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Abstract Background China exhibits a great diversity of ecosystems and abundant cattle resources, with nearly 30 million cattle from 53 indigenous breeds reared in specific geographic regions. To explore the genetic diversity and population structure of Chinese indigenous cattle, a population genetic analysis at both the individual and population levels was conducted and the admixture analysis was performed. We genotyped 572 samples from 20 Chinese indigenous cattle breeds using GeneSeek Genomic Profiler Bovine LD (GGP-LD, 30 K) and downloaded the published data of 77 samples from 4 worldwide commercial breeds genotyped with Illumina BovineSNP50 Beadchip (SNP50, 50 K). Results In principal component analysis (PCA) and neighbour-joining (NJ) tree analysis, samples of the same breeds were grouped together, leading to clear separation from other breeds. And Chinese indigenous cattle were clustered into two groups of southern and northern breeds, originated from Asian Bos indicus lineage and Eurasian Bos taurus lineage, respectively. In STRUCTURE K = 2, a clear transition occurred from the northern breeds to the southern breeds. Additionally, the northern breeds contained a smaller Eurasian taurine (62.5%) descent proportion than that reported previously (more than 90%). In STRUCTURE K = 3, a distinct descent was detected in the southern Chinese breeds, which could reflect a long-term selection history of Chinese indigenous cattle. The results from TreeMix and f3 statistic provided the evidence of an admixture history between southern breeds and northern breeds. Conclusions Consistent with the observed geographical distributions, Chinese indigenous cattle were divided into two genetic clusters, northern indigenous cattle and southern indigenous cattle. Three improved breeds in the northern area also exhibited northern indigenous ancestry. We found that the breeds distributed in the northern China showed more southern lineage introgression than previously reported. Central-located populations appeared to the admixture between southern and northern lineages, and introgression events from European cattle were observed in Luxi Cattle, Qinchuan Cattle and Jinnan Cattle. The study revealed the population structures and levels of admixture pattern among Chinese indigenous cattle, shedding light on the origin and evolutionary history of these breeds.

Abstract Dielectric techniques are fundamental methods for measuring soil water content, and they commonly rely on the conventional laboratory calibration of the dielectric permittivity models between a dielectric constant and water content. As a non‐negligible factor, porosity has been constructed differently in some models as a calibration constant, but the systematic porosity variations during the laboratory model calibration and field applications are not yet well addressed. Based on time‐domain reflectometer laboratory calibration experiments, this study investigated this issue using three preestablished dielectric permittivity models: the Purdue calibration equation (American Society for Testing and Materials model [ASTM]), the complex refractive index model (CRIM), and a piecewise CRIM model (CRIMP). Results demonstrate that a generalized porosity constant used in the calibration would bring in additional structural bias compared with the calibration using variable porosities, and its magnitude varies with the model structure. The deviation of the generalized porosity constant can further amplify the structural bias of ASTM and CRIM for soils with low clay content, but it is insensitive for the soils with high clay content due to the overwhelming role of model structure error. Only the model CRIMP with a “perfect” model structure can effectively cope with the systematic porosity variation and keep a stable built‐in capability for estimating calibration constants from readily available soil data. These findings highlight ignoring porosity variation should not be taken for granted for calibrating and applying the preestablished models.

The human brain shows neuroplastic adaptations caused by motor skill training. Of note, there is little known about the plastic architecture of the whole-brain network in resting state. The purpose of the present study was to detect how motor training affected the density distribution of whole-brain resting-state functional connectivity (FC). Resting-state functional magnetic resonance imaging data was assessed based on a comparison of fast-ball student athletes (SA) and non-athlete healthy controls (NC). The voxel-wise data-driven graph theory approach, global functional connectivity density (gFCD) mapping, was applied. Results showed that the SA group exhibited significantly decreased gFCD in brain regions centered at the left triangular part of the inferior frontal gyrus (IFG), extending to the opercular part of the left IFG and middle frontal gyrus compared to the NC group. In addition, findings suggested the idea of an increased neural efficiency of athletes’ brain regions associated with attentional–motor modulation and executive control. Furthermore, behavioral results showed that in the SA group, faster executive control reaction time relates to smaller gFCD values in the left IFG. These findings suggested that the motor training would decrease the numbers of FC in IFG to accelerate the executive control with high attentional demands and enable SA to rapidly focus the attention to detect the intriguing target.

The effects of land use and fertilization regimes on soil denitrification potential (SDP) are often investigated, but less is known about the responses of SDP to different types of land use with different fertilization regimes in soils derived from the same parent material. We conducted a study using soil samples from 2 long-term (over 30 years) fertilization experiments to determine the difference in SDP between paddy soil and upland soil derived from the same quaternary red clay parent material. The results showed that the SDP in paddy soil was 6.82 times higher than the upland soil, which was due to the higher abundances of narG, nirS, and nirK genes and nirS-denitrifying bacteria (Bradyrhizobium, Cupriavidus, and Herbaspirillum) in paddy soil. Inorganic fertilization regimes did not significantly affect the SDP of upland soil over the control group, whereas SDP in the NPK and 2NPK treatments in paddy soil were reduced relative the control group by 26.48% and 75.65%, respectively. Compared with the control, the NPKOM treatment consistently yielded the highest SDP in both soils, with 2.47 times and 2.86 times higher for paddy soil and upland soil, respectively. The SDP of paddy soil was significantly correlated with narG and nirS genes mainly regulated by amorphous aluminum oxide, whereas the treatment effect on SDP for upland soil largely depended on differences in nirS-denitrifying bacteria at the genus level (Herbaspirillum, Sulfuritalea, and Cupriavidus) and species level, which were mainly controlled by soil pH. Partial least squares path models further demonstrated that the direct effects of functional genes on SDP were greatest for paddy soil, whereas nirS-denitrifying bacterial communities played a larger role in upland soil. The results presented herein represent a key step toward understanding the mechanisms that govern SDP under land use and long-term fertilization.

BACKGROUND AND AIMS:This paper was primarily devoted to understand the interactions of soil aggregates, organic carbon (C) and carbon cycle enzymes in aggregates under different fertilization managements, aiming to identify the effects of organic and inorganic fertilizer amendments on soil organic C accumulation and the activities of carbon cycle enzymes within aggregates in Vertisol. METHODS:A Vertisol soil following 4-year compost and inorganic fertilizer amendments, i.e. no fertilizer (CK), mineral fertilizer (FR) and 60% compost N plus 40% fertilizer N (FRM), was collected to identify the dynamics of organic C, enzymes activities and their associations with macroaggregation using aggregate fractionation techniques. RESULTS:The organic C content in all FR and FRM treatments was 8.24-41.15% higher than that in CK. An increased amounts of carbon cycle enzymes in aggregates or 0-20 cm bulk soil were also observed in FRM plots. Compared to FR, FRM significantly strengthened the structural stability of macroaggregates and the intimate connection between enzyme activities and macroaggregates. CONCLUSIONS:As a recommended measure, supplementation with organic manure such as compost strengthened the process of mutual promotion between carbon cycle enzymes and macroaggregates, and the synergistic effect would be highly beneficial to soil organic C sequestration.

A 5-year (2012-2016) field experiment was conducted to investigate the impacts of different fertilizer treatments (no fertilizer, mineral and organic fertilizer) on organic carbon and soil water-stable aggregates in a North China Plain Vertisol. Compared with no fertilizer (control), single mineral fertilizer did not significantly (P < 0.01) affect organic carbon content or aggregate mass proportion in bulk soil. Small and large macroaggregate mass proportions increased, but applying organic manure significantly decreased the silt + clay fraction and microaggregates. Organic manure amendment significantly enhanced organic carbon concentrations in aggregates (large macroaggregates, > 2000 μm; small macroaggregates, 2000-250 μm; microaggregates, 53-250 μm; and free silt + clay fraction, < 53 μm) and aggregate subfractions, including intraparticulate organic matter and silt + clay subfractions (< 53 μm). Single mineral fertilizer amendment increased organic carbon concentrations in macroaggregates, particularly intraparticulate organic matter. The results indicated that the organic carbon increase in organic manure-amended soil were possibly due to enhanced silt + clay subfractions, which then promoted macroaggregates formation. Applying organic manure could improve organic carbon sequestration and maintain its stability in aggregates, whereas mineral fertilizer only enhanced organic carbon in large macroaggregates, but with low stability.

In order to solve some problems of lower blastocyst rate of early embryos in vitro and poor quality of blastocyst, application of a microfluidic chip with mechanical stimuli (MS) to imitate microenvironment of fallopian tube, to investigate the effect of a microfluidic chip with mechanical stimuli on the early embryo development in mammalian. We designed and made a novel and simple system to apply MS similar to those generated inside the oviduct to cultured mammalian embryos by using poly(dimethylsiloxiane). Using this microfluidic chip embryo culture device to culture 2-cell stage mouse embryo from in vitro fertilization. There were large differences in development rates of > 4-cell stage, development rates of morula (63.1 ± 12.5% vs. 80.6 ± 17.9%) and blastocyst (58.6 ± 2.3% vs. 73.2 ± 2.8%) between control and microfluidic chip group (p > 0.05). meanwhile, there was large differences in cell numbers of inner cell mass in blastocyst between control and microfluidic chip group (p > 0.05). These results suggested that the culture device can significantly improve blastocyst development rate of embryo in vitro and the quality of the blastocyst in mice, this novel and simple system was feasible to generate and apply MS similar to those generated inside the oviduct to cultured mammalian embryos

Background/Aims: Bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9), which are secreted by oocytes, are important regulators of follicular growth and development and ovarian function. These two factors can regulate the proliferation and apoptosis of cumulus cells via modulation of the Smad signaling pathway. Studies have shown that BMP15 and GDF9 can affect the level of miR-375, whereas the target gene of miR-375 is BMPR2, the type II receptor of BMP15 and GDF9. However, whether or how the BMP15/ GDF9-miR-375-BMPR2 pathway affects the proliferation and apoptosis of bovine cumulus cells through regulation of the Smad signaling pathway remains unclear. Methods: In this study, cumulus cells were first obtained from cumulus-oocyte complexes (COCs). Appropriate concentrations of BMP15 and GDF9 were added during the in vitro culture process. Cell Counting Kit-8 (CCK-8) analyses and flow cytometry were used to determine the effects of BMP15/GDF9 on bovine cumulus cells proliferation and apoptosis. Subsequently, miR-375 mimics, miR-375 inhibitor and BMPR2 siRNA were synthesized and used for transfection experiments. Western Blot analysis was used to detect changes before and after transfection in the expression levels of the BMP15/GDF9 type I receptors ALK4, ALK5 and ALK6; the phosphorylation levels of Smad2/3 and Smad1/5/8, which are key signaling pathway proteins downstream of BMP15/GDF9; the expression levels of PTX3, HAS2 and PTGS2, which are key genes involved in cumulus cells proliferation; and Bcl2/Bax, which are genes involved in apoptosis. Results: The addition of 100 ng/mL BMP15 or 200 ng/mL GDF9 or the combined addition of 50 ng/mL BMP15 and 100 ng/mL GDF9 effectively inhibited bovine cumulus cell apoptosis and promoted cell proliferation. BMP15/GDF9 negatively regulated miR-375 expression and positively regulated BMPR2 expression. High levels of miR-375 and inhibition of BMPR2 resulted in increased expression of ALK4 and decreased expression of PTX3, HAS2 and PTGS2, whereas miR-375 inhibition resulted in the opposite results. BMP15 and GDF9 significantly activated the levels of p-Smad2/3 and p-Smad1/5/8, whereas miR-375 inhibited the levels of p-Smad2/3 and p-Smad1/5/8 by negatively regulating BMPR2 and also led to apoptosis. Conclusion: BMP15 and GDF9 have synergistic effects and can act through miR-375 to affect the expression levels of type I receptor ALK4 and type II receptor BMPR2 and the activation of Smad signaling pathway, which subsequently affected the proliferation, spread and apoptosis of cumulus cells.

m6A is one of the most common and abundant modifications of RNA molecules present in eukaryotes. The methyltransferase complex, consisting of methyltransferase-like 3 (METTL3), METTL14, and WTAP, is responsible for the m6A modification of RNA. WTAP was identified as an mRNA splicing regulator. Its role as a regulatory subunit of the m6A methyltransferase complex in embryonic development remains largely unknown. To investigate the role of WTAP in porcine early embryonic development, si-WTAP was microinjected into porcine parthenogenetic zygotes. WTAP knockdown significantly reduced the blastocyst rate and global m6A levels, but did not affect the cleavage rate. Betaine was supplemented into the in vitro culture (IVC) to increase the m6A levels. Betaine significantly increased the global m6A levels but did not affect the blastocyst rate. Furthermore, the pluripotency genes, including OCT4, SOX2, and NANOG, were downregulated following WTAP knockdown. The apoptotic genes BAX and CASPASE 3 were upregulated, while the anti-apoptotic gene BCL2 was downregulated in WTAP knockdown blastocysts. TUNEL staining revealed that the number of apoptotic cells was significantly increased following WTAP knockdown. Our study indicated that WTAP has an indispensable role in porcine early embryonic development.

Background: Bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9) are members of the transforming growth factor beta (TGF-β) superfamily. Through autocrine and paracrine mechanisms, these two factors can regulate cell differentiation, proliferation, and other functions in the ovary locally. Furthermore, GDF9 and BMP15 play vital roles in follicular growth, atresia, ovulation, fertilization, reproduction, and maintenance. Numerous studies have demonstrated a synergy between BMP15 and GDF9. Studies in humans and mice have indicated that the synergy between BMP15 and GDF9 is primarily mediated by the bone morphogenetic protein type II receptor (BMPR2). The BMP15/GDF9 heterodimer needs to bind to the BMPR2-ALK4/5/7-ALK6 receptor complex to activate the SMAD2/3 signaling pathway. However, it is not clear which genes mediate and regulate the effects of the BMP15/GDF9 proteins on bovine cumulus cells (CCs). Methods: Our earlier study showed that BMPR2 is a gene that is directly targeted and regulated by miR-375. Therefore, we designed and synthesized an miR-375 mimics/inhibitor and regulated BMPR2 expression in bovine CCs by the overexpression or inhibition of miR-375. After the overexpression or inhibition of miR-375, the apoptosis rate of bovine CCs was measured by flow cytometry; changes in critical gene expression were measured by RT-qPCR and western blot assays; and the proliferation of bovine CCs was measured by CCK-8 assay. Results: In bovine CCs, the overexpression of miR-375 resulted in decreased BMPR2 and ALK7 expression, whereas the inhibition of miR-375 caused increased BMPR2 and ALK7 expression. The overexpression of miR-375 attenuated the proliferation ability and significantly increased the apoptosis rate of bovine CCs, whereas the inhibition of miR-375 did not significantly change the proliferation ability or apoptosis rate. Conclusions: BMPR2, a target of miR-375, is regulated by this molecule, thereby affecting expression of BMP15/GDF9 receptors, and the proliferation and apoptosis of bovine CCs.

Aims: This study aims to clarify the effects of polyadenylation status on M-phase promoting factors (MPFs) during in vitro porcine oocyte maturation. Methods: In this study, porcine follicular oocytes from large follicles (> 5 millimeter (mm)) and small follicles (Cyclin B1 and cell division cycle 2 (Cdc2) were determined by real-time quantitative PCR. Immunofluorescence was used to assess spindle formation and chromosome alignment in the examined oocytes. Results: In large-follicle oocytes, the effects of inhibiting polyadenylation caused the percentage of mature to be significantly lower for the treated group than for the untreated group (p in vitro and inhibits Cdc2 polyadenylation. Cyclin B1 plays a significant role in promoting the maturation of large-follicle oocytes. Polyadenylation contributes to the formation of dominant follicles and facilitates the selection of dominant follicles. However, the inhibition of adenylation affected spindle formation-related propulsion and chromosome alignment in both large- and small-follicle oocytes. The first polar body could not be extruded in certain large follicles. Conclusions: 3'-da can significantly improve the rate of small oocyte maturation in vitro, but it can also affect spindle formation-related propulsion and chromosome alignment.

Spatial variability and influential factors are important to evaluate soil organic C (SOC) and the C pool in large areas. In the present study, sampling was conducted from May to November 2011 in Henan Province, a typical agricultural region of Central China, to study the effects of soil properties and anthropogenic factors on SOC variability in cropland. Physicochemical properties of soil samples collected at 280 sites from the surface layer (at a depth of 0–20 cm) were analyzed, and related data about the sampling sites were also collected from the Second State Soil Survey of China, conducted in 1981. Firstly, we found increasing trends in SOC density (SOCD) and SOC pool (SOCP) were obvious from 1981 to 2011, and we concluded that cropland presents great C sequestration potential for the future. Carbon pool ability varied with soil properties: the order of fixed Camount in different soil types was found to be Inceptisols > Luvisols > semihydromorphic soil > Anthrosols, and the average SOCP increased significantly from 1981 to 2011. Secondly, soil bulk density, pH, and returning straw were the key influential factors for SOCD in the past 30 yr. Thirdly, although random factors (returning straw) only explain 29.1% of SOCD variability, more attention should be paid to the factors, because application of returning straw was the most dominant anthropogenic factor, which can be used to improve cropland productivity and C sink capacity within a short period if they are properly managed in the future.

Potassium (K) distribution is horizontally heterogeneous under the conservation agriculture approach of no-till with strip fertilization. The root foraging strategy of wheat for K heterogeneity is poorly understood. In this study, WinRHIZO, microarray, Non-invasive Micro-test Technology (NMT) and a split-root system were performed to investigate root morphology, gene expression profiling and fluxes of K+ and O2 under K heterogeneity and homogeneity conditions. The split-root system was performed as follows: C. LK (both compartments had low K), C. NK (both compartments had normal K), Sp. LK (one compartment had low K) and Sp. NK (the other compartment had normal K). The ratio of total root length and root tips in Sp. NK was significantly higher than that in C. NK, while no significant differences were found between Sp. LK and C. LK. Differential expression genes in C. LK vs. C. NK had opposite responses in Sp. LK vs. C. LK and similar responses in Sp. NK vs. C. NK. Low-K responsive genes, such as peroxidases, mitochondrion, transcription factor activity, calcium ion binding, glutathione transferase and cellular respiration genes were found to be up-regulated in Sp. NK. However, methyltransferase activity, protein amino acid phosphorylation, potassium ion transport, and protein kinase activity genes were found to be down-regulated in Sp. LK. The up-regulated gene with function in respiration tended to increase K+ uptake through improving O2 influx on the root surface in Sp. NK, while the down-regulated genes with functions of K+ and O2 transport tended to reduce K+ uptake on the root surface in Sp. LK. To summarize, wheat roots tended to perform active-foraging strategies in Sp. NK and dormant-foraging strategies in Sp. LK through the following patterns: (1) root development in Sp. NK but not in Sp. LK; (2) low-K responsive genes, such as peroxidases, mitochondrion, transcription factor activity, calcium ion binding and respiration, were up-regulated in Sp. NK but not in Sp. LK; and (3) root K+ and O2 influxes increased in Sp. NK but not in Sp. LK. Our findings may better explain the optimal root foraging strategy for wheat grown with heterogeneous K distribution in the root zone.

Vehicles are developing in the direction of energy-saving and electrification. suspension has been widely developed in the field of vehicles as a key component. Traditional hydraulic energy-supply suspensions dissipate vibration energy as waste heat to suppress vibration. This part of the energy is mainly generated by the vehicle engine. In order to effectively utilize the energy of this part, the energy-regenerative suspension with energy recovery converts the vibrational energy into electrical energy as the vehicle’s energy supply equipment. This article reviews the hydraulically powered suspension of vehicles with energy recovery. The importance of such suspension in vehicle energy recovery is analyzed. The main categories of energy-regenerative suspension are illustrated from different energy recovery methods, and the research status of hydraulic energy-regenerative suspension is comprehensively analyzed. Important factors that affect the shock-absorbing and regenerative characteristics of the suspension system are studied. In addition, some unresolved challenges are also proposed, which provides a reference value for the development of energy-regenerative suspension systems for hybrid new energy vehicles

Although increasing evidences have emerged for responses of soil microorganisms to fertilizations, the knowledge regarding community assemblages that cause variations in composition is still lacking, as well as the possible feedback to soil fertility. Phylogenetic conservatism of species indicates their similar environmental preferences and/or function traits and phylogenetic signals further can infer community assemblages and influenced ecological processes. Here, we calculated the mean pairwise phylogenetic distance and nearest relative index, characterizing phylogenetic signal and the undergone ecological process to evaluate the community assembly of soil bacterial phylotypes in 20-year fertilized soils. The bacterial community assembly is structured by environmental filtering, regardless of fertilization regime. Soil phosphorous (P) availability imposes selection on community assemblage and influences their community turnover among fertilizations. When P nutrient lacks, the effect of environmental filtering becomes stronger, hence bacterial functional traits become more coherent; this process results into increased intraspecific interactions characterized by co-occurrence network analysis. In contrast, when P nutrient becomes abundant, the environmental selection is mitigated; function traits are evened. This process reduces intraspecific interactions and increases carbon sequestration efficiency, which is finally of great favor to the increases in soil fertility. This study has made the first attempt, at the bacterial level, to understand how fertilization affects agroecosystems. When more phylogenetic information on how nutrient cycling-related microbes respond to fertilization becomes available, the systematic knowledge will eventually provide guidance to optimal fertilization strategies.

Recent experimental results showed that the Tg of cured resin scarcely decreased and the impact strength of resins increased by over 50% when a tetrafunctional epoxy named N,N,N',N'-tetraglycidyl-4,4'-diaminodiphenyl ether (TGDDE) was introduced to an appropriate flexible chain from a dimer fatty acid (DFA). In order to understand the reason for this phenomenon, the modification and the chemical structure of the prepolymer together with the curing reaction and the viscoelasticity of the cured resins were studied in detail in the present work. The results indicated that the modification would help the prepolymer improve its molecular mobility. As a result, the resins could be further cured, resulting in the cross-linking density increasing. This is because the curing efficiency was increased, but the tetrafunctional epoxy was not cured completely due to its large steric hindrance. Moreover, the flexibility of some parts of the networks was improved, which was beneficial for the toughness of the cured resins. Therefore, the toughness of the tetrafunctional resin was improved with little influence on the thermal properties when the epoxies were modified with an appropriate content of DFA.

Aims: Our study aims to clarify the effects of demecolcine, alone or in combination with sucrose on bovine oocyte protrusion rate, MAPK1 protein level and c-mos gene expression level. Methods: The effects of the demecolcine concentration, treatment duration, and synergistic effects with sucrose solution on the rate of membrane protrusions of bovine oocytes were investigated. Using real-time fluorescent quantitative PCR, western blot analysis, and immunofluorescence assays, the expression of the maternal c-mos gene, the protein level of mitogen-activated protein kinase 1 (MAPK1), and the change in the localization of spindles and nuclei during the demecolcine treatment were analyzed in bovine oocytes. Results: Treatment of bovine oocytes with both demecolcine (0.6 μg/mL) and sucrose (0.05 M) for 1 h led to the highest rate of membrane protrusions, and synergistic effects were also observed. Real-time fluorescent quantitative PCR analysis revealed that the demecolcine treatment up-regulated the expression of the maternal c-mos gene. Western blot analysis indicated that the demecolcine treatment enhanced the protein level of MAPK1 in bovine oocytes. Immunofluorescence analysis indicated that the spindles and nuclei were localized at the place of the membrane protrusions. Conclusions: The present results suggest that demecolcine might contribute to the activation of the Mos/MAPK pathway and affect spindle structure. These results provide a reference for more efficient generation of enucleated bovine oocytes.

Pot experiments are a low-cost and easy-to-use technique for studies of soil evaporation and plant transpiration in controlled environments. However, little attention has been paid to the applicability of evapotranspiration (ET) measured in pot experiments to the field. The objective of this study was to determine whether a pot experiment can be used for measuring field ET. Evapotranspiration experiments with winter wheat and summer maize were conducted in pots and lysimeters under various water-deficit conditions. The measured ET values in the pot experiments under different water conditions were considerably different from those of the lysimeters. Causes of such differences in ET were analyzed, and a series of corrections were proposed to eliminate the effects of different crop densities, representative areas per plant, and soil moisture conditions on pot experiment results. After these corrections, the discrepancy in the total ET of wheat-maize seasons between pots and lysimeters was greatly reduced from a maximum of 117% to only approximately 10%. The relative mean square errors (RMSEs) for daily ET values also decreased from a maximum value of 4.56 mm to less than 1.5 mm for the wheat season and from a maximum value of 6.02 mm to approximately 2 mm for the maize season. Possible measures were proposed to further improve the accuracy of the corrected ET obtained from pot experiments. In sum, pot experiments can serve as a feasible tool for estimating ET in the field just with a few routine measurements at regions where large-scale weighing lysimeters, an eddy covariance device, and even meteorological data are not available. The proposed corrections can also be used for upscaling small-scale ET measurements to a large scale.

Background: Bone morphogenetic protein-15 (BMP15) is a maternal gene necessary for mammalian reproduction. BMP15 expression increased in oocytes accompanied by follicle growth and development. The function and regulation mechanism of BMP15 in porcine cumulus cell apoptosis process is still unclear now. Methods: In this study, flow cytometry (FCM) was used to analyze the effects of BMP15 with different concentrations to cumulus cell apoptosis. High-throughput sequencing technology was carried out to screen regulatory genes linked closely with BMP15. In order to confirm the function of (MCP-1)/CCL2 and FBN1 in cumulus cell apoptosis, RNA interference (RNAi) method was used to inhibit the expression of (MCP-1)/CCL2 and FBN1. Apoptosis and proliferation of cumulus cell treated with siRNA transfection technology were measured by FCM, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, quantitative real time-PCR (RT-qPCR) and western blotting. Results: The results showed that the apoptosis levels of cumulus cell treated by BMP15 decreased significantly in a dose-dependent manner. The expression of related genes protein 1 (MCP-1)/CCL2 and fibrillin1 (FBN1) were both regulated by BMP15. After transfection, the proliferation of porcine cumulus cells increased significantly and apoptosis of cumulus cells was prevented while FBN1 was silenced after BMP15 treatment. The proliferation of cumulus cells decreased significantly and apoptosis rate of cumulus cells increased significantly while CCL2 was silenced. Conclusion: The results obtained in this study firstly demonstrated that CCL2 and FBN1 are important regulatory factors of BMP15 in preventing cumulus cell apoptosis in porcine ovaries.

Aims: Our study aims to characterize the functions of the ADIPOQ gene in the process of fat deposition of pigs, thereby providing a basis for the use of this gene as a molecular marker for pork quality. Methods: We used healthy Junmu1 piglets less than 7 days of age to establish an in vitro culture system for porcine preadipocytes. Chemically synthesized short hairpin RNAs (shRNA) were transfected into porcine preadipocytes to silence the expression of the ADIPOQ gene. We monitored preadipocyte differentiation and determined the levels of the adipocyte differentiation transcription factors lipoprotein lipase (LPL), peroxisome proliferator-activated receptor γ (PPARγ) and adipocyte fatty acid binding protein (AP2) mRNAs to investigate the effects of ADIPOQ on the differentiation of porcine preadipocytes. Results: After transfection, the mRNA and protein levels of the ADIPOQ gene were significantly decreased (P Conclusions: These results suggest that interference with ADIPOQ gene expression can inhibit the differentiation of porcine preadipocytes.

Chemical composition of soil organic carbon (SOC) is central to soil fertility. We hypothesize that change in SOC content resulting from various long-term fertilization strategies accompanies the shift in SOC chemical structure. This study examined the effect of fertilization strategies along with the time of fertilizer application on the SOC composition by 13C nuclear magnetic resonance (NMR) spectroscopy. The soils (Aquic Inceptisol) subjected to seven fertilizer treatments were collected in 1989, 1999 and 2009, representing 0, 10 and 20 years of fertilization, respectively. The seven fertilizer treatments were (1-3) balanced fertilization with application of nitrogen (N), phosphorus (P) and potassium (K) including organic compost (OM), half organic compost plus half chemical fertilizer (1/2OM), and pure chemical NPK fertilizer (NPK); (4-6) unbalanced chemical fertilization without application of one of the major elements including NP fertilizer (NP), PK fertilizer (PK), and NK fertilizer (NK); and (7) an unamended control (CK). The SOC content in the balanced fertilization treatments were 2.3-52.6% and 9.4-64.6% higher than in the unbalanced fertilization/CK treatments in 1999 and 2009, respectively, indicating significant differences in SOC content with time of fertilizer application between the two treatment groups. There was a significantly greater proportion of O-alkyl C and a lower proportion of aromatic C in the balanced fertilization than in unbalanced fertilization/CK treatments in 1999, but not in 2009, because their proportions in the former treatments approached the latter in 2009. Principal component analysis further showed that the C functional groups from various fertilization strategies tended to become compositionally similar with time. The results suggest that a shift in SOC chemical composition may be firstly dominated by fertilization strategies, followed by fertilization duration.

BACKGROUND: One important protein family that functions in nucleotide excision repair (NER) factors is the SNF2 family. A newly identified mouse ERCC6-like gene, Ercc6l (excision repair cross-complementing rodent repair deficiency, complementation group 6-like), has been shown to be another developmentally related member of the SNF2 family. METHODOLOGY/PRINCIPAL FINDINGS: In this study, Sika deer Ercc6l cDNA was first cloned and then sequenced. The full-length cDNA of the Sika deer Ercc6l gene is 4197 bp and contains a 3732 bp open reading frame that encodes a putative protein of 1243 amino acids. The similarity of Sika deer Ercc6l to Bos taurus Ercc6l is 94.05% at the amino acid sequence level. The similarity, however, is reduced to 68.42-82.21% when compared to Ercc6l orthologs in other mammals and to less than 50% compared to orthologs in Gallus gallus and Xenopus. Additionally, the expression of Ercc6l mRNA was investigated in the organs of fetal and adult Sika deer (FSD and ASD, respectively) by quantitative RT-PCR. The common expression level of Ercc6l mRNA in the heart, liver, spleen, lung, kidney, and stomach from six different developmental stages of 18 Sika deer were examined, though the expression levels in each organ varied among individual Sika deer. During development, there was a slight trend toward decreased Ercc61 mRNA expression. The highest Ercc6l expression levels were seen at 3 months old in every organ and showed the highest level of detection in the spleen of FSD. The lowest Ercc6l expression levels were seen at 3 years old. CONCLUSIONS/SIGNIFICANCE: We are the first to successfully clone Sika deer Ercc6l mRNA. Ercc6l transcript is present in almost every organ. During Sika deer development, there is a slight trend toward decreased Ercc61 mRNA expression. It is possible that Ercc6l has other roles in embryonic development and in maintaining the growth of animals.

ABSTRACT In this study, our goal was to clarify the role of proteasomal subunit α-1 (PSMA1) in both the differentiation of preadipocytes and the accumulation of lipids in adipocytes. Preadipocytes from healthy one-day-old calves were collected, isolated, and cultured in vitro. The expression pattern of the PSMA1 gene was explored during the differentiation of bovine preadipocytes firstly. Then, the expression of the PSMA1 gene was inhibited by transfection of a chemically synthesized small interfering RNA (siRNA) before differentiation. After induction of differentiation, the mRNA levels of key regulating genes involved in preadipocyte differentiation and the lipid content of mature adipocytes with and without inhibition of PSMA1 were detected by qRT-PCR and oil red O staining, respectively. The data showed that PSMA1 mRNA was differentially expressed during the differentiation of bovine preadipocytes under normal culture conditions in vitro. The expression level of peroxisome proliferator-activated receptor gamma (PPARγ), CCAAT enhancer-binding protein alpha (C/EBPα), and lipoprotein lipase (LPL) were significantly decreased in the transfected PSMA1-siRNA group compared with those in the control group, and the mRNA levels of the preadipocyte factor-1 (Pref-1) were significantly upregulated in the transfected PSMA1-siRNA group compared with those in the control group. In addition, significantly fewer lipid droplets were formed by adipocytes transfected with PSMA1-siRNA than by the negative control group (adipocytes transfected with NC-siRNA). Therefore, PSMA1 plays an important role in differentiation and lipid deposition.

Introduction: Ammonia oxidizing archaea (AOA) and ammonia oxidizing bacteria (AOB) have been proven to be key microorganisms driving the ammonia oxidation process. However, under different fertilization practices, there is a lack of research on the impact of interaction between predators and AOA or AOB on nitrogen cycling at the multi-trophic level. Methods: In this study, a network-oriented microscopic culture experiment was established based on four different long-term fertilization practices soils. We used the nitrification inhibitors 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxide-3-oxyl (PTIO) and 3, 4-Dimethylpyrazole phosphate (DMPP) inhibited AOA and AOB, respectively, to explore the impact of interaction between protists and AOA or AOB on nitrogen transformation. Results: The results showed that long-term nitrogen application promoted the potential nitrification rate (PNR) and nitrous oxide (N2O) emission, and significantly increased the gene abundance of AOB, but had no obvious effect on AOA gene abundance. DMPP significantly reduced N2O emission and PNR, while PTIO had no obvious effect on them. Accordingly, in the multi-trophic microbial network, Cercozoa and Proteobacteria were identified as keystone taxa of protists and AOB, respectively, and were significantly positively correlated with N2O, PNR and nitrate nitrogen. However, Nitrososphaerota archaeon as the keystone species of AOA, had an obvious negative linkage to these indicators. The structural equation model (SEM) showed that AOA and AOB may be competitors to each other. Protists may promote AOB diversity through direct trophic interaction with AOA. Conclusion: The interaction pattern between protists and ammonia-oxidizing microorganisms significantly affects potential nitrification rate and N2O emission, which has important implications for soil nitrogen cycle. [ABSTRACT FROM AUTHOR]