Toxicity of monoterpenes from Pinus radiata cortical oleoresin to Diplodia pinea spores1

The effect on spore germmation and germ tube growth of Diplodia pinea produced by (+)- α-pinene, (?)-β-pinene, (+)-Δ³-carene, (?)-limonene, β-phellandrene, and terpinolene was determined over a range of vapour concentrations. Spore germination was most inhibited by A³-carene; germ tube growth was inhibited by all these monoterpenes except (?)-limonenc and ß-phellandrene. In contrast to the low toxicity of (?)-limonene, (+)-limonene (not known to be present in Pinus radiata cortical oleoresin) was extremely toxic to both spore germination and germ tube growth. The results arc discussed in rotation to an earlier observation that trees free of dieback in the field had high levels of certain monoterpenes.     

The growth-promoting activity of terpenoids on wood-decomposing fungi

Linear growth of wood-decomposing fungi on an agar medium was usually retarded, but in some cases enhanced, by terpenoids in gaseous form. Increased dry matter production was demonstrated in some Coniophora, Peniophora, Stereum and Polyporus species exposed to the vapours of longifolene, α-pinene, β-pinene, Δ-3-carene, limonene or a mixture of cuparene and thujopsene. The positive response to longifolene in some cases seemed to involve a more efficient utilisation of the glucose in the medium.     

Individual variations in monoterpenes released from Cryptomeria japonica and Pinus thunbergii needles

Variations in the compositions of low boiling point (LBP) monoterpenes in needle samples from 99 sugi (Cryptomeria japonica) and 100 kuromatsu (Pinus thunbergii) trees were investigated using a headspace technique. Considerable variations in the proportions of monoterpenes were revealed in both species. In sugi, the proportions of sabinene and α-pinene in the total LBP monoterpenes, ranging from 8.8% to 73.3% and from 14.5% to 73.3%, respectively, showed enormous variations among nine monoterpenes. The proportions of 3-carene and limonene, ranging from 0.1% to 29.5% and from 0.2% to 20.4%, respectively, also showed very specific variations. In kuromatsu, the proportions of β-pinene and α-pinene in the total LBP monoterpenes, ranging from 26.5% to 66.3% and from 18.7% to 46.9%, respectively, showed considerable variations among ten monoterpenes. The proportions of myrcene and 1,8-cineole, ranging from 0.9% to 18.5% and from 0.8% to 12.3%, respectively, also showed specific variations. Part of this article was presented at the 50th Annual Meeting of the Japan Wood Research Society, Kyoto, April, 2000, and the 13th Annual Meeting of the Chugoku Shikoku Branch of the Japan Wood Research Society, Okayama, September 2001     

红脂大小蠹诱导油松树干挥发性单萜相对含量的变化

使用Porapak Q吸附剂,采用动态顶空的吸附方法,连续抽取11个不同阶段的对照、损伤与虫害油松的挥发物成分.运用GC、GC-MS与图谱峰面积进行挥发物成分定性与单萜相对含量的定量分析.与对照和损伤油松相比,虫害油松含有高比率的(+)-α-蒎烯(超过90％)和月桂烯,但(+)-3-蒈烯,(-)-β-蒎烯,莰烯和柠檬烯含量较低.(+) -3-蒈烯含量对照和损伤油松高于虫害油松(低于4％).(+)-3-蒈烯含量从损伤4天后的最低水平逐渐恢复到对照油松中含量水平.(-)-β-蒎烯和柠檬烯在不同处理的油松中含量相近,但在接入雌雄虫后3个处理间存在显著差异.尽管目前不确定我国的红脂大小蠢是否在危害油松后释放信息素,但研究表明红脂大小蠢可能会利用各种单萜成分细节上的变化作为化学信号,从而实现寄主的选择和种群聚集.本研究对于揭示油松抗性机制,完善红脂大小蠢的监测与防控技术体系具有重要意义.     

Low-density Ceratocystis polonica inoculation of Norway spruce (Picea abies) triggers accumulation of monoterpenes with antifungal properties

Among the most devastating pests of Norway spruce (Picea abies) are the European spruce bark beetle (Ips typographus) and the associated pathogenic blue-stain fungus Ceratocystis polonica. Following attack and colonization, the beetle and the fungus must cope with induced host chemical defenses, such as monoterpenes that are generally thought to be toxic to both symbionts. The goal of this study was to better understand the response of Norway spruce following C. polonica inoculation at low density that does not overwhelm the tree and to identify monoterpenes mobilized toward the fungus. We inoculated healthy mature trees and monitored monoterpene profiles 2, 3, and 5 months post-inoculation. We also exposed three different C. polonica strains to the most abundant or significantly up-regulated monoterpenes to determine differences in monoterpene toxicity and resistance among strains. Total monoterpene levels, including limonene, were increased at 2 and 3 months after inoculation and had dropped after 5 months. In in vitro assays, all monoterpenes were inhibitory to C. polonica. Limonene and β-pinene were the most potent inhibitors of fungal growth. The extent of inhibition varied between the three strains tested. These results showed a defense response of Norway spruce to C. polonica, in which limonene may play a critical role in inhibiting the spread of the fungus. We also showed that differences between strains of C. polonica must be taken into account when assessing the role of the fungus in this bark beetle–symbiont system.     

Studies on ectomycorrhizae of pine

Infection of feeder roots of Pinus echinata by the ectomycorrhizal fungus, Pisolithus tinctorius, resulted in an approximate 40-fold increase in the level of 3-carene, and Ccnococcum graniforme induced an approximate 30-fold increase in β-phellandrene. Vapors of these volatile compounds affected the vegetative growth of different ectomycorrhizal and root pathogenic fungi to different degrees in laboratory experiments.     

Association between temperature,relative humidity and concentration of volatile organic compounds from wooden furniture in a model room

Abstract

Emissions of volatile organic compounds (VOCs) from wooden furniture in a child's room were investigated. A model room was set up in a 12?m³ test chamber, and in parallel, a scaled-down version was run in a 1?m³ test chamber. The latter had a steady climate of 23°C/50% relative humidity (RH), while the temperature and humidity conditions were varied in the 12?m³ chamber between 18 and 28°C, and 30 and 80% RH. Compounds found were α-pinene, β-pinene, 3-carene, limonene and hexanal. Variations in the climate showed that there is an association between temperature and room concentrations of all found VOCs. Changes in RH showed effect on hexanal concentration, and all monoterpene concentrations remained unaffected.     

Differential effects of plant ontogeny and damage type on phloem and foliage monoterpenes in jack pine (Pinus banksiana)

Coniferous trees have both constitutive and inducible defences that deter or kill herbivores and pathogens. We investigated constitutive and induced monoterpene responses of jack pine (Pinus banksiana Lamb.) to a number of damage types: a fungal associate of the mountain pine beetle (Dendroctonus ponderosae Hopkins), Grosmannia clavigera (Robinson-Jeffrey & R.W. Davidson); two phytohormones, methyl jasmonate (MJ) and methyl salicylate (MS); simulated herbivory; and mechanical wounding. We only included the fungal, MJ and mechanical wounding treatments in the field experiments while all treatments were part of the greenhouse studies. We focused on both constitutive and induced responses between juvenile and mature jack pine trees and differences in defences between phloem and needles. We found that phytohormone applications and fungal inoculation resulted in the greatest increase in monoterpenes in both juvenile and mature trees. Additionally, damage types differentially affected the proportions of individual monoterpenes: MJ-treated mature trees had higher myrcene and β-pinene than fungal-inoculated mature trees, while needles of juveniles inoculated with the fungus contained higher limonene than MJ- or MS-treated juveniles. Although the constitutive monoterpenes were higher in the phloem of juveniles than mature jack pine trees, the phloem of mature trees had a much higher magnitude of induction. Further, induced monoterpene concentrations in juveniles were higher in phloem than in needles. There was no difference in monoterpene concentration between phytohormone applications and G. clavigera inoculation in mature trees, while in juvenile trees MJ was different from both G. clavigera and simulated herbivory in needle monoterpenes, but there was no difference between phytohormone applications and simulated herbivory in the phloem.     

Analyses of BioVOCs variation related to vegetation predominance in the natural park of ampezzo dolomites, UNESCO world heritage area of dolomites

The determination of emitted Biogenic Volatile Compounds (BioVOCs) profile via Field Air Analysis (FAA) — Solid Phase Micro Extraction (75 μm Carboxen/PDMS portable field sampler) was performed using GC-MS. The six sampling sites located in the Natural Park of Ampezzo Dolomites, part of the UNESCO’s World Heritage List, are characterized by different predominating vegetation. The ubiquitous compound hexanal was chosen as internal standard, in order to compare the amounts of the different compound in the different sites. The main terpenes identified were α-pinene, β-pinene, δ-3-carene and d-limonene, while 1,8 cineole was the highest abundant oxygenated terpene. In the site next the national road a higher amount of benzene derivatives (benzene, toluene, o-xylene, p-xylene and ethylbenzene) were observed. In the site relatively close to the road a marked decrease of benzene derivatives was observed. In the sites close to conifers (i.e. Fir, Pine and Larch) the terpenic content observed was higher than in the site close to the mixed forest (mainly large-leaf and some conifers). Some terpenes were observed also in the high-quote meadow site, although of lower abundance than in the other sites. The highest abundance of terpenes was observed in the Pinus cembra (Swiss Pine) site.     

Comparison of host volatiles under different physiological states and electroantennogram responses of Semanotus bifasciatus （Motschulsky） to these volatiles

Based on an analytical TCT-GC/MS method, volatile compounds of branches of Platycladus orientalis under three physiological states （healthy, weak and baited trees） were analyzed. The results showed that α-pinene, β-phellandrene, 3-careen, limonene and thujopsene were released under all three physiological states. There were other kinds of volatiles released from branches of weak P. orientalis trees, such as camphene, γ-terpinene, （＋）-4-carene and cedrene, but these four volatiles compounds were not released from the branches of healthy P. orientalis. Volatile components from the physically damaged branches of P. orientalis had changed. β-terpinene, mycrene, isocaryophillene and caryophyllene were identified in both healthy and weak branches of P. orientalis. Only cedrene was identified in the baited wood. Electroantennogram （EAG） tests were conducted with nine volatile compounds from the branches of P. orientalis. Antennnae of male and female Semanotus bifasciatus （Motschulsky） were highly sensitive to these nine volatile compounds. Cedrene elicited the strongest response, whereas β-terpinene the lowest. Males and females showed no signifi-cant difference in their EAG responses to the nine volatile compounds.     

Tree-to-tree and clone-to-clone variations of monoterpenes emitted from needles of hinoki (Chamaecyparis obtusa)

Variations in the composition of low boiling point (LBP) monoterpenes emitted from needle samples of 150 hinoki (Chamaecyparis obtuse) trees (30 strains, each with five clones) native to Shimane Prefecture, Japan, were investigated using a headspace technique. The assays revealed considerable proportional variations especially in the amount of sabinene, which ranged from 24% to 78% of the total LBP monoterpenes. The proportions of α-pinene, myrcene, and limonene negatively correlated with that of sabinene overall. In particular, the proportion of limonene showed clear negative correlation with that of sabinene (r = −0.98). Differences in the proportion of sabinene among five clones in each strain were less than 15% in 22 out of 30 strains, indicating that monoterpene composition is constitutively steady in most strains. In a few strains, however, considerable variation in the composition was observed among clones.     

Essential oil from the leaves of Cryptomeria japonica acts as a silverfish (Lepisma saccharina) repellent and insecticide

This is the first article to report the evaluation of a natural product used as an antisilverfish agent. Silverfish (Lepisma saccharina), primitive wingless insects, feed on a variety of materials, including paper, cotton, starch, and cereals. They can be a problem in libraries and other places where books, documents, and papers are stored. In this pilot study, the essential oil from leaves of Cryptomeria japonica was investigated to test its properties as a silverfish repellent and insecticide. The results from a repellency bioassay show that the essential oil significantly repelled silverfish. The repellent activity was 80% at a dosage of 0.01 mg/cm³. When silverfish were exposed to a concentration of 0.16 mg/cm³ of essential oil, they were killed within 10h. The chemical composition of essential oil, the emissions from a test chamber, and the residue left on filter papers previously soaked with the essential oil in a chamber were analyzed by gas chromatography-mass spectrometry. The components of the essential oil were found to be: elemol (18.22%), 16-kaurene (11.63%), 3-carene (9.66%), sabinene (9.37%), 4-terpineol (9.06%), β-eudesmol (5.70%), α-pinene (5.62%), and limonene (5.26%). Only some constituents of the essential oil compounds collected by solid-phase microextraction were found to be emitted in the test chamber. The main constituents were: 3-carene (21.03%), p-cymene (10.95%), limonene (9.49%), β-myrcene (9.39%), γ-terpinene (9.10%), α-terpinene (8.57%), and 4-terpineol (7.97%).     

Effects of oleoresins and monoterpenes on in vitro growth of fungi associated with pine decline in the Southern United States

As a means of exploring pine resistance to root disease and declines, the effects of host plant secondary metabolites on the growth of root colonizing fungi associated with three diseases/declines of southern pines – loblolly pine decline, littleleaf disease and annosum root rot were tested. The associated fungi –Leptographium huntii, L. serpens, L. terebrantis, L. procerum, Heterobasidion annosum and Phytophthora cinnamomi– were grown in saturated atmospheres or in direct contact with, pure monoterpenes and crude oleoresin collected from the four southern pines (Pinus taeda, P. eschinata, P. palustris and P. elliotti) for 7 day. Fungal growth was measured at 3, 5 and 7 day. Root‐infecting fungi differed significantly in sensitivity to crude oleoresin and pure monoterpenes. All fungi tested were inhibited, to some extent, by the resins tested. H. annosum and P. cinnamomi were strongly inhibited by all the monoterpenes tested. The ophiostomatoid fungi were significantly less affected by the compounds tested. L. huntii and L. serpens were less inhibited by monoterpenes than either L. terebrantis or L. procerum. These fungal growth studies show that the kind and amount of secondary metabolite produced by the host plant have a profound effect on tree pathogens. Alterations of tree physiology may have implications for defenses against tree pathogens as well as to the ecology and management of forest ecosystems. Difference in incidence of root disease observed in the field may be explained by the ability of the fungus to tolerate these host defense mechanisms.     

The essential oil of Eucalyptus tereticornis, and its constituents α- and β-pinene,potentiate acetylcholine-induced contractions in isolated rat trachea

The effects of the essential oil of Eucalyptus tereticornis (EOET), especially the effects of its constituents α- and β-pinene, were studied on rat trachea in vitro. In tracheal rings, EOET, α- or β-pinene potentiated the contractions induced by acetylcholine (ACh). Contractions induced by K⁺ (60 mM) were also potentiated by α- and β-pinene, but were reduced by EOET. Our findings show that EOET has myorelaxant effects on rat airways, but potentiates ACh-induced contractions. Monoterpenes α- and β-pinene are involved in its potentiating actions, but are not responsible for its myorelaxant effects. A putative inhibition of the acetylcholinesterase enzyme is involved.     

-蒎烯合成龙脑研究

在固体超强酸催化作用下β-蒎烯和无水草酸平稳地发生酯化反应,生成草酸龙脑酯;加入氢氧化钠溶液发生皂化反应得到龙脑。酯化反应采用程序升温的方式（65℃、1h,75℃、4h,90℃、1h）,皂化反应时间为1h。在上述反应条件下,当催化剂SO4^2-／ZrO2用量为7％时（以β-蒎烯质量计,下同）,粗龙脑产率为62．0％,产品中正、异龙脑比为13．6：48．7;当催化剂S2O8^2-／ZrO2用量为7％时,粗龙脑产率为66．7％,正、异龙脑比为11．5：49．5;这两种催化剂对正龙脑选择性比较差。当用稀土复合固体超强酸SO4^2-／ZrO2-CeO2为催化剂,用量3％时,粗龙脑产率为56．1％,但对正龙脑的选择性高于其他催化剂,正、异龙脑比为46．0：23．2。     

Implication of limonene and linalyl acetate in cytotoxicity induced by bergamot essential oil in human neuroblastoma cells

Bergamot (Citrus bergamia, Risso et Poiteau) essential oil (BEO) is a widely used plant extract showing anxiolytic, analgesic and neuroprotective effects in rodents; also, BEO activates multiple death pathways in cancer cells. Despite detailed knowledge of its chemical composition, the constituent/s responsible for these pharmacological activities remain largely unknown. Aim of the present study was to identify the components of BEO implicated in cell death. To this end, limonene, linalyl acetate, linalool, γ-terpinene, β-pinene and bergapten were individually tested in human SH-SY5Y neuroblastoma cultures at concentrations comparable with those found in cytotoxic dilutions of BEO. None of the tested compounds elicited cell death. However, significant cytotoxicity was observed when cells were cotreated with limonene and linalyl acetate whereas no other associations were effective. Only cotreatment, but not the single exposure to limonene and linalyl acetate, replicated distinctive morphological and biochemical changes induced by BEO, including caspase-3 activation, PARP cleavage, DNA fragmentation, cell shrinkage, cytoskeletal alterations, together with necrotic and apoptotic cell death. Collectively, our findings suggest a major role for a combined action of these monoterpenes in cancer cell death induced by BEO.     

Loblolly pine (Pinus taeda L.) essential oil yields affected by environmental and physiological changes

ABSTRACT

Loblolly pine (Pinus taeda L.) is one of the most abundant timber species in the United States and needles from this species contain essential oils (EOs) with antibacterial properties. Needles from trees of one loblolly pine clone were assayed for the EOs α-pinene, β-pinene, terpineol, limonene, and caryophyllene across a growing season. Results showed strong positive correlations among EOs, except for caryophyllene, with Pearson’s correlation values ranging from .66–.96. Simultaneously, physiological attributes of tree stomatal conductance, water stress, and environmental attributes including soil moisture, photosynthetic active radiation (PAR), temperature, wind speed, and relative humidity were measured. The plants modulated the EO concentrations as a function of a changing environment season. The collection month had the largest effect on EO yields and highest yields were during the beginning of the growing season. Decreases in EOs were evident as the growing season progressed and as plants exhibited greater water stress, temperatures, solar radiation, and less stomatal conductance and soil moisture. Loblolly pine’s main constituents of EOs were tightly linked and were influenced by seasonal changes (i.e., month); still, environmental/physiological attributes exhibited significant effects on α-pinene, β-pinene, and limonene concentrations. Early spring conditions with ample water were most conducive to high concentrations of EOs.     

Tropical red gums — a source of 1,8-cineole-rich Eucalyptus oil

Tropical provenances of Eucalyptus camaldulensis Dehnh. and E. tereticornis Smith were studied, in their natural habitat in Australia and in a 3.75-year-old progeny trial in Zimbabwe, for their potential to produce medicinal-grade essential oils. Substantial interand intra-specific variation in the contents of five prominent monoterpenes, 1,8-cineole, -pinene, -pinene, limonene and p-cymene was found. Plantations of E. camaldulensis established in the wet/dry tropics using seed from Petford in northern Queensland offer immediate potential for oil production. Other widely-planted Queensland provenances with oil potential are Gilbert River Bridge E. camaldulensis and, after some genetic improvement, E. tereticonis from Morehead and Kennedy Rivers. Select individual trees at Petford provide oil of enhanced quality, at about double the yield of average trees.     

Neuropharmacological activities of phytoncide released from <Emphasis Type="Italic">Cryptomeria japonica</Emphasis>

Forest-air bathing and walking (shinrin-yoku) is beneficial to human heath. In this study the phytoncide (volatile compounds) released from Cryptomeria japonica plantation forest was characterized by using solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS). The main volatile compounds were α-pinene (19.35%), β-myrcene (16.98%), d-limonene (15.21%), and γ-muurolene (7.42%). Furthermore, the neuropharmacological activity of the essential oils from leaves of C. japonica (ECJ) was evaluated by several animal behavior tests. ECJ could prolong the sleeping phase of ICR (imprinting control region) mice in the pentobarbital-induced sleeping time model. Furthermore, both EJC and one of its monoterpenes, d-limonene, possessed potent anxiolytic and analgesic activities based on the results obtained from elevated plus maze and writhing tests. The volatile compounds released from C. japonica provide relaxing and stress-relieving effects on mice, and further study on the effect of phytoncide on humans is worthwhile.     

Field attraction of Cerambyx welensii to fermentation odors and host monoterpenes

Cerambyx welensii Küster is one of the greatest threats to Quercus suber L. and Quercus ilex L. in Spain. Efficient control methods for this species are currently being tested, and the use of traps baited with food-related products for their monitoring and control has been proposed. Cerambyx welensii adults prefer to colonize old or decayed trees, in which cortical exudates frequently release fermentation odors. Fermentation-related compounds, such as ethanol, have been cited as attractants to various hardwood woodborers (as Cerambyx cerdo L.), used either alone or sometimes in combination with other host odors. In this work, we conducted field experiments to ascertain whether chemical blends mimicking fermentation odors were attractive for C. welensii. To this end, we used three types of traps (multiple funnel, cross-vane and single-funnel) baited with six synthetic blends including one or more fermentation-related chemicals (ethanol, ethyl acetate, 3-methyl-1-butanol, acetaldehyde, 2-methyl-1-propanol and 2-methyl-1-butanol), various food-related products (beer, melon and red wine mixed with peach juice) and host monoterpenes (α-pinene, β-pinene, limonene, α-phellandrene and camphene). At high release rates (5–11 g/day), various synthetic blends proved attractive irrespective of trap type. Our results contribute to understand the impact of volatile organic compounds in host selection by woodborers infesting deciduous trees. The results hold promise for optimizing trapping strategies to control C. welensii within an integrated pest management framework.