Interspecific hybridization of Eucalyptus as a potential tool to improve the bioactivity of essential oils against permethrin-resistant head lice from Argentina

The essential oils extracted from Eucalyptus grandis, Eucalyptus camaldulensis, Eucalyptus tereticornis, and the hybrids E. grandisxE. camaldulensis, and E. grandisxE. tereticornis were analyzed by GC-MS, and evaluated for their fumigant and repellent effects on permethrin-resistant head lice. Fumigant activity of both hybrids was higher than that for pure species. E. grandisxE. tereticornis and E. grandisxE. camaldulensis showed KT50 values of 12.99 and 13.63min, respectively. E. grandis, E. camaldulensis, and E. tereticornis showed KT50 values of 25.57, 35.01, and 31.31, respectively. A simple regression analysis revealed a significant correlation between KT50 data and % of 1,8-cineole in these essential oils. Repellency varied from 47.80+/-16% to 80.69+/-6% for the five Eucalyptus essential oils tested. Interspecific hybridization improves the pediculicidal activity of Eucalyptus essential oils.     

Chemical composition and fumigant toxicity of the essential oils from 16 species of Eucalyptus against Haematobia irritans (Diptera: Muscidae) adults

Oils extracted from various species of Eucalyptus (Eucalyptus badjensis Beuzev & Welch, Eucalyptus badjensis x Eucalyptus nitens, Eucalyptus benthamii variety dorrigoensis Maiden & Cambage, Eucalyptus botryoides Smith, Eucalyptus dalrympleana Maiden, Eucalyptus fastigata Deane & Maiden, Eucalyptus nobilis L.A.S. Johnson & K. D. Hill, Eucalyptus polybractea R. Baker, Eucalyptus radiata ssp. radiata Sieber ex Spreng, Eucalyptus resinifera Smith, Eucalyptus robertsonii Blakely, Eucalyptus rubida Deane & Maiden, Eucalyptus smithii R. Baker, Eucalyptus elata Dehnh, Eucalyptus fraxinoides Deane & Maiden, E. obliqua L'Hér) were obtained by hydrodistillation. The chemical composition of essential oils was determined by gas chromatography coupled to mass spectrometry. Essential oils were mainly composed of 1,8-cineole, alpha-pinene, alpha-terpineol, 4-terpineol, and p-cymene. Vapors from these essential oils and their major components were found to be toxic to Haematobia irritans (L.) (Diptera: Muscidae) adults. An aliquot of each oil was placed in a cylindrical test chamber, and the number of knocked down flies was recorded as a function of time. Knockdown time 50% was then calculated. Results showed that essential oil of E. polybractea had the highest knockdown activity of 3.44 min. A correlation was observed between the content of 1,8-cineole in the Eucalyptus essential oils and the corresponding toxic effect.     

Monoterpene engineering in a woody plant Eucalyptus camaldulensis using a limonene synthase cDNA

Metabolic engineering aimed at monoterpene production has become an intensive research topic in recent years, although most studies have been limited to herbal plants including model plants such as Arabidopsis. The genus Eucalyptus includes commercially important woody plants in terms of essential oil production and the pulp industry. This study attempted to modify the production of monoterpenes, which are major components of Eucalyptus essential oil, by introducing two expression constructs containing Perilla frutescens limonene synthase ( PFLS ) cDNA, whose gene products were designed to be localized in either the plastid or cytosol, into Eucalyptus camaldulensis . The expression of the plastid-type and cytosol-type PFLS cDNA in transgenic E. camaldulensis was confirmed by real-time polymerase chain reaction (PCR). Gas chromatography with a flame ionization detector analyses of leaf extracts revealed that the plastidic and cytosolic expression of PFLS yielded 2.6- and 4.5-times more limonene than that accumulated in wild-type E. camaldulensis , respectively, while the ectopic expression of PFLS had only a small effect on the emission of limonene from the leaves of E. camaldulensis. Surprisingly, the high level of PFLS in Eucalyptus was accompanied by a synergistic increase in the production of 1,8-cineole and α-pinene, two major components of Eucalyptus monoterpenes. This genetic engineering of monoterpenes demonstrated a new potential for molecular breeding in woody plants.     

Acaricidal and repellent effects of myrtacean essential oils and their major constituents against Tetranychus urticae (Tetranychidae)

Nineteen plant essential oils (EOs) extracted from the family Myrtaceae growing in Australia were screened for their acaricidal and repellent activities against two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), in the laboratory by dipping method and choice- and no-choice tests. Acaricidal bioassays showed that five EOs of Callistemon viminalis, Eucalyptus bicostata, Eucalyptus maidenii, Eucalyptus sideroxylm and Eucalyptus approximans significantly increased the mortality of female adult mites and decreased the total number of eggs. In a choice test, Callistemon sieberi, E. bicostata, Eucalyptus ovata, E. sideroxylm, Eucalyptus mannifera, Eucalyptus dives, Eucalyptus elata, Eucalyptus condonocarpa, Kunzea ericoides, Melaleuca armillaris and Melaleuca fulgens demonstrated good repellency to the mite. In another test, C. sieberi, E. bicostata, E. mannifera, E. condonocarpa, K. ericoides, M. armillaris, and M. fulgens EOs decreased the egg production of the females significantly. In the acaricidal and repellent tests, E. bicostata and E. sideroxylon EOs showed both acaricidal and repellent effects (choice test) and decreased the number of eggs on treated bean leaves. The gas chromatograph/mass spectroscopy analyses revealed that the major components of E. bicostata and E. sideroxylon were 1,8-cineole, limonene, and α-pinene. The 1,8-cineole and limonene showed significant repellent effects on the mites, resulting in reduced numbers of eggs in the choice test. Hence, EOs of E. bicostata and E. sideroxylon and limonene and 1,8-cineole may be potential agents to be used in the sustainable management of T. urticae.     

Variation in volatile leaf oils of seven eucalyptus species harvested from Zerniza arboreta (Tunisia)

Leaves of seven species of the genus Eucalyptus L'Hér., viz., E. cladocalyx F. Muell., E. citriodora Hook., E. diversicolor F. Muell., E. fasciculosa F. Muell., E. grandis W. Hill, E. ovata Labill., and E. botryoides Sm., were harvested from Zerniza arboreta (region of Sejnene, northwest of Tunisia) in June 2007. Of the latter species, leaves were collected from trees having two origins, Morocco and Italy. Hydrodistillation of the dried leaves provided essential oils in yields varying from 0.4±0.0 to 3.3±0.1%, according to the species. E. citriodora had the highest mean percentage of essential oil amongst the species examined, whereas the lowest one was obtained for E. botryoides originating from Morocco. Analysis by GC (RI) and GC/MS allowed the identification of 140 compounds, representing 92.5 to 99.4% of the total oil composition. The contents of the different samples varied according to the species. The main components were 1,8-cineole (2), followed by α-pinene (1), p-cymene, borneol, α-terpineol, cryptone, spathulenol, trans-pinocarveol (4), bicyclogermacrene (5), caryophyllene oxide, and β-phellandrene. Principal components analysis and hierarchical cluster analysis separated the eight Eucalyptus leaf essential oils into five groups, each constituting a chemotype.     

Fumigant toxicity of plant essential oils to Thrips palmi (Thysanoptera: Thripidae) and Orius strigicollis (Heteroptera: Anthocoridae)

The fumigant toxicity of 92 plant essential oils to adult Thrips palmi Karny (Thysanoptera: Thripidae) and Orius strigicollis Poppius (Heteroptera: Anthocoridae) was examined by using a vapor phase toxicity bioassay and compared with those of dichlorvos, emamectin benzoate, spinosad, and thiamethoxam, four commonly used insecticides. Responses varied according to oil type and insect species. As judged by 24-h LC50 values, pennyroyal oil (2.63 mg/liter air) was the most toxic fumigant and was 23.6-fold more toxic than dichlorvos (62.09 mg/liter air) against adult T. palmi. Potent fumigant toxicity (LC50, 11.03-19.21 mg/liter air) was observed in armoise, basil, cedarleaf, coriander, cypress, howood, hyssop, marjoram, myrtle, niaouli, rosemary, and sage (Dalmatia) oils. Neither emamectin benzoate, spinosad, nor thiamethoxam exhibited fumigant action. Against adult O. strigicollis, dichlorvos (LC50, 6.3 x 10(-6) mg/liter air) was the most toxic fumigant, whereas the LC50 values of the 13 essential oils ranged from 17.29 to 158.22 mg/liter air. O. strigicollis was 1.4-22.1 times less susceptible than T. palmi to the essential oils. The essential oils described merit further study as potential fumigants for the control of T. palmi in greenhouses.     

Phytochemical analysis and in vitro antiviral activities of the essential oils of seven Lebanon species

The chemical composition of the essential oils of Laurus nobilis, Juniperus oxycedrus ssp. oxycedrus, Thuja orientalis, Cupressus sempervirens ssp. pyramidalis, Pistacia palaestina, Salvia officinalis, and Satureja thymbra was determined by GC/MS analysis. Essential oils have been evaluated for their inhibitory activity against SARS-CoV and HSV-1 replication in vitro by visually scoring of the virus-induced cytopathogenic effect post-infection. L. nobilis oil exerted an interesting activity against SARS-CoV with an IC(50) value of 120 microg/ml and a selectivity index (SI) of 4.16. This oil was characterized by the presence of beta-ocimene, 1,8-cineole, alpha-pinene, and beta-pinene as the main constituents. J. oxycedrus ssp. oxycedrus oil, in which alpha-pinene and beta-myrcene were the major constituents, revealed antiviral activity against HSV-1 with an IC(50) value of 200 microg/ml and a SI of 5.     

Essential oils from Azorean Laurus azorica

The essential oils isolated from leaves of ten and from unripe berries of eight populations of Laurus azorica (Seub.) Franco, collected on five islands of the Azorean archipelago, were analysed by GC and GC-MS. All oil samples were dominated by their monoterpene fraction (60-94%), alpha-pinene (15-37%) and 1,8-cineole (12-31%) being the main components of the leaf oils, while trans-beta-ocimene (27-45%) and alpha-pinene (12-22%) were the main components of the oils from the berries. The sesquiterpene fractions of the oils ranged from 3 to 17% and the main components were beta-caryophyllene (traces-8%) and beta-elemene (traces-3%) both in the leaf and berry oils. Some phenylpropanoid components were also present, in total amounting to 17%, trans-cinnamyl acetate (215% of the leaf oils) being the main component of this fraction. Cluster analysis of the enantiomeric composition of alpha- and beta-pinene in the oils from the leaves clearly showed two groups, one constituted by the two populations growing on the island S. Jorge, and the other constituted by the remaining populations.     

Fumigant toxicity of Eucalyptus intertexta, Eucalyptus sargentii and Eucalyptus camaldulensis against stored-product beetles

Abstract:  Three essential oils extracted from the species of the family Myrtaceae found in Kashan (centre regions of Iran) were shown to have potent fumigant toxicity against three major stored-product insects: Callosobruchus maculatus (Fab.), Sitophilus oryzae (L.) and Tribolium castaneum (Herbst). These were the essential oils from Eucalyptus intertexta R.T. Baker, Eucalyptus sargentii Maiden and Eucalyptus camaldulensis Dehnh. The mortality of 1- to 7-day-old adults of the insect pests increased with concentration from 37 to 926  μ l/l air and with exposure time from 3 to 24 h. The LC₅₀ values to the selected essential oils were between 2.55 and 3.97  μ l/l air for C. maculatus , 6.93 and 12.91  μ l/l for S. oryzae and 11.59 and 33.50  μ l/l air for T. castaneum . Fumigant effects of these essential oils were considered to warrant further research into their potential for commercial use.     

Fumigant Toxicity of Plant Essential Oils to Plutella xylostella (Lepidoptera: Yponomeutidae) and Cotesia glomerata (Hymenoptera: Braconidae)

The fumigant toxicity of 66 plant essential oils to Plutella xylostella (L.) larvae and Cotesia glomerata (L.) adults was examined using a vapor-phase toxicity bioassay and compared with that of dichlorvos. Responses varied according to oil and insect species used. Based on 24 h LD₅₀ values, pennyroyal oil [10.77 mg/filter paper (4.25 cm diameter)] was the most toxic fumigant, followed by rosemary and sage (Dalmatin) oils (15.15 mg/paper). Potent fumigant toxicity was also produced from armoise, buchu leaf, cedarleaf, coriander, eucalyptus, howood, lavender, myrtle, niaouli, peppermint, and rosewood oils (LD₅₀, 21.29–27.31 mg/paper). All essential oils were less effective than dichlorvos (LD₅₀, 0.52 mg/paper). Against adult C. glomerata, dichlorvos (LD₅₀, 0.03 mg/paper) was the most toxic fumigant, whereas the LD₅₀ values of the 14 essential oils ranged from 1.59 to 8.51 mg/paper. Based on selective toxicity ratio (STR, P. xylostella LD₅₀/C. glomerata LD₅₀), the 14 essential oils (STR, 2.5–14.5) are more selective than dichlorvos (STR, 17.3). The essential oils tested merit further study as potential fumigants for the control of P. xylostella in greenhouses because of their selective toxicity to adult C. glomerata and their much greater activity as a fumigant.     

Anti-fungal activity of essential oils of Ceylon Eucalyptus species for the control of Fusarium solani and Sclerotium rolfsii

The essential oils were extracted from the leaves of Eucalyptus microcorys, Eucalyptus grandis and Eucalyptus robusta which were grown in Sri Lanka and their major chemical compounds were determined. 1,8-Cineole and α-pinene were identified as major aroma compounds in these oils. In this study, the anti-fungal activity of essential oils of E. microcorys, E. grandis and E. robusta, ethanol extract of E. microcorys and 1,8-cineole were evaluated against Sclerotium rolfsii, a fungi responsible for leaf spot disease of indoor plants and Fusarium solani, a fungi responsible for dry rot diseases of potato by poisoned food technique, and minimum inhibitory concentrations (MICs) were determined. The essential oils from three Eucalyptus species showed significant inhibitory effect against S. rolfsii and F. solani than the ethanol extract of E. microcorys. Of treatments, the essential oil of E. grandis showed the best anti-fungal activity with the MIC values of less than 0.1% for S. rolfsii and 0.5% for F. solani. The MICs of the oils of E. microcorys and E. robusta were between 0.3–0.5% against S. rolfsii and 0.5–0.75% for F. solani. The 1,8-cineole did not exhibit inhibition activity as much of Eucalyptus essential oils and hence, it can be assumed that minor chemical components of the oils contribute to the growth inhibition of the tested fungi. This is the first report of anti-fungal activity of Sri Lankan oils of E. microcorys, E. grandis and E. robusta and ethanol extract of E. microcorys against S. rolfsii and F. solani. These findings would be useful for the designing of natural fungicide for agriculture- and food-based industries.     

Contact toxicity, feeding reduction, and repellency of essential oils from three plants from the ginger family (Zingiberaceae) and their major components against Sitophilus zeamais and Tribolium castaneum

The essential oils from rhizomes of Alpinia conchigera Griff, Zingiber zerumbet Smitt, Curcuma zedoaria (Berg.) Roscoe; their major compounds (camphene, camphor, 1,8-cineole, alpha-humulene, isoborneol, alpha-pinene, beta-pinene and terpinen-4-ol); and synthetic essential oils comprised of mixtures of major pure compounds in the same ratios as the extracted essential oils were tested for contact, feeding reduction, and repellency against Sitophilus zeamais Motschulsky and Tribolium castaneum (Herbst) adults. Via topical applications, the three extracted oils had similar toxicity against S. zeamais (LD50 fiducial limits: 18-24 microg oil/mg insect). T. castaneum had similar sensitivity to all three oils (35-58 microg/mg), and it was less sensitive than S. zeamais. The LD50 values of synthetic A. conchigera and synthetic Z. zerumbet oils were similar to those of their corresponding extracted essential oils. The synthetic C. zedoaria oils showed lower contact toxicity than the extracted C. zedoaria oils to both insects. Sitophilus zeamais and T. castaneum were sensitive to terpinen-4-ol and isoborneol in contact toxicity tests. In antifeedant tests, the three extracted oils were able to decrease the consumption of flour disks, especially Z. zerumbet oils, whereas both insect species could feed on the flour disks treated with three synthetic essential oils. Only terpinen-4-ol deterred feeding in both insects. In repellency tests, A. conchigera oils at highest concentration repelled S. zeamais and T. castaneum. None of the synthetic essential oils repelled S. zeamais (315 microl/cm2) and T. castaneum (31 microl/cm2) Only terpinen-4-ol showed repellent activity against both insects.     

The effects of Eucalyptus terpenes on hepatic cytochrome P450 CYP4A, peroxisomal Acyl CoA oxidase (AOX) and peroxisome proliferator activated receptor alpha (PPARalpha) in the common brush tail possum (Trichosurus vulpecula)

Eucalyptus leaves contain a high proportion of essential oils comprising of a complex mixture of monoterpenes such as 1,8-cineole, alpha-pinene, d-limonene and p-cymene. In this study, hepatic levels of microsomal lauric acid hydroxylase and peroxisomal cyanide-intensive palmitoyl coenzyme A oxidative activities were examined in livers of possums given an artificial diet consisting of the above monoterpenes for 10 days. These values were compared with those of possums fed a control diet containing only fruits and cereals. Peroxisomal cyanide-intensive palmitoyl coenzyme A oxidative activity was significantly higher in livers of treated possums relative to that of control possums (2.96+/-0.93 vs. 0.98+/-0.88 nmol/mg protein per min, P<0.01) (mean+/-S.D., n=4). A small increase in microsomal lauric acid hydroxylase activity was observed in the treated possum in comparison with the control group (4.40+/-0.85 vs. 3.60+/-0.48 nmol/mg protein per min) (mean+/-S.D., n=4). A higher NAD/ NADP ratio was observed in treated possums as compared with control possums (4.73+/-0.65 vs. 3.51+/-0.64 nmol/mg protein per min, P<0.05) (mean+/-S.D., n=4). No other statistically significant differences in pyridine nucleotide contents were found between control and treated possums. Northern blot analysis of mRNA from rat, human, terpene treated and control possum livers, using the corresponding koala cDNA probes, detected a more intense acyl CoA oxidase (AOX) mRNA band in livers of terpene fed possums. Negligible differences in the intensity of CYP4A and PPARalpha mRNA bands were observed between the two groups. These data suggest that Eucalyptus terpenes elevate hepatic AOX expression in possums.     

Sensitivity of Aedes aegypti adults (Diptera: Culicidae) to the vapors of Eucalyptus essential oils

Vapors of essential oils extracted from various species of Eucalyptus (E. gunnii, E. tereticornis, E. grandis, E. camaldulensis, E. dunnii, E. cinerea, E. saligna, E. sideroxylon, E. globulus ssp. globulus, E. globulus ssp. maidenii, E. viminalis and the hybrids E. grandis × E. tereticornis and E. grandis × E. camaldulensis) and their major components were found to be toxic to Aedes aegypti adults, the yellow fever mosquito.An aliquot of each oil was placed in a cylindrical test chamber and the number of knocked-down mosquitoes was recorded as function of time. Knockdown time 50% was then calculated. Results showed that E. viminalis had the fastest knockdown time at of 4.2 min, on the same order as dichlorvos, a standard knockdown agent. A correlation was observed between the content of 1,8-cineole in the Eucalyptus essential oils and the corresponding toxic effect.The correlation between KT₅₀ values and calculated vapor pressures of the essential oil components showed that the fumigant activity of simple organic compounds in insects is correlated with their volatility.     

Comparative GC–FID and GC–MS analysis of the mono and sesquiterpene secondary metabolites produced by the field grown and micropropagated plants of <Emphasis Type="Italic">Artemisia amygdalina</Emphasis> Decne

The essential oil obtained by hydrodistillation from the leaves of micropropagated plants of Artemisia amygdalina was analyzed by capillary GC–FID and GC–MS and compared with that obtained from the leaves of field growing parent plants. The oil yield from the micropropagated plants was lower (0.05% v/w) than the oil yield obtained from field-grown plants (0.2% v/w). The major constituents of the field-grown plants were p-cymene (21.0%), 1,8-cineole (24.9%), α-terpineol (5.9%), β-caryophyllene (4.7%), germacrene D (4.0%), while as the major constituents from the micropropagated plants were p-cymene (11.3%),1,8-cineole (10.2%), borneol (7.9%), α-longipinene (5.5%), α-copaene (5.5%) and β-caryophyllene (17%). The essential oil from field-grown plant was dominated by the presence of oxygenated monoterpenes (41.5%), monoterpene hydrocarbons (35.9%) and sesquiterpene hydrocarbons (16.3%) while as the essential oil of micropropagated plants was characterized by sesquiterpene hydrocarbons (40.0%), oxygenated monoterpenes (25.2%) and monoterpene hydrocarbons (21.6%).     

三种桉叶油化学成分研究

通过水蒸汽蒸油试验,发现已贮藏半年多的风干桉叶,仍有较高的含油量。云南省弥勒产的直杆桉(Eucalyptus maidenii)得油率为4.14％,蓝桉(E.globulus)为3.5％,福建省惠安产的窿缘桉(E.exserta)为1.2％,桉叶油生产厂也可以通过贮藏的风干桉叶生产桉叶油。通过气相色谱和质谱分析,从直杆桉叶油中鉴定出44个组份,蓝桉叶油中鉴定出35个组份,窿缘桉叶油中鉴定出26个组份。三种桉叶油单萜部分的组份基本一致,只是相对含量各有不同。桉油素含量,以直杆桉最高为68.02％,其次是蓝桉为67.54％,窿缘桉最低为34.33％。直杆桉、蓝桉可以怍材、油两用树种,窿缘桉不宜作油用树种。     

桉叶素生物合成研究进展

作为桉叶油的主要成分,桉叶素是具有多种生物活性的单萜化合物,被广泛应用于药品、食品及化妆品等领域。桉叶油主要从桉树叶提取,该过程耗费大量人力及自然资源,且容易污染环境。近年来,随着微生物代谢工程与合成生物学的快速发展,加上越来越多萜类生物合成途径得到解析,为桉叶素的绿色生产提供了新的途径。对桉叶素的生物合成途径、桉叶素合酶的结构与功能及近年来桉叶素的微生物合成进行了综述,并对利用微生物代谢工程合成桉叶素等单萜化合物的瓶颈问题及解决方案进行了探讨和归纳,为构建高产桉叶素等单萜微生物工程菌株提供参考。     

Evaluation of extracts and oils of tick-repellent plants from Sweden

Abstract. Leaves of Myrica gale Linnaeus (Myricaceae), Rhododendron tomentosum (Stokes) H. Harmaja (formerly Ledum palustre Linnaeus: Ericaceae) and Artemisia absinthium Linnaeus (Asteraceae) were extracted with organic solvents of different polarities and the essential oils of leaves were obtained by steam distillation. The extracts or oils were tested in the laboratory for repellency against host-seeking nymphs of Ixodes ricinus Linnaeus (Acari: Ixodidae). Rhododendron tomentosum oil, 10%, diluted in acetone, exhibited 95% repellency; R. tomentosum and A. absinthium extracts in ethyl acetate, > 70% repellency; A. absinthium extract in hexane, approximately 62% repellency; and M. gale oil, 10%, approximately 50% repellency on I. ricinus nymphs. Compounds in the leaf extracts or in the oils were collected by solid phase microextraction (SPME) and identified by gas chromatography-mass spectrometry (GC-MS) and/or MS. Characteristic volatiles detected from oil or extract of M. gale were the monoterpenes 1,8-cineole, alpha-terpineol, 4-terpineol and thujenol; and of R. tomentosum myrcene and palustrol. Characteristic volatiles from leaf extracts of A. absinthium were sabinene, oxygenated monoterpenes, e.g. thujenol and linalool, and geranyl acetate. Each plant species synthesized numerous volatiles known to exhibit acaricidal, insecticidal, 'pesticidal' and/or arthropod repellent properties. These plants may be useful sources of chemicals for the control of arthropods of medical, veterinary or agricultural importance.