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Cotton leaf curl disease (CLCuD) is a major constraint for production of cotton (Gossypium sp.) in Northwest India. CLCuD is caused by a monopartite, circular ssDNA virus belonging to the genus Begomovirus in association with betasatellites and alphasatellites, and ttransmitted by a whitefly vector (Bemisia tabaci). To explore the genetic variability in betasatellites and alphasatellite associated with the CLCuD-begomovirus complex in Northwest India. A survey was conducted for successive three years of 2014 to 2016 and twig samples from symptomatic and healthy cotton plants randomly were collected. Total plant DNAs were isolated, subjected to rolling circle amplification (RCA), cloning and sequencing. Full-length genome of 12 betasatellites and 13 alphasatellites, those were obtained in the present study, were analyzed. Sequence analysis showed that all the present betasatellites shared 85-99 percent nucleotide identity (PNI) among themselves and 84-95 PNI with other members of Cotton leaf curl Multan betasatellite (CLCuMB) and fell into one genogroup along with CLCuMB. But in close observation the present betasatellites clustered into two phylogenetic subgroups under single CLCuMB. The present alphasatellites showed 72-100 PNI among themselves and fell under three alphasatellite species, Gossypium Darwinii symptomless alphasatellite (GDarSLA), Cotton leaf curl Multan alphasatellite (CLCuMA) and Cotton leaf curl Burewala alphasatellite (CLCuBuA). In the recombination analysis, all the present betasatellites and alphasatellites were found to be recombinants involving intra species recombination in betasatellite, and interspecies recombination in alphasatellite species. The present study indicated that the betasatellite and alphasatellite molecules associated with CLCuD-begomovirus complex in Northwest India are genetically diverse., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2025 Biswas et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Animals in urban areas often encounter novel and potentially stressful conditions. It is important to understand how wildlife cope with anthropogenic disturbance. To investigate this specific adaptation we live-trapped squirrels in two study sites in Warsaw: a forest reserve and an urban park and we estimated stress responses at three levels: long-term and medium-term stress (the level of stress hormones, i.e. cortisol and cortisone concentrations, in hair and feces) and acute reaction to human-induced stress (measured during handling with the aid of the three indices: breath rate, struggle rate, and vocalization). According to GLMM models no difference in the stress hormones level was found between the two populations. The only differences in cortisol concentrations clearly depended on the season, i.e. being higher in autumn and winter comparying to other seasons. There was no influence of sex, or reproductive status on stress hormones. Forest squirrels had significantly higher breath rates, suggesting they were more stressed by handling. There was no difference in the struggle rate between study areas, this index was mostly affected by season (i.e. being highest in winter). First-trapped squirrels vocalized less than during the subsequent trappings. Assumingly, during the first, and more stressful trapping, squirrels used 'freezing' and/or little vocalization, while during next captures they used alarm calls to warn conspecifics. Overall, we showed that the two squirrel populations differed only in terms of their breath rate. This suggests that they did not differ in medium-term and long-term stress in general, but they can differ in acute response to handling. This also suggests that both populations were similarly affected by environmental factors. The lack of clear effects may also be due to population heterogeneity. Thus, in order to assess the effects of anthropogenic stressors a broader range of indicators and diverse analytical methods, including behavioral analyses, should be employed., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Beliniak et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Phytophthora pluvialis is an oomycete that primarily infects Pinus radiata and Pseudotsuga menziesii causing the destructive foliar disease red needle cast (RNC). Recent observations show that P. pluvialis can also infect western hemlock inducing resinous cankers. High-throughput and reproducible infection assays are integral to find key information on tree health and oomycete pathogenicity. In this protocol, we describe the propagation and spore induction of P. pluvialis, followed by detached needle assays for verification and quantification of virulence of P. pluvialis in P. radiata needles. These needle assays can be employed for high-throughput screening of tree needles with diverse genetic backgrounds. In downstream analysis, Quantitative PCR (qPCR) was utilized to assess relative gene expression, as exemplified by candidate RxLR effector protein PpR01. Additional techniques like RNA sequencing, metabolomics, and proteomics can be combined with needle assays and can offer comprehensive insights into P. pluvialis infection mechanisms., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Eccersall et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Rehabilitation of injured or immature individuals has become an increasingly used conservation and management tool. However, scientific evaluation of rehabilitations is rare, raising concern about post-release welfare as well as the cost-effectiveness of spending scarce financial resources. Over the past 20 years, events of juvenile Eurasian lynx presumably orphaned have been observed in many European lynx populations. To guide the management of orphaned lynx, we documented survival, rehabilitation and fate after the release and evaluated the potential relevance of lynx orphan rehabilitation for population management and conservation implications. Data on 320 orphaned lynx was collected from 1975 to 2022 from 13 countries and nine populations. The majority of orphaned lynx (55%) were taken to rehabilitation centres or other enclosures. A total of 66 orphans were released back to nature. The portion of rehabilitated lynx who survived at least one year after release was 0.66. Release location was the best predictor for their survival. Of the 66 released lynx, ten have reproduced at least once (8 females and 2 males). Conservation implications of rehabilitation programmes include managing genetic diversity in small, isolated populations and reintroducing species to historical habitats. The lynx is a perfect model species as most reintroduced populations in Central Europe show significantly lower observed heterozygosity than most of the autochthonous populations, indicating that reintroduction bottlenecks, isolation and post-release management have long-term consequences on the genetic composition of populations. The release of translocated orphans could be a valuable contribution to Eurasian lynx conservation in Europe. It is recommended to release orphans at the distribution edge or in the frame of reintroduction projects instead of a release in the core area of a population where it is not necessary from a demographic and genetic point of view. Rehabilitation programmes can have conservation implications that extend far beyond individual welfare benefits., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Molinari-Jobin et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Wood is difficult for most animals to digest due to large amounts of indigestible polymers, but some wood-feeding insects are considered to be able to utilize it as food with the aid of microbial symbionts. Most members of flower longicorn beetles (Coleoptera: Cerambycidae: Lepturinae) feed on nectar and pollen of flowers as adults and wood as larvae. In some lepturines, associations with yeasts are known: female adults possess fungus-storing organs (termed mycetangia) at ovipositors, and larvae also possess such organs (termed mycetomes) in their midguts to carry the associated yeasts. Despite the high diversity of Lepturinae in the world, lepturine-yeast associations, such as the consistency of associated yeasts among the beetle's developmental stages and ecological function of yeast symbionts, have been poorly documented. Here, we investigated the yeast symbiont of the Japanese common lepturine Leptura ochraceofasciata. X-ray computed microtomography revealed that a pair of tube-like, S-shaped mycetangia was located at the basal part of the ovipositor and that a muscle bundle joined the apex of the mycetangium to spiculum ventrale of sternum VIII. All female adults harbored only one yeast species, Scheffersomyces insectosa, in the mycetangia. All larvae harbored S. insectosa exclusively in the mycetomes. Scheffersomyces insectosa was also recovered from surfaces of eggs. Scheffersomyces insectosa assimilated wood-associated sugars including xylose, cellobiose, and xylan in culture. These results suggest the intimate association between L. ochraceofasciata and S. insectosa: S. insectosa is transmitted from the mother to offspring during oviposition and may be related to larval growth in wood., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Kishigami et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Leguminous trees and saltbushes provide potential alternatives to conventional feeds to overcome feed deficiency in arid and semi-arid countries. However, these plants are rich in antinutritional factors that have adverse effects on rumen microbiota and the host- animal. Some rumen microbiota detoxifies plants' secondary metabolites; thus, understanding plant-microbe interaction in the rumen could improve the plants' utilization. This study investigated the bacterial colonization and degradation of non-extracted and extracted tanniniferous plants: Atriplex halimus, Acacia saligna, and Leucaena leucocephala, in the rumen of three fistulated camels at 6 and 12 hours. The results showed that these plants have high nutritional value and tannins contents. The rumen degradation and microbial diversity of plant-attached bacteria varied according to plant type and phenols' extraction. Atriplex and leucaena showed higher microbial diversity at 6 and 12h, respectively. Bacteroidetes and Firmicutes were the main bacterial phyla, and the main genera were Prevotella, RC9&#95;gut&#95;group, Butyrivibrio that overrepresented in non-extracted plants (P<0.05). Fibrobacteres and Anaerovibrio showed sensitivity to plant toxins and Ruminococcus attached to plants with lower tannins. Several bacterial genera in the camel rumen have the potential to resist antinutritional factors in fodder plants, which could be used to improve the performance of grazing animals., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Rabee et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Tree planting programs now need to consider climate change increasingly, therefore, the resistance to pests plays an essential role in enabling tree adaptation to new ranges through tree population movement. The weevil Pissodes strobi (Peck) is a major pest of spruces and substantially reduces lumber quality. We revisited a large Interior spruce provenance/progeny trial (2,964 genotypes, 42 families) of varying susceptibility, established in British Columbia. We employed multivariate mixed linear models to estimate covariances between, and genetic control of, juvenile height growth and resistance traits. We performed linear regressions and ordinal logistic regressions to test for impact of parental origin on growth and susceptibility to the pest, respectively. A significant environmental component affected the correlations between resistance and height, with outcomes dependent on families. Parents sourced from above 950 m a.s.l. elevation negatively influenced host resistance to attacks, probably due to higher P. engelmannii proportion. For the genetic contribution of parents sourced from above 1,200 m a.s.l., however, we found less attack severity, probably due to a marked mismatch in phenologies. This clearly highlights that interspecific hybrid status might be a good predictor for weevil attacks and delineates the boundaries of successful spruce population movement. Families resulting from crossing susceptible parents generally showed fast-growing trees were the most affected by weevil attacks. Such results indicate that interspecific 'hybrids' with a higher P. glauca ancestry might be genetically better equipped with an optimized resource allocation between defence and growth and might provide the solution for concurrent improvement in resistance against weevil attacks, whilst maintaining tree productivity., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2022 Klápště et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Peatlands account for 15 to 30% of the world's soil carbon (C) stock and are important controls over global nitrogen (N) cycles. However, C and N concentrations are known to vary among peatlands contributing to the uncertainty of global C inventories, but there are few global studies that relate peatland classification to peat chemistry. We analyzed 436 peat cores sampled in 24 countries across six continents and measured C, N, and organic matter (OM) content at three depths down to 70 cm. Sites were distinguished between northern (387) and tropical (49) peatlands and assigned to one of six distinct broadly recognized peatland categories that vary primarily along a pH gradient. Peat C and N concentrations, OM content, and C:N ratios differed significantly among peatland categories, but few differences in chemistry with depth were found within each category. Across all peatlands C and N concentrations in the 10-20 cm layer, were 440 ± 85.1 g kg-1 and 13.9 ± 7.4 g kg-1, with an average C:N ratio of 30.1 ± 20.8. Among peatland categories, median C concentrations were highest in bogs, poor fens and tropical swamps (446-532 g kg-1) and lowest in intermediate and extremely rich fens (375-414 g kg-1). The C:OM ratio in peat was similar across most peatland categories, except in deeper samples from ombrotrophic tropical peat swamps that were higher than other peatlands categories. Peat N concentrations and C:N ratios varied approximately two-fold among peatland categories and N concentrations tended to be higher (and C:N lower) in intermediate fens compared with other peatland types. This study reports on a unique data set and demonstrates that differences in peat C and OM concentrations among broadly classified peatland categories are predictable, which can aid future studies that use land cover assessments to refine global peatland C and N stocks., Competing Interests: There are no competing interests, (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

Lignocellulosic biomass such as barley straw is a renewable and sustainable alternative to traditional feeds and could be used as bioenergy sources; however, low hydrolysis rate reduces the fermentation efficiency. Understanding the degradation and colonization of barley straw by rumen bacteria is the key step to improve the utilization of barley straw in animal feeding or biofuel production. This study evaluated the hydrolysis of barley straw as a result of the inoculation by rumen fluid of camel and sheep. Ground barley straw was incubated anaerobically with rumen inocula from three fistulated camels (FC) and three fistulated sheep (FR) for a period of 72 h. The source of rumen inoculum did not affect the disappearance of dry matter (DMD), neutral detergent fiber (NDFD). Group FR showed higher production of glucose, xylose, and gas; while higher ethanol production was associated with cellulosic hydrolysates obtained from FC group. The diversity and structure of bacterial communities attached to barley straw was investigated by Illumina Mi-Seq sequencing of V4-V5 region of 16S rRNA genes. The bacterial community was dominated by phylum Firmicutes and Bacteroidetes. The dominant genera were RC9&#95;gut&#95;group, Ruminococcus, Saccharofermentans, Butyrivibrio, Succiniclasticum, Selenomonas, and Streptococcus, indicating the important role of these genera in lignocellulose fermentation in the rumen. Group FR showed higher RC9&#95;gut&#95;group and group FC revealed higher Ruminococcus, Saccharofermentans, and Butyrivibrio. Higher enzymes activities (cellulase and xylanase) were associated with group FC. Thus, bacterial communities in camel and sheep have a great potential to improve the utilization lignocellulosic material in animal feeding and the production of biofuel and enzymes., Competing Interests: The authors have declared that no competing interests exist.

The choice of revegetating via direct seeding or planting nursery-grown seedlings influences the potential stresses suffered by seedlings such as herbivory and drought. The outcome of the balance between both revegetation methods may ultimately depend on how species identity and traits such as seed and seedling size interact with environmental conditions. To test this, we will conduct a continental-scale experiment consisting of one mini-experiment replicated by multiple participants across Europe. Each participant will establish a site with seeded and planted individuals of one or more native, locally growing oak (Quercus) species; the selection of this genus aims to favour continental-scale participation and to allow testing the response of a widely distributed genus of broad ecological and economic relevance. At each site, participants will follow the present protocol for seed collection, seeding in the field, nursery cultivation, outplanting, protection against herbivores, site maintenance, and measurement of seedling performance and environmental variables. Each measurement on each species at each site will produce one effect size; the data will be analysed through mixed-effects meta-analysis. With this approach we will assess the main effect of revegetation method, species, plant functional traits, and the potential effect of site-specific effect moderators. Overall, we will provide a continental-scale estimate on the seeding vs. planting dilemma and analyse to what extent the differences in environmental conditions across sites, seed size, functional traits, and the phylogenetic relatedness of species can account for the differences in the effect of revegetation method on seedling performance across study sites and species., Competing Interests: The authors have declared that no competing interests exist.

Fall armyworm (FAW; Spodoptera frugiperda), an exotic moth which recently invaded Africa, is a highly destructive pest of cereals especially maize a highly valued staple crop in Nigeria. The use of natural enemies such as predators or parasitoids for FAW control is more economically viable and environmentally safer than currently recommended synthetic insecticides. Natural enemies to combat the pest have not yet been reported in Nigeria. An exploration for the pests' natural enemies was undertaken by collecting FAW eggs and larvae from maize fields. These were reared in the laboratory for emergence, identification and efficacy as natural enemies. This yielded Euplectrus laphygmae (Hymenoptera: Eulophidae); Telenomus remus (Hymenoptera: Platygastridae) and Trombidium sp. (Acari.: Trombidiidae). Cotesia or Apanteles spp. were inferred to occur since Stictopisthus sp. (Hym.: Ichneumonidae), a secondary parasitoid, that attacks cocoons of Microgasterinae (e.g. Cotesia, Apanteles etc.) also emerged. Species of yet-to-be identified predators were also observed in various niches of maize plants. A positive relationship was found between FAW instar and the number of E. laphygmae eggs/instar ranging, on average, from 1.5 on second instar to 5.5 on fourth instars hosts. Parasitism rate of T. remus on FAW eggs was 100%. Parasitic mite infestation resulted in increasing paleness, reduced feeding, growth and movement as well as death of FAW 1st instars. Thus, the occurrence of FAW natural enemies in Nigeria calls for advocacy campaign to incorporate their use into integrated pest management strategies that attract and allow natural enemies to thrive for FAW management., Competing Interests: The authors have declared that no competing interests exists.

The Common or Brown Garden Snail, Cornu aspersum, is an invasive land snail that has successfully colonized a diverse range of global environments. Like other invasive land snails, it is a significant pest of a variety of agricultural crops, including citrus, grapes and canola. Cornu aspersum secretes a mucus trail when mobile that facilitates locomotion. The involvement of the trail in conspecific chemical communication has also been postulated. Our study found that anterior tentacle contact with conspecific mucus elicited a significant increase in heart rate from 46.9 to 51 beats per minute. In order to gain a better understanding of the constituents of the trail mucus and the role it may play in snail communication, the protein and volatile components of mucus trails were investigated. Using two different protein extraction methods, mass spectrometry analysis yielded 175 different proteins, 29 of which had no significant similarity to any entries in the non-redundant protein sequence database. Of the mucus proteins, 22 contain features consistent with secreted proteins, including a perlucin-like protein. The eight most abundant volatiles detected using gas chromatography were recorded (including propanoic acid and limonene) and their potential role as putative pheromones are discussed. In summary, this study has provided an avenue for further research pertaining to the role of trail mucus in snail communication and provides a useful repository for land snail trail mucus components. This may be utilized for further research regarding snail attraction and dispersal, which may be applied in the fields of agriculture, ecology and human health., Competing Interests: The authors have declared that no competing interests exist. The commercial funding provided by the GRDC does not alter our adherence to PLOS ONE policies on sharing data and materials.

The linden tree (Tilia spp.) is a popular tree for landscaping and urban environments in central and northwest European countries, and it is one of the most popular in cities in Poland. Ectomycorrhizal fungi form a symbiosis with many urban tree species and protect the host plant from heavy metals and against salinity. The aim of this study was to characterise the ECM fungal community of urban linden trees along the tree damage gradient. The study was performed on two sites located in the centre of the city of Gdańsk, in northern Poland. The vitality assessment of urban linden trees was made according to Roloff's classification. Tree damage classes were related to soil characteristics using principal component analysis. The five ectomycorrhizal fungal species were shared among all four tree damage classes, and Cenococcum geophilum was found to be the most abundant and frequent ectomycorrhizal fungal species in each class. Soil samples collected in the vicinity of trees belonging to the R0 class had significantly lower pH Na, Cl and Pb content than other soils. Our knowledge of ectomycorrhizal communities in urban areas is still limited, and these findings provide new insights into ectomycorrhizal distribution patterns in urban areas., Competing Interests: The authors have declared that no competing interests exist.

Sugi (Cryptomeria japonica D. Don) is an important conifer used for afforestation in Japan. As the genome of this species is 11 Gbps, it is too large to assemble within a short timeframe. Transcriptomics is one approach that can address this deficiency. Here we designed a workflow consisting of three stages to de novo assemble transcriptome using Oases and Trinity. The three transcriptomic stage used were independent assembly, automatic and semi-manual integration, and refinement by filtering out potential contamination. We identified a set of 49,795 cDNA and an equal number of translated proteins. According to the benchmark set by BUSCO, 87.01% of cDNAs identified were complete genes, and 78.47% were complete and single-copy genes. Compared to other full-length cDNA resources collected by Sanger and PacBio sequencers, the extent of the coverage in our dataset was the highest, indicating that these data can be safely used for further studies. When two tissue-specific libraries were compared, there were significant expression differences between male strobili and leaf and bark sets. Moreover, subtle expression difference between male-fertile and sterile libraries were detected. Orthologous genes from other model plants and conifer species were identified. We demonstrated that our transcriptome assembly output (CJ3006NRE) can serve as a reference transcriptome for future functional genomics and evolutionary biology studies., Competing Interests: The authors have declared that no competing interests exist.

The rapid growth of industrial and agricultural activities in Malaysia are leading to the impairment of most of the rivers in recent years through realising various trace metals. This leads to toxicity, particularly when the toxic has entered the food chain. Perak River is one of the most dynamic rivers for the Malaysian population. Therefore, in consideration of the safety issue, this study was conducted to assess the concentration of such metals (Cd, Cu, Zn, Fe, and Pb) in the muscles of most widely consumed fish species (Barbonymus schwanenfeldii, Puntius bulum, Puntius daruphani, Hexanematichthys sagor, Channa striatus, Mystacoleucus marginatus, and Devario regina) from different locations of Perak River, Malaysia by employing inductively coupled plasma optical emission spectroscopy (ICP-OES). Among the trace metals, Fe and Cd were found to be the highest (29.33-148.01 μg/g) and lowest (0.16-0.49 μg/g) concentration in all of the studied species, respectively. Although the estimated daily intakes (μg/kg/day) of Cd (0.65-0.85), Fe (79.27-352.00) and Pb (0.95-12.17) were higher than their reference, the total target hazard quotients values suggested that the local residents would not experience any adverse health effects from its consumption. In contrast, the target cancer risk value suggested that all fish species posed a potential cancer risk due to Cd and cumulative cancer risk values, strongly implying that continuous consumption of studied fish species would cause cancer development to its consumers., Competing Interests: The authors have declared that no competing interests exist.

Genus Trichogramma Westwood is being utilized in various biological control programme worldwide. Trichogramma spp. are egg parasitoids which lay their eggs inside the host eggs. More than 240 species are known, of which, 45 species are recorded from India. It is quite difficult to identify and differentiate among the species of Trichogramma due to smaller size. In present study, we hypothesized a methodology to identify the Trichogramma species based on potential morphometric characteristics. The males and females of five different species of Trichogramma namely, T. brassicae, T. chilotraeae, T. danaidiphaga, T. danausicida and T. dendrolimi were morphometrically analysed. A total of 33 characters in males and 27 in females were taken into consideration for morphometric identification and analysis using principal component method. It was observed that few characters of male like body length, length of flagellar hair, maximum marginal hair length of fore wing, genitalia characters including aedeagus length, distance between chelate structure and gonoforceps, and others were identified as important morphometric characters. Similarly, in females, ovipositor length, body and head length, eye width, antennal club width and number of setae in forewings were found important for taxonomic identification. Male and female of each species are clearly identified from high definition microscopic images., Competing Interests: The authors have declared that no competing interests exist.

Background: The presupposition of genomic selection (GS) is that predictive accuracies should be based on population-wide linkage disequilibrium (LD). However, in species with large, highly complex genomes the limitation of marker density may preclude the ability to resolve LD accurately enough for GS. Here we investigate such an effect in two conifer species with ~ 20 Gbp genomes, Douglas-fir (Pseudotsuga menziesii Mirb. (Franco)) and Interior spruce (Picea glauca (Moench) Voss x Picea engelmannii Parry ex Engelm.). Random sampling of markers was performed to obtain SNP sets with totals in the range of 200-50,000, this was replicated 10 times. Ridge Regression Best Linear Unbiased Predictor (RR-BLUP) was deployed as the GS method to test these SNP sets, and 10-fold cross-validation was performed on 1,321 Douglas-fir trees, representing 37 full-sib F1 families and on 1,126 Interior spruce trees, representing 25 open-pollinated (half-sib) families. Both trials are located on 3 sites in British Columbia, Canada., Results: As marker number increased, so did GS predictive accuracy for both conifer species. However, a plateau in the gain of accuracy became apparent around 10,000-15,000 markers for both Douglas-fir and Interior spruce. Despite random marker selection, little variation in predictive accuracy was observed across replications. On average, Douglas-fir prediction accuracies were higher than those of Interior spruce, reflecting the difference between full- and half-sib families for Douglas-fir and Interior spruce populations, respectively, as well as their respective effective population size., Conclusions: Although possibly advantageous within an advanced breeding population, reducing marker density cannot be recommended for carrying out GS in conifers. Significant LD between markers and putative causal variants was not detected using 50,000 SNPS, and GS was enabled only through the tracking of relatedness in the populations studied. Dramatically increasing marker density would enable said markers to better track LD with causal variants in these large, genetically diverse genomes; as well as providing a model that could be used across populations, breeding programs, and traits., Competing Interests: No competing interests.

Annual trunk increments are essential for short-term analyses of the response of trees to various factors. For instance, based on annual trunk increments, it is possible to develop and calibrate forest growth models. We investigated the possibility of estimating annual trunk increments from the terrestrial structure from motion (SfM) photogrammetry. Obtaining the annual trunk increments of mature trees is challenging due to the relatively small growth of trunks within one year. In our experiment, annual trunk increments were obtained by two conventional methods: measuring tape (perimeter increment) at heights of 0.8, 1.3, and 1.8 m on the trunk and increment borer (diameter increment) at a height of 1.3 m on the trunk. The following tree species were investigated: Fagus sylvatica L. (beech), Quercus petraea (Matt.) Liebl. (oak), Picea abies (L.) H. Karst (spruce), and Abies alba Mill (fir). The annual trunk increments ranged from 0.9 cm to 2.4 cm (tape/perimeter) and from 0.7 mm to 3.1 mm (borer/diameter). The data were collected before- and after-vegetation season, besides the data collection increment borer. When the estimated perimeters from the terrestrial SfM photogrammetry were compared to those obtained using the measuring tape, the root mean square error (RMSE) was 0.25-1.33 cm. The relative RMSE did not exceed 1% for all tree species. No statistically significant differences were found between the annual trunk increments obtained using the measuring tape and terrestrial SfM photogrammetry for beech, spruce, and fir. Only in the case of oak, the difference was statistically significant. Furthermore, the correlation coefficient between the annual trunk increments collected using the increment borer and those derived from terrestrial SfM photogrammetry was positive and equal to 0.6501. Terrestrial SfM photogrammetry is a hardware low-demanding technique that provides accurate three-dimensional data that can, based on our results, even detect small temporal tree trunk changes., Competing Interests: The authors have declared that no competing interests exist.

The ever-increasing vehicle counts have resulted in a significant increase in air pollution impacting human and natural ecosystems including trees, and physical properties. Roadside plantations often act as a first defense line against the vehicular emissions to mitigate the impacts of pollutants. However, they are themselves vulnerable to these pollutants with varying levels of tolerance capacity. This demands a scientific investigation to assess the role of roadside plantation for better management and planning for urban sprawl where selected trees could be grown to mitigate the impacts of harmful pollutants. The present study assesses the impacts of vehicular emissions on the adaptation and mitigation potential of two important roadside tree species i.e. Grevillea robusta and Mangifera indica planted along roadsides in the capital city of Uttarakhand. Uttarakhand is one of the Indian Western Himalayan State and its capital city is situated on the foothills of Himalaya. The adaptation and mitigation potential were evaluated by studying the response of pollutants on the functional traits which drive the physiology of the trees. The CO2 assimilation rate, transpiration rate, stomatal conductance, water use efficiency (WUE), air pollution tolerance index (APTI), copper and proline accumulation, dust removal efficiency (DRE), leaf thickness and cooling created by plantation were studied to evaluate the response of trees exposed to roadside traffics. To compare the influence of pollutants, traits of trees grown in a control site with few or absence of vehicular movement were compared with the roadside trees. The control site represented part of a reserve forest where human interference is controlled and human-induced activities are prohibited. The vehicular frequency was found to modulate tree characteristics. The tree characteristics representing WUE, APTI, proline and copper accumulation, leaf thickness, cooling impact, and DRE were enhanced significantly, while the decreased CO2 assimilation rate was observed near roadside trees compared to the control site. We found both of the species to perform well to be used as one of the potential species for roadside and urban greening. However, there is a need to assess the potential of other species in reference to the present study., Competing Interests: The authors have declared that no competing interests exist.

Development of genome-wide resources for application in genomic selection or genome-wide association studies, in the absence of full reference genomes, present a challenge to the forestry industry, where longer breeding cycles could benefit from the accelerated selection possible through marker-based breeding value predictions. In particular, large conifer megagenomes require a strategy to reduce complexity, whilst ensuring genome-wide coverage is achieved. Using a transcriptome-based reference template, we have successfully developed a high density exome capture genotype-by-sequencing panel for radiata pine (Pinus radiata D.Don), capable of capturing in excess of 80,000 single nucleotide polymorphism (SNP) markers with a minor allele frequency above 0.03 in the population tested. This represents approximately 29,000 gene models from a core set of 48,914 probes. A set of 704 SNP markers capable of pedigree reconstruction and differentiating individual genotypes were tested within two full-sib mapping populations. While as few as 70 markers could reconstruct parentage in almost all cases, the impact of missing genotypes was noticeable in several offspring. Therefore, 60 sets of 110 randomly selected SNP markers were compared for both parentage reconstruction and clone differentiation. The performance in parentage reconstruction showed little variation over 60 iterations. However, there was notable variation in discriminatory power between closely related individuals, indicating a higher density SNP marker panel may be required to elucidate hidden relationships in complex pedigrees., Competing Interests: The authors LGN and MRJr are affiliated with RAPiD Genomics LLC and undertook probe design with input from the remaining authors, and dataset generation. The study paid for the dataset generation as a third-party service provider. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

A novel approach was employed to load α-Fe2O3 and α-FeOOH onto sewage sludge biochar (SBC) with the purpose of efficient nickel (Ni) removal. A high Ni(II) adsorption capacity of 35.50 mg·g-1 in 100 ppm Ni(II) solution with 10 mg modified sewage sludge biochar (MSBC) was achieved. The adsorption kinetic and isotherm were fitted well by the pseudo-second-order model and the Langmuir model, respectively. The optimal pH was found around a neutral pH of 7. The adsorption mechanisms of Ni(II) onto MSBC were described as the synergistic effects of electrostatic attraction, ion exchange, inner-sphere complexation and co-precipitation. The initial rapid adsorption phenomenon could be attributed to electrostatic attraction and ion exchange, and then inner-sphere complexation and co-precipitation acted as a crucial role in the following step. The remarkable performance of MSBC provides an effective waste utilization approach to simultaneous sewage sludge recycle and Ni removal from aqueous solution., Competing Interests: The authors have declared that no competing interests exist.

Understanding the epidemiology of infectious diseases in a host population is a major challenge in forestry. Radiata pine plantations in New Zealand are impacted by a foliar disease, red needle cast (RNC), caused by Phytophthora pluvialis. This pathogen is dispersed by water splash with polycyclic infection affecting the lower part of the tree canopy. In this study, we extended an SI (Susceptible-Infectious) model presented for RNC to analyse the key epidemiological drivers. We conducted two experiments to empirically fit the extended model: a detached-needle assay and an in vivo inoculation. We used the detached-needle assay data to compare resistant and susceptible genotypes, and the in vivo inoculation data was used to inform sustained infection of the whole plant. We also compared isolations and real-time quantitative PCR (qPCR) to assess P. pluvialis infection. The primary infection rate and the incubation time were similar for susceptible and resistant genotypes. The pathogen death rate was 2.5 times higher for resistant than susceptible genotypes. Further, external proliferation of mycelium and sporangia were only observed on 28% of the resistant ramets compared to 90% of the susceptible ones. Detection methods were the single most important factor influencing parameter estimates of the model, giving qualitatively different epidemic outputs. In the early stages of infection, qPCR proved to be more efficient than isolations but the reverse was true at later points in time. Isolations were not influenced by the presence of lesions in the needles, while 19% of lesioned needle maximized qPCR detection. A primary infection peak identified via qPCR occurred at 4 days after inoculation (dai) with a secondary peak observed 22 dai. Our results have important implications to the management of RNC, by highlighting the main differences in the response of susceptible and resistant genotypes, and comparing the most common assessment methods to detect RNC epidemics., Competing Interests: The authors have declared that no competing interests exist.

Genomic selection is a proven technology in animal and plant breeding to accelerate genetic gain, but as yet is to be fully realised in forest tree breeding. This paper examines, through stochastic simulation, the potential benefits of genomic selection (GS) over forward selection (FS) in a typical conifer breeding program. Methods of speeding the deployment of selected material were also considered, including top-grafting onto mature seed orchard ortets, using additional replicates of clones in archives for crossing, and embryogenesis and clonal propagation. Genetic gain per generation was found to increase considerably when the size of the training population was larger (800 c.f. 3000 clones), or when the heritability was higher (0.2 c.f. 0.5). The largest genetic gain, of 24% was achieved where large training populations (3000 clones) and high heritability traits (0.5) were combined. The accuracy of genomic breeding values (GEBVs) increased with the increase in the number of clones in the training population, the heritability of the trait and the density of the SNP markers. Calculated accuracies of simulated GEBVs and genetic gain per unit of time suggested that 2000 clones in the training population is the minimum size for effective genomic selection for conifers. Compared with forward selection, genomic selection with 2000 clones in the training population, and a 60K SNP panel, an increase of 1.58 mm per year in diameter-at-breast-height (DBH) and 2.44 kg/m3 per year for wood density can be expected. After one generation (9-years), this would be equivalent to 14.23 mm and 21.97 kg/m3 for DBH and wood density respectively. Deploying clones of the selected individuals always resulted in higher additional genetic gain than deploying progeny/seedlings. Deploying genetic material selected from genomic selection with top-grafting for early coning appeared to be the best option. Application of genomic selection to conifer breeding programs, combined with deployment tools such as top-grafting and embryogenesis are powerful tools to speed the delivery of genetic gain to the forest., Competing Interests: The authors have declared that no competing interests exist.

Estimating animal abundance is essential for research, management and conservation purposes. Although reliable methods exist to estimate absolute density for populations with individually marked animals, robust relative abundance indices (RAIs) may allow to track changes in population size when individual identification is not possible. Their performance, however, needs be thoroughly evaluated. We investigated the relative performance of several common faeces-based and camera-based RAIs for estimating small-scale variation in red fox abundance, a mesopredator of high relevance for management, in two different study areas. We compared precision, cost and performance of the methods in capturing relationships with covariates of local abundance. Random transect-based RAIs had a low mean, a comparatively high coefficient of variation and a high proportion of zeros, prohibiting or impeding analysis in relation to environmental predictors. Rectangular scat plots and transects along linear landscape features had an intermediate amount of zeros while retaining a high precision, but were less sensitive to local variation in abundance related to environmental predictors and required a large field effort. Camera trap-based RAIs yielded low to intermediate precision, but were more sensitive to small-scale variation in relative abundance than faeces-based methods. Camera traps were the most expensive methods for an initial monitoring session, but required the lowest field effort, were cheapest in the long run and were the least susceptible to observer bias and detection error under a robust sampling protocol. Generally, faeces count-based RAIs appear more suitable for studies that aim to compare local abundance between several study sites of equal landscape composition under constant detection probability. Camera traps provide more flexible data for studies that require accounting for influences of landscape composition on local abundance and are more cost-effective for long-term or continuous monitoring and more suitable to achieve high replication. Accordingly, the choice of the most suitable method and plot design is context-dependent., Competing Interests: The authors have declared that no competing interests exist.

Pollinosis caused by Japanese cedar (Cryptomeria japonica) is a widespread social problem in Japan. To date, 23 male-sterile C. japonica trees have been selected in Japan to address pollinosis, from which four male-sterility loci (MS1, MS2, MS3, and MS4) have been identified from test crossing results. For efficient breeding of male-sterile C. japonica trees, more male-sterile individuals and individuals heterozygous for male-sterile genes are required. Therefore, we aimed to develop DNA markers for marker-assisted selection of four types of male-sterile genes from populations without a family structure. First, for four families exhibiting segregation of each male-sterile locus (MS1, MS2, MS3, and MS4), genome-wide single-nucleotide polymorphism and insertion/deletion (indel) genotyping was performed using the Axiom myDesign Targeted Genotyping Array method. Four high-density linkage maps for mapping the MS1, MS2, MS3, and MS4 families were constructed, which included 4923, 1722, 1896, and 2247 markers, respectively. In these maps, 15, 4, 2, and 2 markers were located 0.0, 3.3, 1.1, and 0.0 cM from the MS1, MS2, MS3, and MS4 loci, respectively. Second, for the markers located 0.0 cM from a male-sterile locus (i.e., MS1 and MS4), to clarify the most tightly linked markers, we calculated the prediction rate of male-sterile gene genotypes from marker genotypes for 78 trees. The markers with the highest prediction rates were AX-174127446 (0.95) for MS1 and AX-174121522 (1.00) for MS4. The AX-174121522 marker was considered to be suitable for selecting trees homozygous or heterozygous for the MS4 gene from plus-trees without a pollination test, which requires a large amount of time and effort. The nearest markers to the male-sterile loci found in this study may facilitate the isolation of male-sterile genes in C. japonica via combination with the draft genomic sequence that is currently being collated., Competing Interests: The authors have declared that no competing interests exist.

There is a wide diversity of bioinformatic tools available for the assembly of next generation sequence and subsequence variant calling to identify genetic markers at scale. Integration of genomics tools such as genomic selection, association studies, pedigree analysis and analysis of genetic diversity, into operational breeding is a goal for New Zealand's most widely planted exotic tree species, Pinus radiata. In the absence of full reference genomes for large megagenomes such as in conifers, RNA sequencing in a range of genotypes and tissue types, offers a rich source of genetic markers for downstream application. We compared nine different assembler and variant calling software combinations in a single transcriptomic library and found that Single Nucleotide Polymorphism (SNPs) discovery could vary by as much as an order of magnitude (8,061 SNPs up to 86,815 SNPs). The assembler with the best realignment of the packages trialled, Trinity, in combination with several variant callers was then applied to a much larger multi-genotype, multi-tissue transcriptome and identified 683,135 in silico SNPs across a predicted 449,951 exons when mapped to the Pinus taeda ver 1.01e reference., Competing Interests: The authors have declared that no competing interests exist.

Twenty-eight clonal trials of radiata pine planted across Australia and New Zealand were used to investigate genetic variation and genotype by environment (G×E) interaction for diameter-at-breast-height (DBH), height and Dothistroma resistance (DO&#95;R). The average narrow-sense heritabilities were 0.11, 0.21 and 0.30 while the average broad-sense heritabilities were 0.27, 0.34 and 0.40 for DBH, height and Dothistroma resistance, respectively. Dothistroma resistance was assessed as the percentage of needles that were not affected by Dothistroma needle blight. G×E interactions were analysed using an approximate reduced factor analytic model. Apparent G×E interactions were estimated for DBH, height and Dothistroma resistance. Estimates of G×E interactions and their standard errors were strongly influenced by the level of connectivity between trials, in terms of common clones and common parents. When there was sufficient connectivity between trials (more than 30% common clones between trials), a high level of G×E interaction was found for DBH and height but not for Dothistroma resistance. In two simulated clonal trials planted in two environments, low connectivity between environments resulted in a lower estimated genetic correlation between environments with an increased standard error. These results suggest that the number of clones in common between clonal trials is a key factor for inclusion in future experimental designs for estimating G×E interaction. When designing clonal trials for use in multiple environments for accurately estimating the level of G×E, if the resource for creating connectivity between environments is limited, at least 30% of the clones need to be in common between environments., Competing Interests: This manuscript was funded by Scion Core Funding and Funding from Forest Genetics Ltd. Authors Mike Carson and Sue Carson were working at Forest Genetics Ltd during the course of the study. Authors Mike Carson and Sue Carson are affiliated to Carson Associates Ltd and author Yongjun Li is affiliated to Agriculture Victoria. There are no patents, products in development or marketed products to declare. This does not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials.

Large mammals are often a source of conflict, and consequently there has been increasing interest in close encounters with them. Knowledge of wildlife responses to human disturbance is crucial for the management of increasing and expanding populations of large animals. We investigated flight initiation distance (FID) and aggressive responses of the European bison-the largest terrestrial mammal of Europe-to human disturbance in the Białowieża Forest (NE Poland). When encountered by humans, bison usually flee. Aggression was observed in only 0.4% of approach attempts. Mean FID was 77±46 m and was influenced by habitat, sex, and supplementary feeding intensity. Females showed greater timidity than males, FID was lower in forest than in open habitats, and supplementary feeding caused a drop in FID. In 84.5% of all documented aggression cases, bison attacks were provoked by humans approaching too close to the bison or by deliberate scaring them. Males were more aggressive than females, and attacked mainly during the rut, while females attacked during the winter and calving. Bison attacked in built-up areas significantly more often than expected. The mean critical distance of attacks was 21±2 m. Most attacks took the form of a short chase preceded by warning signs. Goring was observed in 22.7% of all aggression cases and no fatalities were recorded. Our study shows that bison are not dangerous animals and only manifest aggression in response to prolonged disturbance at close ranges. The education of people and recommendations for minimum approach distances should ensure a low rate of disturbance and safety when encountering large mammals., Competing Interests: The authors have declared that no competing interests exist.

False banana /Ensete ventricosum [Welw.] Cheesman/ is exploited as a food crop in Ethiopia where it represents an important staple food. The plant is harvested and large amounts of biomass residues are originated, mainly from the pseudo stem (i.e., fiber bundles obtained from the leaf sheaths after being scrapped to produce starchy food) and the inflorescence stalk. These materials were studied in relation to their summative chemical composition, composition of lignin, lipophilic and polar extracts. Moreover, their structural characteristics, in view of their valorization, were scrutinized. The analytical studies were performed with the aid of FTIR, GC/MS, Py-GC/MS and SEM. The fiber bundles are aggregates of mainly long and slender fibers with low ash, extractives and lignin contents (3.8%. 4.4% and 10.5% respectively) and high holocellulose and α-cellulose contents (87.5% and 59.6% respectively). The hemicelluloses in the fibers are mostly highly acetylated xylans and the lignin is of the H-type (H:G:S, 1:0.7:0.8). This lignin composition is in line with the FTIR peaks at 1670 cm-1 and 1250 cm-1.The inflorescence stalk has high ash content (12.3% in the main stalk and 24.6% in fines) with a major proportion of potassium, high extractives (25.9%), and low lignin and α-cellulose contents (5.8% and 17.9% respectively). The stalk includes numerous starch granules in the cellular structure with the predominant presence of parenchyma. The potential valorization routes for these materials are clearly different. The fiber bundles could be used as a fiber source for paper pulp production with the possibility of a prior hemicelluloses removal while the inflorescence stalk has nutritional value for food and fodder. Furthermore, it can also be used for sugar fermentation products., Competing Interests: The authors have declared that no competing interests exist.

There are a few common species and many rare species in a biological community or a multi-species collection in given space and time. This hollow distribution curve is called species abundance distribution (SAD). Few studies have examined the patterns and dynamics of SADs during the succession of forest communities by model selection. This study explored whether the communities in different successional stages followed different SAD models and whether there existed a best SAD model to reveal their intrinsic quantitative features of structure and dynamics in succession. The abundance (the number of individuals) of each vascular plant was surveyed by quadrat sampling method from the tree, shrub and herb layers in two typical communities (i.e., the evergreen needle- and broad-leaved mixed forest and the monsoon evergreen broad-leaved forest) in southern subtropical Dinghushan Biosphere Reserve, South China. The sites of two forest communities in different successional stages are both 1 ha in area. We collected seven widely representative SAD models with obviously different function forms and transformed them into the same octave (log2) scale. These models are simultaneously confronted with eight datasets from four layers of two communities, and their goodness-of-fits to the data were evaluated by the chi-squared test, the adjusted coefficient of determination and the information criteria. The results indicated that: (1) the logCauchy model followed all the datasets and was the best among seven models; (2) the fitness of each model to the data was not directly related to the successional stage of forest community; (3) according to the SAD curves predicted by the best model (i.e., the logCauchy), the proportion of rare species decreased but that of common ones increased in the upper layers with succession, while the reverse was true in the lower layers; and (4) the difference of the SADs increased between the upper and the lower layers with succession. We concluded that the logCauchy model had the widest applicability in describing the SADs, and could best mirror the SAD patterns and dynamics of communities and their different layers in the succession of forests. The logCauchy-modeled SADs can quantitatively guide the construction of ecological forests and the restoration of degraded vegetation.

Species response to climate change is influenced by predictable (selective) and unpredictable (random) evolutionary processes. To understand how climate change will affect present-day species, it is necessary to assess their adaptive potential and distinguish it from the effects of random processes. This will allow predicting how different genotypes will respond to forecasted environmental change. Space for time substitution experiments are an elegant way to test the response of present day populations to climate variation in real time. Here we assess neutral and putatively adaptive variation in 11 populations of Festuca rubra situated along crossed gradients of temperature and moisture using molecular markers and phenotypic measurements, respectively. By comparing population differentiation in putatively neutral molecular markers and phenotypic traits (QST-FST comparisons), we show the existence of adaptive differentiation in phenotypic traits and their plasticity across the climatic gradient. The observed patterns of differentiation are due to the high genotypic and phenotypic differentiation of the populations from the coldest (and wettest) environment. Finally, we observe statistically significant covariation between markers and phenotypic traits, which is likely caused by isolation by adaptation. These results contribute to a better understanding of the current adaptation and evolutionary potential to face climate change of a widespread species. They can also be extrapolated to understand how the studied populations will adjust to upcoming climate change without going through the lengthy process of phenotyping.

The coniferous forest tree Douglas-fir (Pseudotsuga menziesii) is native to the pacific North America, and is increasingly planted in temperate regions worldwide. Nitrogen (N) metabolism is of great importance for growth, resistance and resilience of trees. In the present study, foliar N metabolism of adult trees of three coastal and one interior provenance of Douglas-fir grown at two common gardens in southwestern Germany (Wiesloch, W; Schluchsee, S) were characterized in two subsequent years. Both the native North American habitats of the seed sources and the common garden sites in Germany differ in climate conditions. Total and mineral soil N as well as soil water content were higher in S compared to W. We hypothesized that i) provenances differ constitutively in N pool sizes and composition, ii) N pools are affected by environmental conditions, and iii) that effects of environmental factors on N pools differ among interior and coastal provenances. Soil water content strongly affected the concentrations of total N, soluble protein, total amino acids (TAA), arginine and glutamate. Foliar concentrations of total N, soluble protein, structural N and TAA of trees grown at W were much higher than in trees at S. Provenance effects were small but significant for total N and soluble protein content (interior provenance showed lowest concentrations), as well as arginine, asparagine and glutamate. Our data suggest that needle N status of adult Douglas-fir is independent from soil N availability and that low soil water availability induces a re-allocation of N from structural N to metabolic N pools. Small provenance effects on N pools suggest that local adaptation of Douglas-fir is not dominated by N conditions at the native habitats.

Wind turbines represent a source of hazard for bats, especially through collision with rotor blades. With increasing technical development, tall turbines (rotor-swept zone 50-150 m above ground level) are becoming widespread, yet we lack quantitative information about species active at these heights, which impedes proposing targeted mitigation recommendations for bat-friendly turbine operation. We investigated vertical activity profiles of a bat assemblage, and their relationships to wind speed, within a major valley of the European Alps where tall wind turbines are being deployed. To monitor bat activity we installed automatic recorders at sequentially increasing heights from ground level up to 65 m, with the goal to determine species-specific vertical activity profiles and to link them to wind speed. Bat call sequences were analysed with an automatic algorithm, paying particular attention to mouse-eared bats (Myotis myotis and Myotis blythii) and the European free-tailed bat (Tadarida teniotis), three locally rare species. The most often recorded bats were the Common pipistrelle (Pipistrellus pipistrellus) and Savi's pipistrelle (Hypsugo savii). Mouse-eared bats were rarely recorded, and mostly just above ground, appearing out of risk of collision. T. teniotis had a more evenly distributed vertical activity profile, often being active at rotor level, but its activity at that height ceased above 5 ms-1 wind speed. Overall bat activity in the rotor-swept zone declined with increasing wind speed, dropping below 5% above 5.4 ms-1. Collision risk could be drastically reduced if nocturnal operation of tall wind turbines would be restricted to wind speeds above 5 ms-1. Such measure should be implemented year-round because T. teniotis remains active in winter. This operational restriction is likely to cause only small energy production losses at these tall wind turbines, although further analyses are needed to assess these losses precisely.

Documenting the scale and intensity of fine-scale spatial genetic structure (FSGS), and the processes that shape it, is relevant to the sustainable management of genetic resources in timber tree species, particularly where logging or fragmentation might disrupt gene flow. In this study we assessed patterns of FSGS in three species of Dipterocarpaceae (Parashorea tomentella, Shorea leprosula and Shorea parvifolia) across four different tropical rain forests in Malaysia using nuclear microsatellite markers. Topographic heterogeneity varied across the sites. We hypothesised that forests with high topographic heterogeneity would display increased FSGS among the adult populations driven by habitat associations. This hypothesis was not supported for S. leprosula and S. parvifolia which displayed little variation in the intensity and scale of FSGS between sites despite substantial variation in topographic heterogeneity. Conversely, the intensity of FSGS for P. tomentella was greater at a more topographically heterogeneous than a homogeneous site, and a significant difference in the overall pattern of FSGS was detected between sites for this species. These results suggest that local patterns of FSGS may in some species be shaped by habitat heterogeneity in addition to limited gene flow by pollen and seed dispersal. Site factors can therefore contribute to the development of FSGS. Confirming consistency in species' FSGS amongst sites is an important step in managing timber tree genetic diversity as it provides confidence that species specific management recommendations based on species reproductive traits can be applied across a species' range. Forest managers should take into account the interaction between reproductive traits and site characteristics, its consequences for maintaining forest genetic resources and how this might influence natural regeneration across species if management is to be sustainable.

In declining birches (Betula sp.) from different European stands affected by the "birch leaf-roll disease" (BLRD) a novel virus is identified by means of RNA-Seq virome analysis. The virus represents a new member in the genus Badnavirus, family Caulimoviridae, tentatively named Birch leaf roll-associated virus (BLRaV) and it is the first badnavirus found to infect birch. Complete genome sequences (7,862-7,864 nucleotides) of three viral isolates of Finnish and German origin have been determined. The virus sequences show a typical badnavirus organization with three major open reading frames (ORFs) and a fourth potential ORF overlapping with the end of ORF3. ORFs 1-2-3 show low level of amino acid identity to the corresponding proteins encoded by other badnaviruses, reaching a maximum of 44% identity (ORF3). Grapevine vein-clearing virus appears as the closest badnavirus when considering the polymerase region. So far, we can exclude evidence for presence of endogenous BLRaV elements in the birch genome, while evidence for the episomal activity of BLRaV is provided. The viral population holds significant haplotype diversity, while co-infection by different BLRaV variants are observed in single hosts. BLRaV presence is associated with the BLRD in both silver (B. pendula) and downy birch (B. pubescens). These results challenge the earlier hypothesis of a causal role of Cherry leaf roll virus in BLRD. Further work is now needed to finally prove that BLRaV is the causal agent for the BLRD.

The brown marmorated stink bug, Halyomorpha halys, is an agricultural and urban pest that has become widely established as an invasive species of major concern in the USA and across Europe. This species forms large aggregations when entering diapause, and it is often these aggregations that are found by officials conducting inspections of internationally shipped freight. Identifying the presence of diapausing aggregations of H. halys using their emissions of volatile organic compounds (VOCs) may be a potential means for detecting and intercepting them during international freight inspections. Headspace samples were collected from aggregations of diapausing H. halys using volatile collection traps (VCTs) and solid phase microextraction. The only compound detected in all samples was tridecane, with small amounts of (E)-2-decenal found in most samples. We also monitored the release of defensive odors, following mechanical agitation of diapausing and diapause-disrupted adult H. halys. Diapausing groups were significantly more likely to release defensive odors than diapause-disrupted groups. The predominant compounds consistently found from both groups were tridecane, (E)-2-decenal, and 4-oxo-(E)-2-hexenal, with a small abundance of dodecane. Our findings show that diapausing H. halys do release defensive compounds, and suggest that volatile sampling may be feasible to detect H. halys in freight.

Land tenure and tenure security are among the most important factors determining the viability and success of Reducing Emissions from Deforestation and Forest Degradation (REDD+) initiatives. The premise of the present paper is that territorial conflicts lead to forest loss and compromise the successful implementation of REDD+. Within this context, the main objectives of this paper are to (i) document, relying on participatory methods, the extent to which land conflicts drive deforestation and (ii) reflect on the legal context of REDD+ examining if, from an Indigenous perspective, it offers tools to resolve such conflicts. We used the Upper Bayano Watershed in eastern Panama as a case study of complex land tenure dynamics, and their effects on forest conservation in the context of REDD+. Combining a range of participatory methods including participatory mapping and forest carbon stock assessment, we estimated the consequences of land invasions on forest carbon stocks. Our analysis shows that invasions of Indigenous territories amounted to 27.6% of the total deforestation for the period of 2001-2014. The situation is of paramount concern in the Embera territory of Majé where 95.4% of total deforestation was caused by colonist invaders. Using and validating the maps made freely available by the Global Forest Change initiative of the University of Maryland, we then developed a reference level for the watershed and carried out a back of the envelop estimation of likely REDD+ revenue, showing its potential to bring much needed income to Indigenous communities striving to protect their forest estate. Our analysis of current legislation in Panama highlights confusion and important legal voids and emphasizes the strong links between land tenure, carbon ownership, and territorial invasions. The options and shortcoming of implementing REDD+ in Indigenous territories is discussed in the conclusion taking our legal review into account.

The Rajah Brooke's Birdwing, Trogonoptera brookiana, is a large, iconic butterfly that is facing heavy commercial exploitation and habitat loss. Males of some subspecies exhibit puddling behavior. A method of conservation monitoring was developed for subspecies albescens in Ulu Geroh, Peninsular Malaysia, where the males consistently puddle in single-species aggregations at stable geothermal springs, reaching well over 300 individuals when the population is at its highest. Digital photography was used to conduct counts of numbers of males puddling. The numbers of birdwings puddling were significantly correlated with counts of birdwings in flight, but were much higher. The numbers puddling during the peak hour were correlated with numbers puddling throughout the day and could be predicted using the numbers puddling at an alternative hour, enabling flexibility in the time of counts. Average counts for three images taken at each puddle at three peak hours between 1400-1600 hours over 2-3 days were used as a monthly population index. The numbers puddling were positively associated with higher relative humidity and brightness during monitoring hours. Monthly counts of birdwings from monitoring of puddles over a period of two years are presented. The minimum effort required for a monitoring program using counts of puddling males is discussed, as well as the potential of using the method to monitor other species of puddling butterflies.

Every year, there are millions of documented vehicle collisions involving cervids across Europe and North America. While temporal patterns in collision occurrence are relatively well described, few studies have targeted deer behaviour as a critical component of collision prevention. In this study, we investigated weekly and daily patterns in road crossing behaviour in roe deer. Using road crossing events and movement data obtained from GPS telemetry, we employed mixed-effect models to explain frequency and timing of crossings at five road segments by a number of predictors including traffic volume, deer movement activity and the presence of wildlife warning reflectors. We analysed 13,689 road crossing events by 32 study animals. Individual variation in crossing frequency was high but daily patterns in crossing events were highly consistent among animals. Variation in the intensity of movement activity on a daily and seasonal scale was the main driver of road crossing behaviour. The seasonal variation in crossing frequency reflected differences in movement activity throughout the reproductive cycle, while daily variation in the probability to cross exhibited a clear nocturnal emphasis and reflected crepuscular activity peaks. The frequency of road crossings increased as a function of road density in the home-range, while traffic volume only exerted marginal effects. Movement activity of roe deer in our study coincided with commuter traffic mainly in the early morning and late afternoon during winter and during periods of high spatial activity such as the rut. Both timing and frequency of crossing events remained unchanged in the presence of reflectors. Our results emphasise the importance of behavioural studies for understanding roe deer vehicle-collision patterns and thus provide important information for collision prevention. We suggest that mitigation of collision risk should focus on strategic seasonal measures and animal warning systems targeting drivers.

Accurate inference of relatedness between individuals in breeding population contributes to the precision of genetic parameter estimates, effectiveness of inbreeding management and the amount of genetic progress delivered from breeding programs. Pedigree reconstruction has been proven to be an efficient tool to correct pedigree errors and recover hidden relatedness in open pollinated progeny tests but the method can be limited by the lack of parental genotypes and the high proportion of alien pollen from outside the breeding population. Our study investigates the efficiency of sib-ship reconstruction in an advanced breeding population of Eucalyptus nitens with only partially tracked pedigree. The sib-ship reconstruction allowed the identification of selfs (4% of the sample) and the exploration of their potential effect on inbreeding depression in the traits studied. We detected signs of inbreeding depression in diameter at breast height and growth strain while no indications were observed in wood density, wood stiffness and tangential air-dry shrinkage. After the application of a corrected sib-ship relationship matrix, additive genetic variance and heritability were observed to increase where signs of inbreeding depression were initially detected. Conversely, the same genetic parameters for traits that appeared to be free of inbreeding depression decreased in size. It therefore appeared that greater genetic variance may be due, at least in part, to contributions from inbreeding in these studied populations rather than a removal of inbreeding as is traditionally thought.

Snakehead fishes of the family Channidae are predatory freshwater teleosts from Africa and Asia comprising 38 valid species. Snakeheads are important food fishes (aquaculture, live food trade) and have been introduced widely with several species becoming highly invasive. A channid barcode library was recently assembled by Serrao and co-workers to better detect and identify potential and established invasive snakehead species outside their native range. Comparing our own recent phylogenetic results of this taxonomically confusing group with those previously reported revealed several inconsistencies that prompted us to expand and improve on previous studies. By generating 343 novel snakehead coxI sequences and combining them with an additional 434 coxI sequences from GenBank we highlight several problems with previous efforts towards the assembly of a snakehead reference barcode library. We found that 16.3% of the channid coxI sequences deposited in GenBank are based on misidentifications. With the inclusion of our own data we were, however, able to solve these cases of perpetuated taxonomic confusion. Different species delimitation approaches we employed (BIN, GMYC, and PTP) were congruent in suggesting a potentially much higher species diversity within snakeheads than currently recognized. In total, 90 BINs were recovered and within a total of 15 currently recognized species multiple BINs were identified. This higher species diversity is mostly due to either the incorporation of undescribed, narrow range, endemics from the Eastern Himalaya biodiversity hotspot or the incorporation of several widespread species characterized by deep genetic splits between geographically well-defined lineages. In the latter case, over-lumping in the past has deflated the actual species numbers. Further integrative approaches are clearly needed for providing a better taxonomic understanding of snakehead diversity, new species descriptions and taxonomic revisions of the group.

Current understanding of litterfall and fine root dynamics in temperate forests is limited, even though these are the major contributors to carbon and nutrient cycling in the ecosystems. In this study, we investigated litterfall and fine root biomass and production in five deciduous and four coniferous forests at the Gwangneung Experimental Forest in Korea. We used ingrowth cores to measure fine root production and root turnover rate. The litterfall was separated into leaves, twigs, and others, and then leaves were further separated according to species. Annual litterfall mass was not significantly different between the years, 360 to 651 g m-2 in 2011 and 300 to 656 g m-2 in 2012. Annual fine root (<5 mm) production was significantly higher in 2012 (421 to 1342 g m-2) than in 2011 (99 to 872 g m-2). Annual litterfall mass was significantly different among the stands, while fine root production did not statistically differ among the stands. The average fine root turnover rate, calculated by dividing the annual fine root production by the maximum standing fine root biomass, was 1.65 for deciduous forests and 1.97 for coniferous forests. Fine root production constituted 18-44% of NPP, where NPP was the sum of woody biomass production, litterfall production, and fine root production. Belowground production was a greater fraction of NPP in more productive forests suggesting their greater carbon allocation belowground.

Diameter at breast height (DBH), wood density (DEN) and predicted modulus of elasticity (PME) are considered as 'key traits' (KT) in the improvement in radiata pine breeding programmes in New Zealand. Any other traits which are also of interest to radiata pine breeders and forest growers are called 'non-key traits' (NKTs). External resin bleeding (ERB), internal checking (IC), number of heartwood rings (NHR) are three such non-key traits which affect wood quality of radiata pine timber. Economic importance of the KTs and NKTs is hard to define in radiata pine breeding programmes due to long rotation period. Desired-gain index (DGIs) and robust selection were proposed to incorporate NKTs into radiata pine breeding programme in order to deal with the uncertainty of economic importance. Four desired-gain indices A-D were proposed in this study. The desired-gain index A (DGI-A) emphasized growth and led to small decrease in ERB and small increase in IC and NHR. The expected genetic gains of all traits in the desired-gain index B (DGI-B) were in the favourable directions (positive genetic gains in the key traits and negative genetic gains in the non-key traits). The desired-gain index C (DGI-C) placed emphasis on wood density, leading to favourable genetic gain in the NKTs but reduced genetic gains for DBH and PME. The desired-gain index D (DGI-D) exerted a bit more emphasis on the non-key traits, leading large favourable reduction in the non-key traits and lower increase in the key traits compared with the other DGIs. When selecting both the key traits and the non-key traits, the average EBVs of six traits were all in the same directions as the expected genetic gains except for DBH in the DGI-D. When the key traits were measured and selected, internal checking always had a negative (favourable) genetic gain but ERB and NHR had unfavourable genetic gain in the most of time. After removing some individuals with high sensitivity to the change of economic weights, robust desired-gain selection made genetic gains of all the key and non-key traits to move a little bit toward unfavourable directions in the four indices. It is concluded that desired-gain index combined with robust selection concept is an efficient way for selecting the key and non-key traits in radiata pine breeding programmes.

The rapid spread and diversification of outdoor recreation can impact on wildlife in various ways, often leading to the avoidance of disturbed habitats. To mitigate human-wildlife conflicts, spatial zonation schemes can be implemented to separate human activities from key wildlife habitats, e.g., by designating undisturbed wildlife refuges or areas with some level of restriction to human recreation and land use. However, mitigation practice rarely considers temporal differences in human-wildlife interactions. We used GPS telemetry data from 15 red deer to study the seasonal (winter vs. summer) and diurnal (day vs. night) variation in recreation effects on habitat use in a study region in south-western Germany where a spatial zonation scheme has been established. Our study aimed to determine if recreation infrastructure and spatial zonation affected red deer habitat use and whether these effects varied daily or seasonally. Recreation infrastructure did not affect home range selection in the study area, but strongly determined habitat use within the home range. The spatial zonation scheme was reflected in both of these two levels of habitat selection, with refuges and core areas being more frequently used than the border zones. Habitat use differed significantly between day and night in both seasons. Both summer and winter recreation trails, and nearby foraging habitats, were avoided during day, whereas a positive association was found during night. We conclude that human recreation has an effect on red deer habitat use, and when designing mitigation measures daily and seasonal variation in human-wildlife interactions should be taken into account. We advocate using spatial zonation in conjunction with temporal restrictions (i.e., banning nocturnal recreation activities) and the creation of suitable foraging habitats away from recreation trails.

The development of timber tracking methods based on genetic markers can provide scientific evidence to verify the origin of timber products and fulfill the growing requirement for sustainable forestry practices. In this study, the origin of an important Dark Red Meranti wood, Shorea platyclados, was studied by using the combination of seven chloroplast DNA and 15 short tandem repeats (STRs) markers. A total of 27 natural populations of S. platyclados were sampled throughout Malaysia to establish population level and individual level identification databases. A haplotype map was generated from chloroplast DNA sequencing for population identification, resulting in 29 multilocus haplotypes, based on 39 informative intraspecific variable sites. Subsequently, a DNA profiling database was developed from 15 STRs allowing for individual identification in Malaysia. Cluster analysis divided the 27 populations into two genetic clusters, corresponding to the region of Eastern and Western Malaysia. The conservativeness tests showed that the Malaysia database is conservative after removal of bias from population subdivision and sampling effects. Independent self-assignment tests correctly assigned individuals to the database in an overall 60.60-94.95% of cases for identified populations, and in 98.99-99.23% of cases for identified regions. Both the chloroplast DNA database and the STRs appear to be useful for tracking timber originating in Malaysia. Hence, this DNA-based method could serve as an effective addition tool to the existing forensic timber identification system for ensuring the sustainably management of this species into the future.

We studied a community of frugivorous Lepidoptera in the lowland rainforest of Papua New Guinea. Rearing revealed 122 species represented by 1,720 individuals from 326 woody plant species. Only fruits from 52% (171) of the plant species sampled were attacked. On average, Lepidoptera were reared from 1 in 89 fruits and a kilogram of fruit was attacked by 1.01 individuals. Host specificity of Lepidoptera was notably low: 69% (33) of species attacked plants from >1 family, 8% (4) fed on single family, 6% (3) on single genus and 17% (8) were monophagous. The average kilogram of fruits was infested by 0.81 individual from generalist species (defined here as feeding on >1 plant genus) and 0.07 individual from specialist species (feeding on a single host or congeneric hosts). Lepidoptera preferred smaller fruits with both smaller mesocarp and seeds. Large-seeded fruits with thin mesocarp tended to host specialist species whereas those with thick, fleshy mesocarp were often infested with both specialist and generalist species. The very low incidence of seed damage suggests that pre-dispersal seed predation by Lepidoptera does not play a major role in regulating plant populations via density-dependent mortality processes outlined by the Janzen-Connell hypothesis.

The increase in the deer population observed in recent decades has strongly impacted forest regeneration and the forest itself. The reduction in the quality of raw wood material, as a consequence of deer-mediated damage, constitutes a significant burden on forest owners. The basis for the commencement of preventive actions in this setting is the understanding of the populations and behaviors of deer in their natural environment. Although multiple studies have been carried out regarding this subject, only a few suggested topography as an important factor that may influence the distribution and intensity of deer-mediated damage. The detailed terrain models based on LiDAR data as well as the data on damage caused by deer from the State Forests database enabled thorough analyses of the distribution and intensity of damage in relation to land form in this study. These analyses were performed on three mountain regions in Poland: the Western Sudety Mountains, the Eastern Sudety Mountains, and the Beskidy Mountains. Even though these three regions are located several dozen to several hundred kilometers apart from each other, not all evaluated factors appeared common among them, and therefore, these regions have been analyzed separately. The obtained results indicated that the forest damage caused by deer increased with increasing altitude above 1000 m ASL. However, much larger areas of damage by deer were observed at elevations ranging from 401 to 1000 m ASL than at elevations below 400 m ASL. Moreover, the locations of damage (forest thickets and old stands) indicated that red deer is the species that exerts the strongest pressure on forest ecosystems. Our results show the importance of deer foraging behavior to the structure of the environment., Competing Interests: The authors have declared that no competing interests exist.

Little is known about edaphic selection pressures as drivers of contrasting white spruce ectomycorrhizal fungal community structure and diversity in the Canadian boreal forest. We hypothesized that community composition differs among the four sites sampled-nursery, mining site, forest edge, and natural forest. Ectomycorrhizal (ECM) fungal community structure and diversity was studied at the four locations with soil fertility gradient through morpho-molecular and phylogenetic analyses in relationships with rhizospheric soil chemical properties. 41 different species were identified. Mining site had a significantly different species composition than the surrounding environments. Soil pH and percentage of roots colonized by ECM fungi increased while soil P, N, Fe, C, K, Mg, Al, Ca, and Na contents declined across the soil fertility gradient: nursery → natural forest → forest edge → mining site. Contrary to the preference of acid soils by ECM fungi, a few ecologically adapted to high pH, poor soil chemical fertility, and low organic matter content colonize white spruce roots on the non-acidogenic mining site, allowing natural regeneration of white spruce seedlings. Other ECM fungi are adapted to high fertigation level of commercial nursery. This study clearly shows the contrasting difference in white spruce ectomycorrhizal fungal community structure and diversity driven by edaphic selection pressures., Competing Interests: The authors have declared that no competing interests exist.