Comprehensive Time-Course Transcriptome Reveals the Crucial Biological Pathways Involved in the Seasonal Branch Growth in Siberian Elm (Ulmus pumila).

Timber, the most prevalent organic material on this planet, is the result of a secondary xylem emerging from vascular cambium. Yet, the intricate processes governing its seasonal generation are largely a mystery. To better understand the cyclic growth of vascular tissues in elm, we undertook an extensive study examining the anatomy, physiology, and genetic expressions in Ulmus pumila. We chose three robust 15-year-old elm trees for our study. The cultivars used in this study were collected from the Inner Mongolia Autonomous Region in China and nurtured in the tree farm of Shandong Normal University. Monthly samples of 2-year-old elm branches were taken from the tree from February to September. Marked seasonal shifts in elm branch vascular tissues were observed by phenotypic observation: In February, the cambium of the branch emerged from dormancy, spurring growth. By May, elms began generating secondary xylem, or latewood, recognized by its tiny pores and dense cell structure. From June to August, there was a marked increase in the thickness of the secondary xylem. Transcriptome sequencing provides a potential molecular mechanism for the thickening of elm branches and their response to stress. In February, the tree enhanced its genetic responses to cold and drought stress. The amplified expression of CDKB, CYCB, WOX4, and ARF5 in the months of February and March reinforced their essential role in the development of the vascular cambium in elm. Starting in May, the elm deployed carbohydrates as a carbon resource to synthesize the abundant cellulose and lignin necessary for the formation of the secondary wall. Major genes participating in cellulose (SUC and CESA homologs), xylan (UGD, UXS, IRX9, IRX10, and IRX14), and lignin (PAL, C4H, 4CL, HCT, C3H, COMT, and CAD) biosynthetic pathways for secondary wall formation were up-regulated by May or/and June. In conclusion, our findings provided a foundation for an in-depth exploration of the molecular processes dictating the seasonal growth of elm timber.

Effect of the metal ion-induced carbonylation modification of mitochondrial membrane channel protein VDAC on cell vitality, seedling growth and seed aging.

Introduction: Seeds are the most important carrier of germplasm preservation. However, an irreversible decrease in vigor can occur after the maturation of seeds, denoted as seed aging. Mitochondrion is a crucial organelle in initiation programmed cell death during seed aging. However, the underlying mechanism remains unclear. Methods: Our previous proteome study found that 13 mitochondria proteins underwent carbonylation modification during the aging of Ulmus pumila L. (Up) seeds. This study detected metal binding proteins through immobilized metal affinity chromatography (IMAC), indicating that metal binding proteins in mitochondria are the main targets of carbonization during seed aging. Biochemistry, molecular and cellular biology methods were adopted to detect metal-protein binding, protein modification and subcellular localization. Yeast and Arabidopsis were used to investigate the biological functions in vivo. Results and discussion: In IMAC assay, 12 proteins were identified as Fe2+/Cu2+/Zn2+ binding proteins, including mitochondrial voltage dependent anion channels (VDAC). UpVDAC showed binding abilities to all the three metal ions. His204Ala (H204A) and H219A mutated UpVDAC proteins lost their metal binding ability, and became insensitive to metal-catalyzed oxidation (MCO) induced carbonylation. The overexpression of wild-type UpVDAC made yeast cells more sensitive to oxidative stress, retarded the growth of Arabidopsis seedlings and accelerated the seed aging, while overexpression of mutated UpVDAC weakened these effects of VDAC. These results reveal the relationship between the metal binding ability and carbonylation modification, as well as the probable function of VDAC in regulating cell vitality, seedling growth and seed aging.

Integrated Analysis of miRNAome and Transcriptome Identify Regulators of Elm Seed Aging.

After maturity, seed vigor irreversibly decreases. Understanding the underlying mechanism is important to germplasm preservation. MicroRNAs (miRNAs) play vital regulatory roles in plants. However, little is known about how miRNAs regulate seed aging. Here, elm (Ulmus pumila L.) seeds of three aging stages were subjected to a multi-omics analysis including transcriptome, small RNAome and degradome, to find regulators of seed aging. In the small RNAome, 119 miRNAs were identified, including 111 conservative miRNAs and eight novel miRNAs specific to elm seeds, named upu-miRn1-8. A total of 4900 differentially expressed genes, 22 differentially expressed miRNAs, and 528 miRNA-target pairs were identified during seed ageing. The target genes were mainly involved in the processing of proteins in the endoplasmic reticulum, metabolism, plant hormone signal transduction, and spliceosome. The expression of several DEGs and miRNAs were verified by qRT-PCR. The degradome data showed the exact degradation sites of upu-miR399a on ABCG25, and upu-miR414a on GIF1, etc. The dual-luciferase assay verified the negative regulation of upu-miR399a on ABCG25 and upu-miR414a on GIF1 in tobacco leaves. This study outlined the regulation network of mRNA, miRNA and miRNA-target genes during seed aging, which is helpful in integrating the regulation mechanisms of seed vigor at the transcriptional and post-transcriptional levels.

Condensed tannins from Ulmus pumila L. leaves induce G2/M phase arrest and apoptosis via caspase-cascade activation in TFK-1 cholangiocarcinoma cells.

Condensed tannins the polyphenolic compounds that are widespread in plants have been proved to have antitumor potential. Here, we purified the bioactive condensed tannins from leaves of Ulmus pumila L. and explored their structural characteristics, antitumor effect on TFK-1 cholangiocarcinoma cells as well as the related potential mechanism. The UV-Vis, FT-IR spectroscopy, ESI-Full-MS, and thiolysis-HPLC-ESI-MS demonstrated that U. pumila condensed tannins (UCTs) consisted essentially of procyanidins with epicatechin as the main flavan-3-ol extension unit. The UCTs could significantly reduce the survival rate of human cholangiocarcinoma TFK-1, SK-CHA-1, and MZ-CHA-1 cells with the better inhibitory effect on TFK-1 cell proliferation. Flow cytometric assay showed that UCTs affected TFK-1 survival by G2/M phase arrest and inducing apoptosis in a dose-dependent manner. In addition, a total of 6592 differentially expressed genes (DEGs), consisting of 94 upregulated and 6498 downregulated DEGs, were identified between untreated and UCTs-treated TFK-1 cells using RNA-seq technology. Enrichment analysis based on the KEGG database revealed that these DEGs were closely associated with cell cycle and p53 apoptotic signaling pathways. Furthermore, qRT-PCR confirmed that treatment of UCTs to TFK-1 cells caused significant changes in the expression of cyclin E, cdc25 A, cytochrome c, caspase-3, and caspase-8. These results indicated that UCTs exhibited the growth inhibition effect on TFK-1 cells possibly via G2/M cell cycle arrest and activation of caspase-cascade to induce apoptosis, and had potential as an anti-cholangiocarcinoma drug for further development. PRACTICAL APPLICATIONS: Ulmus pumila L. as a valuable tree species has been widely used in fields of medicine and food. Condensed tannins, the polyphenolic compounds widespread in plants, have been proved to have antitumor potential and be safe to normal cells. In this study, the condensed tannins from leaves of U. pumila (UCTs) remarkably suppressed cholangiocarcinoma (CCA) cell viability possibly via G2/M cell cycle arrest and activation of caspase-cascade to induce apoptosis. The results provided evidence for the application of UCTs as a potential therapeutic drug for CCA tumor.

Comparative Transcriptome Analysis Reveals Candidate Genes and Pathways for Potential Branch Growth in Elm (Ulmus pumila) Cultivars.

Wood plays a vital role in human life. It is important to study the thickening mechanism of tree branches and explore the mechanism of wood formation. Elm (Ulmus pumila) is a strong essential wood, and it is widely used in cabinets, sculptures, and ship making. In the present study, phenotypic and comparative transcriptomic analyses were performed in U. pumila fast- (UGu17 and UZuantian) and slow-growing cultivars (U81-07 and U82-39). Phenotypic observation showed that the thickness of secondary xylem of 2-year-old fast-growing branches was greater compared with slow-growing cultivars. A total of 9367 (up = 4363, down = 5004), 7159 (3413/3746), 7436 (3566/3870), and 5707 (2719/2988) differentially expressed genes (DEGs) were identified between fast- and slow-growing cultivars. Moreover, GO and KEGG enrichment analyses predicted that many pathways were involved in vascular development and transcriptional regulation in elm, such as "plant-type secondary cell wall biogenesis", "cell wall thickening", and "phenylpropanoid biosynthesis". NAC domain transcriptional factors (TFs) and their master regulators (VND1/MYB26), cellulose synthase catalytic subunits (CESAs) (such as IRX5/IRX3/IRX1), xylan synthesis, and secondary wall thickness (such as IRX9/IRX10/IRX8) were supposed to function in the thickening mechanism of elm branches. Our results indicated that the general phenylpropanoid pathway (such as PAL/C4H/4CL) and lignin metabolism (such as HCL/CSE/CCoAOMT/CCR/F5H) had vital functions in the growth of elm branches. Our transcriptome data were consistent with molecular results for branch thickening in elm cultivars.

Stomatal opening ratio mediates trait coordinating network adaptation to environmental gradients.

A trait coordination network is constructed through intercorrelations of functional traits, which reflect trait-based adaptive strategies. However, little is known about how these networks change across spatial scales, and what drivers and mechanisms mediate this change. This study bridges that gap by integrating functional traits related to plant carbon gain and water economy into the coordination network of Siberian elm (Ulmus pumila), a eurybiont that survives along a 3800 km environmental gradient from humid forest to arid desert. Our results demonstrated that both stomatal density and stomatal size reached a physiological threshold at which adjustments in these traits were not sufficient to cope with the increased environmental stress. Network analysis further revealed that the mechanism for overcoming this threshold, the stomatal opening ratio, gratio , was represented by the highest values for centrality across different spatial scales, and therefore mediated the changes in the trait coordination network along environmental gradients. The mediating roles manifested as creating the highest maximum theoretical stomatal conductance (gsmax ) but lowest possible gratio for pathogen defense in humid regions, while maintaining the gratio 'sweet spot' (c. 20% in this region) for highest water use efficiency in semihumid regions, and having the lowest gsmax and highest gratio for gas exchange and leaf cooling in arid regions. These results suggested that the stomatal traits related to control of stomatal movement play fundamental roles in balancing gas exchange, leaf cooling, embolism resistance and pathogen defense. These insights will allow more accurate model parameterization for different regions, and therefore better predictions of species' responses to global change.

Cytology, transcriptomics, and mass spectrometry imaging reveal changes in late-maturation elm (Ulmus pumila) seeds.

During seed maturation, the seed deposits storage compounds (starches, oils, and proteins), synthesizes defense compounds, produces a seed coat, initiates embryo dormancy, and becomes desiccated. During the late-maturation stage, seed storage compound contents and compositions change dramatically. Although maturation has been extensively studied in model species and crops, it remains less well characterized in woody perennial plants. In this study, we conducted morphological and cytological observations, transcriptome profiling, and chemical constituent analysis of elm (Ulmus pumila L.) seeds during the late-maturation stage. Light and electron microscopy revealed that closely packed yet discrete lipid bodies frequently surrounded the densely stained protein bodies, and the protein bodies became irregular or even partially disintegrated at the end of seed development. RNA-seq detected substantial transcriptome changes during the late-maturation stage, and pathway enrichment analysis showed that the differentially expressed genes were associated with phenylpropanoid biosynthesis, starch and sucrose metabolism, plant-pathogen interactions, and hormone signal transduction. Furthermore, we used mass spectrometry imaging to detect the relative intensity and spatial distribution of fatty acids, phospholipids, and waxes in elm seeds. Our findings provide a framework for understanding the changes in cytological features and chemical composition during the final stage of elm seed development, and a detailed reference for seed development in woody plants.

Revegetation of coal mine degraded arid areas: The role of a native woody species under optimum water and nutrient resources.

Ecological restoration of coal mine degraded soils across arid and semi-arid environments worldwide remains particularly challenging. We used a combination of greenhouse and field experiments to assess the potential role of a woody species, Ulmus pumila, in the restoration of degraded soils associated with coal-mining activities in the northwest China. We investigated how various combinations of water-nitrogen-phosphorus (W-N-P) resources affect multiple growth parameters in U. pumila. We found that several plant growth traits significantly improved with W-N applications, regardless of P inputs. Moderate-to-highest W-N-P doses increased net photosynthesis and transpiration rates, water use efficiency, stomatal conductance, chlorophyll and carotenoid contents under greenhouse conditions. A combination of high W together with low N-P applications led to high relative water content and net photosynthetic rates under field conditions. Increasing of N-P doses under W-shortage condition, aided U. pumila to enhance osmotic adjustments by increasing contents of proline and soluble sugar and also boost the activity of superoxide dismutase, peroxidase and catalase in leaf tissues to reduce accumulation of reactive oxygen species and malondialdehyde content in all conditions of greenhouse and field. Our study is the first to assess the optimum W-N-P resources in U. pumila and demonstrate that optimum growth performance could be obtained under W supplements corresponding to 90 mm year-1, N and P at 110 and 45 kg ha-1, respectively, under field condition. These findings can have far reaching implications for vegetation restoration of degraded areas associated with coal-mining activities across arid and semi-arid regions worldwide.

Prevalence and risk factors of allergic rhinitis and asthma in the southern edge of the plateau grassland region of northern China: A cross-sectional study.

BACKGROUND: The prevalence rates of allergic rhinitis (AR) and asthma in the border region of China may be different from those in the central region of plateau grasslands. A survey was performed to investigate the prevalence and risk factors for AR, asthma, and AR combined with asthma among adults (age ≥ 20 years) residing in the southern border of plateau grasslands in northern China. METHODS: From May to August 2018, a cross-sectional survey was completed by subjects that were selected using a cluster random sampling method. The subjects completed a questionnaire and were administered skin prick tests (SPTs). Risk factors for AR, asthma, and AR combined with asthma were examined by multivariate logistic regression analyses. RESULTS: A total of 1815 adult subjects in the selected region completed study. The prevalence rates of physician-diagnosed AR, asthma, and AR combined with asthma were 13.9% (253), 9.8% (177), and 2.9% (52), respectively. Among the patients with AR, 20.6% were found to have concurrent asthma; among the patients with asthma, 29.4% were found to have concurrent AR. Artemisia and Humulus pollen were the most common sensitizing pollen types. Approximately 70% of subjects with AR and <30% of asthma patients were sensitized to Artemisia and Humulus pollen. Symptoms of AR and asthma mainly appeared during August. A multivariable logistic regression analysis identified sensitization pollen as an independent risk factor for both AR and AR combined with asthma (AR: OR = 16.23, 95% CI: 10.15-25.96; AR combined with asthma: OR = 6.16, 95% CI: 1.28-29.66). An age >40 years old, family history of asthma, moderate-to-severe AR, adverse food reactions, and mold allergies were independent risk factors for AR combined with asthma. CONCLUSIONS: This study identified the prevalence rates of AR and asthma in the southern borders of the plateau grassland in northern China (>1500 m above sea level). Sensitization pollen is an independent risk factor for AR and AR combined with asthma.

Characteristics and functions of glyceraldehyde 3-phosphate dehydrogenase S-nitrosylation during controlled aging of elm and Arabidopsis seeds.

Seed aging is the gradual decline in seed vigor, during which programmed cell death (PCD) occurs. The functions of nitric oxide (NO) are exerted through protein S-nitrosylation, a reversible post-translational modification. During seed aging, more than 80 proteins are S-nitrosylated, but the particular role of individual proteins is unknown. Here, we showed that the S-nitrosylation level of glyceraldehyde 3-phosphate dehydrogenase (UpGAPDH) in elm (Ulmus pumila L.) seeds increased after controlled deterioration treatment. UpGAPDH was S-nitrosylated at Cys154 during S-nitrosoglutathione (GSNO) treatment, and its oligomerization was triggered both in vitro and in elm seeds. Interestingly, UpGAPDH interacted with the mitochondrial voltage-dependent anion channel in an S-nitrosylation-dependent way. Some UpGAPDH-green fluorescent protein in Arabidopsis protoplasts co-localized with mitochondria during the GSNO treatment, while the S-nitrosylation-defective UpGAPDH C154S-GFP protein did not. Seeds of oxUpGAPDH lines showed cell death and lost seed vigor rapidly during controlled deterioration treatment-triggered seed aging, while those overexpressing S-nitrosylation-defective UpGAPDH-Cys154 did not. Our results suggest that S-nitrosylation of UpGAPDH may accelerate cell death and seed deterioration during controlled deterioration treatment. These results provide new insights into the effects of UpGAPDH S-nitrosylation on protein interactions and seed aging.

Gene Coexpression Network Analysis Indicates that Hub Genes Related to Photosynthesis and Starch Synthesis Modulate Salt Stress Tolerance in Ulmus pumila.

Ulmus pumila L. is an excellent afforestation and biofuel tree that produces high-quality wood, rich in starch. In addition, U. pumila is highly adaptable to adverse environmental conditions, which is conducive to its utilization for vegetating saline soils. However, little is known about the physiological responses and transcriptional regulatory network of U. pumila under salt stress. In this study, we exposed five main cultivars in saline-alkali land (Upu2, 5, 8, 11, and 12) to NaCl stress. Of the five cultivars assessed, Upu11 exhibited the highest salt resistance. Growth and biomass accumulation in Upu11 were promoted under low salt concentrations (<150 mM). However, after 3 months of continuous treatment with 150 mM NaCl, growth was inhibited, and photosynthesis declined. A transcriptome analysis conducted after 3 months of treatment detected 7009 differentially expressed unigenes (DEGs). The gene annotation indicated that these DEGs were mainly related to photosynthesis and carbon metabolism. Furthermore, PHOTOSYNTHETIC ELECTRON TRANSFERH (UpPETH), an important electron transporter in the photosynthetic electron transport chain, and UpWAXY, a key gene controlling amylose synthesis in the starch synthesis pathway, were identified as hub genes in the gene coexpression network. We identified 25 and 62 unigenes that may interact with PETH and WAXY, respectively. Overexpression of UpPETH and UpWAXY significantly increased the survival rates, net photosynthetic rates, biomass, and starch content of transgenic Arabidopsis plants under salt stress. Our findings clarify the physiological and transcriptional regulators that promote or inhibit growth under environmental stress. The identification of salt-responsive hub genes directly responsible for photosynthesis and starch synthesis or metabolism will provide targets for future genetic improvements.

Neuroprotective activity of Ulmus pumila L. in Alzheimer's disease in rats; role of neurotrophic factors.

Alzheimer's disease (AD) is one of the most prevalent neurodegenerative disorders which affects the hippocampus and cortical neurons leading to impairment of cognitive ability. Treatment of AD depends mainly on acetylcholinesterase inhibitors, however, a novel therapeutic approach is introduced based on the maintenance of neuronal viability and functionality exerted through neurotrophic factors. In the current study, Ulmus pumila L. leaves alcoholic extract was investigated for its neuroprotective activity in AlCl3-induced AD in rats. Rats were orally treated with AlCl3 (17 mg/kg) for 4 weeks followed by U. pumila extract (150 mg/kg b.wt.) for another 6 weeks. Treatment of neuro-intoxicated rats with U. pumila extract resulted in a significant regulation in neurotrophic factors; brain derived neurotrophic factor and transforming growth factor-ß and pro-inflammatory cytokine; TNF. It also induced an elevation in serum levels of monoamine neurotransmitters; norepinephrine, dopamine and serotonin and a decline in brain acetlycholinesterase activity. U. pumila extract also showed potent antioxidant activity as indicated by the declined malondialdehyde and elevated reduced glutathione, catalase and super oxide dismutase levels in AD rats' brains. Histological improvement was detected in the cerebral cortex, the hippocampus and striatum of the treated rats. The phytochemical analysis of U. pumila extract revealed high contents of flavonoids and phenolics and the major compounds were isolated and chemically characterized. Additionally, U. pumila extract and the isolated compounds exerted a prominent activity in in-vitro acetylcholinesterase inhibition assay with kaempferol-3-O-ß-glucoside being the most potent compound showing IC50 of 29.03 ± 0.0155 µM. A molecular docking study indicated high affinity of kaempferol-3-O-ß-robinobioside on acetylcholine esterase binding site with estimated binding free energy of -8.26 kcal/mol.

A Non-Invasive Analysis of Seed Vigor by Infrared Thermography.

This paper establishes robust regression models for fast and efficient estimation of seed vigor based on high-resolution infrared thermography. High seed quality is of great significance for agricultural and silvicultural purposes, and seed vigor is a crucial agent of seed quality. In this study, we used the non-invasive technology of infrared thermal imaging to analyze seed vigor of Ulmus pumila L. and Oryza sativa L. Temperatures of young age and aged seeds during thermal decay were monitored over time. We found that the thermal decay dynamics of U. pumila seeds were highly differential among seeds with differential vigor. Furthermore, a regression model was developed to estimate seed vigor based on its thermal decay dynamics. Similarly, a close relationship was also found between thermal decay processes and seed vigor in O. sativa. These results suggest that infrared thermography can be widely applied in non-invasive examination of seed vigor and allows fast and efficient seed screening for agricultural and silvicultural purposes in the future.

Inhibitory Effects of AF-343, a Mixture of Cassia tora L., Ulmus pumila L., and Taraxacum officinale, on Compound 48/80-Mediated Allergic Responses in RBL-2H3 Cells.

The purpose of this study was to determine the antiallergic effects of AF-343, a mixture of natural plant extracts from Cassia tora L., Ulmus pumila L., and Taraxacum officinale, on rat basophilic leukemia (RBL-2H3) cells. The inhibitory effects on cell degranulation, proinflammatory cytokine secretion, and reactive oxygen species (ROS) production were studied in compound 48/80-treated RBL-2H3 cells. The bioactive compounds in AF-343 were also identified by HPLC-UV. AF-343 was found to effectively suppress compound 48/80-induced b-hexosaminidase release, and interleukin (IL)-4 and tumor necrosis factor-a (TNF-a) production in RBL-2H3 cells. In addition, AF-343 exhibited DPPH free radical scavenging effects in vitro (half-maximal inhibitory concentration (IC50) = 105 µg/mL) and potently inhibited compound 48/80-induced cellular ROS generation in a 2',7'-dichlorofluorescein diacetate (DCFH-DA) assay. Specifically, treatment with AF-343 exerted stronger antioxidant effects in vitro and antiallergic effects in cells than treatment with three single natural plant extracts. Furthermore, AF-343 was observed to contain bioactive compounds, including catechin, aurantio-obtusin, and chicoric acid, which have been reported to elicit antiallergic responses. This study reveals that AF-343 attenuates allergic responses via suppression of b-hexosaminidase release, IL-4 and TNF-a secretion, and ROS generation, perhaps through mechanisms related to catechin, aurantio-obtusin, and chicoric acid. The results indicate that AF-343 can be considered a treatment for various allergic diseases.

Seasonal dynamics of microbial diversity in the rhizosphere of Ulmus pumila L. var. sabulosa in a steppe desert area of Northern China.

The seasonal dynamics of microbial diversity within the rhizosphere of Ulmus pumila L. var. sabulosa in the hinterland of the Otindag Sandy Land of China were investigated using high-throughput sequencing of bacterial 16S rRNA genes and fungal ITS region sequences. A significant level of bacterial and fungal diversity was observed overall, with detection of 7,676 bacterial Operational Taxonomic Units (OTUs) belonging to 40 bacteria phyla and 3,582 fungal OTUs belonging to six phyla. Proteobacteria, Actinobacteria, and Firmicutes were the dominant bacterial phyla among communities, while Ascomycota, Basidiomycota, and Zygomycota were the dominant phyla of fungal communities. Seasonal changes influenced the α-diversity and ß-diversity of bacterial communities within elm rhizospheres more than for fungal communities. Inferred functional analysis of the bacterial communities identified evidence for 41 level two KEGG (Kyoto Encyclopedia of Genes and Genomes) orthology groups, while guild-based analysis of the fungal communities identified eight ecological guilds. Metabolism was the most prevalent bacterial functional group, while saprotrophs prevailed among the identified fungal ecological guilds. Soil moisture and soil nutrient content were important factors that affected the microbial community structures of elm rhizospheres across seasons. The present pilot study provides an important baseline investigation of elm rhizosphere microbial communities.

The Reason for Growth Inhibition of Ulmus pumila 'Jinye': Lower Resistance and Abnormal Development of Chloroplasts Slow Down the Accumulation of Energy.

Ulmus pumila 'Jinye', the colorful leaf mutant of Ulmus pumila L., is widely used in landscaping. In common with most leaf color mutants, U. pumila 'Jinye' exhibits growth inhibition. In this study, U. pumila L. and U. pumila 'Jinye' were used to elucidate the reasons for growth inhibition at the physiological, cellular microstructural, and transcriptional levels. The results showed that the pigment (chlorophyll a, chlorophyll b, and carotenoids) content of U. pumila L. was higher than that of U. pumila 'Jinye', whereas U. pumila 'Jinye' had a higher proportion of carotenoids, which may be the cause of the yellow leaves. Examination of the cell microstructure and RNA sequencing analysis showed that the leaf color and growth inhibition were mainly due to the following reasons: first, there were differences in the structure of the thylakoid grana layer. U. pumila L. has a normal chloroplast structure and clear thylakoid grana slice layer structure, with ordered and compact thylakoids. However, U. pumila 'Jinye' exhibited the grana lamella stacking failures and fewer thylakoid grana slice layers. As the pigment carrier and the key location for photosynthesis, the close stacking of thylakoid grana could combine more chlorophyll and promote efficient electron transfer promoting the photosynthesis reaction. In addition, U. pumila 'Jinye' had a lower capacity for light energy absorption, transformation, and transportation, carbon dioxide (CO2) fixation, lipopolysaccharide biosynthesis, auxin synthesis, and protein transport. The genes related to respiration and starch consumption were higher than those of U. pumila L., which indicated less energy accumulation caused the growth inhibition of U. pumila 'Jinye'. Finally, compared with U. pumila 'Jinye', the transcription of genes related to stress resistance all showed an upward trend in U. pumila L. That is to say, U. pumila L. had a greater ability to resist adversity, which could maintain the stability of the intracellular environment and maintain normal progress of physiological metabolism. However, U. pumila 'Jinye' was more susceptible to changes in the external environment, which affected normal physiological metabolism. This study provides evidence for the main cause of growth inhibition in U. pumila 'Jinye', information for future cultivation, and information on the mutation mechanism for the breeding of colored leaf trees.

Grazing Altered the Pattern of Woody Plants and Shrub Encroachment in a Temperate Savanna Ecosystem.

Ulmus pumila-dominated temperate savanna is an important tree-grass complex ecosystem in the Otindag sand land, northern China. To date, few investigations have been undertaken on the spatial patterns and structure of this ecosystem and its driving factors under different grazing pressures. The objective of our study therefore is to explore whether grazing has affected the population structure/pattern of woody plants and shrub encroachment in a temperate savanna ecosystem. Results indicate that species richness and seedlings decreased with increasing grazing pressure. An increase in grazing pressure did not significantly affect adult-tree density, but it hindered the normal regeneration of U. pumila seedlings, further inducing population decline. U. pumila seedlings had a more significant aggregated distribution than juvenile or adult trees. The adult and juvenile trees had an aggregated distribution at the small scale and a random distribution at the large scale. Shrubs also showed a significant aggregated distribution. No clear effect on the spatial patterns of adult trees was observed; however, there was a noticeable effect for juveniles and seedlings under different grazing pressures. U. pumila seedlings had a positive association with their juveniles and Spiraea aquilegifolia, but a negative association with Caragana microphylla. Shrub encroachment occurred with decreasing grazing pressures. In conclusion, overgrazing led to the decline of U. pumila population, but the decrease in grazing pressure increased shrub encroachment in the temperate savanna ecosystem. Moderate grazing management may be a better way to enhance the stability of U. pumila population and reduce shrub encroachment.

Stigmellamultispicata Rociene. & Stonis, an Asian leafminer on Siberian elm, now widespread in eastern North America (Lepidoptera, Nepticulidae).

Stigmellamultispicata Rociene & Stonis, 2014, previously known from the single male holotype from Primorye, Russia, is reported as a new invasive species mining leaves of Siberian elm, Ulmuspumila L., in eastern North America. Both adults and leafmines have been reported from many sites as unidentified Nepticulidae since 2010. Crucial for the identification was a match of the DNA barcode of a single larva collected on Ulmuspumila in Beijing with adults from North America. The single larva constitutes a new record for China. Stigmellamultispicata is closely related to the European S.ulmivora (Fologne, 1860), feeding likewise on Ulmus, but differs in details of external morphology and genitalia, particularly in the female, where S.multispicata has a remarkable elongated narrow ovipositor, suitable for oviposition in underside hairy leaf vein axils, where all mines start. In North America S.multispicata is the only Ulmus-feeding nepticulid with green larvae. Currently the species is known from USA: Illinois, Indiana, Iowa, Maryland, Massachusetts, Minnesota, New York, Ohio, Tennessee, Wisconsin, and Canada: Ontario and Québec. In Sagaponack, on Long Island, New York, larvae have been reported to occur en masse on Siberian elms from at least two sites. The current distribution could be reconstructed thanks also to many online photographs from observation websites. The species is redescribed, with the first descriptions of female, larva, and leafmine, and compared with S.ulmivora, which is fully redescribed. The two native North American nepticulid Ulmus leafminers, S.apicialbella (Chambers, 1873) and Ectoedemiaulmella (Braun, 1912), are diagnosed and new provincial and state records are provided. A key to linear mines on Ulmus in North America is provided. We suspect that trade of live plants through nurseries played a role in the sudden spread of this invasive species.

The characterization, selenylation and anti-inflammatory activity of pectic polysaccharides extracted from Ulmus pumila L.

The specific objective of this study was to investigate characterization, selenylation, and anti-inflammatory activities of pectic polysaccharides extracted from Ulmus pumila L. (PPU). Four different monosaccharides were found in PPU, including galacturonic acid, galactose, rhamnose, and glucose. FT-IR spectra indicated that pectic polysaccharides were successfully extracted from Ulmus pumila L., and were probably low methoxyl pectin. GC-MS and NMR analysis of PPU suggested the major monosaccharide of PPU was α-1,4-linked galacturonic acid with α-1,2-linked rhamnose as the backbone and glucose or galactose residues as branches at C-3 and C-4 positions of rhamnose. Selenylation of PPU was synthesized by 0.2 and 0.4% of sodium selenites. Selenized-PPU (Se-PPU) inhibited LPS-induced nitric oxide production in RAW 264.7 cells, and increasing selenium content enhanced anti-inflammatory properties of PPU. Therefore, Se-PPU can be used as a potential source of bioactive compounds for nutraceuticals and pharmaceutical applications.

Biological traits and the complex of parasitoids of the elm pest Orchestes steppensis (Coleoptera: Curculionidae) in Xinjiang, China.

The flee-weevil Orchestes steppensis Korotyaev (Coleoptera: Curculionidae) is a steppe eastern Palaearctic species, notable as a serious pest of elms (Ulmus spp., Ulmaceae), by feeding on the leaves (adults) or mining them heavily (larvae), especially of Ulmus pumila L. in Xinjiang, China. We have corrected the previous misidentifications of this weevil in China as O. alni (L.) or O. mutabilis Boheman and demonstrated that it is likely to be an invasive species in Xinjiang. Prior to this study, natural enemies of O. steppensis were unknown in Xinjiang. Resulting from field investigation and rearing in the laboratory during 2013-2016, seven parasitoid species were found to be primary and solitary, attacking larval and pupal stages of the host weevil. Pteromalus sp. 2 is the dominant species and also is the most competitive among the seven parasitoids, which could considered to be a perspective biological control agent of O. steppensis. Yet, the current control of this pest by the local natural enemies in Xinjiang is still currently inefficient, even though in 2016 parasitism was about 36% on U. pumila in Urumqi, so the potential for a classical biological control program against it needs to be further investigated, including an assessment of its parasitoids and other natural enemies in the native range of O. steppensis. The presented information on the natural enemies of this weevil can be also important for a potential classical biological control program against it in North America (Canada and USA), where it is a highly damaging and rapidly spreading invasive species.