential oil are showed (Tables 2 and 3). and E. gunnii), slightly susceptible (E. grandis and E. saligna), Qualitative differences in the infestation rate of G. brim- moderately susceptible (E. grandis × E. tereticornis and blecombei were observed in relation to the chemical com- E. grandis × E. camaldulensis), and highly susceptible (E. position of Eucalyptus essential oils. The most infested camaldulensis and E. tereticornis). species of Eucalyptus (E. tereticornis and E. camaldulensis) E. dunnii, E. globulus ssp. maidenii, E. globulus ssp. share several components, which are not found in other globulus, E. viminalis, E. cinerea, E. sideroxylon,andE. gunnii species: 𝛼-thujene (1.99–1.70%); sabinene (1.26–1.05%); 𝛼- were not infested by G. brimblecombei (0 lerps/individual). terpinene (<1%); 𝛽-phellandrene (22.64–16.34%); linalool 4 Journal of Insects

Table 3: Chemical composition of essential oils from 13 species of Eucalyptus, expressed as a relative percentage of the total area of the chromatogram (TIC). a: main constituents of essential oils, determined with CG-MS; b: numbers represent the botanical names; 1: E. grandis × E. tereticornis;2:E. grandis × E. camaldulensis;3:E. tereticornis;4:E. camaldulensis;5:E. saligna;6:E. cinerea;7:E. dunnii;8:E. gunnii;9: E. globulus ssp. maidenii;10:E. globulus ssp. globulus; 11: E. sideroxylon;12:E. viminalis;13:E. grandis; c: data from Lucia et al., 2008 [16]; d: data from Lucia et al., 2007 [17].

Eucalyptus speciesb Oil constituentsa 1c 2c 3c 4c 5c 6c 7c 8c 9c 10c 11c 12c 13d 𝛼-Thujene — — 1.99 1.7 — — — <1—— ——— 𝛼-Pinene 22.8 30.65 1.35 <1 12.95 4.94 5.45 4.06 5.84 11.09 1.95 1.16 52.71 Camphene — <1——————————— 𝛽-Pinene 3.29 1.1 — — — — — — — — — — — Sabinene — — 1.26 1.05 — — — — — — — — — Myrcene — — 1.54 1.21 — <1—<1 <1 <1—<1— 𝛼-Phellandrene — — 9.44 6.45 — — 1.37 7.05 — — — <1— Linalool acetate — 1.43 — — — — — — — — — — — 𝛼-Terpinene — — <1 <1————————— p-Cymene 1.92 2.77 14.51 17.93 21.25 1.63 4.43 12.28 2.31 — 1.77 1.93 9.7 𝛽-Phellandrene — — 22.64 16.34 — — — — — — — — — 1,8-Cineole 63.04 49.65 18.59 19.13 34.02 79.84 48.48 17.95 77.91 76.66 91.27 85.03 18.38 cis-Ocimene — — — — — — <11.14—— — — — 𝛾-Terpinene — — 2.25 1.87 20.13 — 12.96 3.35 4.82 <1—— 5 𝛼-Terpinolene — — — <1——<1—————— Linalool — — <13.08— — — — — — — — — p-Menthane-2-en-1-ol — — <1 <1————————— Fenchol <1 <1——————————— Cryptone — — 4.14 5.71 — — — — — — — — — 𝛼-Campholene aldehyde <1 <1——1.15———————1.18 trans-Pinocarveol 1.22 2.75 — — <1———————1.93 Borneol 1 1.42 — — <1———————2.95 4-Terpineol <1 <1 5.83 6.73 2.63 <1 3.56 1.86 1.58 <1 <1 <11.04 𝛼-Terpineol 2.55 4.05 1.59 — 3.45 3.79 3.59 1.38 6.04 1.45 2.61 1.75 5.67 𝛼-Terpineol acetate — — — — — 8.2 5.14 — — 3.98 — — — trans-Caryophyllene — — — — — — — 2.12 — — — — — Aromadendrene — — — — — — 3.40 1.21 — 1.11 — 1.98 — 𝛼-Humulene — — — — — — — <1—— ——— Alloaromadendrene — — — — — — <1 1.67 — — — — — trans-Carveol — — — — <1———————— Carvacrol — — — — <1———————— 4-Isopropylbenzaldehyde — — 1.32 2.6 — — — — — — — — — Phellandral — — <11.45— — — — — — — — — 𝛾-Elemene — — <1—————————— Caryophyllene oxide — — 1.69 — — — — — — — — — — Epiglobulol — — — — — — <1 <1—— —1.24— Spathulenol <1 <1 6.83 7.32 — — — 12.28 — — — — — Globulol <1 — — — — — 4.15 <1—— —2.49— Unidentified compounds 1.56 2.66 2.01 2.96 2.43 1.22 6.89 16.20 0.7 1.7 1.64 1.85 1.43

(<1–3.08%); p-menth-2-en-1-ol (<1%); cryptone (4.14–5.71%); were infested with G. brimblecombei.Thesimpleregres- 4-isopropylbenzaldehyde (1.32–2.60%); and phellandral (<1– sion analysis showed a statistically significant relationship 1.45%). between infestation rate and the following components: 𝛼- 2 The regression analysis between infestation rate and the phellandrene (𝐹:453;DG:5;𝑃 < 0.01; 𝑅 :99),𝛽-phe- 2 relative concentration of several compounds of essential llandrene (𝐹:829;DG:5;𝑃 < 0.01; 𝑅 :99).Themultiplere- oils was based only on the six Eucalyptus species that gression analyses showed a significant relationship between Journal of Insects 5 infestation rate and the relative concentration of 1,8-cineole It has been previously demonstrated that the most sus- 2 (𝑃 < 0.01); due to the values of 𝐹 (3450.5) and 𝑅 (99.9) ceptible species are E. camaldulensis and E. nitens;meanwhile of the new model (𝑃value model: 0.0003), it was decided to in E. grandis, E. globulus,andE. viminalis the susceptibility include it. The equation of the fitted model is as follows: is between low and medium [20]. The species that are number of lerps = −9.39 + 0.55 ∗ 1,8-cineole (%) + 58.19 ∗ commercially reared in Brazil and highly susceptible to the 𝛽-phellandrene (%) − 92.39 ∗ 𝛼-phellandrene (%). infestation are E. camaldulensis and E. tereticornis,both The statistics of each compound were as follows: 𝛼- known as “red Eucalyptus”; E. grandis and E. saligna are phellandrene (SE: 15.3; 𝑇: −6.02; 𝑃 < 0.05), 𝛽-phellandrene partially susceptible [5, 8]. (SE: 6.31; 𝑇:9.21;𝑃 < 0.05), and 1,8-cineole (SE: 0.13; 𝑇:4.17; The immatures of G. brimblecombei are covered by a wax 2 𝑃 < 0.05). The 𝑅-squared (𝑅 ) statistic indicates that the layer called lerp. The immatures use this lerp as protection model as fitted explains 99.9% of the variability in number from climatic conditions. This adaptation could be respon- of lerps. The adjusted 𝑅-squared statistic, which is more sible for the increment in the concentration of the essential suitable for comparing models with different numbers of oils under the lerp. When the insects are exposed to these vapors, the volatile substances enter through the spiracles independent variables, is 99.9%. The standard error of the and subsequently are transported to various tissues by the estimate shows the standard deviation of the residuals to be tracheas and tracheoles net arriving, finally, at their action 4.45.Themeanabsoluteerror(MAE)of1.82istheaverage site [21]. These kind of substances also circulate through the value of the residuals. The Durbin-Watson (DW) statistic hemolymph free, in solution, or absorbed by proteins, and tests the residuals to determine if there is any significant spreads throughout the body of the [22]. correlation based on the order in which they occur in your 𝑆 = 2.4 The fumigant toxicity of Eucalyptus essential oils against data file. Since the DW value ( ) is greater than 1.4, different insects was attributed to 1,8-cineole: Musca domes- there is not any significant autocorrelation in the residuals. tica Linnaeus [23], Pediculus humanus capitis De Geer [24], In determining whether the model can be simplified, notice 𝑃 Haematobia irritans Linnaeus [25], Aedes aegypti Linnaeus that the highest value on the independent variables is 0.05, [16], and Blattella germanica Linnaeus [26]. The correlation belonging to 1,8-cineole. between fumigant toxicity and vapor pressures (i.e., volatil- ity) of the components present in the essential oil was in 4. Discussion accordance with previous studies [27]. The toxic effect of volatile substances penetrating through the insect spiracles, According to these results, the composition of the essen- as part of the respiratory process, is strongly associated with tial oils in Eucalyptus should play an important role in their rate of volatility [28]. Consequently, Rice and Coats theinfestation with G. brimblecombei.Bothchemicaland [29] evaluated the knockdown effect of 22 monoterpenes biological insecticides have been used in an attempt to control against M. domestica and found a polynomial correlation infestations of the red gum lerp psyllid. Contact insecticides between their fumigant toxicity and their corresponding areknowntobeineffectiveastheimmaturesareprotected volatility, leading to the conclusion that the toxicity becomes by their shields (i.e., lerps) [1, 2]. Systemic insecticides have greater when the monoterpene increases in volatility. The been used with some success, but their use is of limited value fast knockdown effect produced by 1,8-cineole in head lice, inplantationforestryduetothehighcost. P. humanu s capiti s ,suggeststhatthismonoterpeneprobably The performance of adult psyllids was previously studied has a neurotoxic effect. The toxic action of monoterpenoids by in no-choice experiments on juvenile and adult leaves of may be mediated by pathways such as GABA receptors [30] Eucalyptus globulus anditwasconcludedthattheepicuticular and octopamine receptors [31, 32]. For such reason, the wax of juvenile leaves plays an important role in resistance partition properties of these compounds play an important to G. brimblecombei;howeveritwassuggestedthatother role in the penetration and distribution, until they reach unknown factors are probably involved [18]. The blend the action site. Therefore, the low infestation with G. brim- composition of essential oils could be, probably, a cause blecombei on some Eucalyptus species could be related to of tolerance of Eucalyptus species to G. brimblecombei.The the fumigant toxicity of 1,8-cineole present in the essential differences in the chemical composition could be responsible oils. for the female oviposition choice, explaining the different The interspecific hybrids of Eucalyptus have been used in infestation rates. E. gunnii, despite its low content of 1,8- forestry for decades [33]. The main areas planted with hybrids cineole, remained also uninfested. The chemical composition on a large scale are in Brazil, Congo, China, Indonesia, and of the essential oil of E. gunnii is similar to E. tereticornis South Africa. However, there are also small plantations in and E. camaldulensis, except for the lack of 𝛽-phellandrene other countries in Asia (e.g., Philippines, Vietnam, Thailand, in the first one. Besides, in comparison to the other species, and Malaysia) and South America (e.g., Argentina, Chile, E. gunnii has 16.20% of unidentified compounds. One of these Paraguay, and Uruguay) [34]. Besides growth, the main might be involved in the plant-insect interaction, preventing traits which have been improved by hybridization include the infestation by G. brimblecombei. propagation, coppicing, frost, drought and salt resistance, Different behavioral responses for an oviposition choice wood density, resistance to pests, and pulp yield [35]. On were observed in other insects that affect Eucalyptus.For the other hand, Eucalyptus hybridization has been used as a example, Mnesampela privata Guen´ ee´ females use terpenes technique to improve the quality and quantity of essential oils as chemical clues for oviposition choice by the female [19]. [36]. 6 Journal of Insects

5. Conclusions de eucalipto e em Eucalyptus camaldulensis sob diferentes temperaturas,” in Tesis de Mestre em Agronomia (Protec¸ao˜ de The lerp abundance was significantly lower in those species Plantas),p.51,FaculdadedeCienciasˆ Agronomicasˆ da UNESP, of Eucalyptus with high essential oil yield and very high Botucatu, Brazil, 2004. content of 1,8-cineole. The terpenes 𝛼-and𝛽-phellandrene [9]J.J.W.Coppen,“Eucalyptus oils. Flavours and fragrances of are the major components in the most infested Eucalyptus plant origin,” in Non-Wood Forest Products,pp.37–52,FAO, species. The low infestation of G. brimblecombei in those Rome,Italy,1995. Eucalyptus species rich in 1,8-cineole could be attributed to [10] M. Marco and L. Harrand, “El mejoramiento genetico´ forestal the toxicity of this component for the immatures that live y su aporte al desarrollo del sector forestal regional,” in Infor- under the lerp. For the hybrid species, the results suggest macion´ Forestal. EEA, INTA Concordia,vol.14,pp.55–61,1999. that the low infestation rate with respect to the pure species [11] E. J. Mac´ıas-Samano,´ Mediacion´ Semioqu´ımica Entre Insectos could be due to the increment of 1,8-cineole in the essential Descortezadores y Arboles´ de Con´ıferas. Relaciones Qu´ımicas oils. These findings are important because they give a clue Entre Organismos: Aspectos Basicos´ y Perspectivas de su Apli- about the susceptibility and tolerance to the attack of the cacion´ , Instituto de Ecolog´ıa, UNAM, 2001. psyllid in relation to the blend of terpenes in the essential [12] K. F. Raffa, B. H. Aukema, N. Erbilgin, K. D. Klepzig, and K. F. oil. Yield, profile, and chemical composition of essential oils Wallin, “Interactions among conifer terpenoids and bark beetles are genetically controlled by different enzymes related to across multiple levels of scale: an attempt to understand links terpene biosynthesis [37, 38]. 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