Terpenoids are involved in the expression of systemic-induced resistance in Austrian pine.

Terpenoids are defense metabolites that are induced upon infection or wounding. However, their role in systemic-induced resistance (SIR) is not known. Here, we explored the role of terpenoids in this phenomenon at a very early stage in the interaction between Austrian pine and the tip blight and canker pathogen Diplodia pinea. We induced Austrian pine saplings by either wounding or inoculating the lower stems with D. pinea. The seedlings were then challenged after 12 h, 72 h, or 10 days with D. pinea on the stem 15 cm above the induction. Lesion lengths and terpenoids were quantified at both induction and challenge locations. Key terpenoids were assayed for antifungal activity in in vitro bioassays. SIR increased with time and was correlated with the inducibility of several compounds. α-Pinene and a cluster of ß-pinene, limonene, benzaldehyde, dodecanol, and n-dodecyl acrylate were positively correlated with SIR and were fungistatic in vitro, while other compounds were negatively correlated with SIR and appeared to serve as a carbon source for D. pinea. This study shows that, overall, terpenoids are involved in SIR in this system, but their role is nuanced, depending on the type of induction and time of incubation. We hypothesize that some, such as α-pinene, could serve in SIR signaling.

Effects of graphene on morphology, microstructure and transcriptomic profiling of Pinus tabuliformis Carr. roots.

Graphene has shown great potential for improving growth of many plants, but its effect on woody plants remains essentially unstudied. In this work, Pinus tabuliformis Carr. bare-rooted seedlings grown outdoors in pots were irrigated with a graphene solution over a concentration range of 0-50 mg/L for six months. Graphene was found to stimulate root growth, with a maximal effect at 25 mg/L. We then investigated root microstructure and carried out transcript profiling of root materials treated with 0 and 25 mg/L graphene. Graphene treatment resulted in plasma-wall separation and destruction of membrane integrity in root cells. More than 50 thousand of differentially expressed genes (DEGs) were obtained by RNA sequencing, among which 6477 could be annotated using other plant databases. The GO enrichment analysis and KEGG pathway analysis of the annotated DEGs indicated that abiotic stress responses, which resemble salt stress, were induced by graphene treatment in roots, while responses to biotic stimuli were inhibited. Numerous metabolic processes and hormone signal transduction pathways were altered by the treatment. The growth promotion effects of graphene may be mediated by encouraging proline synthesis, and suppression of the expression of the auxin response gene SMALL AUXIN UP-REGULATED RNA 41 (SAUR41), PYL genes which encode ABA receptors, and GSK3 homologs.

Transcriptome-Wide Identification of WRKY Transcription Factors and Their Expression Profiles under Different Types of Biological and Abiotic Stress in Pinus massoniana Lamb.

Pinus massoniana Lamb, an economically important conifer tree, is widely distributed in China. WRKY transcription factors (TFs) play important roles in plant growth and development, biological and abiotic stress. Nevertheless, there is little information about the WRKY genes in P. massoniana. By searching for conserved WRKY motifs in transcriptomic RNA sequencing data for P. massoniana, 31 sequences were identified as WRKY TFs. Then, phylogenetic and conserved motif analyses of the WRKY family in P. massoniana, Pinus taeda and Arabidopsis thaliana were used to classify WRKY genes. The expression patterns of six PmWRKY genes from different groups were determined using real-time quantitative PCR for 2-year-old P. massoniana seedings grown in their natural environment and challenged by phytohormones (salicylic acid, methyl jasmonate, or ethephon), abiotic stress (H2O2) and mechanical damage stress. As a result, the 31 PmWRKY genes identified were divided into three major groups and several subgroups based on structural and phylogenetic features. PmWRKY genes are regulated in response to abiotic stress and phytohormone treatment and may participate in signaling to improve plant stress resistance. Some PmWRKY genes behaved as predicted based on their homology with A. thaliana WRKY genes, but others showed divergent behavior. This systematic analysis lays the foundation for further identification of WRKY gene functions to aid further exploration of the functions and regulatory mechanisms of PmWRKY genes in biological and abiotic stress in P. massoniana.

Comparative Transcriptome Analysis of Pine Trees Treated with Resistance-Inducing Substances against the Nematode Bursaphelenchus xylophilus.

The pinewood nematode (PWN) Bursaphelenchus xylophilus causes pine wilt disease, which results in substantial economic and environmental losses across pine forests worldwide. Although systemic acquired resistance (SAR) is effective in controlling PWN, the detailed mechanisms underlying the resistance to PWN are unclear. Here, we treated pine samples with two SAR elicitors, acibenzolar-S-methyl (ASM) and methyl salicylic acid (MeSA) and constructed an in vivo transcriptome of PWN-infected pines under SAR conditions. A total of 252 million clean reads were obtained and mapped onto the reference genome. Compared with untreated pines, 1091 and 1139 genes were differentially upregulated following the ASM and MeSA treatments, respectively. Among these, 650 genes showed co-expression patterns in response to both SAR elicitors. Analysis of these patterns indicated a functional linkage among photorespiration, peroxisome, and glycine metabolism, which may play a protective role against PWN infection-induced oxidative stress. Further, the biosynthesis of flavonoids, known to directly control parasitic nematodes, was commonly upregulated under SAR conditions. The ASM- and MeSA-specific expression patterns revealed functional branches for myricetin and quercetin production in flavonol biosynthesis. This study will enhance the understanding of the dynamic interactions between pine hosts and PWN under SAR conditions.

Rising [CO2] effect on leaf drought-induced metabolome in Pinus pinaster Aiton: Ontogenetic- and genotypic-specific response exhibit different metabolic strategies.

Rising atmospheric CO2 concentrations ([CO2]) together with water deficit can influence ecological interactions of trees through an array of chemically driven changes in plant leaves. In four drought stressed Pinus pinaster genotypes, grown under two levels of atmospheric [CO2] (ambient (aCO2) and enriched (eCO2)) the metabolome of adult and juvenile needles was analyzed to know if the metabolic responses to this environmental situation could be genotype-dependent and vary according to the stage of needle ontogeny. Drought had the highest incidence, followed by needle ontogeny, being lower the eCO2 effect. The eCO2 reduced, eliminated or countered the 50 (adult needles) - 44% (juvenile) of the drought-induced changes, suggesting that CO2-enriched plants could perceived less oxidative stress under drought, and proving that together, these two abiotic factors triggered a metabolic response different from that under single factors. Genotype drought tolerance and ontogenetic stage determined the level of metabolite accumulation and the plasticity to eCO2 under drought, which was mainly reflected in antioxidant levels and tree chemical defense. At re-watering, previously water stressed plants showed both, reduced C and N metabolism, and a "drought memory effect", favoring antioxidants and osmolyte storage. This effect showed variations regarding genotype drought-tolerance, needle ontogeny and [CO2], with remarkable contribution of terpenoids. Chemical defense and drought tolerance were somehow linked, increasing chemical defense during recovery in the most drought-sensitive individuals. The better adaptation of trees to drought under eCO2, as well as their ability to recover better from water stress, are essential for the survival of forest trees.

Using morphological attributes for the fast assessment of nutritional responses of Buddhist pine (Podocarpus macrophyllus [Thunb.] D. Don) seedlings to exponential fertilization.

Culturing slowly growing tree seedlings is a potential approach for managing the conflict between the increasing demand for ornamental stock and the decreasing area of farmlands due to urbanization. In this study, Buddhist pine (Podocarpus macrophyllus [Thunb.] D. Don) seedlings were raised in multishelves with light-emitting diode lighting in the spectrum of 17:75:8 (red:green:blue) at 190-320 µmol m-2 s-1 with controlled temperature and relative humidity at 19.5°C and 60%, respectively. Seedlings were fed by exponential fertilization (EF) (nitrogen [N]-phosphorus [P]2O5-K2O, 10-7-9) at eight rates of 0 (control), 20 (E20), 40 (E40), 60 (E60), 80 (E80), 100 (E100), 120 (E120), and 140 (E140) mg N seedling-1 for four months through 16 fertilizer applications. The nutritional responses of Buddhist pine seedlings can be identified and classified into various stages in response to increasing doses, up to and over 120 N seedling-1. Morphological traits, i.e., the green color index and leaf area (LA) obtained by digital analysis and the fine root growth, all remained constant in response to doses that induced steady nutrient loading. LA had a positive relationship with most of the nutritional parameters. A dose range between 60 and 120 mg N seedling-1 was recommended for the culture of Buddhist pine seedlings. At this range of fertilizer doses, measuring the leaf area through digital scanning can easily and rapidly indicate the inherent nutrient status of the seedlings.

Increased endogenous indole-3-acetic acid:abscisic acid ratio is a reliable marker of Pinus massoniana rejuvenation.

Pinus massoniana is a recalcitrant tree species for rooting in vitro. We rejuvenated 26-year-old P. massoniana trees by successive grafting. Rooting rates of rejuvenated shoots were > 83.1% after rooting induction. We compared endogenous levels of indole-3-acetic acid (IAA), abscisic acid (ABA), gibberellins (GAs) and zeatin-riboside (ZR), and the rhizogenesis ability of axillary shoots of mature and rejuvenated materials in vitro, i.e., somaplants and grafts. Enhancement of the rooting ability of mature materials in vitro following somatic embryogenesis or repeated grafting onto juvenile rootstocks was accompanied by increased IAA and GAs levels, and by decreased ABA levels in scions used as starting material for micropropagation in vitro. Successive subcultures did not influence the rooting ability of shoots from untreated mature material. Rooting ability of shoots in vitro, however, gradually increased with subculture frequency during repeated subculturing in grafting materials. The IAA:ABA ratio in shoots in vitro after grafting five times, and consequently capable of root organogenesis, was higher than in shoots of untreated mature material incapable of root organogenesis in vitro. A high IAA:ABA ratio was detected in scions of somaplants that were capable of rooting in vitro despite subculture times. We found that the endogenous IAA:ABA ratio is a reliable marker for the recovery of root organogenesis in vitro after rejuvenating treatments for mature P. massoniana trees.

Effects of Urban-rural Atmospheric Environment on Heavy Metal Accumulation and Resistance Characteristics of Pinus tabulaeformis in Northern China.

The washed and unwashed current (C) and previous year (C + 1) needles, branches and top soils of Pinus tabulaeformis trees were sampled at five sites Haitai industrial district (HT), Puji River (PJ), Fukang road (FK), Residential area (RA) and Baxian Mountain (BX) in Tianjin along an urban-rural gradient and analyzed for heavy metals (Cu, Mn, Zn, Pb and Cd) concentrations via ICP. C + 1 needles generally had higher Mn, Pb, Cd than C needles while the opposite was for Cu and Zn. Total Cu, Zn, Pb, Cd in soils peaked at HT and decreased at RA and BX. Heavy metals were generally higher in the unwashed needles than the washed needles at all sites. Meanwhile MDA, soluble sugar and free proline concentration in needles were increased with the increasing of heavy metal contents along the urban-rural gradient, further correlated with the heavy metal contents.

Ectomycorrhizal fungi may not act as a barrier inhibiting host plant absorption of heavy metals.

Whether the huge external hyphal system of ectomycorrhizae that promotes host plants' acquisition of water and nutrients can selectively inhibit their transport of heavy metals at the same time remains unclear. In this experiment, we designed and conducted two types of soil-pot test to clarify the effects of EMF on the absorption and transport of copper (Cu) and cadmium (Cd) by host Pinus thunbergii seedlings. In the root-bag test, external hyphae took the initiative into the Cu/Cd-contaminated bulk soil, absorb and transport Cu and Cd to the rhizosphere soils and further transport it to the shoots of the host plants. Inoculation with EMF also promoted the uptake of nutrients by host plants, thereby increasing their biomass and improving Cu/Cd tolerance compared with non-inoculated plants. Inoculation with EMF species with higher Cu or Cd tolerance generated more phytostabilization and phytoextraction of Cu or Cd by host plants. In a short-term exposure test, inoculation with EMF accelerated the absorption of Cu and Cd by P. thunbergii within 12 h of Cu or Cd irrigation. Therefore, we concluded that EMF do not act as a barrier inhibiting the absorption of heavy metals by host plants, but rather promote this absorption. Improving the plant's nutritional status and promoting growth, diluting heavy metal concentrations, thereby reducing the toxic effects of heavy metals on host plants. These results provide the theoretical basis for the application of EMF in plant-microbial combinations for the phytostabilization and phytoextraction of heavy metal-contaminated soils.

Metabolic response to elevated CO2 levels in Pinus pinaster Aiton needles in an ontogenetic and genotypic-dependent way.

Global climate changes involve elevated atmospheric [CO2], fostering the carbon allocation to tree sink tissues, partitioning it into metabolic pathways. We use metabolomics analysis in adult and juvenile needles of four Pinus pinaster genotypes exposed to two levels of growth [CO2]: ambient (400 µmol mol-1) and enriched (800 µmol mol-1), to know if the metabolic responses are genotype-dependent and vary according to the stage of needle ontogeny. The eCO2-induced changes in the needle metabolomes are more significant in secondary metabolism pathways and especially meaningful in juvenile needles. The heteroblasty has important consequences in the expression of the metabolome, and on the plasticity to CO2, determining the level of specific metabolite accumulation, showing an interdependence between adult and juvenile needles. The P. pinaster needle metabolomes also show clear quantitative differences linked to genotype, as well as regarding the metabolic response to eCO2, showing both, common and genotype-specific biochemical responses. Thus, the changes in flavonol levels are mainly genotype-independent, while those in terpenoid and free fatty acids are mainly genotype-dependent, ratifying the importance of genotype to determine the metabolic response to eCO2. To understand the adaptation mechanisms that tree species can develop to cope with eCO2 it is necessary to know the genetically distinct responses within a species to recognize the CO2-induced changes from the divergent approaches, what can facilitate knowing also the possible interrelation of the physiological and metabolic responses. That could explain the controversial effects of eCO2 on the carbon-based metabolite in conifers, at the inter- and intra-specific level.

Tissue accumulation patterns and concentrations of potassium, phosphorus, and carboxyfluorescein translocated from pine seed to the root.

MAIN CONCLUSION: Potassium (K), phosphorous (P), and carboxyfluorescein (CF) accumulate in functionally distinct tissues within the pine seedling root cortex. Seedlings of Pinus pinea translocate exogenous CF and endogenous K and P from the female gametophyte/cotyledons to the growing radicle. Following unloading in the root tip, these materials accumulate in characteristic spatial patterns. Transverse sections of root tips show high levels of P in a circular ring of several layers of inner cortical cells. K and CF are minimal in the high P tissue. In contrast, high levels of K and CF accumulate in outer cortical cells, and in the vascular cylinder. These patterns are a property of living tissue because they change after freeze-thaw treatment, which kills the cells and results in uniform distribution of K and P. K concentration can be reduced to undetectable levels by incubation of roots in 100 mM NaCl. Inductively coupled plasma optical emission spectrometry (ICP-OES) analysis and scanning electron microscopy (SEM)/energy-dispersive X-ray spectroscopy (EDS) of root segments both reliably determine K and P concentrations.

Single-generation effects on terpenoid defenses in lodgepole pine populations following mountain pine beetle infestation.

The recent mountain pine beetle outbreak in western Canada provides an opportunity to study the selection and heritability of tree defenses. We examined terpenoid-based defenses of seedling lodgepole pines which were offspring of mature trees subjected to high levels of mountain pine beetle selective pressure. Seedlings were grown from one of three types of cones: old cones on live trees; young cones on live trees; and cones on trees killed by beetles. Offspring thus represented crosses of non-surviving (NS) x surviving (S), S x S, and NS x NS parents, respectively. Methyl jasmonate was used to induce a defensive reaction in the seedlings. Seed source had a significant effect on levels of ten different terpenes, but not on total terpene concentrations. When the seedlings were grouped by location and treatment type, the seedlings of different cone types could be almost entirely distinguished by terpene profiles.

Limited evidence for CO2 -related growth enhancement in northern Rocky Mountain lodgepole pine populations across climate gradients.

Forests sequester large amounts of carbon annually and are integral in buffering against effects of global change. Increasing atmospheric CO2 may enhance photosynthesis and/or decrease stomatal conductance (gs ) thereby enhancing intrinsic water-use efficiency (iWUE), having potential indirect and direct benefits to tree growth. While increasing iWUE has been observed in most trees globally, enhanced growth is not ubiquitous, possibly due to concurrent climatic constraints on growth. To investigate our incomplete understanding of interactions between climate and CO2 and their impacts on tree physiology and growth, we used an environmental gradient approach. We combined dendrochronology with carbon isotope analysis (δ13 C) to assess the covariation of basal area increment (BAI) and iWUE over time in lodgepole pine. Trees were sampled at 18 sites spanning two climatically distinct elevation transects on the lee and windward sides of the Continental Divide, encompassing the majority of lodgepole pine's northern Rocky Mountain elevational range. We analyzed BAI and iWUE from 1950 to 2015, and explored correlations with monthly climate variables. As expected, iWUE increased at all sites. However, concurrent growth trends depended on site climatic water deficit (CWD). Significant growth increases occurred only at the driest sites, where increases in iWUE were strongest, while growth decreases were greatest at sites where CWD has been historically lowest. Late summer drought of the previous year negatively affected growth across sites. These results suggest that increasing iWUE, if strong enough, may indirectly benefit growth at drier sites by effectively extending the growing season via reductions in gs . Strong growth decreases at high elevation windward sites may reflect increasing water stress as a result of decreasing snowpack, which was not offset by greater iWUE. Our results imply that increasing iWUE driven by decreasing gs may benefit tree growth in limited scenarios, having implications for future carbon uptake potential of semiarid ecosystems.

UV-B-induced forest sterility: Implications of ozone shield failure in Earth's largest extinction.

Although Siberian Trap volcanism is considered a primary driver of the largest extinction in Earth history, the end-Permian crisis, the relationship between these events remains unclear. However, malformations in fossilized gymnosperm pollen from the extinction interval suggest biological stress coinciding with pulsed forest decline. These grains are hypothesized to have been caused by enhanced ultraviolet-B (UV-B) irradiation from volcanism-induced ozone shield deterioration. We tested this proposed mechanism by observing the effects of inferred end-Permian UV-B regimes on pollen development and reproductive success in living conifers. We find that pollen malformation frequencies increase fivefold under high UV-B intensities. Surprisingly, all trees survived but were sterilized under enhanced UV-B. These results support the hypothesis that heightened UV-B stress could have contributed not only to pollen malformation production but also to deforestation during Permian-Triassic crisis intervals. By reducing the fertility of several widespread gymnosperm lineages, pulsed ozone shield weakening could have induced repeated terrestrial biosphere destabilization and food web collapse without exerting a direct "kill" mechanism on land plants or animals. These findings challenge the paradigm that mass extinctions require kill mechanisms and suggest that modern conifer forests may be considerably more vulnerable to anthropogenic ozone layer depletion than expected.

Contrasting acclimation abilities of two dominant boreal conifers to elevated CO2 and temperature.

High latitude forests will experience large changes in temperature and CO2 concentrations this century. We evaluated the effects of future climate conditions on 2 dominant boreal tree species, Pinus sylvestris L. and Picea abies (L.) H. Karst, exposing seedlings to 3 seasons of ambient (430 ppm) or elevated CO2 (750 ppm) and ambient temperatures, a + 4 °C warming or a + 8 °C warming. Pinus sylvestris responded positively to warming: seedlings developed a larger canopy, maintained high net CO2 assimilation rates (Anet ), and acclimated dark respiration (Rdark ). In contrast, carbon fluxes in Picea abies were negatively impacted by warming: maximum rates of Anet decreased, electron transport was redirected to alternative electron acceptors, and thermal acclimation of Rdark was weak. Elevated CO2 tended to exacerbate these effects in warm-grown Picea abies, and by the end of the experiment Picea abies from the +8 °C, high CO2 treatment produced fewer buds than they had 3 years earlier. Treatments had little effect on leaf and wood anatomy. Our results highlight that species within the same plant functional type may show opposite responses to warming and imply that Picea abies may be particularly vulnerable to warming due to low plasticity in photosynthetic and respiratory metabolism.

Chitosan-based nanoparticles of avermectin to control pine wood nematodes.

Pine wood nematode disease is a most devastating disease of pine trees. Avermectin (AVM) is a widely used bio-nematocide which can effectively to kill the pine wood nematode (PWN). However, its poor solubility in water and rapid photolysis are responsible for its poor bioavailability, which causes environmental pollution because of excessive applied rates. Here, a simple electrostatic interaction method was used to encapsulate AVM within nanoparticles composed of poly-γ-glutamic acid (γ-PGA) and chitosan (CS). The loading capacity of the resulting AVM-CS/γ-PGA nanoparticles was as much as 30.5%. The encapsulation of AVM within these nanoparticles reduced its losses by more than 20.0% through photolysis. An in vitro test showed that the rate of release of AVM from the nanoparticles was dependent on the ambient pH, with rapid release occurring in an alkaline environment. The mortality rate of nematodes which were treated with 1ppm of AVM content of AVM-CS/γ-PGA was 98.6% after 24h, while one of free AVM was only 69.9%. In addition, FITC-labeled CS/γ-PGA nanoparticles (FITC-CS/γ-PGA) showed that the nanoparticles could enrich in intestines and head of nematodes. All of these results showed that those nanoparticles of AVM are a potential multifunctional formulation to control the pest and reduce environment pollution.

Profiling methyl jasmonate-responsive transcriptome for understanding induced systemic resistance in whitebark pine (Pinus albicaulis).

KEY MESSAGE: RNA-seq analysis on whitebark pine needles demonstrated that methyl jasmonate (MeJA)-triggered transcriptome re-programming substantially overlapped with defense responses against insects and fungal pathogens in Pinus species, increasing current knowledge regarding induced systemic resistance (ISR) to pathogens and pests in whitebark pine. Many whitebark pine populations are in steep decline due to high susceptibility to mountain pine beetle and the non-native white pine blister rust (WPBR). Resistance, including induced systemic resistance (ISR), is not well characterized in whitebark pine, narrowing the current options for increasing the success of restoration and breeding programs. Exogenous jasmonates are known to trigger ISR by activating the plant's immune system through regulation of gene expression to produce chemical defense compounds. This study reports profiles of whitebark pine needle transcriptomes, following methyl jasmonate (MeJA) treatment using RNA-seq. A MeJA-responsive transcriptome was de novo assembled and transcriptome profiling identified a set of differentially expressed genes (DEGs), revealing 1422 up- and 999 down-regulated transcripts with at least twofold change (FDR corrected p < 0.05) in needle tissues in response to MeJA application. GO analysis revealed that these DEGs have putative functions in plant defense signalling, transcription regulation, biosyntheses of secondary metabolites, and other biological processes. Lineage-specific expression of defense-related genes was characterized through comparison with MeJA signalling in model plants. In particular, MeJA-triggered transcriptome re-programming substantially overlapped with defense responses against WPBR and insects in related Pinus species, suggesting that MeJA may be used to improve whitebark pine resistance to pathogens/pests. Our study provides new insights into molecular mechanisms and metabolic pathways involved in whitebark pine ISR. DEGs identified in this study can be used as candidates to facilitate identification of genomic variation contributing to host resistance and aid in breeding selection of elite genotypes with better adaptive fitness to environmental stressors in this endangered tree species.

Phosphite shifts physiological and hormonal profile of Monterey pine and delays Fusarium circinatum progression.

Fusarium circinatum is the causal agent of pitch canker disease affecting Pinus spp. and Pseudotsuga menziesii worldwide. Under strict quarantine measures, alternative approaches for disease control are necessary. Phosphite (Phi) salts are known for their fungicidal activity and as plant resistance elicitors; however, its potential is yet to be acknowledged in the Pinus-F. circinatum model. The main aim of this study was to assess whether the application of a Phi-based commercial formulation would delay the progression of the pitch canker on Pinus radiata plants, and on the in vitro fungal growth. In vitro assays were performed using different Phi concentrations (1% and 4%) and a non-treated control (0%), and repeated in vivo using inoculated and non-inoculated plants. Plant physiological parameters and hormonal content were evaluated. Phi was effective at inhibiting in vitro mycelial growth in a dose dependent manner. Regardless of fungal inoculation, Phi application induced positive effects on plant performance, despite phytotoxic effects found at 4%. Fusarium circinatum infection led to a reduction in gas exchange and chlorophyll fluorescence (Fv/Fm and φPSII), while proline and hormone (JA, ABA and SA) levels increased. Phi was effective in delaying disease symptom development in a dose dependent manner, concurrent with in vitro observations: gas exchange and chlorophyll fluorescence (Fv/Fm) were unaffected; proline, MDA and ABA decreased; electrolyte leakage and total soluble sugars increased. This suggests a direct (pathogen growth inhibition) and indirect (host defense priming) action of Phi, showing that Phi represents a potential strategy to control F. circinatum infection.

Sorption and desorption characteristics of methyl bromide during and after fumigation of pine (Pinus radiata D. Don) logs.

BACKGROUND: The sorption and desorption characteristics of methyl bromide (MB) were determined during and after fumigation of recently harvested pine (Pinus radiata D. Don) logs. The effects of dose (48 or 120 g m-3 ), degree of bark cover (0, 50 or 100%) and end-grain sealing (sealed or unsealed) on sorption and desorption were determined over time. RESULTS: Sorption of MB was proportional to the dose applied and dependent on the amount of end-grain sealed. After 16 h, an average of 70.7 ± 2.5% of the initial concentration remained in the treated space when end-grains were sealed, whereas only 47.3 ± 2.5% remained when unsealed. During aeration, MB was released from logs, initially ranging from 2.8 to 8.8 g · h m-3 , depending on the treatment. The rate of desorption quickly decreased during aeration. CONCLUSION: The surface area of a log is the most important factor influencing MB sorption and desorption rates, with greater surface area resulting in greater (de)sorption rates. Sorption data can now be combined with insect toxicity data to estimate a minimum effective dose of MB for further evaluation, while desorption data can be combined with fumigant plume modelling to assess worker safety. © 2016 Society of Chemical Industry.

Proteomic analysis of the venom and venom sac of the woodwasp, Sirex noctilio - Towards understanding its biological impact.

UNLABELLED: The European horntail woodwasp, Sirex noctilio, is an invasive insect that attacks conifer hosts, particularly Pinus species. Venom injected by female S. noctilio, together with its symbiotic fungus, damages the normal physiology of Pinus, leading to death of the tree. To identify the proteinaceous components in the venom and uncover the interplay between venom proteins and tree proteins, clarification of the overall profile of proteins produced in the venom gland apparatus was carried out in this work. The venom sac proteome utilised in-solution digested in either a natural or deglycosylated state, prior to nanoHPLC LTQ-Orbitrap under CID/ETD mode. Here, we report the identification of 1454 and 1225 proteins in venom and sac, respectively, with 410 mutual proteins. Approximately 90 proteins were predicted to be secretory, of which 8 have features characteristic of toxins. Chemosensory binding proteins were also identified. Gene ontology and KEGG pathway analysis were employed to predict the protein functions and biological pathways in venom and sac. Protein-protein interaction (PPI) analysis suggested that one-step responses represent the majority of the Sirex-Pinus PPIs, and the proteins representing network hub nodes could be of importance for the development of pest management strategies. SIGNIFICANCE: The woodwasp Sirex noctilio is an invasive species in many parts of the world, including Australia and North America, where it is considered within the top 10 most serious forest insects. Where they have been introduced, the female woodwasps attack living pine trees, causing significant economic losses. Central to this destruction is the woodwasp's life cycle requirement to bore a hole to deposit eggs and a toxic mucus that disables the tree's network for transporting water and nutrients, yet aids in larval survival. Here we specifically examine the mucus gland apparatus and its contents, revealing the protein components that together with 'noctilisin' facilitate this complex association. The identification of chemosensory binding proteins further supports a role for the woodwasp ovipositor as an instrument for early stages of host tree selection. These findings could provide important clues towards the development of novel control tools against this pest.