Weak responses of pine sawfly larvae to high needle flavonoid concentrations in scots pine

Responses of sawfly larvae (Hymenoptera, Diprionidae) to the flavonoid taxifolin glucoside in their host plant were studied in a laboratory experiment. Larvae ofNeodiprion sertifer andDiprion pini were raised from egg hatch to cocoon spinning on two Scots pine (Pinus sylvestris) chemotypes, one without needle taxifolin glucoside (–) and the other containing 2–4% taxifolin glucoside (+). The (+) chemotype had somewhat lower concentrations of needle terpenoids (resin acids) than the (–) chemotype. Current-year needles had higher taxifolin glucoside concentrations than mature needles. There were no differences in survival or body size betweenN. sertifer larvae that fed on the (+) chemotype and those that fed on the (–) chemotype. FemaleD. pini larvae raised on (+) needles developed 6% more slowly than larvae fed (–) needles. The results from this study are contrary to earlier findings showing that flavonoid glucosides have strong negative effects on insect performance. Possible explanations for the different outcomes are discussed.     

Assessment and Implications of Intraspecific and Phenological Variability in Monoterpenes of Scots Pine (<Emphasis Type="Italic">Pinus sylvestris</Emphasis>) Foliage

Scots pine populations contain individuals with widely differing amounts and composition of monoterpenes and exist as one of two chemotypes: with or without Δ³-carene. We investigated the significance for ecological studies of two types of variation in monoterpenes: (1) the inherent variability in the concentration of monoterpenes or their relative amounts in needles of seedlings, saplings, and mature trees; and (2) phenological variation in developing needles. The relative composition of needle monoterpenes in 5-year-old saplings changed during the needle development period until the final composition was reached upon needle maturity. Changes in composition depended on chemotype. Needles of the “no-Δ³-carene” chemotype had higher absolute concentrations of α-pinene, β-pinene, camphene, and total monoterpenes than “Δ³-carene” chemotype. For the “Δ³-carene” chemotype, the relative concentration of Δ³-carene during the needle growing season and immediately after emergence of seedlings was higher compared to that reached at needle maturity. Repeated removal of single needles (at weekly intervals during growth) from 5-year-old saplings did not influence the composition of monoterpenes. Within a natural Scots pine dominated woodland, 18% of mature Scots pines (N = 574) belonged to the “no-Δ³-carene” chemotype. Chemotypic variation within populations means that the statistical power with which differences in monoterpene concentrations can be detected is lower when sampling from the whole population compared to sampling within chemotypes. Reduction of this background variation and accounting for chiral variation if present, would significantly aid efficiency, interpretation, and understanding of processes in chemical and ecological research. One method for achieving this is the screening of plants for chemotypes before the establishment of experiments or field sampling regimes. We present a summary of suitable analytical methods for needle tissue that facilitates this prior screening.     

Drought Stress Alters the Concentration of Wood Terpenoids in Scots Pine and Norway Spruce Seedlings

Drought is known to have an impact on the resistance of conifers to various pests, for example, by affecting resin flow in trees. Little is known, however, about the quantitative and qualitative changes in resin when trees are growing in low moisture conditions. We exposed Scots pine (Pinus sylvestris L.) and Norway spruce (Picea abies (L.) Karst.) seedlings to medium and severe drought stress for two growing seasons and analyzed the monoterpenes and resin acids in the main stem wood after two years of treatment. In addition to secondary chemistry, we measured the level of nutrients in the needles and the growth response of seedlings. After the first year of treatment, drought stress did not affect the growth of seedlings, but in the second year, shoot growth was retarded, especially in Scots pine. In both conifer species, severe drought increased the concentrations of several individual monoterpenes and resin acids. Total monoterpenes and resin acids were 39 and 32% higher in severe drought-treated Scots pine seedlings than in the controls, and 35 and 45% higher in Norway spruce seedlings. In Scots pine needles, the concentrations of nitrogen and phosphorus increased, while magnesium and calcium decreased compared to controls. In Norway spruce needles, nutrient concentrations were not affected. The results suggest that drought stress substantially affects both the growth of conifers and the chemical quality of the wood. We discuss the potential trade-off in growth and defense of small conifer seedlings.     

Oleoresin Chemistry Mediates Oviposition Behavior and Fecundity of a Tree-Killing Bark Beetle

Many herbivores are sensitive to the secondary chemistry of their host plants. However, the influence of pine secondary chemicals (monoterpenes) on bark beetle fitness is poorly understood. We tested the hypothesis that the monoterpene composition of the phloem oleoresin of ponderosa pine, Pinus ponderosa var scopulorum, mediates rates of host acceptance, oviposition behavior, and fecundity of the western pine beetle, Dendroctonus brevicomis. We performed reciprocal rearing experiments, controlling for the monoterpene composition (chemotype) of host material. We tested the effects of two geographically interspersed host chemotypes on beetles with unknown (wild) and known (reared F₁) chemical histories. Host chemotype and insect chemical history did not affect rates of acceptance of host material by female beetles. Insect chemical history affected egg gallery construction, and beetles constructed egg galleries that were on average 24.3% longer when reared in host material that was chemically similar to their natal host material. However, mean egg gallery lengths did not differ between host chemotypes. Insect chemical history also influenced fecundity: F₁ beetles produced 52.7% more offspring on average when reared in host material that was chemically similar to their natal host. Our experiments demonstrate that the chemical history of bark beetles mediates egg gallery construction and fecundity, but not host acceptance. This implicates chemical history as a more important factor than host chemotype in the oviposition behavior and fecundity of D. brevicomis.     

Qualitative and quantitative variation in monoterpene co-occurrence and composition in the essential oil of Thymus vulgaris chemotypes

Thymus vulgaris has a chemical polymorphism with six different chemotypes that show marked spatial segregation in nature. Although some populations have a single chemotype in majority, many have two or three chemotypes. In this study we analyze the quantitative variation among T. vulgaris populations in the percentage of oil composed of the dominant monoterpene(s) for each chemotype. In general, phenolic chemotypes (thymol and carvacrol), which occur at the end of the biosynthetic chain, have a significantly lower proportion of their oil composed of their dominant monoterpene than nonphenolic chemotypes (geraniol, -terpineol, and linalool). This is due to the presence of high amounts of precursors (-terpinene and paracymene) in the oil of phenolic chemotypes. The essential oil of the nonphenolic thuyanol chemotype has four characteristic monoterpenes that together make up a lower proportion of the oil than the single dominant monoterpene of the other nonphenolic chemotypes. For all chemotypes, the percentage composition of the dominant monoterpene decreased significantly at sites where the chemotype is not the majority type. This decrease is correlated with a significant increase in either the proportion of the two precursors for the thymol chemotype or the monoterpenes characteristic of the other chemotypes at the site. The latter result suggests that a plant with dominant genes is responsible for the production of different monoterpenes can produce several molecules.     

Identification and Field Activity of a Male-Produced Aggregation Pheromone in the Pine Sawyer Beetle, <Emphasis Type="Italic">Monochamus galloprovincialis</Emphasis>

The pine sawyer beetle, Monochamus galloprovincialis, is a pest of pine trees in Europe and North Africa. Previously considered a secondary pest of stressed and dying trees, it is now receiving considerable attention as a vector of the pine wood nematode, Bursaphelenchus xylophilus, the causal agent of a lethal wilting disease in susceptible species of pines. Adult beetles are attracted to traps baited with a kairomone blend consisting of a host volatile, α-pinene, and two bark beetle pheromone components, ipsenol and 2-methyl-3-buten-2-ol. More recently it has been shown that mature male M. galloprovincialis produce a pheromone that attracts mature females in a laboratory bioassay. Here, volatiles were collected from mature male and female M. galloprovincialis, and a compound produced specifically by mature males was identified as 2-undecyloxy-1-ethanol from its gas chromatographic retention times, its mass spectrum, and by comparison with synthetic standards. The naturally-derived and synthetic compounds elicited electroantennographic responses from both females and males. Sealed polyethylene vials and polyethylene sachets were shown to be effective dispensers with zero-order release, the latter giving a higher release rate than the former. In two field tests, multiple-funnel traps baited with synthetic 2-undecyloxy-1-ethanol caught both female and male M. galloprovincialis, with higher catches at the higher release rate. This compound also synergized the attractiveness of the kairomone blend, the combined mixture catching 80–140% more beetles than the sum of the catches to each bait separately and luring up to two beetles/trap/d in a moderate-density population. We conclude that 2-undecyloxy-1-ethanol is a male-produced aggregation pheromone of M. galloprovincialis. This is the first example of a sex-specific compound in the cerambycid subfamily Lamiinae with significant behavioral activity in the field at a range sufficient to make it a useful trap bait. The possible roles of this pheromone in the chemical ecology of M. galloprovincialis and its potential use in pine wilt disease management are discussed.     

Effect of Water Stress and Fungal Inoculation on Monoterpene Emission from an Historical and a New Pine Host of the Mountain Pine Beetle

The mountain pine beetle (Dendroctonus ponderosae, MPB) has killed millions of lodgepole pine (Pinus contorta) trees in Western Canada, and recent range expansion has resulted in attack of jack pine (Pinus banksiana) in Alberta. Establishment of MPB in the Boreal forest will require use of jack pine under a suite of environmental conditions different from those it typically encounters in its native range. Lodgepole and jack pine seedlings were grown under controlled environment conditions and subjected to either water deficit or well watered conditions and inoculated with Grosmannia clavigera, a MPB fungal associate. Soil water content, photosynthesis, stomatal conductance, and emission of volatile organic compounds (VOCs) were monitored over the duration of the six-week study. Monoterpene content of bark and needle tissue was measured at the end of the experiment. β-Phellandrene, the major monoterpene in lodgepole pine, was almost completely lacking in the volatile emission profile of jack pine. The major compound in jack pine was α-pinene. The emission of both compounds was positively correlated with stomatal conductance. 3-Carene was emitted at a high concentration from jack pine seedlings, which is in contrast to monoterpene profiles of jack pine from more southern and eastern parts of its range. Fungal inoculation caused a significant increase in total monoterpene emission in water deficit lodgepole pine seedlings right after its application. By 4 weeks into the experiment, water deficit seedlings of both species released significantly lower levels of total monoterpenes than well watered seedlings. Needle tissue contained lower total monoterpene content than bark. Generally, monoterpene tissue content increased over time independent from any treatment. The results suggest that monoterpenes that play a role in pine-MPB interactions differ between lodgepole and jack pine, and also that they are affected by water availability.     

The Influence of Pine Volatile Compounds on the Olfactory Response by <Emphasis Type="BoldItalic">Neodiprion sertifer</Emphasis> (Geoffroy) Females

Females of the pine sawfly Neodiprion sertifer (Hymenoptera Diprionidae) usually avoid Pinus pinea trees as host plants. In contrast, this sawfly species is highly attracted by P. sylvestris and P. nigra trees. Here, we investigated which pine volatiles might mediate this behavior by in situ sampling experiments and olfactometer laboratory tests. Volatiles emitted from P. pinea, P. sylvestris, and P. nigra foliage were sampled by solid-phase microextraction (SPME). Analysis of these volatiles by coupled gaschromatography/mass spectrometry revealed that the relative amounts of the compounds emitted by the three species were significantly different. A discriminant analysis showed that the amounts of limonene and myrcene significantly contributed to the species-specific volatile patterns. Pinus pinea emitted higher relative amounts of limonene than the other pine species. Pinus sylvestris emitted the highest relative amounts of myrcene. When testing the response of N. sertifer females to these pine terpenoids in an olfactometer bioassay, a low amount of limonene was attractive, while a repellent effect was evident when higher amounts were used. The sawfly females showed no significant olfactory response to myrcene. These data suggest that low relative amounts of limonene have a significant function in attracting N. sertifer females, while high amounts might contribute to avoidance of a tree.     

Gallery initiation byTomicus piniperda (Coleoptera: Scolytidae) on scots pine trees baited with host volatiles

In a trapping experiment, wood chips from freshly cut Scots pine trees attracted flying adults of the bark beetleTomicus piniperda (L.) Healthy Scots pine trees, which usually are not attacked byT. piniperda, were baited with chips from freshly cut trees. In other experiments trees were baited with a mixture of (–)--pinene, (+)-3-carene, and terpinolene; the individual monoterpenes; or with ethanol alone. All baited treatments were attacked byT. piniperda. Most of the attacks resulted in short egg galleries and in larval galleries which were only a few millimeters in length. In trees from which a new generation of beetles emerged, net reproduction was well below 1.0. Unbaited control trees remained unattacked or received a few isolated attacks.     

Pine Weevil (Hylobius abietis) Antifeedants from Lodgepole Pine (Pinus contorta)

Pine weevils (Hylobius abietis) fed less on bark of lodgepole pine (Pinus contorta) than on bark of Scots pine (P. sylvestris). Two pine weevil antifeedants, ethyl trans-cinnamate and ethyl 2,3-dibromo-3-phenyl-propanoate, were isolated from bark of lodgepole pine. These two compounds significantly reduced pine weevil feeding in a laboratory bioassay. In field assays, the second compound significantly decreased pine weevil damage on planted seedlings. Ethyl 2,3-dibromo-3-phenylpropanoate has not previously been reported as a natural product.     

Chemical analysis of volatiles emitted by Pinus svlvestris after induction by insect oviposition

Gas chromatography – mass spectrometry analyses of the headspace volatiles of Scots pine (Pinus sylvestris) induced by egg deposition of the sawfly Diprion pini were conducted. The odor blend of systemically oviposition-induced pine twigs, attractive for the eulophid egg parasitoid Chrysonotomyia ruforum, was compared to volatiles released by damaged pine twigs (control) that are not attractive for the parasitoid. The mechanical damage inflicted to the control twigs mimicked the damage by a sawfly female prior to egg deposition. The odor blend released by oviposition-induced pine twigs consisted of numerous mono- and sesquiterpenes, which all were also present in the headspace of the artificially damaged control twigs. A quantitative comparison of the volatiles from oviposition-induced twigs and controls revealed that only the amounts of (E)--farnesene were significantly higher in the volatile blend of the oviposition-induced twigs. Volatiles from pine twigs treated with jasmonic acid (JA) also attract the egg parasitoid. No qualitative differences were detected when comparing the composition of the headspace of JA-treated pine twigs with the volatile blend of untreated control twigs. JA-treated pine twigs released significantly higher amounts of (E)--farnesene. However, the JA treatment induced a significant increase of the amount of further terpenoid components. The release of terpenoids by pine after wounding, egg deposition, and JA treatment is discussed with special respect to (E)--farnesene.     

Magnetic Resonance Imaging of Water Distribution in Welded Woods

This study was performed for a better understanding of water effect on welded wood and improving its water resistance. In this article, we have also attempted to demonstrate the feasibility of using Magnetic Resonance Imaging technology to study water movement in welded woods. Water distribution in welded woods of Scots pine (Pinus sylvestris) and beech (Fagus sylvatica) was investigated by Magnetic Resonance Imaging. Axial specimens were cut from beech and sapwood of Scots pine in longitudinal direction of wood grain. Two pieces of each wood species were welded together by a linear vibration machine. Sub-samples measuring 30 mm × 20 mm × 100 mm were cut from the welded specimens for Magnetic Resonance Imaging. The results showed that weldline of Scots pine was more resistant to water than weldline of beech. Pine joint was still holding after 40 h immersion in water, while a rapid wetting of the beech joint resulted in breakage of the joint in even less than an hour. This preliminary study also showed that MRI is a powerful tool to measure water distribution in welded woods and highlighted the potential of this technique to enhance understanding of wood welding.     

The Lodgepole × Jack Pine Hybrid Zone in Alberta, Canada: A Stepping Stone for the Mountain Pine Beetle on its Journey East Across the Boreal Forest?

Historical data show that outbreaks of the tree killing mountain pine beetle are often preceded by periods of drought. Global climate change impacts drought frequency and severity and is implicated in the range expansion of the mountain pine beetle into formerly unsuitable habitats. Its expanded range has recently reached the lodgepole × jack pine hybrid zone in central Alberta, Canada, which could act as a transition from its historical lodgepole pine host to a jack pine host present in the boreal forest. This field study tested the effects of water limitation on chemical defenses of mature trees against mountain pine beetle-associated microorganisms and on beetle brood success in lodgepole × jack pine hybrid trees. Tree chemical defenses as measured by monoterpene emission from tree boles and monoterpene concentration in needles were greater in trees that experienced water deficit compared to well-watered trees. Myrcene was identified as specific defensive compound, since it significantly increased upon inoculation with dead mountain pine beetles. Beetles reared in bolts from trees that experienced water deficit emerged with a higher fat content, demonstrating for the first time experimentally that drought conditions benefit mountain pine beetles. Further, our study demonstrated that volatile chemical emission from tree boles and phloem chemistry place the hybrid tree chemotype in-between lodgepole pine and jack pine, which might facilitate the host shift from lodgepole pine to jack pine.     

Differences in attraction to semiochemicals present in sympatric pine shoot beetles,Tomicus minor andT. piniperda

The chemical ecology of host- and mate-finding in the pine shoot beetles,Tomicus minor andT. piniperda, was studied in southern Sweden. Beetles were collected in the field from defined attack phases on Scots pine. Using gas chromatography-mass spectroscopy, a number of oxygen-containing monoterpenes, e.g., 3-carene-10-ol, myrtenol,trans-verbenol, and verbenone, were identified from hindgut extracts of both sexes of both species. Compared toT. minor,T. piniperda contained additional compounds and in larger amounts. The amounts were highest in both species at the time when the beetles had bored into contact with the resin-producing xylem-phloem tissue. The synthesis of (1S,6R)-3-carene-10-ol by photooxidatipn of (+)-(1S,6R)-3-carene is described. In comparative electroantennogram (EAG) measurements on males and females of both species, the most active of the tested compounds wastrans-verbenol. Laboratory bioassays of walking beetles showed thatT. piniperda was attracted to uninfestèd pine logs.T. minor was more strongly attracted to pine logs infested with females than to uninfested pine logs, indicating a female-produced aggregation pheromone. Field tests confirmed thatT. piniperda was strongly attracted to pine logs. The attraction ofT. minor to logs was significant only when logs were combined with racemictrans-verbenol and (1S,6R)-3-carene-10-ol.T. minor was also attracted to a combination of these monoterpene alcohols alone. We suggest that host and mate location inT. piniperda is achieved by means of a kairomone composed of host monoterpenes, whileT. minor utilizes a primitive pheromone synergized by host odors. Evolution of host colonization strategies of the two beetles are discussed.Coleoptera: Scolytidae.This study was made within the project Odour signals for control of pest insects.     

Aggregation pheromone components of two species ofPissodes weevils (Coleoptera: Curculionidae) Isolation,identification, and field activity

Two related volatile compounds were identified from each of two species ofPissodes bark weevils and implicated as components of their aggregation pheromones. Grandisol (cis-2-isopropenyl-1-methylcyclobutaneethanol), and its corresponding aldehyde, grandisal, were isolated from males of bothP. strobi andP. approximatus and were found in the abdomens and hindguts of the respective species. In field tests synthetic grandisol and grandisal together with odors from cut pine acted synergistically in attracting both sexes ofP. approximatus. This response was similar to that elicited by maleP. approximatus feeding on cut pine. Males and females of natural populations ofP. strobi were more responsive to caged males feeding on leaders of white pine than they were to leaders alone. The combination of grandisol, grandisal, and leaders was less attractive than males on leaders, but more attractive than leaders alone. From isolation of pheromone components at different times of the year, it was determined that males of both species produced grandisol and grandisal only at times when cohort females were reproductively mature.     

Comparative Investigation of the Volatile Urinary Profiles in Different <Emphasis Type="Italic">Phodopus</Emphasis> Hamster Species

Stir bar sorptive extraction method was used for investigation of the urinary volatile profiles in male and female Phodopus campbelli and Phodopus sungorus hamsters. Additionally, female Phodopus roborowsky urinary profiles were characterized. A quantitative analytical approach allowed comparisons of 17 selected compounds in urine. Results showed that campbelli and sungorus species show similar urinary volatile profiles for males and females. Differences appeared only in concentrations. Several unique compounds, such as pyrazine derivatives, were found to be gender- and age-specific. P. roborowsky females exhibited a completely different urinary volatile profile from campbelli and sungorus females, featuring a unique set of substituted quinoxalines.     

Detection of Pinosylvins in Solid Wood of Scots Pine Using Fourier Transform Raman and Infrared Spectroscopy

Abstract

Diffuse reflectance Fourier transform infrared (DRIFT) and near infrared (NIR) FT-Raman spectroscopy were used to detect pinosylvins in the wood of Pinus sylvestris L. trees. NIR FT-Raman spectroscopy offered the possibility of revealing pinosylvins simply by visual inspection of Raman spectra whereas DRIFT spectra needed a more complicated evaluation. Pinosylvin and resin acids from Scots pine were examined as to the possibility of their being the cause of observed spectral differences between sapwood and heartwood. Since pinosylvins are important compounds for the decay resistance of Scots pine wood, the detection of pinosylvins with Raman spectroscopy might be used to assess durability of wood products.     

Resistance of western white pine to feeding and oviposition byPissodes strobi peck in western canada

Forced and choice feeding and oviposition experiments were conducted by caging overwinteredPissodes strobi on lateral branches or leaders of sympatric Engelmann spruce and/or western white pine trees in Manning Provincial Park, British Columbia, or on transplanted, potted trees in Burnaby, British Columbia. Whereas femaleP. strobi preferred to feed on 1-year-old Engelmann spruce lateral branches in both the forced- and choice-feeding experiments, male weevils failed to discriminate between this host and western white pine. Forced-feeding experiments that utilized 1-year-old leaders demonstrated that western white pine is an acceptable host forP. strobi feeding, but that the native host, Engelmann spruce, is preferred under choice conditions. In all experiments, female weevils oviposited only on Engelmann spruce. These results indicate that the releasing stimulus for feeding is present in the bark of both conifer species and suggests that a separate releasing stimulus for oviposition is absent from western white pine.Coleoptera: Curculionidae.Research supported by Canadian Forestry Service Science Subvention Grant, National Science Foundation (USA) Grant No. BMS-7413643, and National Research Council (Canada) Grant No. A3881.     

Pheromonal Mediation of Intraseasonal Declines in the Attractivity of Female Red-Sided Garter Snakes, Thamnophis sirtalis parietalis

During the breeding season, female red-sided garter snakes (Thamnophis sirtalis parietalis) produce and express a sexual attractiveness pheromone that elicits male courtship behavior. Composed of a homologous series of saturated and monounsaturated methyl ketones, this pheromone is expressed in female skin lipids. Recent studies have shown that the sexual attractivity of unmated female garter snakes declines as the breeding season progresses. Here, we investigated whether temporal changes in the quantity and/or quality of the female sexual attractiveness pheromone are responsible for the observed loss of attractivity. Female red-sided garter snakes were collected immediately following spring emergence and held under natural conditions for the duration of the breeding season. Behavioral experiments confirmed that unmated females become significantly less attractive to males within two weeks of emergence from hibernation. Additionally, these females had lower estradiol concentrations at two weeks post-emergence. Subsequent chemical analyses revealed qualitative variation between the pheromone profiles of newly emerged females and those of females at two weeks post-emergence. Together, these results support the hypothesis that changes in the female sexual attractiveness pheromone are responsible for declining post-emergence female attractivity in garter snakes.     

Defoliation-induced responses in peroxidases,phenolics, and polyamines in scots pine (Pinus sylvestris L.) needles

Effects of artificial defoliation on defensive needle chemistry in Scots pine (Pinus sylvestris L.) were evaluated with particular emphasis on peroxidases, phenolic compounds, soluble sugars, polyamines, and foliar nitrogen levels. The study was carried out on a nutrient-poor Scots pine stand with 8- to 25-year-old trees. Defoliation treatment consisted of repeated defoliation in two successive years with respective control trees. Defoliation was done before needle flushing by removing all mature needles. Guaiacol peroxidase activity increased in the needles after the first defoliation. The difference between treatments diminished towards autumn, and disappeared before the second defoliation in the next summer. After the second defoliation, the activities showed a similar trend. Apparently, peroxidases are involved in inducible chemical changes and recovery reactions that occur in the intact needles shortly after defoliation. After the second defoliation, total nitrogen concentration in the current year needles was about 20% lower, and free putrescine (a polyamine) concentration was 40% lower in the defoliated trees than in control needles. These changes indicate a loss of nitrogen due to defoliation. Specific phenolic compounds such as quercitrin, (+)-catechin, and two catechin derivatives increased in current year needles in response to defoliation. Accumulation of starch and sucrose in the current year needles of repeatedly defoliated trees may imply decreased assimilate transport. The results are indicative that changes in needle phytochemistry in response to defoliation accompany changes in needle nitrogen metabolism.