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Abstract Background Caloric value is an important indicator of grassland ecosystem function, but the response of caloric value to nitrogen (N) addition and mowing is still unclear. We explored the adaptive changes of plant caloric value and energy standing crop along a N addition gradient after six-year NH4NO3 addition and mowing treatments in an Inner Mongolian temperate meadow steppe in northern China. Results We found that the response of plant caloric value to N addition at different organizational levels was diverse. The caloric value of legumes increased linearly with N addition rates. The caloric value of grasses exhibited a non-linear response trend, initially increasing followed by saturation or decrease, with a N response threshold present. Due to the dominance of grass species, the caloric value at the community level followed a similar pattern to that of the grasses along the N addition gradient. Under mowing, the caloric value of plants at each organizational level increased and usually mowing enhanced the N response threshold. Amongst these, the N response threshold of Leymus chinensis increased from 3.302 to 5.443 g N m−2 yr−1, grasses increased from 4.414 to 5.746 g N m−2 yr−1, and community increased from 5.373 to 9.216 g N m−2 yr−1. Under non-mowing treatment, the N response thresholds of the most dominant species, Leymus chinensis, and community energy standing crop were 10.001 and 15.119 g N m−2 yr−1, respectively. Under mowing, the energy standing crops showed a linear increasing trend. Conclusions N response thresholds of plant caloric value and energy standing crop vary at different organizational levels (community > functional group > species). The results reveal varying regulatory capabilities of plants on the ecological environment at different organizational levels. These findings enhance our understanding of plant-environment interactions in grassland ecosystems under N deposition from an energy perspective, which is of great significance to clarify the response mechanism of grassland ecosystem structure and function to N deposition.

Herbaceous plants contribute greatly to plant diversity and play an important role in regulating the structure and functioning of forest ecosystems. Comparing with woody species, herbs exhibit higher sensitivity to seasonal changes from spring-growth to autumn-mortality. Despite extensive research on understory herb diversity (e.g., abundance and richness) in forests, how the underlying mechanisms change across seasons remain unclear, especially in temperate forests. We explored the seasonal dynamics of herb diversity (here abundance and richness) by surveying herbs of 174 quadrats (1 m × 1 m) in a 25-ha (500 m × 500 m) temperate forest in spring, summer, and autumn of 2022, respectively. We examined the effects of overstory trees (diversity, composition, and three-dimensional (3D) structural complexity detected by LiDAR), topography (elevation, slope, and aspect), and microsite conditions (light availability, soil nutrients, and soil water content) on herb diversity in different seasons. We found a similar seasonal pattern for herb abundance and richness, i.e., both decreased from spring to summer and then to autumn, but the driving mechanisms differed remarkably across seasons. Specifically, the importance of overstory trees, topography, and microsite conditions on herb abundance vs. richness varied inversely across seasons. For herb abundance, the importance of overstory trees (primarily 3D structural complexity) increased from spring to autumn, while the importance of topography decreased. Conversely, for herb richness, the importance of overstory trees decreased from spring to autumn, while the importance of topography increased. Microsite conditions governed spatial variations of herb abundance in all seasons but only in autumn for herb richness, with importance of soil nutrients remaining relatively constant while that of soil water content and light availability changed greatly across seasons. Our results demonstrate substantial seasonal variations in driving mechanisms of herb diversity in the temperate forest. These differentiated responses of herb abundance and richness to different drivers across seasons further highlight the significance of considering seasonal variations of the mechanisms underlying understory plant diversity in forest biodiversity conservation.

Abstract Background Despite the crucial role of nitrogen (N) availability in carbon (C) cycling in terrestrial ecosystems, soil organic C (SOC) mineralization in different sizes of soil aggregates under various land use types and their responses to N addition is not well understood. To investigate the responses of soil C mineralization in different sized aggregates and land use types to N addition, an incubation experiment was conducted with three aggregate-size classes (2000, 250, and 53 μm) and two land use types (a Chinese fir plantation and a paddy land). Results Cumulative C mineralization of the

Leaf is essential for plant growth and development; however, the relative importance of environmental filtering and phylogeny in determining leaf size across different spatial scales in grassland ecosystems remains poorly explored. We used transect methodology to explore the spatial variation in leaf size and its underlying mechanisms in grasslands along three topographically and floristically distinctive plateaus in northern China. We measured leaf size of a total of 1192 grassland species in the Tibetan Plateau (TP, temperature limited), Loess Plateau (LP, soil-nutrient limited), and Mongolia Plateau (MP, precipitation limited) along three transects encompassing meadow, typical, and desert steppes. Leaf size ranged from 0.01 to 258.16 cm2, with an average of 5.54 cm2. The smallest leaves were measured in the TP. At the vegetation association level, the largest leaves were present in the meadow steppe, followed by those in typical and desert steppes, irrespective of the plateau. Unexpectedly, phylogeny had a negligible effect on the spatial variation in leaf size in the grasslands. Leaf size was positively correlated with growing-season temperature and precipitation but negatively correlated with ultraviolet (UV) radiation, suggesting that environmental filtering plays a more important role in affecting leaf size than phylogeny. Furthermore, leaf size in the TP and MP was mainly affected by precipitation and UV radiation, respectively, whereas that in the LP was affected by temperature, precipitation, soil nutrients and UV light. Specifically, our results underscored the importance of environmental filtering rather than phylogeny in determining plant leaf size and shed light on the unexpected role of UV radiation in contributing to leaf size variations in these plateaus. Our study provides novel insights into the response of plants to global change, especially in plateaus, alpine zone, and high-latitude areas.

In temperate perennial grasslands, the recruitment/regeneration of aboveground vegetation predominantly relies on belowground bud bank. Thus, understanding how belowground bud bank density and composition respond to global changes is essential to explain and predict plant community dynamics and ecosystem functions under global change context. The belowground bud bank and aboveground vegetation under the simulated precipitation changes and nitrogen deposition (by watering and N addition) were investigated in the temperate semiarid steppe of Inner Mongolia, China. N addition decreased total bud density but facilitated aboveground productivity, however, watering itself and its combination with N addition had little influence on total bud density. Different bud bank types showed specific responses to watering and N addition, especially, buds of grasses and forbs responded in an opposite way. Overall, shoot abundance were positively related to bud density, and showed similar responses with those of bud banks, especially for forbs, which are more sensitive to global changes. Our results imply that the changes in belowground bud bank might affect subsequent responses of plant communities to precipitation change and/or nitrogen deposition under future global change scenarios.

Anthropogenic input of reactive nitrogen (N) is an environmental problem that threatens the diversity and stability of belowground ecosystems. Soil nematodes are abundant in soil and are occurring at multiple trophic levels in the soil food web. However, how N deposition affects the composition and stability of soil nematode community is largely unknown. Here, we investigated the response of soil nematode community composition to N deposition at different sampling seasons and also estimated the stability of nematode community in a semiarid grassland in northern China. We found that the addition of N not only reduced the diversity of the nematode community, but also reduced the temporal stability of nematode community. The stability of different nematode trophic groups had different responses to N addition, and the community of plant parasites was more stable than the other trophic groups at a relatively higher N addition level. Moreover, soil pH was closely correlated with the stability of bacterivores, fungivores and predators-omnivores and the diversity of nematode community under different N additions. Our results highlight that N additions indirectly influence the synchrony and the stability of nematode community through change in soil nematode abundance and richness, and the variations of nematode community stability under different N additions are closely related to soil pH. These changes in nematode community composition and stability will eventually influence soil ecosystem function and nutrient cycling through biotic interactions in the soil food web.

Let {r(n)}n≥0 be the Rudin-Shapiro sequence, and let ρ(n):=max{∑j=ii+n−1r(j)∣i≥0}+1 be the abelian complexity function of the Rudin-Shapiro sequence. In this note, we show that the function ρ(n) has many similarities with the classical summatory function Sr(n):=∑i=0nr(i). In particular, we prove that for every positive integer n, 3≤ρ(n)n≤3. Moreover, the point set {ρ(n)n:n≥1} is dense in [3,3].

Soil microbial processes are crucial for understanding the ecological functions of arid and semi-arid lands which occupy approximately 40% of the global terrestrial ecosystems. However, how soil microbial metabolic activities may change across a wide aridity gradient in drylands remains unclear. Here, we investigated three soil microbial metabolic indices (soil organic carbon (SOC)-based microbial respiration, metabolic quotient, and microbial biomass as a proportion of total SOC) and the degree of carbon limitation for microbial respiration along a 3,200 km transect with a wide aridity gradient. The aridity gradient was customarily expressed using the aridity index (AI) which was calculated as the ratio of mean annual precipitation to mean annual evaporation, therefore, a lower AI value indicated a higher degree of aridity. Our results showed non-linear relationships between AI values and the metabolic indices with a clear aridity threshold for each of the three metabolic indices along the aridity gradient, respectively (AI = 0.13 for basal respiration, AI = 0.17 for metabolic quotient, and AI = 0.17 for MBC:SOC ratio). These metabolic indices linearly declined when AI was above the thresholds, but did not show any clear patterns when AI was below the thresholds. We also found that soil microbial respiration was highly limited by available carbon substrates at locations with higher primary production and relatively lower level of water limitation when AI was above the threshold, a counter-intuitive pattern that microbes were more starved in ecosystems with more substrate input. However, the increasing level of carbon limitation did correspond to the declining trend of the three metabolic indices along the AI gradient, which indicates that the carbon limitation influences microbial metabolism. We also found that the ratio of microbial biomass carbon to SOC in arid regions (AI < 0.2) with extremely low precipitation and primary production were not quantitatively related to SOC content. Overall, our results imply that microbial metabolism is distinctively different in arid lands than in non-arid lands.

Plant biomass allocation between below- and above-ground parts can actively adapt to the ambient growth conditions and is a key parameter for estimating terrestrial ecosystem carbon (C) stocks. To investigate how climatic variations affect patterns of plant biomass allocation, we sampled 548 plants belonging to four dominant genera (Stipa spp., Cleistogenes spp., Agropyron spp., and Leymus spp.) along a large-scale (2500 km) climatic gradient across the temperate grasslands from west to east in northern China. Our results showed that Leymus spp. had the lowest root/shoot ratios among the each genus. Root/shoot ratios of each genera were positively correlated with mean annual temperature (MAT), and negatively correlated with mean annual precipitation (MAP) across the transect. Temperature contributed more to the variation of root/shoot ratios than precipitation for Cleistogenes spp. (C4 plants), whereas precipitation exerted a stronger influence than temperature on their variations for the other three genera (C3 plants). From east to west, investment of C into the belowground parts increased as precipitation decreased while temperature increased. Such changes in biomass allocation patterns in response to climatic factors may alter the competition regimes among co-existing plants, resulting in changes in community composition, structure and ecosystem functions. Our results suggested that future climate change would have great impact on C allocation and storage, as well as C turnover in the grassland ecosystems in northern China.

The growth rate hypothesis (GRH) proposes that higher growth rate (the rate of change in biomass per unit biomass, μ) is associated with higher P concentration and lower C:P and N:P ratios. However, the applicability of the GRH to vascular plants is not well-studied and few studies have been done on belowground biomass. Here we showed that, for aboveground, belowground and total biomass of three study species, μ was positively correlated with N:C under N limitation and positively correlated with P:C under P limitation. However, the N:P ratio was a unimodal function of μ, increasing for small values of μ, reaching a maximum, and then decreasing. The range of variations in μ was positively correlated with variation in C:N:P stoichiometry. Furthermore, μ and C:N:P ranges for aboveground biomass were negatively correlated with those for belowground. Our results confirm the well-known association of growth rate with tissue concentration of the limiting nutrient and provide empirical support for recent theoretical formulations.

Nitrogen (N) enrichment resulting from anthropogenic activities has greatly changed the composition and functioning of soil communities. Nematodes are one of the most abundant and diverse groups of soil organisms, and they occupy key trophic positions in the soil detritus food web. Nematodes have therefore been proposed as useful indicators for shifts in soil ecosystem functioning under N enrichment. Here, we monitored temporal dynamics of the soil nematode community using a multi-level N addition experiment in an Inner Mongolia grassland. Measurements were made three years after the start of the experiment. We used structural equation modeling (SEM) to explore the mechanisms regulating nematode responses to N enrichment. Across the N enrichment gradient, significant reductions in total nematode abundance, diversity (H' and taxonomic richness), maturity index (MI), and the abundance of root herbivores, fungivores and omnivores-predators were found in August. Root herbivores recovered in September, contributing to the temporal variation of total nematode abundance across the N gradient. Bacterivores showed a hump-shaped relationship with N addition rate, both in August and September. Ammonium concentration was negatively correlated with the abundance of total and herbivorous nematodes in August, but not in September. Ammonium suppression explained 61% of the variation in nematode richness and 43% of the variation in nematode trophic group composition. Ammonium toxicity may occur when herbivorous nematodes feed on root fluid, providing a possible explanation for the negative relationship between herbivorous nematodes and ammonium concentration in August. We found a significantly positive relationship between fungivores and fungal phospholipid fatty acids (PLFA), suggesting bottom-up control of fungivores. No such relationship was found between bacterivorous nematodes and bacterial PLFA. Our findings contribute to the understanding of effects of N enrichment in semiarid grassland on soil nematode trophic groups, and the cascading effects in the detrital soil food web.

CoNiCrAlY alloys, commonly used as the bond coat of thermal barrier coatings (TBCs), are often exposed to high temperatures of about 1000 °C, the evolution of mechanical and thermal properties is critical to the reliability and durability of TBCs. In this paper, the hardness and thermal properties (weight change and heat flow) of CoNiCrAlY alloys at temperatures within 1000 °C were evaluated by indentation and TGA-DSC thermal analysis. It was found that hardness and thermal properties change gradually within 600 °C but change sharply above 600 °C, exhibiting an approximately synchronized evolution. An illustrative explanation was given for the correlation in terms of the Boltzmann energy distribution theory of microscopic particles. Based on these findings, an idea of inter-calibration between the two evolutions was proposed.

The shift from natural mixed broadleaved forests to pure coniferous plantations results in soil degradation and the unsustainable development of plantations due to the simple stand structure and low species diversity. Thinning can practically sustain the forest structure and promote the regeneration and growth of broadleaved trees in these pure coniferous plantations. The growth of regenerated broadleaved trees is closely related to leaf ecological stoichiometry, which is strongly restricted by environmental factors such as light, soil moisture, and nutrients after thinning. However, the temporal effects of thinning on leaf C:N:P stoichiometry are still not well understood, which constrains our understanding of implementing thinning in coniferous plantations to promote the regeneration and growth of broadleaved species, and further forming the mixed larch-broadleaf forests. Here, we compared canopy openness (i.e., light availability) and the soil and leaf stoichiometry for regenerated broadleaved trees in larch (Larix keampferi) plantations in short-term (1–3 years), medium-term (4–9 years), and long-term (≥10 years) periods after thinning, taking natural mixed broadleaved forests as a control in Northeast China. The results showed that the temporal effects of thinning were not significant with respect to soil C concentrations, but significant with respect to soil C:P and N:P ratios. The regenerated broadleaved trees adjusted their leaf N concentrations and C:N ratios in response to the changed environmental conditions after thinning over time. The responses of soil and leaf stoichiometry to thinning and their significant correlation indicated a strong interaction between the soil and understory regeneration following thinning. Thus, thinning affects the soil and leaf stoichiometry of regenerated trees over time. These findings provide new insights into the conversion of pure coniferous plantations into mixed larch-broadleaf forests by controlling thinning intervals. [ABSTRACT FROM AUTHOR]

ABSTRACT Coronaviruses (CoVs) encode nonstructural proteins 1–16 (nsps 1–16) which form replicase complexes that mediate viral RNA synthesis. Remdesivir (RDV) is an adenosine nucleoside analog antiviral that inhibits CoV RNA synthesis. RDV resistance mutations have been reported only in the nonstructural protein 12 RNA-dependent RNA polymerase (nsp12-RdRp). We here show that a substitution mutation in the nsp13-helicase (nsp13-HEL A335V) of the betacoronavirus murine hepatitis virus (MHV) that was selected during passage with the RDV parent compound confers partial RDV resistance independently and additively when expressed with co-selected RDV resistance mutations in the nsp12-RdRp. The MHV A335V substitution did not enhance replication or competitive fitness compared to WT MHV and remained sensitive to the active form of the cytidine nucleoside analog antiviral molnupiravir (MOV). Biochemical analysis of the SARS-CoV-2 helicase encoding the homologous substitution (A336V) demonstrates that the mutant protein retained the ability to associate with the core replication proteins nsps 7, 8, and 12 but had impaired helicase unwinding and ATPase activity. Together, these data identify a novel determinant of nsp13-HEL enzymatic activity, define a new genetic pathway for RDV resistance, and demonstrate the importance of surveillance for and testing of helicase mutations that arise in SARS-CoV-2 genomes. IMPORTANCE Despite the development of effective vaccines against COVID-19, the continued circulation and emergence of new variants support the need for antivirals such as RDV. Understanding pathways of antiviral resistance is essential for surveillance of emerging variants, development of combination therapies, and for identifying potential new targets for viral inhibition. We here show a novel RDV resistance mutation in the CoV helicase also impairs helicase functions, supporting the importance of studying the individual and cooperative functions of the replicase nonstructural proteins 7–16 during CoV RNA synthesis. The homologous nsp13-HEL mutation (A336V) has been reported in the GISAID database of SARS-CoV-2 genomes, highlighting the importance of surveillance of and genetic testing for nucleoside analog resistance in the helicase.

In this study, molecular dynamics simulation is adopted to investigate the friction and wear mechanism of Fe-based amorphous composited coatings. It was found that the ceramic phase can improve plasticity of the composite coatings to reduce wear. In addition, when the Al2O3 content is 15 wt%, due to the stress concentration between ceramic phase and amorphous phase, the viscosity flow in the amorphous phase results in plastic deformation. Through this, the best wear resistance has been obtained. In addition, with the increase of compression depth, the number of atoms accumulated constantly increases, and the degree of plastic deformation becomes higher in the composite coatings. Finally, a high sliding speed results in the reduction of the wear resistance of coatings.

Background and Aims Understanding the changes in below-ground bud bank density and composition along a climatic gradient is essential for the exploration of species distribution pattern and vegetation composition in response to climatic changes. Nevertheless, investigations on bud banks along climatic gradients are still scarce. The below-ground bud bank is expected to be reduced in size in arid conditions, and costly, bud-bearing organs with long spacers would be replaced by more compact forms with buds that are better protected than those found in moist conditions. Methods How total bud density and composition (different bud bank types) change with aridity (calculated value 0·43-0·91), mean annual precipitation (MAP; 93-420mm) and mean annual temperature (MAT; -1·51 to 6·93 °C) was tested at 21 sites along a 2500-km climatic gradient in the temperate steppe of northern China. Key Results The relationship between below-ground bud bank density and precipitation/aridity was fundamentally changed along the climatic gradient. Bud bank density increased first and then decreased along the aridity and MAP gradients, the turning points being aridity=0·67 and MAP=260mm, respectively, while it decreased consistently with increasing MAT. The proportion of the bud bank that comprised tiller buds or dicotyledonous herb buds fluctuated along the aridity gradient. The proportion of rhizome buds was higher at relatively moist sites (aridity <0·75), while that of bulb buds was higher at drier sites (aridity >0·75). Conclusions Belowground bud bank density decreases towards the dry, hot end of the climatic gradient. Based on the distribution of bud types along the climatic gradient, bulb buds and tiller buds of tussock grasses seem to be more resistant to environmental stress than rhizome buds. The dominance of annual species and smaller bud banks in arid region implies that plant reproductive strategies and vegetation composition will be shifted in scenarios of increased drought under future climate change. [ABSTRACT FROM AUTHOR]

Transition from forest to rubber (Hevea brasiliensis Muell. Arg.) plantation has occurred in tropical China for decades. Rubber has been planted on 1 million ha to provide raw materials to the rubber industry. The role of various-aged rubber plantations in carbon (C) sequestration remains unclear. The biomass C accumulation including latex C and C distribution in soil of five different-aged stands (7, 13, 19, 25 and 47 years old) were examined. The total biomass C stock (TBC) and total net primary productivity (NPPtotal), whether with or without latex C, had a close quadratic relationship with stand age. Regardless of stand age, around 68% of the C was stored in aboveground biomass, and NPPlatex contributed to approximately 18% of C sequestration. Soil organic carbon stock in the 100-cm depth remained relatively stable, but it lost about 16.8 Mg ha-1 with stand age. The total ecosystem C stock (TEC) across stands averaged 159.6, 174.4, 229.6, 238.1 and 291.9Mg ha-1, respectively, of which more than 45% was stored in the soil. However, biomass would become the major C sink rather than soil over a maximal rubber life expectancy. Regression analysis showed that TEC for rubber plantation at 22 years is comparable to a baseline of 230.4 Mg ha-1 for tropical forest in China, and would reach the maximum value at around 54 years. Therefore, rubber plantation can be considered as alternative land use without affecting net forest ecosystem C storage. In addition to the potential C gains, a full set of ecosystem and economic properties have to be quantified in order to assess the trade-offs associated with forest-to-rubber transition. [ABSTRACT FROM AUTHOR]

The low temperature solid deposition characteristics of cold spraying technology are conducive to the remanufacturing/repair of oxidation sensitive titanium alloy in the atmosphere. Ti6Al4V alloy deposits prepared by cold spray (CS) usually show porous microstructure due to insufficient plastic deformation of Ti6Al4V alloy particles. The interfacial bonding and the microstructural evolution mechanisms of Ti6Al4V particles during cold spray are still unclear. In this study, the Ti6Al4V deposits were prepared by cold spray with nitrogen as process gas at a temperature of 800 °C and a pressure of 4 MPa. The micron CT results show that the porosity is 8.25%. The deformation ultrafine grain zone and un-deformation martensitic laths zone at the bonding interface between Ti6Al4V particles were observed. The EBSD results show that the grain orientation signal was only detected at the edge of the particles deformation area and the 23% of grain sizes detected by EBSD are less than 500 nm, which indicates that the nanocrystallization induced by dynamic recrystallization occurs at the inter-particle boundaries between cold-sprayed Ti6Al4V particles. The TEM and SADPs demonstrate a gradient structure consisting of equiaxed nanograins zone, crushing dendrites transition zone and original martensitic laths zone from the interface to the core of particles.

Aims We aimed to quantify the variation of leaf δ13C along an arid and semi-arid grassland transect in northern China. We also evaluated the effects of environmental factors (i.e. precipitation, temperature and altitude) on the spatial variation of leaf δ13C in northern grasslands and Tibetan Plateau, China. Method We sampled leaves of plant species belonging to three herb genera (Stipa spp., Leymus spp. and Cleistogenes spp.) and three shrub genera (Caragana spp., Reaumuria spp. and Nitraria spp.) for carbon isotope analysis from 50 locations along a 3200-km arid and semiarid grassland transect in northern China. Leaf δ13C data in Tibetan Plateau and northern grasslands in China were also compiled from studies in literature. Important Findings Along the transect, leaf δ13C for C3 plants ranged from -28.0 to -23.3, and from -16.3 to -13.8 for C4 plant Cleistogenes spp. The change in leaf δ13C ranged from -0.26 to -3.51 with every 100 mm increase of annual precipitation, and leaf δ13C of shrubs (Nitraria spp., Reaumuria spp. and Caragana spp.) responded more markedly to climatic factors (precipitation and temperature) than that of herbs (Stipa spp., Leymus spp. and Cleistogenes spp.), indicating higher sensitivity of shrub δ13C to climatic changes. The most important factor regulating spatial variations of leaf δ13C in Tibetan Plateau was altitude, while it was precipitation in northern grasslands. Our results suggested that shrubs are more adapted to increasing drought in arid and semi-arid grassland. Controls of environmental factors on leaf δ13C depended on the most limiting factors in arid grassland (precipitation) and Tibetan grasslands (atmospheric CO2 concentration). [ABSTRACT FROM AUTHOR]

Aims Conversion of secondary forests to pure larch plantations is a common management practice driven by the increasing demand for timber production in Northeast China, resulting in a reduction in soil nutrient availability after a certain number of years following conversion. Nutrient resorption prior to leaf senescence was related to soil fertility, an important nutrient conservation strategy for plants, being especially significant in nutrient-poor habitats. However, the seasonal dynamics of leaf nutrients and nutrient resorption in response to secondary forest conversion to larch plantations is not well understood. Methods A comparative experiment between larch plantations (Larix spp.) and adjacent secondary forests (dominant tree species including Quercus mongolica, Acer mono, Juglans mandshurica and Fraxinus rhynchophylla) was conducted. We examined the variations in leaf nutrient (macronutrients: N, P, K, Ca and Mg; micronutrients: Cu and Zn) concentrations of these tree species during the growing season from May to October in 2013. Nutrient resorption efficiency and proficiency were compared between Larix spp. and the broadleaved species in the secondary forests. Important Findings Results show that the seasonal variation of nutrient concentrations in leaves generally exhibited two trends, one was a downward trend for N, P, K, Cu and Zn, and another was an upward trend for Ca and Mg. The variations in foliar nutrient concentrations were mainly controlled by the developmental stage of leaves rather than by tree species. Resorption of the observed seven elements varied among the five tree species during leaf senescence. Nutrient resorption efficiency varied 6-75% of N, P, K, Mg, Cu and Zn, while Ca was not retranslocated in the senescing leaves of all species, and Mg was not retranslocated in Larix spp. Generally, Larix spp. tended to be more efficient and proficient (higher than 6-30% and 2-271% of nutrient resorption efficiency and resorption proficiency, respectively) in resorbing nutrients than the broadleaved species in the secondary forests, indicating that larch plantations had higher leaf nutrient resorption and thus nutrient use efficiency. Compared with Larix spp., more nutrients would remain in the leaf litter of the secondary forests, indicating an advantage of secondary forests in sustaining soil fertility. In contrast, the larch plantation would reuse internal nutrients rather than lose nutrients with litter fall and thus produce a positive feedback to soil nutrient availability. In summary, our results suggest that conversion from secondary forests to pure larch plantations would alter nutrient cycling through a plantmediated pathway. [ABSTRACT FROM AUTHOR]

Patterns of variation in plant and animal diversity along precipitation gradients have been extensively studied, but much less is known about how and to what extent precipitation affects the biogeographic distribution of microbial diversity in arid areas across large spatial scales. Here we collected soils from 54 sites along a 3700 km transect covering a wide range of grassland ecosystems with distinct aridity gradients. We quantified the bacterial community diversity and the effects of climate, edaphic parameter and geographic distance on the bacterial community structure using high-throughput 16S rRNA gene sequencing. Of the 35 phyla detected, 6 were dominant: Actinobacteria, Acidobacteria, Alphaproteobacteria, Deltaproteobacteria, Bacteroidetes and Planctomycetes. Aridity was a major factor influencing bacterial diversity, community composition and taxon abundance. Although the pattern of bacterial species richness is markedly different from that of plant species richness, most soil bacteria were endemic to particular bioregions like macro-organisms. Community similarity significantly declined with environmental distance and geographic distance (r = -0.579 and -0.773, respectively). Geographic distance (historical contingencies) contributed more to bacterial community variation (36.02%) than combined environmental factors (24.06%). Overall, our results showed that geographic distance and climatic factors concurrently govern bacterial biogeographic patterns in arid and semi-arid grassland. [ABSTRACT FROM AUTHOR]

Experimental investigation was conducted to explore the formation conditions and provide new insights into the formation mechanisms of the unexplained intermixing phenomenon observed at the substrate-coatings interface of cold sprayed materials. The results indicate that the formation of intermixing interface significantly depends on the extent of plastic deformation at the coating-substrate interface, with severe deformation creating favorable conditions for the intermixing interface. Two factors have been identified to be critical for inducing the severe interfacial plastic deformation: low deposition efficiency and material properties. During low deposition efficiency cold spraying, most of the accelerated particles rebound after impact while inducing accumulative plastic deformation and thus intermixing at the coating-substrate interface. Considering the material properties, the coating materials must have sufficiently high density to attain enough kinetic energy for creating substantial plastic deformation of the first coating layer and the substrate upon their impact. Also, the substrate materials cannot be too hard so that plastic deformation can be induced. Based on the experimental analyses, the hypothesis of the intermixing interface formation mechanism is proposed in this paper.

Recombination is proposed to be critical for coronavirus (CoV) diversity and emergence of SARS-CoV-2 and other zoonotic CoVs. While RNA recombination is required during normal CoV replication, the mechanisms and determinants of CoV recombination are not known. CoVs encode an RNA proofreading exoribonuclease (nsp14-ExoN) that is distinct from the CoV polymerase and is responsible for high-fidelity RNA synthesis, resistance to nucleoside analogues, immune evasion, and virulence. Here, we demonstrate that CoVs, including SARS-CoV-2, MERS-CoV, and the model CoV murine hepatitis virus (MHV), generate extensive and diverse recombination products during replication in culture. We show that the MHV nsp14-ExoN is required for native recombination, and that inactivation of ExoN results in decreased recombination frequency and altered recombination products. These results add yet another critical function to nsp14-ExoN, highlight the uniqueness of the evolved coronavirus replicase, and further emphasize nsp14-ExoN as a central, completely conserved, and vulnerable target for inhibitors and attenuation of SARS-CoV-2 and future emerging zoonotic CoVs.

Loss of functional diversity has been demonstrated to have a variety of impacts on ecosystem functioning. However, most studies have been implemented in artificially assembled communities by removing the original vegetation and seeding with desired species or functional group compositions. Such approaches could significantly disturb belowground biomass, especially roots, making it difficult to examine belowground responses to diversity manipulations. To circumvent this issue, plant diversity gradients were established by in situ removal of aboveground biomass of different plant functional groups (PFGs) in a typical steppe, and belowground processes related to roots and soil were examined. Root nutrient pools exhibited contrasting patterns, with the phosphorus (P) pool decreasing linearly upon increased PFG removal while the nitrogen (N) pool had a hump-shaped response. Soil NO increased while net N mineralization decreased with PFG removal. In contrast, soil P showed little response to PFG removal. Furthermore, both the identity and number of PFG removed had a significant influence on root and soil properties. The results of this study showed that loss of a combination of PFGs was important in natural grassland, and an approach with minimal influence on belowground processes is promising in studying diversity loss effects in natural ecosystems. [ABSTRACT FROM AUTHOR]

Determining the variation in roots traits within a grass root system is important for understanding the role of fine roots in carbon and nutrient cycling in grassland ecosystems, where the majority of biomass and litter accumulation occur belowground. However, few studies have been conducted in this regard. In this study, the structural and chemical traits of shoot-derived and root-derived roots were examined in three perennial grasses- Cleistogenes squarrosa, Achnatherum sibiricum and Stipa grandis-aiming to explore structural differences, responses to nitrogen and water addition in different types of roots and their correlations with aboveground plant nitrogen. Our results showed significant differences between these two root types, with root-derived roots having higher N concentration, tissue density, and specific root length, but lower C: N and diameter than shoot-derived roots. Trait relationships between root N concentration and tissue density for the two root types differed from that reported among species. These traits in different types of roots were insensitive to resource addition. Furthermore, N concentration in shoot-derived roots was more strongly linked to aboveground plant N concentration than root-derived roots. The results of this study demonstrate structural differences within the root system that may reflect functional heterogeneity in grass roots. [ABSTRACT FROM AUTHOR]

ABSTRACT Emerging coronaviruses (CoVs) cause severe disease in humans, but no approved therapeutics are available. The CoV nsp14 exoribonuclease (ExoN) has complicated development of antiviral nucleosides due to its proofreading activity. We recently reported that the nucleoside analogue GS-5734 (remdesivir) potently inhibits human and zoonotic CoVs in vitro and in a severe acute respiratory syndrome coronavirus (SARS-CoV) mouse model. However, studies with GS-5734 have not reported resistance associated with GS-5734, nor do we understand the action of GS-5734 in wild-type (WT) proofreading CoVs. Here, we show that GS-5734 inhibits murine hepatitis virus (MHV) with similar 50% effective concentration values (EC50) as SARS-CoV and Middle East respiratory syndrome coronavirus (MERS-CoV). Passage of WT MHV in the presence of the GS-5734 parent nucleoside selected two mutations in the nsp12 polymerase at residues conserved across all CoVs that conferred up to 5.6-fold resistance to GS-5734, as determined by EC50. The resistant viruses were unable to compete with WT in direct coinfection passage in the absence of GS-5734. Introduction of the MHV resistance mutations into SARS-CoV resulted in the same in vitro resistance phenotype and attenuated SARS-CoV pathogenesis in a mouse model. Finally, we demonstrate that an MHV mutant lacking ExoN proofreading was significantly more sensitive to GS-5734. Combined, the results indicate that GS-5734 interferes with the nsp12 polymerase even in the setting of intact ExoN proofreading activity and that resistance can be overcome with increased, nontoxic concentrations of GS-5734, further supporting the development of GS-5734 as a broad-spectrum therapeutic to protect against contemporary and emerging CoVs. IMPORTANCE Coronaviruses (CoVs) cause severe human infections, but there are no approved antivirals to treat these infections. Development of nucleoside-based therapeutics for CoV infections has been hampered by the presence of a proofreading exoribonuclease. Here, we expand the known efficacy of the nucleotide prodrug remdesivir (GS-5734) to include a group β-2a CoV. Further, GS-5734 potently inhibits CoVs with intact proofreading. Following selection with the GS-5734 parent nucleoside, 2 amino acid substitutions in the nsp12 polymerase at residues that are identical across CoVs provide low-level resistance to GS-5734. The resistance mutations decrease viral fitness of MHV in vitro and attenuate pathogenesis in a SARS-CoV animal model of infection. Together, these studies define the target of GS-5734 activity and demonstrate that resistance is difficult to select, only partial, and impairs fitness and virulence of MHV and SARS-CoV, supporting further development of GS-5734 as a potential effective pan-CoV antiviral.

ABSTRACT The coronavirus (CoV) RNA genome is the largest among the single-stranded positive-sense RNA viruses. CoVs encode a proofreading 3′-to-5′ exoribonuclease within nonstructural protein 14 (nsp14-ExoN) that is responsible for CoV high-fidelity replication. Alanine substitution of ExoN catalytic residues [ExoN(-)] in severe acute respiratory syndrome-associated coronavirus (SARS-CoV) and murine hepatitis virus (MHV) disrupts ExoN activity, yielding viable mutant viruses with defective replication, up to 20-fold-decreased fidelity, and increased susceptibility to nucleoside analogues. To test the stability of the ExoN(-) genotype and phenotype, we passaged MHV-ExoN(-) 250 times in cultured cells (P250), in parallel with wild-type MHV (WT-MHV). Compared to MHV-ExoN(-) P3, MHV-ExoN(-) P250 demonstrated enhanced replication and increased competitive fitness without reversion at the ExoN(-) active site. Furthermore, MHV-ExoN(-) P250 was less susceptible than MHV-ExoN(-) P3 to multiple nucleoside analogues, suggesting that MHV-ExoN(-) was under selection for increased replication fidelity. We subsequently identified novel amino acid changes within the RNA-dependent RNA polymerase and nsp14 of MHV-ExoN(-) P250 that partially accounted for the reduced susceptibility to nucleoside analogues. Our results suggest that increased replication fidelity is selected in ExoN(-) CoVs and that there may be a significant barrier to ExoN(-) reversion. These results also support the hypothesis that high-fidelity replication is linked to CoV fitness and indicate that multiple replicase proteins could compensate for ExoN functions during replication. IMPORTANCE Uniquely among RNA viruses, CoVs encode a proofreading exoribonuclease (ExoN) in nsp14 that mediates high-fidelity RNA genome replication. Proofreading-deficient CoVs with disrupted ExoN activity [ExoN(-)] either are nonviable or have significant defects in replication, RNA synthesis, fidelity, fitness, and virulence. In this study, we showed that ExoN(-) murine hepatitis virus can adapt during long-term passage for increased replication and fitness without reverting the ExoN-inactivating mutations. Passage-adapted ExoN(-) mutants also demonstrate increasing resistance to nucleoside analogues that is explained only partially by secondary mutations in nsp12 and nsp14. These data suggest that enhanced resistance to nucleoside analogues is mediated by the interplay of multiple replicase proteins and support the proposed link between CoV fidelity and fitness.

Most RNA viruses lack the mechanisms to recognize and correct mutations that arise during genome replication, resulting in quasispecies diversity that is required for pathogenesis and adaptation. However, it is not known how viruses encoding large viral RNA genomes such as the Coronaviridae (26 to 32 kb) balance the requirements for genome stability and quasispecies diversity. Further, the limits of replication infidelity during replication of large RNA genomes and how decreased fidelity impacts virus fitness over time are not known. Our previous work demonstrated that genetic inactivation of the coronavirus exoribonuclease (ExoN) in nonstructural protein 14 (nsp14) of murine hepatitis virus results in a 15-fold decrease in replication fidelity. However, it is not known whether nsp14-ExoN is required for replication fidelity of all coronaviruses, nor the impact of decreased fidelity on genome diversity and fitness during replication and passage. We report here the engineering and recovery of nsp14-ExoN mutant viruses of severe acute respiratory syndrome coronavirus (SARS-CoV) that have stable growth defects and demonstrate a 21-fold increase in mutation frequency during replication in culture. Analysis of complete genome sequences from SARS-ExoN mutant viral clones revealed unique mutation sets in every genome examined from the same round of replication and a total of 100 unique mutations across the genome. Using novel bioinformatic tools and deep sequencing across the full-length genome following 10 population passages in vitro, we demonstrate retention of ExoN mutations and continued increased diversity and mutational load compared to wild-type SARS-CoV. The results define a novel genetic and bioinformatics model for introduction and identification of multi-allelic mutations in replication competent viruses that will be powerful tools for testing the effects of decreased fidelity and increased quasispecies diversity on viral replication, pathogenesis, and evolution.