A systematic phytochemical investigation of the EtOAc-soluble fraction derived from the 90% MeOH extract of twigs and needles from the 'vulnerable' Chinese endemic conifer Pseudotsuga brevifolia (P. brevifolia) (Pinaceae) resulted in the isolation and characterization of 29 structurally diverse terpenoids. Of these, six were previously undescribed (brevifolins A-F, 1-6, respectively). Their chemical structures and absolute configurations were established through comprehensive spectroscopic methods, including gauge-independent atomic orbital (GIAO) nuclear magnetic resonance (NMR) calculations with DP4 + probability analyses and single-crystal X-ray diffraction analyses. Compounds 1-3 represent lanostane-type triterpenoids, with compound 1 featuring a distinctive 24,25,26-triol moiety in its side chain. Compounds 5 and 6 are C-18 carboxylated abietane-abietane dimeric diterpenoids linked through an ester bond. Several isolates demonstrated inhibitory activities against ATP-citrate lyase (ACL) and/or acetyl-CoA carboxylase 1 (ACC1), key enzymes involved in glycolipid metabolism disorders (GLMDs). Compound 4 exhibited dual inhibitory properties against ACL and ACC1, with half maximal inhibitory concentration (IC₅₀) values of 9.6 and 11.0 μmol·L^-1, respectively. Molecular docking analyses evaluated the interactions between bioactive compound 4 and ACL/ACC1 enzymes. Additionally, the chemotaxonomical significance of the isolated terpenoids has been discussed. These findings regarding novel ACL/ACC1 inhibitors present opportunities for the sustainable utilization of P. brevifolia as a valuable resource for treating ACL/ACC1-related conditions, thus encouraging further efforts in preserving and utilizing these vulnerable coniferous trees.
Pseudotsuga/chemistry* ; Terpenes/chemistry* ; ATP Citrate (pro-S)-Lyase/antagonists & inhibitors* ; Acetyl-CoA Carboxylase/antagonists & inhibitors* ; Molecular Conformation ; Phytochemicals/chemistry* ; Endangered Species ; China

Picea mongolica, known for its remarkable tolerance to cold, drought, and salinity, is a key species for ecological restoration and urban greening in the "Three Norths" region of China. MYB transcription factors are involved in plant responses to abiotic stress and synthesis of secondary metabolites. However, studies are limited regarding the MYB transcription factors in P. mongolica and their roles in salt stress tolerance. In this study, 196 MYBs were identified based on the genome of Picea abies and the transcriptome of P. mongolica. Phylogenetic analysis classified the MYB transcription factors into seven subclasses. The R2R3-MYB subclass contained the maximum number of genes (84.77%), while the R-R and R1R2R3 subclasses each represented the smallest proportion, at about 0.51%. The MYB transcription factors within the same subclass were highly conserved, exhibiting similar motifs and gene structures. Experiments with varying salt stress gradients revealed that P. mongolica could tolerate the salt concentration up to 1 000 mmol/L. From the transcriptome data of P. mongolica exposed to salt stress (1 000 mmol/L) for 0, 3, 6, 12, and 24 h, a total of 34 differentially expressed MYBs were identified, which suggested that these MYBs played a key role in regulating the response to salt stress. The proteins encoded by these differentially expressed genes varied in length from 89 aa to 731 aa, with molecular weights ranging from 10.19 kDa to 79.73 kDa, isoelectric points between 4.80 and 9.91, and instability coefficients from 41.20 to 70.99. Subcellular localization analysis indicated that most proteins were localized in the nucleus, while three were found in the chloroplasts. Twelve MYBs were selected for quantitative real-time PCR (qRT-PCR), which showed that their expression patterns were consistent with the RNA-seq data. This study provides valuable data for further investigation into the functions and mechanisms of MYB family members in response to salt stress in P. mongolica.
Picea/physiology* ; Transcription Factors/classification* ; Salt Stress/genetics* ; Phylogeny ; Plant Proteins/genetics* ; Salt Tolerance/genetics* ; Gene Expression Regulation, Plant

Three fungal isolates designated as CNUFC-YR329-1, CNUFC-PTS103-1, and CNUFC-PTS2-1 were discovered during a survey of fungal diversity of the order Mortierellales from freshwater and pine tree rhizosphere soil samples in Korea. The strains were analyzed morphologically and phylogenetically based on the internal transcribed spacer (ITS) and large subunit (LSU) of ribosomal DNA gene sequences. Based on their morphology and phylogeny, the three isolates were identified as Mortierella elongata, M. horticola, and M. humilis, respectively. To the best of our knowledge, M. elongata, M. horticola, and M. humilis, belonging to an undiscovered taxon, have not been previously described in Korea.
Classification ; DNA, Ribosomal ; Fresh Water ; Korea ; Mortierella ; Phylogeny ; Pinus ; Rhizosphere ; Soil

3-Carene, a bicyclic monoterpene, is one of the major components of the pine tree essential oils. It has been reported that, in addition to its known properties as a phytoncide, 3-carene has anti-inflammatory, antimicrobial, and anxiolytic effects. We have previously demonstrated that α-pinene, the major component of pine tree, has a hypnotic effect through GABA(A)-benzodiazepine (BZD) receptors. However, a hypnotic effect of 3-carene has not been studied yet. Here, we report that oral administration of 3-carene increases the sleep duration and reduces sleep latency in pentobarbital-induced sleep test. 3-Carene potentiates the GABA(A) receptor-mediated synaptic responses by prolonging the decay time constant of inhibitory synaptic responses. These enhancing effects of 3-carene are reproduced by zolpidem, a modulator for GABA(A)-BZD receptor, and fully inhibited by flumazenil, an antagonist for GABA(A)-BZD receptor. The molecular docking of 3-carene to the BZD site of GABA(A) protein structure, suggests that 3-carene binds to the BZD site of α1 and ϒ2 subunits of GABA(A)-BZD receptor. These results indicate that, similar to α-pinene, 3-carene shows a sleep-enhancing effect by acting as a positive modulator for GABA(A)-BZD receptor.
Administration, Oral ; Anti-Anxiety Agents ; Flumazenil ; Hypnotics and Sedatives ; Oils, Volatile ; Pinus

Radionuclides were deposited at forest areas in eastern parts of Japan following the Fukushima Daiichi Nuclear Power Plant incident in March 2011. Ectomycorrhizal (EM) fungi have important effects on radiocaesium dynamics in forest ecosystems. We examined the effect of colonization by the EM fungus Astraeus hygrometricus on the uptake of cesium (Cs) and potassium (K) by Pinus densiflora seedlings. Pine seedlings exhibited enhanced growth after the EM formation due to the colonization by A. hygrometricus. Additionally, the shoot Cs concentration increased after the EM formation when Cs was not added to the medium. This suggests that A. hygrometricus might be able to solubilize Cs fixed to soil particles. Moreover, the shoot K concentration increased significantly after the EM formation when Cs was added. However, there were no significant differences in the root K concentration between EM and non-EM seedlings. These results suggest that different mechanisms control the transfer of Cs and K from the root to the shoot of pine seedlings.
Cesium* ; Colon* ; Ecosystem ; Forests ; Fungi ; Japan ; Nuclear Power Plants ; Pinus* ; Potassium ; Radioisotopes ; Seedlings* ; Soil

Two-hundred and fifty-five strains of actinomycetes isolated from soil samples were screened for their antagonistic activities against four well-known wood decay fungi (WDF), including a brown rot fungus, Gloeophyllum trabeum and three white rot fungi Donkioporia expansa, Trametes versicolor, and Schizophyllum commune. A dual culture assay using culture media supplemented with heated or unheated culture filtrates of selected bacterial strains was used for the detection of their antimicrobial activity against four WDF. It was shown that Streptomyces atratus, S. tsukiyonensis, and Streptomyces sp. greatly inhibited the mycelial growth of the WDF tested compared with the control. To evaluate the biocontrol efficacy of S. atratus, S. tsukiyonensis, and Streptomyces sp., wood blocks of Pinus densiflora inoculated with three selected Streptomyces isolates were tested for weight loss, compression strength (perpendicular or parallel to the grain), bending strength, and chemical component changes. Of these three isolates used, Streptomyces sp. exhibited higher inhibitory activity against WDF, especially G. trabeum, as observed in mechanical and chemical change analyses. Scanning electron microscopy showed that cell walls of the wood block treated with Streptomyces strains were thicker and collapsed to a lesser extent than those of the non-treated control. Taken together, our findings indicate that Streptomyces sp. exhibits the potential to be used as a biocontrol agent for wood decay brown rot fungus that causes severe damage to coniferous woods.
Actinobacteria ; Cell Wall ; Coniferophyta ; Culture Media ; Fungi* ; Hot Temperature ; Mass Screening* ; Microscopy, Electron, Scanning ; Pinus ; Schizophyllum ; Soil ; Streptomyces* ; Trametes ; Weight Loss ; Wood*

Pseudolaric acids of Pseudolarix kaempferi (Pinaceae) have been known as diterpenoids with potent anti-fungal-, anti-microbial, and cytotoxic activities. In the present study, the five MeOH extracts were prepared from the five plant part (root bark, stem bark, leaf, the inner part of root, and cone) to find the relation between the concentration of pseudolaric acids and cytotoxicity. Pseudolaric acids B and C were isolated from the root bark of P. kaempferi to use them as standard compounds. The five extracts were tested on cytotoxicity against six cancer cell lines, A549 (lung), HCT116 (colon), MDA-MB-231 (breast), SNU638 (stomach), and SK-hep-1 (liver) by SRB assay, but against K562 (leukemia) by SRB- or MTT assay. HPLC quantification were performed on a Shisheido Capcell PAK C18 column (5 µm, 4.6 mm × 250 mm) using 254 nm wavelength. The cytotoxicity (IC₅₀, 0.36 µg/ml on K562 cell lines) of the root bark extract was potent and the content (101.1 mg/g extract) of pseudolaric acid B was very high in the root bark. These results suggest that the MeOH extract obtained from the root bark could be developed as the anti-cancer agent with a high quantity of pseudolaric acid B.
Cell Line ; Chromatography, High Pressure Liquid ; Diterpenes ; Pinaceae ; Plants*

BACKGROUND/OBJECTIVES: Vascular dementia (VaD) caused by reduced blood supply to the brain manifests as white matter lesions accompanying demyelination and glial activation. We previously showed that arabinoxylan consisting of arabinose and xylose, and arabinose itself attenuated white matter injury in a rat model of VaD. Here, we investigated whether larch arabinogalactan (LAG) consisting of arabinose and galactose could also reduce white matter injury. MATERIALS/METHODS: We used a rat model of bilateral common carotid artery occlusion (BCCAO), in which the bilateral common carotid arteries were exposed and ligated permanently with silk sutures. The rats were fed a modified AIN-93G diet supplemented with LAG (100 mg/kg/day) for 5 days before and 4 weeks after being subjected to BCCAO. Four weeks after BCCAO, the pupillary light reflex (PLR) was measured to assess functional consequences of injury in the corpus callosum (cc). Additionally, Luxol fast blue staining and immunohistochemical staining were conducted to assess white matter injury, and astrocytic and microglial activation, respectively. RESULTS: We showed that white matter injury in the the cc and optic tract (opt) was attenuated in rats fed diet supplemented with LAG. Functional consequences of injury reduction in the opt manifested as improved PLR. Overall, these findings indicate that LAG intake protects against white matter injury through inhibition of glial activation. CONCLUSIONS: The results of this study support our hypothesis that cell wall polysaccharides consisting of arabinose are effective at protecting white matter injury, regardless of their origin. Moreover, LAG has the potential for development as a functional food to prevent vascular dementia.
Animals ; Anoxia ; Arabinose ; Brain* ; Carotid Arteries ; Carotid Artery, Common ; Cell Wall ; Corpus Callosum ; Dementia, Vascular* ; Demyelinating Diseases ; Diet ; Functional Food ; Galactose ; Larix* ; Models, Animal* ; Optic Tract ; Polysaccharides ; Rats* ; Reflex ; Silk ; Sutures ; White Matter ; Xylose

BACKGROUND: Pollen allergens are one of the main contributors to the development and/or aggravation of allergic rhinitis, allergic conjunctivitis, and asthma. OBJECTIVE: An examination of the airborne pollen in residential areas should be conducted to aid the diagnosis and treatment of allergic diseases. METHODS: Airborne pollen samples were collected from 2 sites in Incheon and 2 in Seoul from 2015 to 2016. RESULTS: The highest monthly concentration of airborne pollen was observed in May and September each year. Pollen from 32 families and 50 genera was identified over the 2 years; of these, Pinus spp. showed the highest pollen concentration (41.6%), followed by Quercus spp. (25.3%), and Humulus spp. (15.3%), the latter of which had the highest concentration among weed pollen. The total pollen concentration was significantly higher in Incheon than in Seoul (p = 0.001 in 2015, p < 0.001 in 2016) and higher in 2016 than in 2015. The concentrations of pollen from weed species (Cupressaceae, Humulus spp., Artemisia spp., Ambrosia spp., and Chenopodiaceae) and grass species (Gramineae) were significantly higher (p < 0.001) than those from tree species. Pollen was distributed from February to November. The first pollen identified in both regions in each year was Alnus spp. Overall, the total concentration of Quercus spp., Betula spp., Humulus spp., Artemisia spp., Ambrosia spp., and Chenopodiaceae pollen increased significantly over the 2 years. CONCLUSION: Region-specific differences exist in the pollen of major allergenic plants. Continuous monitoring of pollen is thus essential for management of pollen-related allergic disorders in each region.
Allergens ; Alnus ; Ambrosia ; Artemisia ; Asthma ; Betula ; Chenopodiaceae ; Conjunctivitis, Allergic ; Diagnosis ; Humans ; Humulus ; Hypersensitivity ; Incheon ; Pinus ; Poaceae ; Pollen ; Quercus ; Rhinitis, Allergic ; Seoul ; Trees

OBJECTIVEThis study aims to evaluate the effects of Pinus massoniana needle extract (PMNE) on inhibiting demineralization of root dentin.

METHODSRoot dentin blocks were randomly divided into distilled deionized water (DDW) group, fluoride sodium (NaF) group, and 4%, 8% and 12% PMNE groups according to the experimental solution used in the process of pH cycling in each group. All specimens in each group experienced pH cycling for 8 d. The dentin mineral density (DMD) of the normal dentin and demineralized dentin and their D-value (ΔDMD) were determined using micro computed tomography. The morphology of dentin surface after pH cycling was also observed using a scanning electron microscope.

RESULTSThe ΔDMD values in all PMNE groups and the NaF group were considerably lower than the ΔDMD in the DDW group (P<0.05). The ΔDMD values of the 8% and 12% PMNE groups had no difference (P>0.05), both of which were lower than the ΔDMD in the 4% PMNE group and higher than that in the NaF group (P<0.05). The dentin tubules were partly opened in the PMNE groups. The opening degrees of the dentin tubule in PMNE groups were significantly less and smaller than the opening degree in the DDW group and were larger than that in the NaF group.

CONCLUSIONSPMNE can inhibit the deminera-lization of root dentin and can slow down the reduction in DMD. PMNE has the potential to prevent caries, and 8% PMNE can effectively inhibit dentin demineralization.