Does Prescribed Burning Affect Leaf Secondary Metabolites in Pine Stands?

Prescribed burning (PB) is gaining popularity as a low-cost forest protection measure that efficiently reduces fuel build-up, but its effects on tree health and growth are poorly understood. Here, we evaluated the impact of PB on plant defenses in Mediterranean pine forests (Pinus halepensis and P. nigra ssp. laricio). These chemical defenses were estimated based on needle secondary metabolites (terpenes and phenolics including flavonoids) and discussed in terms of chlorophyll fluorescence and soil nutrients. Three treatments were applied: absence of burning (control plots); single burns (plots burned once); and repeated burns (plots burned twice). For single burns, we also explored changes over time. In P. laricio, PB tended to trigger only minor modifications consisting exclusively of short-lived increases (observed within 3 months after PB) in flavonoid index, possibly due to the leaf temperature increase during PB. In P. halepensis, PB had detrimental effects on physiological performance, consisting of (i) significant decreases in actual PSII efficiency (ΦPSII) in light-adapted conditions after repeated PB, and (ii) short-lived decreases in variable-to-maximum fluorescence ratio (Fv/Fm) after single PB, indicating that PB actually stressed P. halepensis trees. Repeated PB also promoted terpene-like metabolite production, which increased 2 to 3-fold compared to control trees. Correlations between terpene metabolites and soil chemistry were found. These results suggest that PB impacts needle secondary metabolism both directly (via a temperature impact) and indirectly (via soil nutrients), and that these impacts vary according to species/site location, frequency and time elapsed since last fire. Our findings are discussed with regard to the use of PB as a forest management technique and its consequences on plant investment in chemical defenses.     

Structure–Activity Relationships of Benzoic Acid Derivatives as Antifeedants for the Pine Weevil, Hylobius abietis

Aromatic organic compounds found in the feces of the pine weevil, Hylobius abietis (L.) (Coleoptera: Curculionidae), have been shown to deter feeding behavior in this species, which is a serious pest of planted conifer seedlings in Europe. We evaluated 55 benzoic acid derivatives and a few homologs as antifeedants for H. abietis. Structure–activity relationships were identified by bioassaying related compounds obtained by rational syntheses of functional group analogs and structural isomers. We identified five main criteria of efficiency as antifeedants among the benzoic acid derivatives. By predicting optimal structures for H. abietis antifeedants, we attempted to find a commercial antifeedant to protect conifer seedlings against damage by H. abietis in regenerating forests. New, highly effective antifeedants are methyl 2,4-dimethoxybenzoate, isopropyl 2,4-dimethoxybenzoate, methyl 2-hydroxy-3-methoxybenzoate, methyl (3,5-dimethoxyphenyl)acetate, and methyl (2,5-dimethoxyphenyl)acetate. Of these, methyl 2,4-dimethoxybenzoate and isopropyl 2,4-dimethoxybenzoate have the highest antifeedant indices of all substances tested and are the best candidates for practical applications in order to protect planted seedlings in the field.     

Do Pine Voles (Microtus pinetorum) Use Numerous Types of Olfactory Cues to Discriminate Gender?

Olfactory cues can contain information for discrimination of gender that can affect subsequent social interactions. Social rodents are hypothesized to use more olfactory cues than nonsocial rodents to distinguish males from females. The generality of this hypothesis was tested using the pine vole (Microtus pinetorum), a social vole. We examined nine possible sources of odors. A slide containing an odor from a male and a female was presented to each test subject for 3 min. We recorded the amount of time each test subject spent investigating each odor. Females spent significantly more time investigating male urine and male anogenital odors; however, they did not use any other odor sources to discriminate gender. In contrast, males did not use any odor sources to discriminate gender. Our results do not support the hypothesis that all social rodents use numerous odor sources to discriminate gender. Instead, our results are consistent with the alternative hypothesis that use of odor sources to convey information about gender may differ in rodents that live in different microhabitats.     

Synergistic Effect of Ethanol to α-Pinene in Primary Attraction of the Larger Pine Shoot Beetle, Tomicus piniperda

-Pinene and ethanol were released in the approximate proportions 1:0.1, 1:0.9 and 1:9 (at 21°C). Ethanol, released in the range of 3–279 mg/day, generally synergized the attraction of T. piniperda to -pinene (30 mg/day at 21°C), although attraction to the mixtures varied within and between years. The low release rate of ethanol together with -pinene attracted a significantly higher number of beetles than -pinene alone in 1995, April of 1996, and in 1997. Lures with the medium release rate of ethanol were the most attractive only in March of 1996. The high dose of ethanol significantly synergized attraction to -pinene in 1995 and 1997. The variable attraction of T. pinipeda to ethanol and -pinene at various release rates and proportions may be due to the temperature dependent nature of beetle antennal sensitivity. At ambient temperatures of 10–13°C. T. piniperda was most attracted to the lures with -pinene and high release rates of ethanol, at 14–17°C it was most attracted to those with medium release rates of ethanol, and at 18°C and higher it was most attraacted to those with low release rates of ethanol.     

Wood Material Behavior During Drying: Moisture-Dependent Tensile Stiffness and Strength of Radiata Pine at 70–150°C

Improvements in kiln-drying efficiency and dried timber product quality are difficult to realize given the current lack of knowledge as to how in-kiln conditions impact wood basic properties such as strength and stiffness. In the last decade at Scion, significant effort has been invested in an initiative to evaluate the instantaneous stress-strain, time-dependent creep, and mechanosorptive behavior of radiata pine. The strength and stiffness of longitudinal specimens of radiata pine wood from the inner and outer portions of the stem have been evaluated across a range of low to high moisture contents at temperatures from 70 to 150°C. Results show expected decreases in both yield stress and MOE with increasing temperature and increasing moisture. Inner-wood is generally less rigid and strong than outer-wood. This article will review methodology and experimental results in addition to discussing trends in MOE and yield strength with temperature, moisture content, and sample type.     

Screening and Functional Analysis of the Peroxiredoxin Specifically Expressed in Bursaphelenchus xylophilus—The Causative Agent of Pine Wilt Disease

The pine wood nematode, Bursaphelenchus xylophilus, is the causal agent of pine wilt disease. Accurately differentiating B. xylophilus from other nematodes species, especially its related species B. mucronatus, is important for pine wood nematode detection. Thus, we attempted to identify a specific protein in the pine wood nematode using proteomics technology. Here, we compared the proteomes of B. xylophilus and B. mucronatus using Two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization-time-of-flight/time-of-flight (MALDI-TOF/TOF-MS) technologies. In total, 15 highly expressed proteins were identified in B. xylophilus compared with B. mucronatus. Subsequently, the specificity of the proteins identified was confirmed by PCR using the genomic DNA of other nematode species. Finally, a gene encoding a specific protein (Bx-Prx) was obtained. This gene was cloned and expressed in E. coli. The in situ hybridisation pattern of Bx-Prx showed that it was expressed strongly in the tail of B. xylophilus. RNAi was used to assess the function of Bx-Prx, the results indicated that the gene was associated with the reproduction and pathogenicity of B. xylophilus. This discovery provides fundamental information for identifying B. xylophilus via a molecular approach. Moreover, the purified recombinant protein has potential as a candidate diagnostic antigen of pine wilt disease, which may lead to a new immunological detection method for the pine wood nematode.     

Pine Bark and Green Tea Concentrated Extracts: Antioxidant Activity and Comprehensive Characterization of Bioactive Compounds by HPLC–ESI-QTOF-MS

The consumption of polyphenols has frequently been associated with low incidence of degenerative diseases. Most of these natural antioxidants come from fruits, vegetables, spices, grains and herbs. For this reason, there has been increasing interest in identifying plant extract compounds. Polymeric tannins and monomeric flavonoids, such as catechin and epicatechin, in pine bark and green tea extracts could be responsible for the higher antioxidant activities of these extracts. The aim of the present study was to characterize the phenolic compounds in pine bark and green tea concentrated extracts using high-performance liquid chromatography coupled to electrospray ionization mass spectrometry (HPLC–ESI-QTOF-MS). A total of 37 and 35 compounds from pine bark and green tea extracts, respectively, were identified as belonging to various structural classes, mainly flavan-3-ol and its derivatives (including procyanidins). The antioxidant capacity of both extracts was evaluated by three complementary antioxidant activity methods: Trolox equivalent antioxidant capacity (TEAC), ferric reducing antioxidant power (FRAP) and oxygen radical absorbance capacity (ORAC). Higher antioxidant activity values by each method were obtained. In addition, total polyphenol and flavan-3-ol contents, which were determined by Folin–Ciocalteu and vanillin assays, respectively, exhibited higher amounts of gallic acid and (+)-catechin equivalents.     

Heat Treatment Effect on Lignin and Carbohydrates in Corsican Pine Earlywood and Latewood Studied by PY–GC–MS Technique

Lignin-derived degradation products from non-treated (NT) and heat-treated (T) Corsican pine (Pinus nigra subsp. laricio) obtained by pyrolysis–gas chromatography–mass spectrometry (Py–GC–MS) were investigated, whereby the earlywood (EW) and latewood (LW) parts of the annual ring were considered separately. The data evaluation was done by principal component analysis (PCA) and the Kruskal–Wallis test. There are no differences in the pyrolysis products composition between EW and LW, but NT and T samples were discernible by PCA applied to Py–GC–MS data. Less phenols with longer chains (4-vinylguaiacol, and trans-isoeugenol) than those with shorter chains (guaiacol, 4-methylguaiacol) and an increase of anhydrosugar (AHS) were found among the pyrolysis products after heat treatment. These signs for autocondensation and side chain cleavages in the lignin during heat treatment were more evident in the EW than in the LW and for the crystallization of cellulose. A slight decrease of the carbohydrate/lignin ratio (C/L) after heat treatment indicates a greater degradation of carbohydrates compared to lignin. The relation of pyrolysis products of lignin and mechanical properties of wood was evaluated by regression analysis. An inverse correlation between short-chain phenols and MOE and a direct correlation between long-chain phenols and compression strength was found in case of NT wood, while a weak positive correlation could be observed between short-chain phenols and the density in T wood.     

Enantiomeric Composition of Monoterpene Hydrocarbons in Some Conifers and Receptor Neuron Discrimination of α-Pinene and Limonene Enantiomers in the Pine Weevil, Hylobius abietis

The enantiomeric composition of seven monoterpene hydrocarbons in headspace volatiles of spruce sawdust and seedlings (Picea abies), pine seedlings (Pinus sylvestris), and branches of juniper (Juniperus communis) was determined by gas chromatographic separation on a -Cyclodextrin column. For the six monoterpenes, -pinene, camphene, -pinene, sabinene, limonene, and -phellandrene, both enantiomers were present, whereas for 3-carene only the (+)-configuration was found. The amount of each enantiomer varied considerably both in relation to total amount of all of them, and for the six pairs also in relation to the opposite enantiomer. One olfactory receptor neuron in the pine weevil (Hylobius abietis) showed a strong response to -pinene when stimulated with all four headspace materials via a GC equipped with a DB-WAX column. The same neuron was subsequently tested with repeated stimulations via the GC effluent containing the (+)- or (–)-enantiomer. A marked better response to (+)- than to (–)--pinene was elicited. Another olfactory receptor neuron that responded strongly to limonene when stimulated with the spruce volatiles was tested for enantiomers of limonene. This neuron responded more strongly to (–)- than to (+)-limonene, when stimulated alternately with each of the limonene enantiomers. Discrimination between enantiomers by plant olfactory receptor neurons suggests that the enantiomeric ratios of volatile compounds may be important in host location by the pine weevil.