Identification of key genes involved in lignin and flavonoid accumulation during Tilia tuan seed maturation.

KEY MESSAGE: Transcriptomics and phenotypic data analysis identified 24 transcription factors (TFs) that play key roles in regulating the competitive accumulation of lignin and flavonoids. Tilia tuan Szyszyl. (T. tuan) is a timber tree species with important ecological and commercial value. However, its highly lignified pericarp results in a low seed germination rate and a long dormancy period. In addition, it is unknown whether there is an interaction between the biosynthesis of flavonoids and lignin as products of the phenylpropanoid pathway during seed development. To explore the molecular regulatory mechanism of lignin and flavonoid biosynthesis, T. tuan seeds were harvested at five stages (30, 60, 90, 120, and 150 days after pollination) for lignin and flavonoid analyses. The results showed that lignin accumulated rapidly in the early and middle stages (S1, S3, and S4), and rapid accumulation of flavonoids during the early and late stages (S1 and S5). High-throughput RNA sequencing analysis of developing seeds identified 50,553 transcripts, including 223 phenylpropanoid biosynthetic pathway genes involved in lignin accumulation grouped into 3 clusters, and 106 flavonoid biosynthetic pathway genes (FBPGs) grouped into 2 clusters. Subsequent WGCNA and time-ordered gene co-expression network (TO-GCN) analysis revealed that 24 TFs (e.g., TtARF2 and TtWRKY15) were involved in flavonoids and lignin biosynthesis regulation. The transcriptome data were validated by qRT-PCR to analyze the expression profiles of key enzyme-coding genes. This study revealed that there existed a competitive relationship between flavonoid and lignin biosynthesis pathway during the development of T. tuan seeds, that provide a foundation for the further exploration of molecular mechanisms underlying lignin and flavonoid accumulation in T. tuan seeds.

Total Mineral Content in Raw Materials of Ten Medicinal Plants from Roadside Biotopes of Voronezh Oblast.

The total mineral content was studied in medicinal plants from roadside and railside biotopes of the Voronezh region. Pharmacopoeial plant raw materials of 10 species were evaluated: roots of Taraxacum officinale F.H. Wigg and Arctium lappa L.; herb of Polygonum aviculare L., Artemisia absinthium L., Leonurus quinquelobatus Gilib., and Achillea millefolium L.; leaves of Urtica dioica L. and Plantago major L.; and flowers of Tanacetum vulgare L. and Tilia cordata Mill. Plant raw materials were collected near roads and railways of various types in the periods specified in regulatory documents. The total ash content in plant material was used to determine the minimum allowable distances from various roads and railways for collecting plant material. The minimum allowable distance from heavy-traffic motorways was recommended to be 210 m in the forest zone, 240 m in the forest-steppe zone, and 380 m in the steppe zone. A distance of at least 80 m was recommended for secondary low-speed roads and railways.

Variation in insect herbivory across an urbanization gradient: The role of abiotic factors and leaf secondary metabolites.

Urbanization impacts plant-herbivore interactions, which are crucial for ecosystem functions such as carbon sequestration and nutrient cycling. While some studies have reported reductions in insect herbivory in urban areas (relative to rural or natural forests), this trend is not consistent and the underlying causes for such variation remain unclear. We conducted a continental-scale study on insect herbivory along urbanization gradients for three European tree species: Quercus robur, Tilia cordata, and Fraxinus excelsior, and further investigated their biotic and abiotic correlates to get at mechanisms. To this end, we quantified insect leaf herbivory and foliar secondary metabolites (phenolics, terpenoids, alkaloids) for 176 trees across eight European cities. Additionally, we collected data on microclimate (air temperature) and soil characteristics (pH, carbon, nutrients) to test for abiotic correlates of urbanization effects directly or indirectly (through changes in plant secondary chemistry) linked to herbivory. Our results showed that urbanization was negatively associated with herbivory for Q. robur and F. excelsior, but not for T. cordata. In addition, urbanization was positively associated with secondary metabolite concentrations, but only for Q. robur. Urbanization was positively associated with air temperature for Q. robur and F. excelsior, and negatively with soil nutrients (magnesium) in the case of F. excelsior, but these abiotic variables were not associated with herbivory. Contrary to expectations, we found no evidence for indirect effects of abiotic factors via plant defences on herbivory for either Q. robur or F. excelsior. Additional biotic or abiotic drivers must therefore be accounted for to explain observed urbanization gradients in herbivory and their interspecific variation.

Anatomy and micromorphology of Tilia species from Turkey and its taxonomic implication.

The genus Tilia L. belongs to Malvaceae and it is represented by four species in Turkey. These are T. cordata, T. platyphyllos, T. rubra subsp. caucasica, and T. tomentosa. Tilia species have medicinal importance, especially to treat colds, flu, and coughs, and are also used as sedatives. Inflorescence with bract is recorded in the pharmacopeia as used parts. This study aimed to investigate the differences in leaf and bract characteristics in distinguishing Tilia species. Leaves and bracts were examined anatomically and micromorphologically under light and scanning electron microscopes (LM and SEM). There is no comparative anatomical and micromorphological study of Tilia species from Turkey. Microscopic examination allows for the observation of stomata types, trichomes, and other anatomical structures. These features can vary among species and can aid in differentiation. Features of bract that is medicinally used part were evaluated to find efficient diagnostic characters. The leaves also possess distinct characteristics, and leaf morphology is already used in the identification key in Flora. The emphasis is on identifying the species of medically used drugs. A diagnostic key was created using LM and SEM based on bract features to differentiate taxa for the first time. RESEARCH HIGHLIGHTS: Leaf and bract anatomy of Tilia taxa was studied via LM and SEM. The raw plant material is inflorescence with bract so identification by bract is important for species adulteration. An diagnostic key based on bract features was created for the first time.

Two novel Raoultella species associated with bleeding cankers of broadleaf hosts, Raoultella scottia sp. nov. and Raoultella lignicola sp. nov.

Seventeen Gram-negative, facultatively anaerobic bacterial strains were isolated from bleeding cankers of various broadleaf hosts and oak rhizosphere soil in Great Britain. The strains were tentatively identified as belonging to the genus Raoultella based on 16S rRNA gene sequencing. Multilocus sequence analysis (MLSA), based on four protein-encoding genes (fusA, leuS, pyrG, and rpoB), separated the strains into three clusters within the Raoultella genus clade. The majority of strains clustered with the type strain of Raoultella terrigena, with the remaining strains divided into two clusters with no known type strain. Whole genome sequencing comparisons confirmed these two clusters of strains as belonging to two novel Raoultella species which can be differentiated phenotypically from their current closest phylogenetic relatives. Therefore, two novel species are proposed: Raoultella scottia sp. nov. (type strain = BAC 10a-01-01T = LMG 33072T = CCUG 77096T) and Raoultella lignicola sp. nov. (type strain = TW_WC1a.1T = LMG 33073T = CCUG 77094T).

Potential habitat areas and priority protected areas of Tilia amurensis Rupr in China under the context of climate change.

Introduction: Tilia amurensis Rupr (T. amurensis) is one endangered and national class II key protected wild plant in China. It has ornamental, material, economic, edible and medicinal values. At present, the resources of T. amurensis are decreasing, and the prediction of the distribution of its potential habitat in China can provide a theoretical basis for the cultivation and rational management of this species. Methods: In this study, the R language was used to evaluate 358 distribution records and 38 environment variables. The MaxEnt model was used to predict the potential distribution areas of T. amurensis under the current and future climate scenarios. The dominant environmental factors affecting the distribution of T. amurensis were analyzed and the Marxan model was used to plan the priority protected areas of this species. Results: The results showed that Bio18, Slope, Elev, Bio1, Bio9 and Bio2 were the dominant environmental factors affecting the distribution of T. amurensis. Under the future climatic scenarios, the potential suitable areas for T. amurensis will mainly distribute in the Northeast China, the total suitable area will reduce compared with the current climate scenarios, and the general trend of the centroid of suitable habitat will be towards higher latitudes. The SPF value of the best plan obtained from the priority conservation area planning was 1.1, the BLM value was 127,616, and the priority conservation area was about 57.61×104 km2. The results suggested that climate, soil and topographic factors jointly affected the potential geographical distribution of T. amurensis, and climate and topographic factors had greater influence than soil factors. Discussion: The total suitable area of T. amurensis in China under different climate scenarios in the future will decrease, so more effective protection should be actively adopted.

Investigating the Effect of the Crosslinking Factor on the Properties of Hydrogel Materials Containing Tilia platyphyllos Hydrolate.

The use of natural ingredients in recent years has been of great importance in many industries and medicine. In biomedical applications, hydrogel materials also play a significant role. In view of this, the aim of this study was to synthesize and characterize hydrogel materials enriched with broadleaf linden hydrolate. An important aspect was to carry out a series of syntheses with varying types and amounts of crosslinking agents so as to test the possibility of synthesizing materials with controlled properties. The obtained hydrogels were subjected to detailed physicochemical analysis. The results of the tests confirmed the relationship between the selected properties and the type of crosslinking agent used. A crosslinking agent with a lower molar mass (575 g/mol) results in a material with a compact and strongly crosslinked structure, which is characterized by high surface roughness. The use of a crosslinking agent with a molecular weight of 700 g/mol resulted in a material with a looser-packed polymer network capable of absorbing larger amounts of liquids. The work also proved that regardless of the type of crosslinking agent used, the addition of linden hydrolate provides antioxidant properties, which is particularly important in view of the target biomedical application of such materials.

First report of leaf spot disease caused by Alternaria alternata in Tilia miqueliana in Jiangsu Province, China.

Tilia miqueliana Maxim., a tall deciduous tree in the Malvaceae family, is native to china and is cultivated in the Jiangsu, Zhejiang, Anhui, and Jiangxi provinces as an ornamental plant. T. miqueliana has currently received increased attention because of its value as timber, a source plant for honey, and in formal landscape architecture (Wang et al. 2022). In the last three years, symptoms of leaf spot disease were observed in T. miqueliana fields （9-year-old tree）and a breeding nursery _(1-year-old saplings) in Nanjing, Jiangsu Province, China. Field surveys showed that, the disease incidence was approximately 10% and 40% in the fields and breeding nursery, respectively. Symptoms of leaf spot disease on T. miqueliana appeared as small, circular, or near-circular/irregular black lesions on the upper surfaces of the leaves. As the disease progressed, the spots expanded into irregular shapes with the center turning yellow to black and leaves wilted from margins to centers. In severe cases, the yellow necrotic area extended to the margin, which ultimately led to leaf curling and death. To analyze the presence of the pathogenic microorganism, pieces of leaf at the junction of diseased and healthy tissue (about 3x3 mm) were taken from symptomatic plants, sterilized with 75% ethanol for 1 min and 2% sodium hypochlorite (NaClO) for 2 min, and washed 3 times with ddH2O. The leaf pieces were transferred onto Potato Dextrose Agar (PDA) plates and incubated at 28°C for 3 days in the dark. Colonies were observed and transferred to fresh PDA plates. Sixteen fungal isolates were obtained and one (named D4-2) of them was verified using Koch's assumption. Single spores of D4-2 were cultured in PDA medium, while its initial colony morphology was fluffy white colonies, which then gradually turned dark yellow from the centre to the edge. To further analyze the form of the spores, a hypha was incubated on fresh synthetic low-nutrient agar (SNA) using cellophane and inverted cultivation was performed for 20 days in the dark. Obclavate or obpyriform conidia (10 to 30 × 2 to 18 µm, n=5) were produced in chains and were pale brown to brown in color. The morphology was consistent with that of Alternaria sp.. The molecular identification of the isolated representative fungus D4-2 was conducted via the amplification of the internal transcribed spacer (ITS) (White et al. 1990),translation elongation factor 1 alpha (TEF) (Carbone and Kohn. 1999), glyceraldehyde-3-dehydrogenase (GAPDH) (Berbee et al. 1999) and RNA polymerase II beta subunit (RPB2) using the primer pairs ITS1/ITS4, EF1-728F/EF1-986R, GPD1/GPD2, and RPB2-5F2/fRPB2-7cR (Sung et al. 2007) respectively. All these sequences were deposited in GenBank under accession numbers OP108438 (ITS), OP168372 (RPB2), OQ473880 (TEF), and OP168374 (GAPDH). Based on the GAPDH, RPB2, and TEF regions of D4-2, MEGA-11 was used to construct a neighbor-joining tree (1000 replications) with 31 other known species that had a relatively close evolutionary relationship with species from the Alternaria genus (Woudenberg et al. 2013). The phylogenetic tree indicated 100% bootstrap support between D4-2 and A. alternata. The branches showing the distribution of other species were exactly the same as that of the consensus tree based on the Bayesian 50 % majority rule that represented the Alternaria-complex (Woudenberg et al. 2013). The initial Koch's Postulates was verified using PDA slices with mycelium (pure PDA slices were used as control) that were inverted on the leaves at 25℃ for 3 days before removing slices and observing the phenotype after 7 days. Only leaves infected with D4-2 showed the same symptoms. Moreover, the spore suspension (at a concentration of 107 spores/mL) of D4-2 was applied to the leaves of T. miqueliana seedlings. After 15 days of incubation at 26 ℃ , the leaves showed the same symptoms. Subsequently, the pathogen was reisolated and cultured from these invaded leaves using the same method described above. Morphological and molecular identifications were consistent with those of D4-2. Above all, the pathogenic fungus (D4-2) isolated from T. miqueliana leaves was confirmed to be A. alternata. T. miqueliana is considered to be an endangered species due to the deep dormancy of its seeds that result in a low natural germination rate (Wu and Shen. 2021). At present, research on T. miqueliana mainly focuses on seed germination and breeding technology, and there is hardly any research on the diseases occurring in T. miqueliana. Therefore, we believe that this is the first report of leaf spot disease caused by A. alternata on T. miqueliana in China.

Anthropological significance of Tilia trees in Japan.

Tilia (Malvaceae) is a genus of deciduous trees widespread in the northern hemisphere. Tilia species native to Japan include three endemic species, T. japonica, T. maximowicziana, and T. kiusiana, as well as the more widespread T. mandshurica. Other species were also introduced, the most important being T. miqueliana, brought to Japan with the arrival of Buddhism and planted on temple grounds as Bodaiju. Both historically and at present, Tilia trees are valuable to the people in Japan. Some Tilia trees are considered sacred in both Buddhism and Shinto. They are also prominent ornamental and park trees, albeit less popular in Japan than in Europe. Japanese Tilia spp. are used in the manufacturing of honey, cosmetics, lumber (especially plywood and veneers), and traditional bast cloth. Many Tilia trees are significant hubs in pollinator and mycorrhizal networks, but research on Japanese Tilia ecology is scarce. Despite their importance, Japanese Tilia trees have received less scientific attention in comparison with European Tilia species. The most striking example is T. kiusiana, with virtually no scientific literature regarding the species (save for a series of publications studying its secondary metabolites and potential medical uses). Furthermore, most published resources concerning Tilia in Japan are available only in Japanese, restricting their accessibility. This review seeks to translate, collect, and organize the information available on Japanese Tilia species. By doing so, areas are highlighted where new studies are necessary. A better understanding of these important trees would also be instrumental in their conservation.

Methodological and physiological study of seed dormancy release in Tilia henryana.

Tilia henryana is a rare tree of the Tilia family, found exclusively in China. Its seeds have severe dormancy features that limit its normal conditions of reproduction and renewal. Its seeds have severe dormant characteristics that limit its normal conditions of reproduction and renewal. The Dormancy in T. henryana seeds is a comprehensive dormancy (PY + PD) caused by mechanical and permeability barriers of seed coat and the presence of germination inhibitor in endosperm. L9 (34) orthogonal test was used to determine the best procedure for releasing the dormancy of T. henryana seeds, that is, first treating the seeds with H2SO4 for 15 min, followed by the application of 1 g L-1 GA3, stratification at 5 °C for 45 days, and finally germination at 20 °C, which can achieve a 98% seed germination rate. Large amounts of fat are consumed throughout the dormancy release process. As quantities of protein and starch marginally increase, soluble sugars are continuously decreased. Acid phosphatase and amylase activities increased rapidly, and the combined enzyme activities of G-6-PDH and 6-PGDH related to the PPP were also significantly increased. The levels of GA and ZR continued to increase, while the levels of ABA and IAA gradually decreased, among which GA and ABA changed most rapidly. The total amino acids content continued to decrease. Asp, Cys, Leu, Phe, His, Lys and Arg decreased with dormancy release, while Ser, Glu, Ala, Ile, Pro and Gaba showed an upward trend. The physical dormancy of T. henryana seeds is broken with H2SO4 in order to make the seed coat more permeable, which is a prerequisite for germination. As a result, the seeds can absorb water and engage in physiological metabolic activities, particularly the hydrolysis and metabolism of fat, which supply a significant amount of energy for dormancy release. In addition, rapid variations in the levels of different endogenous hormones and free amino acids, induced by cold stratification and GA3 application, are another important factor promoting the quick physiological activation of seeds and breaking the endosperm barrier.

First Record of the Invasive Scale Insect, Pulvinaria hydrangeae Steinweden, 1946 (Hemiptera: Coccomorpha: Coccidae) in Romania.

Over the last few decades, globalization and global trade have increased the risk of the vehiculation of invasive organisms, which has had multiple negative effects, both economic and ecological. Through this study, we aimed to produce a report on the first record of the invasive scale insect Pulvinaria hydrangeae (Stein. 1946) in Brașov County in central Romania. It was found on two native tree species: sycamore (Acer pseudoplatanus) and linden (Tilia cordata). In this paper, we (i) highlight the list of possible hosts, (ii) provide a general outlook on infestations and (iii) review the control options for this particular pest. Because early detection and quick reporting are the most important actions in the successful management of invasive species, in general, we also provide a synthetic morphological description of the adult female specimens and ovisacs. Due to natural occurrence, our findings highlight the potential risks posed by the infestation of this insect to native tree species belonging to the Acer and Tilia genera. Because of the temperate climate in Romania and the fact that females are wingless, the new infestations will probably be made through the vehiculation of infested plants, rather than through natural spreading. However, because of global warming, the chances of this species surviving during the winter are likely to increase, making northern expansion of the cottony hydrangea scale feasible.

Transfer of Erwinia toletana and Erwinia iniecta to a novel genus Winslowiella gen. nov. as Winslowiella toletana comb. nov. and Winslowiella iniecta comb. nov. and description of Winslowiella arboricola sp. nov., isolated from bleeding cankers on broadleaf hosts.

Following a screening campaign of bleeding cankers of broadleaf hosts in Great Britain, numerous bacterial strains were isolated, identified by 16S rRNA and protein-coding gene sequencing and ultimately classified. During the course of the study, several Gram-negative, facultatively anaerobic strains were isolated from bleeding Platanus x acerifolia (London plane) and Tilia x europaea (common lime) cankers that could not be assigned to an existing species. Partial 16S rRNA gene sequencing placed these strains in the genus Erwinia, as a close phylogenetic relative of Erwinia toletana. In an effort to determine the taxonomic position of the strains, a polyphasic approach was followed including genotypic, genomic, phenotypic, and chemotaxonomic assays. Multilocus sequence analysis based on four protein-coding genes (gyrB, rpoB, infB, and atpD) confirmed the phylogenetic position of the strains as a novel taxon of subgroup 3 of the genus Erwinia, along with E. toletana and E. iniecta, and furthermore, provided support for their reclassification in a novel genus. Whole genome comparisons allowed the delimitation of the novel species and also supported the proposed transfer of subgroup 3 species to a novel genus in the Erwiniaeae. Phenotypically the novel species could be differentiated from E. toletana and E. iniecta, and the novel genus could be differentiated from the closely related genera Erwinia and Mixta. Therefore, we propose (1) the reclassification of E. toletana and E. iniecta in a novel genus, Winslowiella gen. nov., as Winslowiella toletana comb. nov. and Winslowiella iniecta comb. nov., with W. toletana comb. nov. as the type species (type strain A37T = CFBP 6631T = ATCC 700880T = CECT 5263T), and (2) classification of the novel strains as Winslowiella arboricola sp. nov. (type strain BAC 15a-03bT = LMG 32576T = NCPPB 4696T).

Pharmacological Interaction of Quercetin Derivatives of Tilia americana and Clinical Drugs in Experimental Fibromyalgia.

Fibromyalgia (FM) is a pain syndrome characterized by chronic widespread pain and CNS comorbidities. Tilia americana var. mexicana is a medicinal species used to treat anxiety, insomnia, and acute or chronic pain. However, its spectrum of analgesic efficacy for dysfunctional pain is unknown. To investigate a possible therapeutic alternative for FM-type pain, an aqueous Tilia extract (TE) and its flavonoid fraction (FF) containing rutin and isoquercitrin were evaluated alone and/or combined with clinical drugs (tramadol-TRA and pramipexol-PRA) using the reserpine-induced FM model in rats. Chromatographic analysis allowed the characterization of flavonoids, while a histological analysis confirmed their presence in the brain. TE (10-100 mg/kg, i.p.) and FF (10-300 mg/kg, i.p.) produced significant and dose-dependent antihyperalgesic and antiallodynic effects equivalent to TRA (3-10 mg/kg, i.p.) or PRA (0.01-1 mg/kg, s.c.). Nevertheless, the combination of FF + TRA or FF + PRA resulted in an antagonistic interaction by possible competitive action on the serotonin transporter or µ-opioid and D2 receptors, respectively, according to the in silico analysis. Flavonoids were identified in cerebral regions because of their self-epifluorescence. In conclusion, Tilia possesses potential properties to relieve FM-type pain. However, the consumption of this plant or flavonoids such as quercetin derivatives in combination with analgesic drugs might reduce their individual benefits.

Description of three novel species of Scandinavium: Scandinavium hiltneri sp. nov., Scandinavium manionii sp. nov. and Scandinavium tedordense sp. nov., isolated from the oak rhizosphere and bleeding cankers of broadleaf hosts.

While investigating the bacterial populations of environmental samples taken from a mix of healthy and Acute Oak Decline afflicted Quercus robur (pedunculate or English oak) rhizosphere soil samples and swabs of bleeding lesions on Tilia spp. (lime) and Quercus rubra (red oak) trees, several strains belonging to the order Enterobacterales were isolated using selective media and enrichment broth. Seven strains from the Q. robur rhizosphere, three strains from Tilia spp. and one from Q. rubra were investigated, with their taxonomic status determined via a polyphasic taxonomic approach. Initially stains were identified as potential members of the recently described genus Scandinavium, based on the partial sequencing of three housekeeping genes. Further analysis of phenotypic traits, including fatty acid profiles, coupled with 16S rRNA gene and phylogenomic analysis of whole genome sequences were applied to a subset of the strains. Phylogenetic and phylogenomic analysis repeatedly placed the isolates in a monophyletic clade within Scandinavium, with four distinct clusters observed, one of which corresponded to Scandinavium goeteborgense, the type species of the genus. The remaining three clusters could be phenotypically and genotypically differentiated from each other and S. goeteborgense. As such, we describe three novel species of the genus, for which we propose the names Scandinavium hiltneri sp. nov. (type strain H11S7T = LMG 32612T = CCUG 76179T), Scandinavium manionii sp. nov. (type strain H17S15T = LMG 32613T = CCUG 76183T) and Scandinavium tedordense sp. nov. (type strain TWS1aT = LMG 32614T = CCUG 76188T). Additionally, the descriptions of the genus Scandinavium and the type species, S. goeteborgense, are emended.

Brenneria tiliae sp. nov., isolated from symptomatic Tilia × moltkei and Tilia × europaea trees in the UK.

Several strains of a previously undescribed bacterial species were isolated from mature Tilia hybrid trees suffering from bleeding cankers at various geographic locations in the UK. The strains were Gram-negative, facultatively anaerobic, and partial sequencing of the gyrB gene revealed that the strains belong to the genus Brenneria with the closest phylogenetic neighbours being Brenneria corticis and Brenneria nigrifluens. Further investigation using a polyphasic approach was undertaken to determine the taxonomic position of the novel species. Phylogenies based on the 16S rRNA gene and multilocus sequence analysis of partial housekeeping gene sequences of gyrB, rpoB, infB and atpD revealed that the strains formed an independent cluster within the genus Brenneria. The phenotypic and chemotaxonomic assays demonstrated that the strains could be differentiated from the closest relatives. Genome analysis of representative strains revealed in silico DNA-DNA hybridization values below the threshold for species delimitation, although the average nucleotide identity values obtained when compared to B. corticis (95.9-96%) were slightly higher than the suggested cut-off value of 95%. However, as all other data suggests that the strains belong to a novel taxon that can be differentiated from the closest relatives, we propose that the strains represent a novel species in the genus Brenneria, Brenneria tiliae sp. nov. (type strain WC1b.1T=LMG 32575T=NCPPB 4697T).

Protective Effect of Tilia americana var. mexicana Against Kainic Acid-induced Damage in Brain, Liver, and Kidney: Behavioral and Biochemical Changes.

Tiliaamericana var. mexicana (Tilia) possesses anticonvulsant, antioxidant, neuroprotective, and hepatoprotective activities. The spectrum of anticonvulsant activity in status epilepticus models has not been sufficiently explored. We evaluated the effects of ethyl acetate (EAc), and methanol (ME) extracts on kainic acid (KA)-induced seizures by measuring rats'behavior (severity and latency) and lipoperoxidation in different brain areas (cerebellum, brain hemispheres, cortex, and medulla), kidneys, and liver. Male Wistar rats were administered KA (10 mg/kg, i.p.) after three days of pretreatment with Tilia extract (100 mg/kg). The EAc and ME Tilia extracts significantly decreased the severity of phase 1 and phase 2 seizures, respectively. The ME Tilia extract increased the latency to seizure (27 ± 2 min) compared to the control (13 ± 2 min). The ME and EAc Tilia extracts significantly prevented the increased lipid peroxidation caused by KA-induced seizures in the cerebellum, brain hemispheres, cortex, medulla, liver, and kidneys. The vehicle olive oil (OO) also showed anticonvulsant effects, decreasing the severity of seizures to phase 3 and lipoperoxidation levels in the cerebellum, brain hemispheres, cortex, medulla, liver, and kidneys. The anticonvulsant activity of Tilia is mediated by antioxidant effects in central and systemic areas that involve synergistic interactions among the chemical constituents of these extracts (glucosides of quercetin and kaempferol), while vehicle OO showed the same effects, probably due to its constituent oleuropein.

Spray-drying Microencapsulation of an Extract from Tilia tomentosa Moench Flowers: Physicochemical Characterization and in Vitro Intestinal Activity.

Silver linden (Tilia tomentosa Moench, TtM) flowers possess several health-promoting properties, especially at the neurological level, such as intestinal relaxation activity associated with specific flavonols, particularly quercetin and kaempferol derivatives. However, such molecules are susceptible to degradation upon different triggers like heat, light and extreme pH values. To overcome the scarce stability of TtM flowers bioactive molecules and make them suitable for developing functional food and supplements, we applied microencapsulation. Spray-drying microencapsulation of TtM flowers extract was performed using three starch-derived wall materials: maltodextrin 12 DE (MD12) and 19 DE (MD19), and OSA-modified starch (OSA-S). The stability of total phenols, flavanols, and antioxidant capacity was monitored for 70 days under accelerated stress conditions (40 °C/70% RH) by HPLC and spectrophotometric methods, and the intestinal contractile activity was tested in a murine model. In comparison to MD12 and MD19, OSA-S stood out for the higher encapsulation efficiency of quercetin and kaempferol glycosides (+ 36-47% compared to MD12 and + 18-24% compared to MD19) and stability thereof (half-life on average + 30% compared to MD12 and + 51% compared to MD19). The intestinal contractile activity of OAS-S powders resulted comparable to the original extract, indicating that flavonols were biologically active and accessible. Our results underly the potential advantages of OSA-S encapsulated formulation as a functional ingredient for the development of nutraceutical products.

Limited Acclimation in Leaf Morphology and Anatomy to Experimental Drought in Temperate Forest Species.

Drought is a critical and increasingly common abiotic factor that has impacts on plant structures and functioning and is a challenge for the successful management of forest ecosystems. Here, we test the shifts in leaf morpho-anatomical or hydraulic traits and plant growth above ground caused by drought. A factorial experiment was conducted with two gymnosperms (Larix gmelinii and Pinus koraiensis) and two angiosperms (Fraxinus mandshurica and Tilia amurensis), tree species grown under three varying drought intensities in NE China. Considering all the species studied, the plant height (PH), root collar diameter (RCD), and plant biomass (PB) were significantly decreased by drought. The leaf thickness (LT) increased, while the leaf area (LA) decreased with drought intensity. In the gymnosperms, the mesophyll thickness (MT) increased, and the resin duct decreased, while in the angiosperms the palisade mesophyll thickness (PMT), the spongy mesophyll thickness (SMT), and the abaxial (ABE) and adaxial epidermis (ADE) thickness were increased by drought. The correlation analysis revealed that P. koraiensis and F. mandshurica had the higher RMF and total plant biomass, but the least LMF, suggesting drought tolerance. In contrast, the L. gmelinii had the least RMF and higher LMF, suggesting vulnerability to drought. Similarly, T. amurensis had the higher leaf size, which increased the evaporative demand and depleted the soil water quickly relative to the other species. The interrelation among the morpho-anatomical leaf traits was equally affected by drought across all the studied species, suggesting that there is no clear evidence to differentiate the taxa based on drought resistance vs. drought tolerance. Thus, we have identified some easily measurable traits (i.e., LMF, RMF, and PB) which evidenced the seedling's ability to cope with drought and which therefore could be used as proxies in the selection of drought tolerant species for reforestation in the temperate forest.

Establishment and Expansion of Harmonia axyridis Pallas (Coleoptera: Coccinellidae) in Urban Green Areas in the Iberian Peninsula: From 2015 to 2021.

In the Western Palearctic region, Harmonia axyridis (Coleoptera: Coccinellidae) is mainly established in urban areas. In this study, we update its occurrence in urban areas of the Iberian Peninsula and determine its expansion after 2014. Changes in the ladybird species assemblage are also evaluated. We compile information about the records of H. axyridis in Spain from 2015 to 2021. In addition, we sample different locations to determine the relative abundances of the species composing the aphidophagous ladybird complex. The expansion of H. axyridis mainly occurred in Catalonia (Iberian Peninsula), where it is possible to identify two clear areas: one that has already been invaded and another that has not yet been invaded. Harmonia axyridis became the dominant species in the invaded area two years after it was first identified. This dominance is clearly shown on Liriodendron tulipifera and Tilia platyphyllos, where it accounted for more than 75% of the total collected individuals in the last year of the study. In the not-yet invaded area, Adalia bipunctata overcame Oenopia conglobata and became the prevalent species from 2020 onwards, likely due to its regular releases for aphid biocontrol. This study reveals that changes in ladybird species assemblages may not only be caused by invasive species, but also by biological control practices.

The protective effect of Tilia amurensis honey on influenza A virus infection through stimulation of interferon-mediated IFITM3 signaling.

Recently, attention has focused on the prevention and treatment of respiratory viruses including influenza viruses. We evaluated the antiviral effect of Tilia amurensis honey (TH) against influenza A virus in murine macrophages. Influenza A virus infection was reduced following pretreatment with TH. Pretreatment of murine macrophages with TH increased the production and secretion of type-1 interferon (IFN) and proinflammatory cytokines and increased phosphorylation of the type-1 IFN-related proteins, TANK-binding kinase (TBK), and STAT. Moreover, TH increased the expression of IFN-stimulating genes and increased the expression of IFN-inducible transmembrane (IFITM3), a protein that interferes with virus replication and entry. Taken together, these findings suggest that TH suppresses influenza A virus infection by regulating the innate immune response in macrophages. This supports the development of preventive and therapeutic agents for influenza A virus and enhances the economic value of TH.