Fir (Abies balsamea) (Pinales: Pinaceae) needle essential oil enhances the knockdown activity of select insecticides.

Because of the increased interest in plant essential oils (PEO) for both home pest control and personal bite protection, the ability of fir needle (Abies balsamea) oil to synergize the 1-h knockdown and 24-h toxicity of 9 different synthetic insecticides was evaluated. Fir needle oil strongly synergized knockdown of the neonicotinoids, clothianidin, and thiamethoxam (between 16- and 24-fold), as well as natural pyrethrins (12-fold), but had less effect with organophosphates and fipronil. For 24-h mortality, only pirimiphos-methyl was strongly synergized by fir needle oil pretreatment (18-fold). Chemical analysis and testing identified delta-3-carene is the most bioactive constituent, producing synergism similar to that of the whole oil. In fact, this constituent synergized the 24-h mortality of clothianidin to a higher degree than fir needle oil itself (4.9-fold vs. 2.4-fold). Synergism is unlikely to be mediated by effects on the nervous system, as fir needle oil caused no change in mosquito central nervous system firing at 100 ppm and did not synergize an inactive concentration of natural pyrethrins (10 nM). To better understand fir needle oil effects, we evaluated the ability of pretreatment with this oil to impact Aedes aegypti monooxygenase degradation of a model substrate, 7-ethoxycoumarin. Interestingly, both fir needle oil and delta-3-carene caused a significant increase in metabolic degradation of 7-ethoxycoumarin, perhaps indicating they upregulate oxidative metabolic processes. Such an action would explain why fir needle oil enhances knockdown, but not 24-h mortality for most of the insecticides studied here, whereas increased bioactivation would explain the synergism of pirimiphos-methyl toxicity.

Seasonal occurrence of bark and woodboring Coleoptera in stands of Pinus densiflora (Pinales: Pinaceae) and Larix kaempferi (Pinales: Pinaceae) and monitoring method using multifunnel traps baited with pine volatiles.

This study investigated the seasonal occurrence of bark and woodboring Coleoptera in Pinus densiflora (Pinales: Pinaceae), and Larix kaempferi (Pinales: Pinaceae) stands using multifunnel traps baited with pine volatiles in Korea. The number and species of bark and woodboring beetles caught in traps baited with ethanol, α-pinene, and ethanol+α-pinene were compared to determine the effective attractants. In addition, the effects of other pine volatiles, such as (-)-ß-pinene, ß-caryophyllene, (±)-limonene, ß-myrcene, and 3-carene, were investigated. A total of 13,134 woodboring beetles from 150 species were collected from pine and larch stands from 2019 to 2020. Tomicus minor (Hartig) (Coleoptera: Curculionidae) adults were more attracted to traps baited with α-pinene, whereas Xyleborinus saxesenii (Ratzeburg) (Coleoptera: Curculionidae), Cyclorhipidion pelliculosum (Eichhoff) (Coleoptera: Curculionidae), and Phloeosinus pulchellus (Blandford) (Coleoptera: Curculionidae) adults were more attracted to traps baited with ethanol. Hylurgops interstitialis (Chapuis) (Coleoptera: Curculionidae), Shirahoshizo genus group, Rhagium inquisitor (Linne) (Coleoptera: Cerambycidae), and Rhadinomerus maebarai (Voss & Chûjô) (Coleoptera: Curculionidae) were more frequently attracted to traps baited with ethanol+α-pinene than to traps baited with other attractants. The addition of 3-carene to ethanol+α-pinene enhanced the capture of H. interstitialis, R. inquisitor, and Hylobius (Callirus) haroldi (Faust) (Coleoptera: Curculionidae).

Disentangling the chemistry of Australian plant exudates from a unique historical collection.

For thousands of years, the unique physicochemical properties of plant exudates have defined uses in material culture and practical applications. Native Australian plant exudates, including resins, kinos, and gums, have been used and continue to be used by Aboriginal Australians for numerous technical and cultural purposes. A historic collection of well-preserved native Australian plant exudates, assembled a century ago by plant naturalists, gives a rare window into the history and chemical composition of these materials. Here we report the full hierarchical characterization of four genera from this collection, Xanthorrhoea, Callitris, Eucalyptus, and Acacia, from the local elemental speciation, to functional groups and main molecular markers. We use high-resolution X-ray Raman spectroscopy (XRS) to achieve bulk-sensitive chemical speciation of these plant exudates, including insoluble, amorphous, and cross-linked fractions, without the limitation of invasive and/or surface specific methods. Combinatorial testing of the XRS data allows direct classification of these complex natural species as terpenoid, aromatic, phenolic, and polysaccharide materials. Differences in intragenera chemistry was evidenced by detailed interpretation of the XRS spectral features. We complement XRS with Fourier-transform infrared (FT-IR) spectroscopy, gas chromatography­mass spectrometry (GC-MS), and pyrolysis­GC-MS (Py-GC-MS). This multimodal approach provides a fundamental understanding of the chemistry of these natural materials long used by Aboriginal Australian peoples.

An integrated high-resolution mapping shows congruent biodiversity patterns of Fagales and Pinales.

The documentation of biodiversity distribution through species range identification is crucial for macroecology, biogeography, conservation, and restoration. However, for plants, species range maps remain scarce and often inaccurate. We present a novel approach to map species ranges at a global scale, integrating polygon mapping and species distribution modelling (SDM). We develop a polygon mapping algorithm by considering distances and nestedness of occurrences. We further apply an SDM approach considering multiple modelling algorithms, complexity levels, and pseudo-absence selections to map the species at a high spatial resolution and intersect it with the generated polygons. We use this approach to construct range maps for all 1957 species of Fagales and Pinales with data compilated from multiple sources. We construct high-resolution global species richness maps of these important plant clades, and document diversity hotspots for both clades in southern and south-western China, Central America, and Borneo. We validate the approach with two representative genera, Quercus and Pinus, using previously published coarser range maps, and find good agreement. By efficiently producing high-resolution range maps, our mapping approach offers a new tool in the field of macroecology for studying global species distribution patterns and supporting ongoing conservation efforts.

Volatile Oil of Platycladus Orientalis (L.) Franco Leaves Exerts Strong Anti-inflammatory Effects via Inhibiting the IκB/NF-κB Pathway.

This study was designed to investigate the anti-inflammatory effects of volatile oil of Platycladus orientalis (L.) Franco leaves (VOPF) and the underlying molecular mechanisms by using the non-infectious inflammation rat models and infectious inflammation mouse models. Ear swelling and intraperitoneal capillary permeability in mice, and carrageenan-induced toe swelling and cotton ball-induced granuloma in rats were used to reveal anti-inflammatory effects of VOPF. Moreover, the lipopolysaccharide (LPS)-induced mouse model of acute lung injury was used to explore the anti-inflammatory mechanism of VOPF. The results showed that VOPF could significantly inhibit auricular swelling, intraperitoneal capillary permeability in mice, and reduce granuloma swelling and paw swelling in rats. Furthermore, it significantly alleviated the pathological damage of the lung tissue. In addition, VOPF could reduce the contents of IL-1ß and TNF-α and increase the content of IL-10 in the serum. It had little effect on the expression of p65 but reduced the phosphorylation level of p65 and IκB in NF-κB pathway. In conclusion, VOPF has anti-inflammatory effects and the mechanisms involve the down-regulation of the phosphorylation levels of p65 and IκB and blockage of the NF-κB signaling pathway.

Comparisons of Efficiency of Two Formulations of Verbenone (4, 6, 6-trimethylbicyclo [3.1.1] hept-3-en-2-one) for Protecting Whitebark Pine, Pinus albicaulis (Pinales: Pinaceae) From Mountain Pine Beetle (Colopetera: Curculionidae).

Whitebark pine, Pinus albicaulis Engelm., is a subalpine tree endemic to western North America. This species provides multiple ecosystem services and is suffering widespread mortality from mountain pine beetle, Dendroctonus ponderosae Hopkins. Verbenone is a pheromone produced as D. ponderosae feed, and high air concentrations of verbenone deter D. ponderosae from colonizing trees. Synthetic verbenone has been formulated into products used to prevent D. ponderosae from colonizing trees. We compared the ability of verbenone pouches and SPLAT Verb to protect individuals and small stands of P. albicaulis. With individual trees in Montana, all treated trees survived regardless of verbenone formulation and rate, whereas untreated trees suffered 70 and 90% mortality in 2015 and 2016. In plot experiments in California from 2015 to 2017, and Oregon from 2015 to 2018, verbenone was applied to trees spaced ~10 m apart, and survival of small (12.7-23 cm DBH = diameter at 1.37 m height), medium (23.1-33 cm DBH) and large (>33 cm DBH) trees was compared. In California, where >80% of untreated trees survived, pouches increased survival ~2 to 3% and SPLAT Verb increased survival ~4 to 7% regardless of tree size. In Oregon, verbenone pouches and SPLAT Verb performed similarly on medium and small trees, but large trees had greater survival when treated with SPLAT Verb (~93%) than pouches (~82%). Compared to verbenone pouches, SPLAT Verb appears to better protect P. albicaulis from D. ponderosae.

Impact of Hemlock Woolly Adelgid (Hemiptera: Adelgidae) Infestation on the Jasmonic Acid-Elicited Defenses of Tsuga canadensis (Pinales: Pinaceae).

Hemlock woolly adelgid is an invasive piercing-sucking insect in eastern North America, which upon infestation of its main host, eastern hemlock ('hemlock'), improves attraction and performance of folivorous insects on hemlock. This increased performance may be mediated by hemlock woolly adelgid feeding causing antagonism between the the jasmonic acid and other hormone pathways. In a common garden experiments using hemlock woolly adelgid infestation and induction with methyl jasmonate (MeJA) and measures of secondary metabolite contents and defense-associated enzyme activities, we explored the impact of hemlock woolly adelgid feeding on the local and systemic induction of jasmonic acid (JA)-elicited defenses. We found that in local tissue hemlock woolly adelgid or MeJA exposure resulted in unique induced phenotypes, whereas the combined treatment resulted in an induced phenotype that was a mixture of the two individual treatments. We also found that if the plant was infested with hemlock woolly adelgid, the systemic response of the plant was dominated by hemlock woolly adelgid, regardless of whether MeJA was applied. Interestingly, in the absence of hemlock woolly adelgid, hemlock plants had a very weak systemic response to MeJA. We conclude that hemlock woolly adelgid infestation prevents systemic induction of JA-elicited defenses. Taken together, compromised local JA-elicited defenses combined with weak systemic induction could be major contributors to increased folivore performance on hemlock woolly adelgid-infested hemlock.

Spirosoma telluris sp. nov. and Spirosoma arboris sp. nov. isolated from soil and tree bark, respectively.

Two novel strains (HMF3257T and HMF4905T), isolated from freshwater and bark samples, were investigated to determine their relationships within and between species of the genus Spirosoma by using a polyphasic approach. They were aerobic, Gram-stain-negative, non-motile and rod-shaped bacteria. The major fatty acids (>10%) in both strains were identified as summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c) and C16 : 1 ω5c, while strains HMF3257T and HMF4905T contained a moderately high amount of C16 : 0 and iso-C15 : 0, respectively. The predominant respiratory quinone was MK-7 for both strains. In addition to phosphatidylethanolamine and one unidentified glycolipid, the polar lipid profile of strain HMF3257T consisted of three unidentified aminophospholipids, one unidentified aminolipid and two unidentified polar lipids, and that of strain HMF4905T consisted of one unidentified aminophospholipid, two unidentified aminolipids and three unidentified polar lipids. The DNA G+C contents of strains HMF3257T and HMF4905T were 47.2 and 46.4 mol%, respectively. Phylogenetic analysis based on 16S rRNA gene sequences showed that strains HMF3257T and HMF4905T are closely related to Spirosoma migulaei 15J9-8T (97.0 % sequence similarity), while sharing 97.4 % sequence similarity with each other. The average nucleotide identity value between strains HMF3257T and HMF4905T was 81.1 %, and the digital DNA-DNA hybridization value between these two strains was 24.4 %. Based on the above data, strains HMF3257T and HMF4905T represent two novel members within the genus Spirosoma, for which the names Spirosoma telluris sp. nov. and Spirosoma arboris sp. nov. are proposed, respectively. The type strain of S. telluris is HMF3257T (=KCTC 62463T=NBRC 112670T) and type strain of S. arboris is HMF4905T (=KCTC 72779T=NBRC 114270T).

Chimpanzee ranging responses to fruit availability in a high-elevation environment.

Most primates experience seasonal fluctuations in the availability of food resources and face the challenge of balancing energy expenditure with energy gain during periods of resource scarcity. This is likely to be particularly challenging in rugged, montane environments, where available energy is relatively low and travel costs are high. Chimpanzees (Pan troglodytes) show extensive behavioral diversity across study sites. Yet, as most research has focused on low- and mid-elevation sites, little is known on how chimpanzees respond to periods of low fruit availability in harsh montane environments. We use focal follow and phenology data to investigate how fruit availability influences daily path length and monthly home range in chimpanzees living in Nyungwe National Park, a montane forest in Rwanda. Nyungwe chimpanzees decreased their daily travel distances during periods of fruit scarcity. However, this decrease in travel effort did not correspond with a decrease in foraging area. Instead, monthly homes ranges shifted location across the study period. Nyungwe chimpanzees occupy a relatively wide altitudinal range and the shifts in monthly home range location may reflect differences in the altitudinal distribution of food resources. Chimpanzee monthly diet was often dominated by one or two species and each of these species were confined to different elevation zones. One important species, Podocarpus latifolius, grew only at high elevations (2,600-2,950 m) and chimpanzees ranged at the altitudinal peak of their range for 2 consecutive months while feeding on this species. Thus, while high elevations are often thought to be harsh environments for primates, they can be an important part of a species' home range when they provide a refugium for densely distributed, important food species.

Extensive Shifts from Cis- to Trans-splicing of Gymnosperm Mitochondrial Introns.

Hundreds of plant mitogenomes have been sequenced from angiosperms, but relatively few mitogenomes are available from its sister lineage, gymnosperms. To examine mitogenomic diversity among extant gymnosperms, we generated draft mitogenomes from 11 diverse species and compared them with four previously published mitogenomes. Examined mitogenomes from Pinaceae and cycads retained all 41 protein genes and 26 introns present in the common ancestor of seed plants, whereas gnetophyte and cupressophyte mitogenomes experienced extensive gene and intron loss. In Pinaceae and cupressophyte mitogenomes, an unprecedented number of exons are distantly dispersed, requiring trans-splicing of 50-70% of mitochondrial introns to generate mature transcripts. RNAseq data confirm trans-splicing of these dispersed exons in Pinus. The prevalence of trans-splicing in vascular plant lineages with recombinogenic mitogenomes suggests that genomic rearrangement is the primary cause of shifts from cis- to trans-splicing in plant mitochondria.

Statistical Comparison of Trait-Dependent Biogeographical Models Indicates That Podocarpaceae Dispersal Is Influenced by Both Seed Cone Traits and Geographical Distance.

The ability of lineages to disperse long distances over evolutionary timescales may be influenced by the gain or loss of traits adapted to enhance local, ecological dispersal. For example, some species in the southern conifer family Podocarpaceae have fleshy cones that encourage bird dispersal, but it is unknown how this trait has influenced the clade's historical biogeography, or its importance compared with other predictors of dispersal such as the geographic distance between regions. We answer these questions quantitatively by using a dated phylogeny of 197 species of southern conifers (Podocarpaceae and their sister family Araucariaceae) to statistically compare standard, trait-independent biogeography models with new BioGeoBEARS models where an evolving trait can influence dispersal probability, and trait history, biogeographical history, and model parameters are jointly inferred. We validate the method with simulation-inference experiments. Comparing all models, those that include trait-dependent dispersal accrue 87.5% of the corrected Akaike Information Criterion (AICc) model weight. Averaged across all models, lineages with nonfleshy cones had a dispersal probability multiplier of 0.49 compared with lineages with fleshy cones. Distance is included as a predictor of dispersal in all credible models (100% model weight). However, models with changing geography earned only 22.0% of the model weight, and models submerging New Caledonia/New Zealand earned only 0.01%. The importance of traits and distance suggests that long-distance dispersal over macroevolutionary timespans should not be thought of as a highly unpredictable chance event. Instead, long-distance dispersal can be modeled, allowing statistical model comparison to quantify support for different hypotheses.

Forecasting Spring Flight of Ips subelongatus (Coleoptera: Curculionidae: Scolytinae) in Japanese larch Larix kaempferi (Pinales: Pinaceae) Forests in the Republic of Korea.

Ips subelongatus is a species of bark beetle experiencing population outbreaks in Korea. In this study, a predictive model and empirical prediction were used to forecast the spring flight of these beetles in Japanese larch forests. The number of beetles caught in pheromone traps was investigated in larch forests thinned in 2009, 2010, 2012, or 2013. Data from the sites thinned in 2009, 2010, or 2012 were used in the predictive model based on a degree-day model that was validated using data from the site thinned in 2013. The lower threshold temperature for flight (LTF) and a thermal sum for the spring flight of I. subelongatus were estimated. The empirical prediction that beetles initiate their flight when daily maximum temperatures reach 16 or 20°C was tested using daily maximum temperature and the beetles caught. The LTF was estimated as 5.97°C, with 42.95 degree-days required for initiation of spring flight. The median flight dates were estimated with a discrepancy from 1 to 3 d by the predictive model. Using the empirical prediction, differences between the day when daily maximum temperature reached 16 or 20°C and flight peak days ranged from 4 to 45 d. These results demonstrate that the predictive model is more suitable than the empirical prediction for predicting the spring flight of I. subelongatus. Overwintering I. subelongatus adults seem to need to gain a determined thermal sum before initiating spring flight rather than merely waiting for the daily maximum temperature to exceed a critical temperature.

Andean Prumnopitys Andina (Podocarpacae) Fruit Extracts: Characterization of Secondary Metabolites and Potential Cytoprotective Effect.

The fruits from the Chilean Podocarpaceae Prumnopitys andina have been consumed since pre-Hispanic times. Little is known about the composition and biological properties of this fruit. The aim of this work was to identify the secondary metabolites of the edible part of P. andina fruits and to assess their antioxidant activity by means of chemical and cell-based assays. Methanol extracts from P. andina fruits were fractionated on a XAD7 resin and the main compounds were isolated by chromatographic means. Antioxidant activity was determined by means of 2,2-diphenyl-1-picrylhydrazyl radical (DPPH), ferric reducing power (FRAP), trolox equivalent antioxidant capacity (TEAC) and oxygen radical absorbance capacity (ORAC) assays. The cytoprotective activity of the extract against oxidative and dicarbonyl stress was evaluated in human gastric epithelial cells (AGS). The total intracellular antioxidant activity (TAA) of the extract was determined in AGS cells. The inhibition of meat lipoperoxidation was evaluated under simulated gastric digestion conditions. Rutin, caffeic acid ß-glucoside and 20-hydroxyecdysone were identified as major components of the fruit extract. Additional compounds were identified by high-performance liquid chromatography diode-array detector mass spectrometry (HPLC-DAD-MSn) and/or co-injection with standards. Extracts showed dose-dependent cytoprotective effects against oxidative and dicarbonyl-induced damage in AGS cells. The TAA increased with the pre-incubation of AGS cells with the extract. This is the first report on the composition and biological activity of this Andean fruit.

Restoring the Taxol biosynthetic machinery of Aspergillus terreus by Podocarpus gracilior Pilger microbiome, with retrieving the ribosome biogenesis proteins of WD40 superfamily.

Attenuating the Taxol yield of Aspergillus terreus with the subculturing and storage were the technical challenges that prevent this fungus to be a novel platform for industrial Taxol production. Thus, the objective of this study was to unravel the metabolic machineries of A. terreus associated with attenuation of Taxol productivity, and their restoring potency upon cocultivation with the Podocarpus gracilior microbiome. The Taxol yield of A. terreus was drastically reduced with the fungal subculturing. At the 10th subculture, the yield of Taxol was reduced by four folds (78.2 µg/l) comparing to the original culture (268 µg/l), as authenticated from silencing of molecular expression of the Taxol-rate limiting enzymes (GGPPS, TDS, DBAT and BAPT) by qPCR analyses. The visual fading of A. terreus conidial pigmentation with the subculturing, revealing the biosynthetic correlation of melanin and Taxol. The level of intracellular acetyl-CoA influx was reduced sequentially with the fungal subculturing, rationalizing the decreasing on Taxol and melanin yields. Fascinatingly, the Taxol biosynthetic machinery and cellular acetyl-CoA of A. terreus have been completely restored upon addition of 3% surface sterilized leaves of P. gracilior, suggesting the implantation of plant microbiome on re-triggering the molecular machinery of Taxol biosynthesis, their transcriptional factors, and/or increasing the influx of Acetyl-CoA. The expression of the proteins of 74.4, 68.2, 37.1 kDa were exponentially suppressed with A. terreus subculturing, and strongly restored upon addition of P. gracilior leaves, ensuring their profoundly correlation with the molecular expression of Taxol biosynthetic genes. From the proteomic analysis, the restored proteins 74.4 kDa of A. terreus upon addition of P. gracilior leaves were annotated as ribosome biogenesis proteins YTM and microtubule-assembly proteins that belong to WD40 superfamily. Thus, further ongoing studies for molecular cloning and expression of these genes with strong promotors in A. terreus, have been initiated, to construct a novel platform of metabolically stable A. terreus for sustainable Taxol production. Attenuating the Taxol yield of A. terreus with the multiple-culturing and storage might be due to the reduction on main influx of acetyl-CoA, or downregulation of ribosome biogenesis proteins that belong to WD40 protein superfamily.

Reproductive Ontogeny and the Evolution of Morphological Diversity in Conifers and Other Plants.

Biologists often study morphological evolution through form and function relationships. But biological structures can perform multiple functional roles, complicating efforts to understand the evolutionary significance of any one relationship. Plant reproductive organs perform multiple roles in a sequence, however, which provides a unique opportunity to understand how structures evolve to meet multiple functional demands. Using conifers as a study group, we discuss how a shared developmental trajectory links the performance of sequential functional roles. Variation in development among lineages can underlie morphological diversity; pollination-stage seed cones in Pinaceae conifers function similarly but show diverse forms reflecting differences in developmental rate. As cones develop further, the morphologies that they use to perform later functional roles are influenced by the specific developmental patterns used to meet earlier demands, which may ultimately limit morphological diversity. However, we also show how selective pressures relating to the final functional stage (seed dispersal) may influence cone anatomy and morphology over all previous stages, highlighting the complex linkages among form, function, and development. We end by discussing the potential relationships between functional ontogeny and morphological disparity in plant reproductive structures more broadly, suggesting that the complex functional roles associated with seed plant reproduction probably underlie the high disparity in this group.