Differences of stomatal ozone uptake in leaves of mature trees and seedlings of Zelkova serrata.

Ozone uptake through the stomata in tree leaves is an important process for improving air quality by urban trees. Stomatal conductance (gs) is a key determinant of stomatal ozone uptake. The parameterization of gs models for estimating stomatal ozone uptake of trees has mainly been carried out using gs data measured in seedling leaves although the leaf traits may differ between mature trees and seedlings. In the present study, we compared stomatal ozone uptake estimated by gs models parameterised with data from mature trees and seedlings of Zelkova serrata. We measured gs in leaves of mature trees and seedlings of Z. serrata using a leaf porometer for 3-4 growing seasons. The Jarvis-type gs model was parameterised with data from mature trees and seedlings, separately. The maximum gs, and the functions of the seedling gs estimation model regarding the response to air temperature, vapour pressure deficit and atmospheric ozone concentration were the factors inducing lower stomatal ozone uptake. In contrast, the function of the seedling gs estimation model regarding the response to irradiance resulted in a higher estimated stomatal ozone uptake. The estimated stomatal ozone uptake for one growing season (April-September) by the seedling gs estimation model was 27% lower than that by the mature tree gs estimation model. These results indicate that leaf gas exchange traits of Z. serrata were different between mature trees and seedlings, and that estimating ozone uptake in mature tree leaves using a model based on seedling gs measurements results in an underestimation.

The complete chloroplast genome of Ulmus mianzhuensis with insights into structural variations, adaptive evolution, and phylogenetic relationships of Ulmus (Ulmaceae).

BACKGROUND: Ulmus mianzhuensis is an endemic tree species in China with high ornamental and economic value. Currently, little is known regarding its genomic architecture, phylogenetic position, or adaptive evolution. Here, we sequenced the complete chloroplast genome (cp genome) of U. mianzhuensis and further compared the variations in gene organization and structure within Ulmus species to define their genomic evolution, then reconstructed the phylogenomic relationship of 31 related Ulmus species to explore the systematic position of U. mianzhuensis and the utility of cp genome for resolving phylogenetics among Ulmus species. RESULTS: Our results revealed that all the Ulmus species exhibited a typical quadripartite structure, with a large single copy (LSC) region of 87,170 - 88,408 bp, a small single copy (SSC) region of 18,650 - 19,038 bp and an inverted repeat (IR) region of 26,288 - 26,546 bp. Within Ulmus species, gene structure and content of cp genomes were highly conserved, although slight variations were found in the boundary of SC/IR regions. Moreover, genome-wide sliding window analysis uncovered the variability of ndhC-trnV-UAC, ndhF-rpl32, and psbI-trnS-GCU were higher among 31 Ulmus that may be useful for the population genetics and potential DNA barcodes. Two genes (rps15 and atpF) were further detected under a positive selection of Ulmus species. Comparative phylogenetic analysis based on the cp genome and protein-coding genes revealed consistent topology that U. mianzhuensis is a sister group to U. parvifolia (sect. Microptelea) with a relatively low-level nucleotide variation of the cp genome. Additionally, our analyses also found that the traditional taxonomic system of five sections in Ulmus is not supported by the current phylogenomic topology with a nested evolutionary relationship between sections. CONCLUSIONS: Features of the cp genome length, GC content, organization, and gene order were highly conserved within Ulmus. Furthermore, molecular evidence from the low variation of the cp genome suggested that U. mianzhuensis should be merged into U. parvifolia and regarded as a subspecies of U. parvifolia. Overall, we demonstrated that the cp genome provides valuable information for understanding the genetic variation and phylogenetic relationship in Ulmus.

Comparative Transcriptome Sequencing and Endogenous Phytohormone Content of Annual Grafted Branches of Zelkova schneideriana and Its Dwarf Variety HenTianGao.

Zelkova schneideriana is a fast-growing tree species endemic to China. Recent surveys and reports have highlighted a continued decline in its natural populations; therefore, it is included in the Red List of Threatened Species by The International Union for Conservation of Nature. A new variety "HenTianGao" (H) has been developed with smaller plant height, slow growth, and lower branching points. In this study, we attempted to understand the differences in plant height of Z. schneideriana (J) and its dwarf variety H. We determined the endogenous hormone content in the annual grafted branches of both J and H. J exhibited higher gibberellic acid (GA)-19 and trans-Zeatin (tZ) levels, whereas H had higher levels of indole-3-acetic acid (IAA) catabolite 2-oxindole-3-acetic acid (OxIAA), IAA-Glu conjugate, and jasmonic acid (JA) (and its conjugate JA-Ile). The transcriptome comparison showed differential regulation of 20,944 genes enriched in growth and development, signaling, and metabolism-related pathways. The results show that the differential phytohormone level (IAA, JA, tZ, and GA) was consistent with the expression of the genes associated with their biosynthesis. The differences in relative OxIAA, IAA-Glu, GA19, trans-Zeatin, JA, and JA-Ile levels were linked to changes in respective signaling-related genes. We also observed significant differences in the expression of cell size, number, proliferation, cell wall biosynthesis, and remodeling-related genes in J and H. The differences in relative endogenous hormone levels, expression of biosynthesis, and signaling genes provide a theoretical basis for understanding the plant height differences in Z. schneideriana.

Antileishmaniasis effects of monoherbal skin ointment made from bark and leaf of Holoptelea integrifolia (Roxb.) Planch. in patients of cutaneous leishmaniasis.

Prevalence of parasitic skin diseases, cutaneous leishmaniasis is very frequent in Pakistan, especially in Baluchistan where occurrence of phelbotomine sand flies (vector of protozoan parasite of genus Leishmania) is very common. This study was conducted to assess the antileishmaniasis activity of Intericutol plus ointment (2% and 3%) made with ethanolic extract of bark and leaves of Holoptelea integrifolia (Roxb.)Planch. The antileishmaniasis effect was observed by applying the ointments on the lesions of leishmaniasis patients. 150 patients were included in the study that had single and multiple painful wet and dry ulcerative lesions on the exposed area of the body. The patients were divided into two groups of 75 patients each, applying 2% and 3% ointment respectively for 28 days. Dose of 10gm/week was applied topically on wounds. Excellent antileishmaniasis effect was observed with significant quick healing properties in the patients receiving 3% Intericutol plus ointment as compare to 2% showing 89% and 81% cure (p=0.02) respectively. It can be concluded that ointment made from medicinal plant proved to be very effective in treating cutaneous Leishmaniasis.

Dynamic evolution of the plastome in the Elm family (Ulmaceae).

MAIN CONCLUSION: This study compared the plastomes of Ulmaceae allowing analyses of the dynamic evolution, including genome structure, codon usage bias, repeat sequences, molecular mutation rates, and phylogenetic inferences. Ulmaceae is a small family in the order Rosales. This family consists of seven genera, including Ulmus, Zelkova, Planera, Hemiptelea, Phyllostylon, Ampelocera, and Holoptelea. Ulmaceae is an interesting lineage from plant biogeographic, systematic, evolutionary, and paleobotanic perspectives. It is also a good model to investigate the evolution of the plastomes in woody plants. In this study, we sequenced and assembled the complete plastomes of the six Ulmaceae genera to compare genomic structures and reveal the molecular evolutionary patterns. The size of the quadripartite plastomes ranged from 158,290 bp to 161,886 bp. The genomes contained 131 genes, including 87 coding genes, 36 tRNA, and 8 rRNA. The gene number, gene content, and genomic structure were highly consistent among the Ulmaceae genera. Nine variable regions including ndhA intron, ndhF-rpl32, ycf1, psbK-trnS, rps16-trnQ, trnT-trnL, trnT-psbD, trnS-trnG, and rpl32-trnL, were identified in Ulmaceae plastomes according to the nucleotide diversity values. Condon usage was biased among the genes and showed consistent trends in the seven genera. Molecular evolution analyses revealed that most of the genes and all gene groups were under widespread purifying selection. Twelve genes (ccsA, matK, psbH, psbK, rbcL, rpl22, rpl32, rpoA, rps12, rps15, rps16, and ycf2) were under positive selection. Phylogenetic analyses supported that Ulmaceae should be divided into two main clades, such as the temperate clade, including Ulmus, Zelkova, Planera, and Hemiptelea and the tropical clade, including Phyllostylon, Ampelocera and Holoptelea. This study reports the structure and evolutionary characteristics of the Elm family. These new genomic data will benefit assessments of genomic evolution and provide information to elucidate the phylogenetic relationships among Ulmaceae species.

Antinociceptive and anti-inflammatory effects of extract of Celtis iguanaea (Jacq.) Sargent leaves in mice.

The antinociceptive and anti-inflammatory activities of crude ethanolic extract of Celtis iguanaea leaves and their active fractions are reported. The oral treatment with crude ethanolic extract (CEE; 100, 300 or 1000 mg/Kg) inhibited the number of writhings in a dose-dependent manner. The intermediate dose also inhibited formalin-induced nociception in both phases. The oral treatment with dichloromethane fraction (DF; 9 mg/Kg) produced antinociceptive effect in both phases of formalin test; however, the treatment with ethyl acetate fraction (EAF; 16 mg/Kg) reduced pain only in the second phase of this test. The oral treatments with CEE (300 mg/Kg) or DF (9 mg/Kg) reduced the nociception induced by capsaicin and pre-treatment with naloxone did not change these effects. The oral administration of CEE (300 mg/Kg), DF (9 mg/Kg) or ethyl EAF (16 mg/Kg) reduced ear edema, leukocytes migration and myeloperoxidase activity. Furthermore, the oral treatment with CEE (300 mg/Kg) or EAF (16 mg/Kg) reduced the level of Tumor Necrosis Factor - Alpha (TNF-α) in the pleurisy test. In conclusion, the DF showed antinociceptive activity that involves the vanilloid system as well as anti-inflammatory effect and the EAF showed anti-inflammatory activity involving the reduction of TNF-α cytokine.

Effect of Holoptelea integrifolia (Roxb.) Planch. n-Hexane Extract and Its Bioactive Compounds on Wound Healing and Anti-Inflammatory Activity.

The stem bark of Holoptelea integrifolia (Roxb.) Planch. has been applied for the treatment of human cutaneous diseases as well as canine demodicosis in several countries. However, no detailed mechanistic studies have been reported to support their use. In this study, thin-layer chromatography and gas chromatography were used to screen phytochemicals from the fresh stem bark extract of H. integrifolia. We found the two major bioactive compounds, friedelin and lupeol, and their activity on wound healing was further investigated in keratinocytes. Both bioactive compounds significantly reduced wound area and increased keratinocyte migration by increasing matrix metalloproteinases-9 production. Subsequently, we found that the mRNA gene expressions of cadherin 1 and desmoglobin 1 significantly decreased, whereas the gene expression involved in keratinocyte proliferation and homeostasis (keratin-17) increased in compound-treated human immortalized keratinocytes cells. The expression of inflammatory genes (cyclooxygenase-2 and inducible nitric oxide synthase) and pro-inflammatory cytokine genes (tumor necrosis factor-alpha and interleukin-6) was reduced by treatment with n-hexane extract of H. integrifolia and its bioactive compounds. Our results revealed that H. integrifolia extract and its bioactive compounds, friedelin and lupeol, exhibit wound-healing activity with anti-inflammatory properties, mediated by regulating the gene expression involved in skin re-epithelialization.

Floral morphogenesis of Celtis species: implications for breeding system and reduced floral structure.

PREMISE: Celtis is the most species-rich genus of Cannabaceae, an economically important family. Celtis species have been described as wind-pollinated and andromonoecious. However, the andromonoecy of Celtis has been debated because there are reports of monoclinous flowers with non-opening anthers on short filaments. Our objective was to study the floral morphogenesis of Celtis to establish the breeding system and to better understand the developmental patterns that lead to the formation of reduced flowers in the genus. METHODS: Flowers and floral buds of Celtis species were studied using scanning electron microscopy, high-resolution x-ray computed tomography, and light microscopy. RESULTS: All flowers initiate stamens and carpels during early floral development, but either stamens or carpels abort during later stages. Thus, at anthesis, flowers are either functionally pistillate or functionally staminate. In pistillate flowers, stamens abort late and become staminodes with normal-looking anthers. These anthers have no functional endothecium and, in most of the species studied, produce no viable pollen grains. The gynoecium is pseudomonomerous, and its vascularization is similar in the sampled species. In staminate flowers, the gynoecium aborts early resulting in small pistillodes. No vestiges of petals were found. CONCLUSIONS: The species studied are monoecious and not andromonoecious as described earlier. The absence of petals, the carpel and stamen abortion, and the pseudomonomerous gynoecium result in the reduced flowers of Celtis species. The use of high-resolution x-ray computed tomography was essential for a more accurate interpretation of ovary vascularization, confirming the pseudomonomerous structure of the gynoecium.

Selection on fruit traits is mediated by the interplay between frugivorous birds, fruit flies, parasitoid wasps and seed-dispersing ants.

Every organism on Earth must cope with a multitude of species interactions both directly and indirectly throughout its life cycle. However, how selection from multiple species occupying different trophic levels affects diffuse mutualisms has received little attention. As a result, how a given species amalgamates the combined effects of selection from multiple mutualists and antagonists to enhance its own fitness remains little understood. We investigated how multispecies interactions (frugivorous birds, ants, fruit flies and parasitoid wasps) generate selection on fruit traits in a seed dispersal mutualism. We used structural equation models to assess whether seed dispersers (frugivorous birds and ants) exerted phenotypic selection on fruit and seed traits in the spiny hackberry (Celtis ehrenbergiana), a fleshy-fruited tree, and how these selection regimes were influenced by fruit fly infestation and wasp parasitoidism levels. Birds exerted negative correlational selection on the combination of fruit crop size and mean seed weight, favouring either large crops with small seeds or small crops with large seeds. Parasitoids selected plants with higher fruit fly infestation levels, and fruit flies exerted positive directional selection on fruit size, which was positively correlated with seed weight. Therefore, higher parasitoidism indirectly correlated with higher plant fitness through increased bird fruit removal. In addition, ants exerted negative directional selection on mean seed weight. Our results show that strong selection on phenotypic traits may still arise in perceived diffuse species interactions. Overall, we emphasize the need to consider diverse direct and indirect partners to achieve a better understanding of the mechanisms driving phenotypic trait evolution in multispecies interactions.

Velocity of the falling dispersal units in Zelkova abelicea: remarkable evolutionary conservation within the relict tree genus.

PREMISE: Seed dispersal is extremely important for the recovery and restoration of forest communities. Relict tree genus Zelkova possesses a unique dispersal mechanism: mature fruits fall with the entire twig, and the dried leaves that are still attached function as a drag-enhancing appendage, carrying the fruits away from the parent tree. This singular adaptation has never been investigated in Z. abelicea. METHODS: Drop tests with dispersal units and individual fruits of Z. abelicea were performed in controlled conditions to measure their dispersal velocity and to define their flight mode. RESULTS: Zelkova abelicea uses both slowly falling dispersal units with chaotic motion, as well as fast falling individual fruits using a straight path. The falling velocity of Z. abelicea dispersal units is 1.53 m s-1 , which is virtually identical to that of the East Asiatic Z. serrata (1.51 m s-1 ). In contrast, the falling velocity of individual fruits was 2.74 m s-1 (Z. serrata: 5.36 m s-1 ). CONCLUSIONS: Members of the genus Zelkova, growing today in distant regions, show remarkable evolutionary conservation of the velocity and flight mechanics of their dispersal units. This is surprising because the Mediterranean and East Asiatic Zelkova species have been separated at least 15-20 mya. Zelkova abelicea, although growing in the Mediterranean with completely different forest structure and composition, still uses the same dispersal mechanism. The dispersal capacity of the genus Zelkova is less efficient than that of other wind dispersed trees, and it presumably evolved for short-distance ecological spread and not for long-distance biogeographical dispersal.

Comparative physiological mechanisms of arbuscular mycorrhizal fungi in mitigating salt-induced adverse effects on leaves and roots of Zelkova serrata.

Arbuscular mycorrhizal (AM) fungi enhance plant salt tolerance. However, physiological mechanisms of enhanced salt tolerance in leaves and roots of trees rarely have been compared. To reveal the different mechanisms, our study utilized comprehensive analyses of leaves and roots to examine the effects of Funneliformis mosseae on the salinity tolerance of Zelkova serrata. Seedlings of Z. serrata were exposed to four salt levels in a greenhouse with and without F. mosseae inoculation. Treatment comparisons revealed that following F. mosseae inoculation, (1) nutrient deficiency caused by osmotic stress was mitigated by the fungus enhancing nutrient contents (K, Ca, and Mg) in roots and (N, P, K, Ca, and Mg) in leaves, with Ca and K contents being higher in both leaves and roots; (2) mycorrhizas alleviated ion toxicity by maintaining a favorable ion balance (e.g., K+/Na+), and this regulatory effect was higher in leaves than that in roots; and (3) oxidative damage was reduced by an increase in the activities of antioxidant enzymes and accumulation of antioxidant compounds in mycorrhizal plants although the increase differed in leaves and roots. In particular, AM fungus-enhanced catalase activity and reduced glutathione content only occurred in leaves, whereas an enhanced content of reduced ascorbic acid was only noted in roots. Growth, root vitality, leaf photosynthetic pigments, net photosynthetic rate, and dry weight were higher in seedlings with AM fungus inoculation. These results suggest that AM fungus inoculation improved salinity tolerance of Z. serrata, but the physiological mechanisms differed between leaves and roots.

Species delimitation in the East Asian species of the relict tree genus Zelkova (Ulmaceae): A complex history of diversification and admixture among species.

Zelkova species, trees of the elm family (Ulmaceae), are part of the Cenozoic relict flora. In western Eurasia, the genus comprises three species that are restricted to disjunct areas (Z. sicula on Sicily, Z. abelicea on Crete and Z. carpinifolia in Transcaucasia). The situation is different in East Asia, where three species (Z. serrata, Z. schneideriana and Z. sinica) have at least partly overlapping distributions. The phylogenetic and phylogeographic status of these East Asian species is still not well understood, mainly since all previous studies used almost exclusively plant material collected in botanical gardens and were based on very small numbers of individuals. Our study is the first based on 33 natural populations covering all important areas with Zelkova species in the Sino-Japanese Floristic Region. Chloroplast and microsatellite markers were used, and Bayesian analyses were run for both types of markers. East Asian Zelkova species cluster into two groups that partially overlap when comparing the two types of markers. For chloroplast markers, the two groups coincide with all Japanese, some Korean and northern Chinese Z. serrata in one group and all other individuals in the other group, regardless of whether they are attributed to Z. serrata, Z. sinica or Z. schneideriana from Korea, mainland China and Taiwan. At the nuclear level, however, the clustering clearly groups all the Z. serrata individuals together, regardless of whether they are from Japan, Korea or China, and the two other species in a second group. This complex genetic pattern in East Asian Zelkova species is most likely due to a combination of ancient diversification and speciation events and more recent hybridization during the last glacial/interglacial retractions and recolonizations. One of the surprising results of our study concerns the populations from Taiwan, which are genetically similar to Z. schneideriana. Thus, their assignation to a separate taxon (Z. tarokoensis) or to a variety of Z. serrata (Z. serrata var. tarokoensis), as currently reported in all local and national floras, might be in need of revision. Furthermore, our results indicate that the East Asian species are more closely related to Z. carpinifolia than to any other Western European species. Haplotypes of Z. sicula and Z. abelicea (Mediterranean region) as well as those of Z. sinica and Z. schneideriana (East Asia) seem to have diversified more recently. The most ancient haplotypes are found among the western Eurasian Z. carpinifolia and the East Asian Z. serrata. This result is in agreement with the carpinifolia and serrata-like morphotypes commonly found in the fossil record.

Effects of aqueous leaves extract of Holoptelea integrifolia (Roxb) Planch on liver and kidney histopathology of albino rats.

Histopathological studies are an essential element to ascertain comprehensive safety profile of a drug. Unfortunately limited data are available about the toxicity of herbal remedies. Since a popular medicinal plant Holoptelea integrifolia (Roxb) Planch. contains various bioactive molecules, the present study is aimed to assess the histopathological alterations induced by aqueous extract of Holoptelea integrifolia on liver and kidney of wistar albino rat. In this study 60 rats divided in two groups; control and treated with aqueous extract of Holoptelea integrifolia (250mg/kg body weight) for 5 days. Histopathlogical studies by hematoxylin and eosin (H&E) staining were done on the liver and kidney tissues at the end of dosing by using standard procedure. Microscopic examination was then carried out to observe any pathological changes in the animals. The result showed that there is no significant variation in the basic architecture of liver and kidney as compared to control male wistar albino rats. In conclusion, aqueous extract of leaves of H. integrifolia may be safe and nontoxic. Further work on pharmacological aspects is required to evaluate the clinical potential of this plant for different ailments.

Hepatoprotective and neuroprotective flavanes from the fruits of Ulmus pumila L. (Ulmaceae).

Phytochemical study of the EtOAc fraction (active extract) of the fruits of Ulmus pumila L. resulted in the isolation of thirteen flavane derivatives, and they were identified by their precise spectral data and literature. All the compounds (1-13) were obtained from the fruits of U. pumila L. for the first time. Meanwhile, the compounds (1-13) were assayed for their hepatoprotective and neuroprotective activities, respectively. Compounds 1, 2, 5, 7 and 8 (10µM) exhibited remarkable hepatoprotective activities, and compounds 9, 10, and 13 showed significant neuroprotective activities with IC50 values of 4.08, 5.34, and 2.02µM, respectively.

Water-repellent plant surface structure induced by gall-forming insects for waste management.

Many animals and plants have evolved elaborate water-repellent microstructures on their surface, which often play important roles in their ecological adaptation. Here, we report a unique type of water-repellent structure on a plant surface, which develops as an insect-induced plant morphology in a social context. Some social aphids form galls on their host plant, in which they produce large amounts of hydrophobic wax. Excreted honeydew is coated by the powdery wax to form 'honeydew balls', which are actively disposed by soldier nymphs through an opening on their gall. These activities are enabled by a highly water-repellent inner gall surface, and we discovered that this surface is covered with dense trichomes that are not found on normal plant surfaces. The trichomes are coated by fine particles of the insect-produced wax, thereby realizing a high water repellency with a cooperative interaction between aphids and plants. The plant leaves on which the gall is formed often exhibit patchy areas with dense trichomes, representing an ectopic expression of the insect-induced plant morphology. In the pouch-shaped closed galls of a related social aphid species, by contrast, the inner surface was not covered with trichomes. Our findings provide a convincing example of how the extended phenotype of an animal, expressed in a plant, plays a pivotal role in maintaining sociality.

Analytical method for the evaluation of the outdoor air contamination by emerging pollutants using tree leaves as bioindicators.

In this work, an analytical method, based on sonication-assisted extraction, clean-up by dispersive solid-phase extraction and determination by liquid chromatography-tandem mass spectrometry, has been developed and validated for the simultaneous determination of 15 emerging pollutants in leaves from four ornamental tree species. Target compounds include perfluorinated organic compounds, plasticizers, surfactants, brominated flame retardant, and preservatives. The method was optimized using Box-Behnken statistical experimental design with response surface methodology and validated in terms of recovery, accuracy, precision, and method detection and quantification limits. Quantification of target compounds was carried out using matrix-matched calibration curves. The highest recoveries were achieved for the perfluorinated organic compounds (mean values up to 87%) and preservatives (up to 88%). The lowest recoveries were achieved for plasticizers (51%) and brominated flame retardant (63%). Method detection and quantification limits were in the ranges 0.01-0.09 ng/g dry matter (dm) and 0.02-0.30 ng/g dm, respectively, for most of the target compounds. The method was successfully applied to the determination of the target compounds on leaves from four tree species used as urban ornamental trees (Citrus aurantium, Celtis australis, Platanus hispanica, and Jacaranda mimosifolia). Graphical abstract Analytical method for the biomonitorization of emerging pollutants in outdoor air.

Strategy for Screening of Antioxidant Compounds from Two Ulmaceae Species Based on Liquid Chromatography-Mass Spectrometry.

Liquid chromatography-mass spectrometry (LC-MS)-based untargeted metabolomics implies that annotated metabolites can serve as potential markers of the associated bioactivities of plant extracts. Firstly, we selected Aphananthe aspera and Zelkova serrata (Family: Ulmaceae) from 16 Korean plant species based on their distinct principal component analysis (PCA) patterns in LC-MS datasets and antioxidant activity assays. Further, we chose 40% solid-phase extraction (SPE) extracts of the two species displaying the highest antioxidant activities coupled with distinct PCA patterns. Examining the metabolite compositions of the 40% SPE extracts, we observed relatively higher abundances of quercetin, kaempferol, and isorhamnetin O-glucosides for A. aspera, whereas quercetin, isorhamnetin O-glucuronides, and procyanidin dimer were relatively higher in Z. serrata. These metabolites were clearly distinguished in pathway map and displayed strong positive correlations with antioxidant activity. Further, we performed preparative high-performance liquid chromatography (prep-HPLC) analysis coupled with the 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) assay to validate their functional correlations. As a result, quercetin O-sophoroside was determined as the main antioxidant in A. aspera, while isorhamnetin O-glucuronide and procyanidin dimer were the primary antioxidants in Z. serrata. The current study suggests that the LC-MS-based untargeted metabolomics strategy can be used to illuminate subtle metabolic disparities as well as compounds associated with bioactivities.

Anthelmintic effects of indigenous multipurpose fodder tree extracts against Haemonchus contortus.

Condensed tannins (CT) extracted from Balanites aegyptiaca, Tamarindus indica, and Celtis toka browses were used to evaluate their anthelmintic effect on different developmental stages of Haemonchus contortus. To achieve this objective, various serial concentrations of each CT extract of the foliages were used to test adult motility, inhibition of egg hatchability, and larval development. The fodders were selected based on their multipurpose advantage and accessibility to use as fodder for livestock in the low lands of the Gambella region. The fastest and slowest adult motility rate was observed in 2-ml (4 min) and 0.125-ml dose of C. toka, respectively, which is better than that in ivermectin. Egg hatchability inhibition was observed with dose difference within species, but there is no difference between B. aegyptiaca and T. indica. The foliage extracts of the studied browses were observed to inhibit the larvae by 100% at 2 ml, which is similar to ivermectin. There is no significant difference observed in larvae development inhibition between the species and ivermectin (p > 0.05). CT extracts of studied plants have found to own significant anthelmintic activity in a dose-dependent manner. They could serve as anthelmintic economically and eco-friendly after further and series of in vivo experiments.

Chloroplast analysis of Zelkova schneideriana (Ulmaceae): genetic diversity, population structure, and conservation implications.

Zelkova schneideriana is endemic to China and belongs to the Ulmaceae. It is listed as a Near Threatened species in the China Biodiversity Red Data Book. We conducted a phylogeographical study of two chloroplast regions (psbA-trnH and trnG-trnM) in several Chinese Z. schneideriana populations, in order to examine the genetic diversity, population structure, and evolutionary history of the species. In all, 10 haplotypes were detected. The population from Sangzhi, Hunan, had the highest nucleotide diversity (π = 0.00653) and haplotype diversity (HD = 1.000), and should be considered the most suitable population to be protected under an in situ conservation strategy. Seed collections from as many individuals as possible in other populations would preserve the genetic diversity of Z. schneideriana.

Analysis of nuclear microsatellites reveals limited differentiation between Colchic and Hyrcanian populations of the wind-pollinated relict tree Zelkova carpinifolia (Ulmaceae).

UNLABELLED: • PREMISE OF THE STUDY: The Caucasus represents one of the world's biodiversity hotspots and includes the climatic refugia Hyrcan on the southern coast of the Caspian Sea and Colchis on the eastern coast of the Black Sea, where different species survived during the Quaternary climatic oscillations. We evaluated the genetic diversity of the relict tree Zelkova carpinifolia shared between the two refugia and distributed throughout the Caucasus and adjacent areas.• METHODS: Specimens were collected from 30 geographical sites in Azerbaijan, Georgia, Iran, and Turkey and screened for variability at eight nuclear microsatellite loci. The genetic diversity among and within populations was assessed using a set of statistical measures.• KEY RESULTS: We detected 379 different genotypes from a total of 495 individuals with varying degrees of clonal reproduction at the different sites. Low to intermediate levels of genetic diversity were observed at all sites, and strong differentiation between sampling sites was absent. In addition, we observed no clear genetic differentiation between the Colchis and Hyrcan. Bayesian clustering of the genotypes revealed three populations with high levels of admixture between the sampling sites.• CONCLUSIONS: The lack of strong genetic structure of studied populations of Z. carpinifolia contrasts with a previous study based on chloroplast markers and suggests that long-distance pollen dispersal is an important factor of gene flow among populations of Z. carpinifolia. The present study does not reveal any particular site with particularly isolated genotypes that would deserve more attention for conservation purposes than others, although some sites should be considered for further investigation.