Population genetic assessment of Viburnum japonicum in China using ddRAD-seq.

Viburnum japonicum is a rare plant species and endemic to the coastal region of Eastern Asia with extremely small populations. Within mainland China, this species can be only found in narrow habitats of the northeast coastal islands of Zhejiang Province. However, there are scarce conservation genetic studies on V. japonicum, which has limited the effective conservation and management of this rare species. Here, 51 individuals in four natural populations covering the Chinese geographic range of the species were sampled to assess the genetic diversity and population structure. A total of 445,060 high-quality single nucleotide polymorphisms (SNPs) were identified using double digest restriction-site associated sequencing (ddRAD-seq). The overall average values of observed heterozygosity (Ho), expected heterozygosity (He), and average nucleotide diversity (π), were 0.2207, 0.2595, and 0.2741, respectively. The DFS-2 population exhibited the highest level of genetic diversity among all the populations. Genetic differentiation between populations was moderate (F ST = 0.1425), and there was selfing between populations (F IS = 0.1390, S = 24.52%). Of the total genetic variation, 52.9% was found among populations through AMOVA analysis. The Mantel test (r = 0.982, p = 0.030) combined with analyses of the Maximum Likelihood (ML) phylogenetic tree, ADMIXTURE, and principal component analysis (PCA), revealed that populations of V. japonicum were genetically segregated and significantly correlated with their geographical distribution. Our study demonstrated that V. japonicum maintained a medium level of genetic diversity and differentiation with a strong population structure, and the results were mainly affected by its island distribution pattern and self-crossing characteristics. These results provide insights into the genetic diversity and population history of V. japonicum, critical information for conserving and sustainably developing its genetic resources.

[Effects of aridity on carbon, nitrogen, and phosphorus contents in soils and plants of mountain swamps, Zhejiang, China]. / 旱化对浙江山地沼泽湿地土壤与植物碳氮磷含量的影响.

Mountain swamps in Zhejiang Province have been suffered from severe drouhgt threats because of climate change and artificial interruption. Sphagnum bogs and swamps were gradually degraded into arid swamps. However, the effects of arid processes on the C, N, P contents of soils and their stoichiometry in mountain swamps are still unclear. We measured C, N and P concentrations, pH values, and bulk density in the upper 0-60 cm soil layers in the stands of five mountain swamps with the different arid levels. Moreover, the aboveground biomass and the C, N, P concentrations in the crushed plant mixture were also measured. The results showed that the soils of mountain swamps in Zhejiang Province were rich in soil organic carbon (SOC), total nitrogen (TN), but infertile in phosphorus (TP). Aboveground biomass, soil pH, bulk density increased, while SOC, TN, TP, C:N, C:P, N:P decreased with aridity. Soil pH and bulk density had significant negative correlations with SOC, TN, and TP in soils, respectively. The differences in the C, N, P accumulation in the soils were probably associated with litterfall production, the oxygen condition of wetlands, and the degree of plant decomposition at the different types of mountain swamps. In all, arid trends were obvious at the mountain swamps in Zhejiang Province. Soil nutrients, such as C, N, P, deceased, while plant community succeeded from the wet swamp to the mesophyte vegetation with the arid processes. The contents of C, N and P in the plants varied across species, and were not coupled with those in the soils.

Exploring the spatiotemporal changes in carbon storage under different development scenarios in Jiangsu Province, China.

Carbon storage (CS) is closely linked to the global challenge of climate change. Land use/cover (LULC) change is the main factor driving changes in CS, and evaluating the impact of LULC changes on CS is important for carbon balance. Taking Jiangsu Province as an example, we used the Integrated Valuation of Ecosystem Services and Trade-offs model to analyze the spatiotemporal changes in CS during 2000-2015. Then we coupled it with the patch-generating land use simulation model to simulate and predict LULC and CS in 2050 under four different development plans. The results showed that LULC change in Jiangsu Province was manifested mainly as transformation of cropland to construction land (3,485 km2) and cropland to water body (470 km2). The high value area for CS was concentrated mainly in forest land, water body and grassland, whereas the low value area was concentrated mainly in construction land. During 2000-2015, CS decreased by 0.23 Tg, and during 2015-2050, CS was predicted to decrease by 0.16, 1.69, 0.02, and 0.10 Tg under the baseline, fast, slow and harmonious development scenarios. The conversion of a large amount of cropland to construction land was the main cause of CS loss. In all scenarios, the carbon loss was the largest in southern Jiangsu and lowest in central Jiangsu. It is necessary to balance the conflict between economic development and ecological protection during the process of urbanization. This study can provide an important reference for decision makers during the formulation of regional development models and ecological management strategies.

Alleviation of the Byproducts Formation Enables Highly Efficient Biosynthesis of Rosmarinic Acid in Saccharomyces cerevisiae.

Rosmarinic acid as a polyphenolic compound has great values in the pharmaceutical, cosmetic, and food industries. To achieve efficient biosynthesis of rosmarinic acid, the major obstacles such as imbalanced metabolic flux among branching pathways and substrate promiscuity of pathway enzymes should be eliminated. Here, a rosmarinic acid producing Saccharomyces cerevisiae strain was constructed by introducing codon optimized d-lactate dehydrogenase gene mutant (OD-LDHY52A), 4-coumarate CoA ligase gene (OPc4CL2), and rosmarinic acid synthase gene (OMoRAS) into a previously constructed caffeic acid hyper-producer. To identify the metabolic bottleneck, the substrate specificity of OPc4CL2 and OMoRAS was figured out by bioconversion experiments and HPLC-MS/MS analysis. Subsequently, the byproducts formation was alleviated by removing prephenate dehydratase and tuning down the expression level of OPc4CL2. The final strain YRA113-15B produced 208 mg/L rosmarinic acid in a shake-flask culture (a 63-fold improvement over the initial strain), which was the highest rosmarinic acid titer by engineered microbial cells reported to date. This work provides a promising platform for fermentative production of rosmarinic acid and offers a strategy to overcome the intrapathway competition.

Metabolic engineering of Saccharomyces cerevisiae for enhanced production of caffeic acid.

As a natural phenolic acid product of plant source, caffeic acid displays diverse biological activities and acts as an important precursor for the synthesis of other valuable compounds. Limitations in chemical synthesis or plant extraction of caffeic acid trigger interest in its microbial biosynthesis. Recently, Saccharomyces cerevisiae has been reported for the biosynthesis of caffeic acid via episomal plasmid-mediated expression of pathway genes. However, the production was far from satisfactory and even relied on the addition of precursor. In this study, we first established a controllable and stable caffeic acid pathway by employing a modified GAL regulatory system to control the genome-integrated pathway genes in S. cerevisiae and realized biosynthesis of 222.7 mg/L caffeic acid. Combinatorial engineering strategies including eliminating the tyrosine-induced feedback inhibition, deleting genes involved in competing pathways, and overexpressing rate-limiting enzymes led to about 2.6-fold improvement in the caffeic acid production, reaching up to 569.0 mg/L in shake-flask cultures. To our knowledge, this is the highest ever reported titer of caffeic acid synthesized by engineered yeast. This work showed the prospect for microbial biosynthesis of caffeic acid and laid the foundation for constructing biosynthetic pathways of its derived metabolites. KEY POINTS: Genomic integration of ORgTAL, OHpaB, and HpaC for caffeic acid production in yeast. Feedback inhibition elimination and Aro10 deletion improved caffeic acid production. The highest ever reported titer (569.0 mg/L) of caffeic acid synthesized by yeast.

Quantum Transport of the 2D Surface State in a Nonsymmorphic Semimetal.

In a topological semimetal with Dirac or Weyl points, the bulk-boundary correspondence principle predicts a gapless edge mode if the essential symmetry is still preserved at the surface. The detection of such topological surface state has been considered as the fingerprint prove for crystals with nontrivial topological bulk band. On the contrary, it has been proposed that even with symmetry broken at the surface, a new surface band can emerge in nonsymmorphic topological semimetals. The symmetry reduction at the surface lifts the bulk band degeneracies and produces an unusual "floating" surface band with trivial topology. Here, we first report quantum transport probing to ZrSiSe thin flakes and directly reveal transport signatures of this new surface state. Remarkably, though topologically trivial, such a surface band exhibits substantial two-dimensional Shubnikov-de Haas quantum oscillations with high mobility, which signifies a new protection mechanism and may open applications for quantum computing and spintronic devices.

Combinatorial Modulation of Linalool Synthase and Farnesyl Diphosphate Synthase for Linalool Overproduction in Saccharomyces cerevisiae.

Linalool, as a fragrant monoterpene, is an important feedstock for food, pharmaceuticals, and cosmetics industries. Although our previous study had significantly increased linalool production by the directed evolution of linalool synthase and overexpression of the whole mevalonate pathway genes, the engineered yeast strain suffered from dramatically reduced biomass. Herein, a stress-free linalool-producing yeast cell factory was constructed by the combinational regulation of linalool synthase and farnesyl diphosphate synthase instead of multienzyme overexpression. First, the expression level of linalool synthase was successfully enhanced by introducing a N-terminal SKIK tag, which improved linalool production by 3.3-fold. Subsequently, the modular assembly of linalool synthase and dominant negative farnesyl diphosphate synthase via short peptide tags efficiently converted geranyl pyrophosphate to linalool. Additional downregulation of the native farnesyl diphosphate synthase led to the highest reported linalool production (80.9 mg/L) in yeast. This combinatorial modulation strategy may also be applied to the production of other high-value monoterpenes.

Directed evolution of the transcription factor Gal4 for development of an improved transcriptional regulation system in Saccharomyces cerevisiae.

As a well-characterized eukaryotic DNA binding transcription factor, Gal4 has been extensively employed to construct controllable gene expression systems in Saccharomyces cerevisiae. The problem of insufficient inducibility arises in the constructs with multiples genes under control of GAL promoters due to the low expression level and activity of the native Gal4. In order to obtain improved transcriptional regulation systems for multi-gene pathways, Gal4 mutants with improved activity were created by directed evolution. During the preliminary screening, five positive Gal4 variants were isolated based on the lycopene-indicated high-throughput screening method. Analysis of the mutation sites revealed that the majority of positive mutations are localized in the middle homology region with unspecified function, suggesting an important role of this domain in transactivation of Gal4. Through combinatorial site-directed mutagenesis, the best variant Gal4T406A/V413A was obtained, which successfully increased the transcription of PGAL-driven lycopene pathway genes and led to 48 % higher lycopene accumulation relative to the wild-type Gal4. This study demonstrates the viability of modifying Gal4 activity by directed evolution for elevated expression of PGAL-driven genes and therefore enhanced production of the target metabolite.

[Correlation Between Water Purification Capacity and Bacterial Community Composition of Different Submerged Macrophytes].

Eight submerged macrophytes are commonly found in subtropical areas, including Vallisneria natans, Vallisneria denseserrulata, Hydrilla verticillata, Elodea canadensis, Ceratophyllum demersum, Potamogeton malaianus, Potamogeton pectinatus, and Potamogeton maackianus, and these eight macrophytes were selected as research objects. The absorption capacity of nitrogen and phosphorus and water purification ability of submerged macrophytes were compared under indoor static water conditions. Furthermore, combining the bacterial community composition of submerged macrophytes, which was determined by 16S rRNA gene sequencing, the correlation between the water purification ability and the bacterial community of submerged macrophytes was determined. The results showed that all of the submerged macrophytes had obvious purification effects on nitrogen and phosphorus in water. The removal of nitrogen and phosphorus by submerged macrophytes was mainly through plant synergism, and the removal rate of plant absorption and enrichment was low. Among them, the removal rate of nitrogen and phosphorus was the highest in Vallisneria denseserrulata, reaching 91.58% and 96.81%. The self-absorption ability of nitrogen and phosphorus from water of Elodea canadensis and Ceratophyllum demersum was higher than other groups. The plant synergistic purification ability of Vallisneria denseserrulata and Vallisneria natans was the highest. The absolute dominant phyla of eight submerged macrophyte-associated bacteria were Proteobacteria (abundance values were more than 40%). At the genus level, Cupriavidus, Rhodobacter, and Gemmatimonas were the dominant genera for different submerged macrophytes. Most of these bacterial groups were degradable, which may be the main reason for the strong ability of eight submerged macrophytes to purify nitrogen and phosphorus in the water. The LEfSe analysis showed that Vallisneria denseserrulata and Vallisneria natans had the highest number of bacteria with significant differences. Among them, Rhizobiales, Burkholderiales, Flavobacteriales, Alcaligenaceae, Cupriavidus, and Bacillales may be the dominant bacteria to enhance the efficiency of plant purification of the water by Vallisneria denseserrulata. The bacteria of Deinococci, Comamonadaceae, Saprospiraceae, and Hyphomicrobium may be the dominant bacteria to enhance the efficiency of plant purification of the water by Vallisneria natans.

[Spatial and Temporal Variability of CO2 Emissions from the Xin'anjiang Reservoir].

Xin'anjiang Reservoir is the largest reservoir in eastern China, with a surface area of 580 km2 and a mean depth of 30 m. It is in an oligotrophic or mesotrophic state at present. This study measured carbon dioxide (CO2) emissions from the upstream river, the reservoir's main body, and the river downstream of the Xin'anjiang Reservoir to investigate the spatial and seasonal variability of CO2 emissions from the water surface using static floating chambers and gas chromatography. Results showed that the downstream river had, significantly, the highest CO2 emission flux[(1535.00±1447.46) mg·(m2·h)-1], followed by the upstream river[(120.39±135.41) mg·(m2·h)-1]. The reservoir's main body had the lowest flux[(36.65-61.94) mg·(m2·h)-1]. The high CO2 emission flux in the downstream river was probably influenced by turbulence during the discharge periods, which would allow the dissolved CO2 in the hypolimnion before the dam to be released to the atmosphere in the watercourse of the downstream river. However, the CO2 emission flux decreased with distance to the dam, likely because of the drop in strength of the turbulence. Moreover, there was an obvious alternation between CO2 source and CO2 sink in the main body of the reservoir, with CO2 sources in autumn and winter and CO2 sinks in spring and summer. The maximum and minimum CO2 emission values occurred in winter and spring, respectively. Such variability in the CO2 emissions was probably influenced by the bloom of alga in spring and summer, because dissolved CO2 in the water was absorbed by the respiration of alga. However, hydrologic conditions were unstable in the upstream river due to a fast water flow, so alga was difficult to bloom there, and a CO2 source was observed throughout the year, except during April and August. The measurement of the flux from the upstream river, main body, and downstream river required a long period for the investigation of greenhouse gas emissions to avoid underestimating the total CO2 emission from a hydroelectric reservoir system.

Direct Fabrication of Functional Ultrathin Single-Crystal Nanowires from Quasi-One-Dimensional van der Waals Crystals.

Micromechanical exfoliation of two-dimensional (2D) van der Waals materials has triggered an explosive interest in 2D material research. The extension of this idea to 1D van der Waals materials, possibly opening a new arena for 1D material research, has not yet been realized. In this paper, we demonstrate that 1D nanowire with sizes as small as six molecular ribbons, can be readily achieved in the Ta2(Pd or Pt)3Se8 system by simple micromechanical exfoliation. Exfoliated Ta2Pd3Se8 nanowires are n-type semiconductors, whereas isostructural Ta2Pt3Se8 nanowires are p-type semiconductors. Both types of nanowires show excellent electrical switching performance as the channel material for a field-effect transistor. Low-temperature transport measurement reveals a defect level inherent to Ta2Pd3Se8 nanowires, which enables the observed electrical switching behavior at high temperature (above 140 K). A functional logic gate consisting of both n-type Ta2Pd3Se8 and p-type Ta2Pt3Se8 field-effect transistors has also been successfully achieved. By taking advantage of the high crystal quality derived from the parent van der Waals bulk compound, our findings about the exfoliated Ta2(Pd or Pt)3Se8 nanowires demonstrate a new pathway to access single-crystal 1D nanostructures for the study of their fundamental properties and the exploration of their applications in electronics, optoelectronics, and energy harvesting.

Nanoscale Inhomogeneous Superconductivity in Fe(Te1-xSex) Probed by Nanostructure Transport.

Among iron-based superconductors, the layered iron chalcogenide Fe(Te1-xSex) is structurally the simplest and has attracted considerable attention. It has been speculated from bulk studies that nanoscale inhomogeneous superconductivity may inherently exist in this system. However, this has not been directly observed from nanoscale transport measurements. In this work, through simple micromechanical exfoliation and high-precision low-energy ion milling thinning, we prepared Fe(Te0.5Se0.5) nanoflakes with various thicknesses and systematically studied the correlation between the thickness and superconducting phase transition. Our result revealed a systematic thickness-dependent evolution of superconducting transition. When the thickness of the Fe(Te0.5Se0.5) flake is reduced to less than the characteristic inhomogeneity length (around 12 nm), both the superconducting current path and the metallicity of the normal state in Fe(Te0.5Se0.5) atomic sheets are suppressed. This observation provides the first transport evidence for the nanoscale inhomogeneous nature of superconductivity in Fe(Te1-xSex).

High performance field-effect transistor based on multilayer tungsten disulfide.

Semiconducting two-dimensional transition metal chalcogenide crystals have been regarded as the promising candidate for the future generation of transistor in modern electronics. However, how to fabricate those crystals into practical devices with acceptable performance still remains as a challenge. Employing tungsten disulfide multilayer thin crystals, we demonstrate that using gold as the only contact metal and choosing appropriate thickness of the crystal, high performance transistor with on/off ratio of 10(8) and mobility up to 234 cm(2) V(-1) s(-1) at room temperature can be realized in a simple device structure. Furthermore, low temperature study revealed that the high performance of our device is caused by the minimized Schottky barrier at the contact and the existence of a shallow impurity level around 80 meV right below the conduction band edge. From the analysis on temperature dependence of field-effect mobility, we conclude that strongly suppressed phonon scattering and relatively low charge impurity density are the key factors leading to the high mobility of our tungsten disulfide devices.

Linking stoichiometric homeostasis of microorganisms with soil phosphorus dynamics in wetlands subjected to microcosm warming.

Soil biogeochemical processes and the ecological stability of wetland ecosystems under global warming scenarios have gained increasing attention worldwide. Changes in the capacity of microorganisms to maintain stoichiometric homeostasis, or relatively stable internal concentrations of elements, may serve as an indicator of alterations to soil biogeochemical processes and their associated ecological feedbacks. In this study, an outdoor computerized microcosm was set up to simulate a warmed (+5°C) climate scenario, using novel, minute-scale temperature manipulation technology. The principle of stoichiometric homeostasis was adopted to illustrate phosphorus (P) biogeochemical cycling coupled with carbon (C) dynamics within the soil-microorganism complex. We hypothesized that enhancing the flux of P from soil to water under warming scenarios is tightly coupled with a decrease in homeostatic regulation ability in wetland ecosystems. Results indicate that experimental warming impaired the ability of stoichiometric homeostasis (H) to regulate biogeochemical processes, enhancing the ecological role of wetland soil as an ecological source for both P and C. The potential P flux from soil to water ranged from 0.11 to 34.51 mg m(-2) d(-1) in the control and 0.07 to 61.26 mg m(-2) d(-1) in the warmed treatment. The synergistic function of C-P acquisition is an important mechanism underlying C∶P stoichiometric balance for soil microorganisms under warming. For both treatment groups, strongly significant (p<0.001) relationships fitting a negative allometric power model with a fractional exponent were found between n-HC∶P (the specialized homeostatic regulation ability as a ratio of soil highly labile organic carbon to dissolved reactive phosphorus in porewater) and potential P flux. Although many factors may affect soil P dynamics, the n-HC∶P term fundamentally reflects the stoichiometric balance or interactions between the energy landscape (i.e., C) and flow of resources (e.g., N and P), and can be a useful ecological tool for assessing potential P flux in ecosystems.