Deadwood is an important component of forest health and terrestrial carbon storage. However, the diversity of arthropods inhabiting deadwood and the biotic factors influencing decledomposition remain poorly understood, partly due to the challenges associated with studying wood‐dwelling organisms. Ecoacoustic methods—non‐invasive and non‐destructive—offer a promising approach to investigating saproxylic arthropod communities. We tested and optimised ecoacoustic techniques (including recording duration, sensor attachment methods, and soundproofing strategies) for saproxylic detecting abundance and diversity in a multisite experiment spanning Mississippi and Arizona, USA, and Comayagua, Honduras. While linear mixed‐effects models revealed no significant relationships between overall arthropod diversity and ecoacoustic diversity, models incorporating the abundance of particular taxa showed significant associations. Notably, the abundance of Hymenoptera and Blattodea correlated with ecoacoustic diversity measures. Effect size varied, ranging from small (−0.000015 ± 0.000005) to moderate (2.5569 ± 0.7623), depending on the diversity metrics included in the model. This study represents the first application of ecoacoustic methods to access arthropod diversity in deadwood and highlights the potential of these tools for targeting studies of macroarthropods in decomposing wood environments.

US manufacturers are looking to expand the use of cross-laminated timber (CLT) panels into the North American market, including states located in the southeast where termites are important pests. However, there is no current assessment method for determining CLT vulnerability to the highly destructive native termites found in many states across the United States. The impact of damage by these termites is of particularly high interest in areas with suitable climate to their proliferation, such as the southeastern United States. This study evaluated durability of CLT panels and developed a laboratory assay to test susceptibility of this product to termites. Untreated CLT suffered mass losses of up to 5.8% in testing with an average visual rating of 7.2, indicating a moderate to severe attack with 10-30% of the cross section of the product affected by termite intrusion. Recommendations were developed for the inclusion of modifications presented in standardized testing protocols and will be presented to standards organizations. The proposed method may also be applied to evaluate termite resistance of other mass lumber products such as laminated veneer lumber and Glulam.

Soy flour was evaluated as a partial substitute for resin in the manufacture of oriented strand board (OSB), a wood-based composite that often replaces solid lumber and plywood in structural applications in the construction industry. Since the presence of soy could alter OSB biodegradation properties, termite resistance of OSB panels made with 0, 10, and 20 percent of polymeric methylene diphenyl diisocyanate (pMDI) resin substituted with soy flour (OSB0, OSB10, and OSB20, respectively) was investigated. Single choice tests between three types of OSB and southern yellow pine (SYP) solid wood and an OSB choice test (OSB0 vs. OSB10) were evaluated. Results indicated that termites always showed a preference for SYP, with the OSB becoming less palatable when soy flour was present. Percentage weight losses for OSB0, OSB10, and OSB20 were 5.7×, 8.4×, and 8.6× less, respectively, compared with SYP. In the absence of SYP, termites did not differentiate OSB0 from OSB10, with OSB10 showing 1.5× less weight loss compared with OSB0. Visual rating data supported weight loss data, except significantly less damage was only found when the choice paired SYP with OSB made with soy (OSB10 or OSB20). Termite consumption preference for SYP was explained by differences in water absorption kinetics. SYP reached saturation (105% moisture content) within 1 week on moist sand, while moisture content of OSB composites slowly climbed to 79 percent over 4 weeks, never reaching a plateau. Lower moisture content was due to the presence of water-repellent resin and wax in the OSB.

Resistance against infection by the fungus Aspergillus flavus Link in commercial maize (Zea mays L.) is the topic of many studies, but few studies have investigated the effects of A. flavus infection on gene expression levels in ear kernels. A crucial component of gene expression profiling by RT-qPCR is having a reliable set of reference genes that show relatively constant expression across the treatments and phenotypes under study. Currently, however, there is no published information on reference genes suitable for measuring changes in kernel gene expression levels after infection with A. flavus. Thus, in this study, six candidate reference genes (ACT1, β-Tub2, eIF4A2, TATA, EFIα, and GAPDH) were evaluated and ranked according to their expression stability. The genes were amplified from first-strand cDNA samples synthesized from kernels of two susceptible and two resistant maize lines that were either inoculated with A. flavus or water or not inoculated. Three software packages were used to calculate and rank the stability of expression for these genesgeNorm, NormFinder, and BestKeeper. The analysis revealed that the most stable genes to normalize expression levels from maize kernels responding to A. flavus inoculation and wounding were ACT1, EFIα, and eIF4A2.

Bark beetles and root weevils can impact forests through tree death on landscape scales. Recently, subterranean termites have been linked to these beetles via the presence of bluestain fungi (Ascomycota: Ophiostomataceae), which are vectored to trees by beetles. However, only a small subset of bluestain species have been examined. Here, we tested whether termite-bluestain association patterns in the field reflect termite feeding preference in laboratory choice trials. We documented the presence of four bluestain fungi (Leptographium procerum (W.B. Kendr.), L. terebrantis (Barras & Perry), Grosmannia huntii (Rob.-Jeffr.), and G. alacris (T.A. Duong, Z.W. de Beer & M.J. Wingf.) in the roots of 2,350 loblolly pine trees in the southeastern United States and whether termites were present or absent on these roots and paired this with laboratory choice feeding trials. Termites were found 2.5-fold on tree roots with at least one bluestain fungus present than tree roots without bluestain fungi. Although termites in this study and others were associated with L. procerum, L. terebrantis, and marginally G. huntii, termites only showed preferential feeding on wood inoculated with G. huntii in laboratory trials. This suggests that increased termite presence on wood with bluestain fungi may be driven by factors other than increased wood palatability. Termites could thus disproportionately affect wood turnover rates for specific pools (e.g., bark beetle and root weevil attacked trees) and in some cases (e.g., G. huntii) accelerate wood decomposition. This study supports the growing evidence that the association between subterranean termites and bluestain fungi is spatially and taxonomically widespread.

A thorough understanding of microbial communities in the gut of lower termites is needed to develop target-specific and environmentally benign wood protection systems. In this study, the bacterial community from Reticulitermes virginicus was examined by Illumina sequencing of 16S ribosomal RNA (rRNA) spanning the V3 and V4 regions. Prior to library preparation, the termites were subjected to five treatments over an 18-day period: three groups were fed on wood treated with 0.5% chitosan, 25% acetic acid, or water, the fourth group was taken directly from the original collection log, and the fifth group was starved. Metagenomic sequences were analyzed using QIIME 2 to understand the treatments’ effects on the dynamics of the gut bacteria. Four dominant phyla were detected: Bacteroidetes (34.4% of reads), Firmicutes (20.6%), Elusimicrobia (15.7%), and Proteobacteria (12.9%). A significant effect of chitosan treatment was observed in two phyla; Firmicutes abundance was significantly lower with chitosan treatment when compared to other groups, while Actinobacteria was lower in unexposed and starved termites. The results suggest that chitosan treatment not only affects the structure of the microbial community in the gut, but other treatments such as starving also cause shifts in termite gut communities.

In recent years, a bioinformatics method for interpreting genome-wide association study (GWAS) data using metabolic pathway analysis has been developed and successfully used to find significant pathways and mechanisms explaining phenotypic traits of interest in plants. However, the many scripts implementing this method were not straightforward to use, had to be customized for each project, required user supervision, and took more than 24 h to process data. PAST (Pathway Association Study Tool), a new implementation of this method, has been developed to address these concerns. PAST has been implemented as a package for the R language. Two user-interfaces are provided; PAST can be run by loading the package in R and calling its methods, or by using an R Shiny guided user interface. In testing, PAST completed analyses in approximately half an hour to one hour by processing data in parallel and produced the same results as the previously developed method. PAST has many user-specified options for maximum customization. Thus, to promote a powerful new pathway analysis methodology that interprets GWAS data to find biological mechanisms associated with traits of interest, we developed a more accessible, efficient, and user-friendly tool. These attributes make PAST accessible to researchers interested in associating metabolic pathways with GWAS datasets to better understand the genetic architecture and mechanisms affecting phenotypes.

The feasibility of manufacturing cross-laminated timber (CLT) from southern yellow pine (United States grown) treated with micronized copper azole type C (MCA-C) preservative was evaluated. Lumber (2x6 visually graded no. 2 boards) was treated to two retention levels (1.0 and 2.4 kg/m(3)), planed to a thickness of 35 mm, and assembled along with an untreated control group using three adhesive systems following product specifications: melamine formaldehyde (MF), resorcinol formaldehyde (RF), and one-component polyurethane (PUR). Block shear and delamination tests were conducted to examine the bonding performance in accordance with ASTM D905 and ASTM D2559 Standards, respectively. One-way analysis of variance and Kruskal-Wallis H test were conducted to evaluate the effects of preservative retention and adhesive type on block shear strength (BSS) and wood failure percentage (WFP). Regardless of adhesive type, the 1.0 kg/m(3) retention treatment significantly lowered BSS compared to the untreated control. CLT composed of the laminations treated at 2.4 kg/m(3) maintained BSS when PUR and RF were used but not MF. The average WFP of each CLT configuration ranged from 89% to 99%. The untreated CLT specimens did not experience any delamination under accelerated weathering cycles. The delamination rates of the treated specimens assembled using MF and RF increased with the preservative retention level, while PUR provided delamination rates less than 1% to the laminations treated at both levels. These combined data suggest that, under the conditions tested, PUR provided overall better bonding performance than MF and RF for MCA-C treated wood. (C) 2019 Elsevier Ltd. All rights reserved.

Wood is a versatile and sustainable building material but may be vulnerable to fungal decay and insect damage when used outdoors or otherwise subjected to moisture.Pressure treatment with wood preservatives is the most common method of protecting wood from biological deterioration.This publication summarizes characteristics of pressuretreating preservatives and provides guidance for selection of pressure-treated wood for specific applications.It also discusses construction practices, service life expectations, and environmental considerations.The intended audience for this publication is users of pressure-treated wood such as homeowners, builders, contractors, engineers, and architects.

Summary Maize (Zea mays mays) oil is a rich source of polyunsaturated fatty acids (FAs) and energy, making it a valuable resource for human food, animal feed, and bio‐energy. Although this trait has been studied via conventional genome‐wide association study (GWAS), the single nucleotide polymorphism (SNP)‐trait associations generated by GWAS may miss the underlying associations when traits are based on many genes, each with small effects that can be overshadowed by genetic background and environmental variation. Detecting these SNPs statistically is also limited by the levels set for false discovery rate. A complementary pathways analysis that emphasizes the cumulative aspects of SNP‐trait associations, rather than just the significance of single SNPs, was performed to understand the balance of lipid metabolism, conversion, and catabolism in this study. This pathway analysis indicated that acyl‐lipid pathways, including biosynthesis of wax esters, sphingolipids, phospholipids and flavonoids, along with FA and triacylglycerol (TAG) biosynthesis, were important for increasing oil and FA content. The allelic variation found among the genes involved in many degradation pathways, and many biosynthesis pathways leading from FAs and carbon partitioning pathways, was critical for determining final FA content, changing FA ratios and, ultimately, to final oil content. The pathways and pathway networks identified in this study, and especially the acyl‐lipid associated pathways identified beyond what had been found with GWAS alone, provide a real opportunity to precisely and efficiently manipulate high‐oil maize genetic improvement.

Background In recent years, a bioinformatics method for interpreting GWAS data using metabolic pathway analysis has been developed and successfully used to find significant pathways and mechanisms explaining phenotypic traits of interest in plants. However, the many scripts implementing this method were not straightforward to use, had to be customized for each project, required user supervision, and took more than 24 hours to process data. PAST (Pathway Association Study Tool), a new implementation of this method, has been developed to address these concerns. Results PAST is implemented as a package for the R language. Two user-interfaces are provided; PAST can be run by loading the package in R and calling its methods, or by using an R Shiny guided user interface. In testing, PAST completed analyses in approximately one hour by processing data in parallel. PAST has many user-specified options for maximum customization. PAST produces the same results as the previously developed method. Conclusions In order to promote a powerful new method of pathway analysis that interprets GWAS data to find biological mechanisms associated with traits of interest, we developed a more accessible and user friendly tool. This tool is more efficient and requires less knowledge of programming languages to use than previous methods. Moreover, it produces similar results in significantly less time. These attributes make PAST accessible to researchers interested in associating metabolic pathways with GWAS datasets to better understand the genetic architecture and mechanisms affecting phenotype.

The goal of this study was to demonstrate how synchrotron-based X-ray fluorescence microscopy (XFM) can be used to better understand the mechanisms of copper tolerance in wood decay fungi. Copper is a major component in commercial wood preservatives as it is toxic to many wood decay fungi. However, certain fungi are copper tolerant and can attack preservative-treated wood, resulting in structural damage to treated wood members. Here we used large-field XFM to visualize six different elements (K, Ca, Mn, Fe, Cu, and Zn) in the mycelia and wood inoculated with four different species of brown rot wood decay fungi. Wood blocks were partially dipped into a solution of copper sulfate, exposed to fungi in malt extract agar petri dish assays for nine weeks, and then imaged and compared to blocks that were partially dipped in water. The blocks were imaged immediately adjacent to an end-matched control that was placed in malt extract agar petri dish assays for 9 weeks, but not exposed to the fungi so that the differences in the elemental distributions could be directly compared. The colonized wood and mycelia were rich in K, Ca, Mn, and Fe; however, the elements and the spatial distribution in the mycelia and wood differed across fungal species. The most interesting results were the maps showing the copper distribution. While three of the four fungi grew on the copper-rich region of the wood, only one species, Fibroporia radiculosa, dramatically reduced the copper concentration in the region of fungal growth.

Maize (Zea mays mays L.) is a staple crop of economic, industrial, and food security importance. Damage to the growing ears by corn earworm [Helicoverpa zea (Boddie)] is a major economic burden and increases secondary fungal infections and mycotoxin levels. To identify biochemical pathways associated with native resistance mechanisms, a genome‐wide association analysis was performed, followed by pathway analysis using a gene‐set enrichment‐based approach. The gene‐set enrichment exposed the cumulative effects of genes in pathways to identify those that contributed the most to resistance. Single nucleotide polymorphism–trait associations were linked to genes including transcription factors, protein kinases, hormone‐responsive proteins, hydrolases, pectinases, xylogluconases, and the flavonol synthase gene (in the maysin biosynthesis pathway). The most significantly associated metabolic pathways identified included those that modified cell wall components, especially homogalacturonan, wax esters, and fatty acids; those involved in antibiosis, especially 2,4‐dihydroxy‐7‐methoxy‐1,4‐benzoxazin‐3‐one (DIMBOA), flavonoids, and phenolics; and those involved in plant growth, including N uptake and energy production. The pathways identified in this study, and especially the cell wall‐associated pathways, identified here for the first time, provide clues to resistance mechanisms that could guide the identification of new resistant ideotypes and candidate genes for creation of resistant maize germplasm via selection of natural variants or gene editing.

Copper-tolerant brown-rot fungi are able to degrade wood treated with copper or copper-based wood preservatives. This research used quantitative reverse transcriptase polymerase chain reaction to explore what genes of the brown-rot fungus, Fibroporia radiculosa, were expressed when the fungus was overcoming the wood preservatives and decaying the wood. Aryl alcohol oxidase, catalase, oxalate decarboxylase 2, and copper resistance P-type ATPase pump had higher expression on alkaline copper quat type D treated wood compared with week 1. In addition, two genes had high expression at week 5; glycoside hydrolase 5 and glycoside hydrolase 10 when wood strength loss was around 50%. Glyoxylate dehydrogenase had high expression until week 8. This gene might be involved in the production of oxalate. Laccase, oxalate decarboxylase 1 and isocitrate lyase were not differentially expressed, suggesting that these genes were not involved in the decay process of alkaline copper quat-treated wood. These results are important to understand the genes that are involved in the mechanism of copper tolerance and wood decay in F. radiculosa.

Many projects have identified candidate genes for resistance to aflatoxin accumulation or Aspergillus flavus infection and growth in maize using genetic mapping, genomics, transcriptomics and/or proteomics studies. However, only a small percentage of these candidates have been validated in field conditions, and their relative contribution to resistance, if any, is unknown. This study presents a consolidated list of candidate genes identified in past studies or in-house studies, with descriptive data including genetic location, gene annotation, known protein identifiers, and associated pathway information, if known. A candidate gene pipeline to test the phenotypic effect of any maize DNA sequence on aflatoxin accumulation resistance was used in this study to determine any measurable effect on polymorphisms within or linked to the candidate gene sequences, and the results are published here.

Bark beetle outbreaks are increasing in frequency and intensity, generating massive inventories of dead trees globally. During attacks, trees are pre-inoculated with ophiostomatoid fungi via bark beetles, which has been shown to increase termite presence and feeding. These events may, in turn, alter biogeochemical cycles during decomposition. We examined these relationships by experimentally inoculating dead wood with bluestain fungi in a temperate pine forest. Across ten replicate plots, eight 0.5 m-long logs were inoculated with Ophiostoma minus and eight with distilled water. Half of the logs from each inoculation treatment were covered from above with a mesh cage barrier to exclude aboveground beetles while permitting access by belowground decomposers. After 1 year, significant increases in mass (34%) and decreases in moisture content (− 17%) were observed across all treatments, but no consistent changes in density were evident. C concentrations were 12% greater in bark when barriers were present and 17% greater in sapwood when barriers and inoculation fungi were absent. N concentrations were 16% greater in bark for fungal-inoculated logs and 27% greater when barriers were present. C:N ratios in A horizon soils under fungal-inoculated logs were 12% greater. Furthermore, termites were present fourfold more in fungal-inoculated logs than controls and the presence of termites was associated with 6% less C in sapwood and 11% more N in both sapwood and heartwood. Together these results suggest dead wood generated via bark beetle attacks has different biogeochemical responses during initial decomposition phases, which could have implications for the C status in forests following bark beetle outbreaks.

BACKGROUNDChitosan is a derivative form of chitin, which is the major component of exoskeletons of arthropods and the cell walls of fungi. The antimicrobial activity of chitosan against lepidopterans, aphids, fungi and bacteria has been extensively investigated, but only one report on the termiticidal effect of chitosan on termites has been published. In this study, we examined the termiticidal activity of chitosan by exposing single colonies of Reticulitermes flavipes (Kollar) and Reticulitermes virginicus Banks to wood treated with six different concentrations of chitosan solutions. Termite mortality and percent mass loss of wood samples after exposure to termites for 4 weeks were calculated.RESULTSHigh termite mortality (≥ 94%) occurred during exposure of R. flavipes termites to chitosan-treated wood with ≥38 mg g-1 treatment concentrations (≥ 2% chitosan), while <50% termite mortality was observed at lower treatment concentrations (11-15 mg g-1 ; 0.5% and 1% chitosan). For R. virginicus, 100% mortality was observed at all levels of treatment concentrations. A decrease in the percent mass loss of the wood sample was apparent in samples treated with solutions with an increasing chitosan concentration, with a significant difference (P < 0.05) between lower and higher treatment concentrations. Treatment retention in wood samples upon leaching was also determined and showed retention levels of between 0 and 30 mg g-1 chitosan retention.CONCLUSIONThis study investigated the exposure of subterranean termites to chitosan as a wood preservative. The results show that chitosan treatments at sufficiently high loadings could protect wood against termites, preferably under non-leaching conditions. © 2018 Society of Chemical Industry.

The objective of this study was to examine the toxicity of strobilurin fungicides against wood decay fungi in order to assess their potential to act as a co-biocide for copper-based wood protection. Two strobilurin fungicides, Heritage (50% azoxystrobin active ingredient) and Insignia (20% pyraclostrobin active ingredients), and copper sulfate pentahydrate were tested against one white rot fungus (Trametes versicolor) and two brown rot fungi (Gloeophyllum trabeum and Fibroporia radiculosa). SHAM (salicylhydroxamic acid) was also included in the study because it is a known potentiator of strobilurin activity. All treatments were incorporated in an agar-based media and evaluated for their effects on mycelial growth rate. Based on minimum inhibitory concentration values found for F. radiculosa (a copper-tolerant fungus), Insignia was 5.5x more toxic than Heritage. SHAM at 100 ppm, increased toxicity 9x for Heritage and 1.2x for Insignia. In a 20-day toxicity screening study, the four one-compound treatments tested were: copper sulfate (5000 ppm), Heritage (20 ppm), Insignia (20 ppm) and SHAM (100 ppm). The two two-compound treatments were: Heritage + SHAM and Insignia + SHAM. The two three-compound treatments were: copper sulfate + Heritage + SHAM and copper sulfate + Insignia + SHAM. For the two copper-susceptible fungi (T. versicolor and G. trabeum), the treatments that caused complete growth inhibition were copper sulfate alone and the two three-compound treatments (copper sulfate + Heritage + SHAM and copper sulfate + Insignia + SHAM). For F. radiculosa, the two- and three-compound treatments were the most toxic, with maximum daily average growth rates statistically similar to the copper sulfate treatment. A key result, however, was that only the three-compound treatment of copper sulfate + Insignia + SHAM completely inhibited growth of the copper-tolerant fungus. Thus, it appears that pyraclostrobin, which is the active ingredient in Insignia, has greater potential than azoxystrobin to act as a co-biocide for completely inhibiting growth of a copper-tolerant fungus.