Fine roots are essential for water and nutrient uptake in plants, but little is known about the variation in fine root traits and the underlying mechanisms that drive it. Understanding the responses of fine root function traits to changing environmental conditions and the role of fine root traits as drivers of forest ecosystem processes are critical for informing physiological and ecological theory as well as ecosystem management. We measured morphological and physiological traits of fine roots from six soil layers and three diameter classes in Schrenk's spruce (Picea shrenkiana) forests of the Tianshan mountains, China. We found significant effects of nitrogen addition on these morphological and physiological traits, which varied by soil layer and root diameter. Specifically, specific root length (SRL) was higher in medium N addition group (N2) than in control group (N0). Specific root area (SRA) was higher in the control group (N0) than fertilized groups (N1, N2 and N3). Root tissue density (RTD) was higher in low N addition group (N1) than in the other group. Root dry matter content had no significant difference among four treatment groups. SRL, SRA, and RTD of fine roots in different diameter classes were all significantly different between high N addition (N3) and the control (N0) groups. The physiological characteristics of fine roots showed that soluble sugar (SS), fine root vitality (FRV), and tissue water content (TWC) in different soil layers were higher in the control group than in the fertilized groups. While soluble protein (SP), malondialdehyde (MDA) and free proline (FP) were lower in the control group (N0) than in the fertilized groups. In addition, SS, FRV, SP, TWC, FP, and MDA in all N addition treatments groups were significantly different from the control group. Fine root morphological traits were closely related to physiological traits, and added nitrogen inputs change these correlations. Our study confirms that nitrogen addition has specific effects on the morphological and physiological traits of fine roots of Schrenk's spruce, and the effects of N addition vary according to the amount added.

Fine roots (< 2 mm in diameter) play an important role in belowground ecosystem processes, and their physiological ecology is easily altered by nitrogen deposition. To better understand the response of physiological and ecological processes of fine roots to nitrogen deposition, a manipulation experiment was conducted to investigate the effects of exogenous nitrogen addition (control (0 kg ha^-1 a^-1), low (5 kg ha^-1 a^-1), moderate (10 kg ha^-1 a^-1), and high nitrogen (20 kg ha^-1 a^-1)) on the biomass, morphological characteristics, chemical elements and nonstructural carbohydrates of fine roots in a Picea schrenkiana forest. We found that most fine roots were located in the 0-20 cm of soil layer across all nitrogen treatment groups (42.81-52.09% of the total biomass). Compared with the control, the biomass, specific root length and specific root area of the fine roots increased in the medium nitrogen treatment, whereas the fine roots biomass was lower in the high nitrogen treatment than in the other treatments. In fine roots, nitrogen addition promotes the absorption of nitrogen and phosphorus and their stoichiometric ratio, while reducing the content of nonstructural carbohydrates. The content of nonstructural carbohydrates in the small-diameter roots (< 1 mm in diamter) in each nitrogen treatment group was lower than that in the large-diameter roots. Correlation analysis showed that soil carbon and nitrogen were positively correlated with fine root biomass and specific root length and negatively correlated with the nonstructural carbohydrates. Our findings demonstrate that medium nitrogen addition is conducive to the development of fine root morphology, while excessive nitrogen can suppress the growth of root systems.

Tissue-specific gene expression is thought to be one of the major forces shaping mammalian gene order. A recent study that used whole-genome chromosome conformation assays has shown that the mammalian genome is divided into specific topological domains that are shared between different tissues and organisms. Here, we wanted to assess whether gene expression and regulation are involved in shaping these domains and can be used to classify them. We analyzed gene expression and regulation levels in these domains by using RNA-seq and enhancer-associated ChIP-seq datasets for 17 different mouse tissues. We found 162 domains that are active (high gene expression and regulation) in all 17 tissues. These domains are significantly shorter, contain less repeats, and have more housekeeping genes. In contrast, we found 29 domains that are inactive (low gene expression and regulation) in all analyzed tissues and are significantly longer, have more repeats, and gene deserts. Tissue-specific active domains showed some correlation with tissue-type and gene ontology. Domain temporal gene regulation and expression differences also displayed some gene ontology terms fitting their temporal function. Combined, our results provide a catalog of shared and tissue-specific topological domains and suggest that gene expression and regulation could have a role in shaping them.

Human activities have increased the input of nitrogen (N) to forest ecosystems and have greatly affected litter decomposition and the soil environment. But differences in forests with different nitrogen deposition backgrounds. To better understand the response of litter decomposition and soil environment of N-limited forest to nitrogen deposition. We established an in situ experiment to simulate the effects of N deposition on soil and litter ecosystem processes in a Picea schrenkiana forest in the Tianshan Mountains, China. This study included four N treatments: control (no N addition), low N addition (LN: 5 kg N ha^-1 a^-1), medium N addition (MN: 10 kg N ha^-1 a^-1) and high N addition (HN: 20 kg N ha^-1 a^-1). Our results showed that N addition had a significant effect on litter decomposition and the soil environment. Litter mass loss in the LN treatment and in the MN treatment was significantly higher than that in the control treatment. In contrast, the amount of litter lost in the HN treatment was significantly lower than the other treatments. N application inhibited the degradation of lignin but promoted the breakdown of cellulose. The carbon (C), N, and phosphorus (P) contents of litter did not differ significantly among the treatments, but LN promoted the release of C and P. Our results also showed that soil pH decreased with increasing nitrogen application rates, while soil enzyme activity showed the opposite trend. In addition, the results of redundancy analysis (RDA) and correlation analyses showed that the soil environment was closely related to litter decomposition. Soil enzymes had a positive effect on litter decomposition rates, and N addition amplified these correlations. Our study confirmed that N application had effects on litter decomposition and the soil environment in a N-limited P. schrenkiana forest. LN had a strong positive effect on litter decomposition and the soil environment, while HN was significantly negative. Therefore, increased N deposition may have a negative effect on material cycling of similar forest ecosystems in the near future.

The gene regulatory code and grammar remain largely unknown, precluding our ability to link phenotype to genotype in regulatory sequences. Here, using a massively parallel reporter assay (MPRA) of 209,440 sequences, we examine all possible pair and triplet combinations, permutations and orientations of eighteen liver-associated transcription factor binding sites (TFBS). We find that TFBS orientation and order have a major effect on gene regulatory activity. Corroborating these results with genomic analyses, we find clear human promoter TFBS orientation biases and similar TFBS orientation and order transcriptional effects in an MPRA that tested 164,307 liver candidate regulatory elements. Additionally, by adding TFBS orientation to a model that predicts expression from sequence we improve performance by 7.7%. Collectively, our results show that TFBS orientation and order have a significant effect on gene regulatory activity and need to be considered when analyzing the functional effect of variants on the activity of these sequences.

Real-time monitoring of vital sounds from cardiovascular and respiratory systems via wearable devices together with modern data analysis schemes have the potential to reveal a variety of health conditions. Here, a flexible piezoelectret sensing system is developed to examine audio physiological signals in an unobtrusive manner, including heart, Korotkoff, and breath sounds. A customized electromagnetic shielding structure is designed for precision and high-fidelity measurements and several unique physiological sound patterns related to clinical applications are collected and analyzed. At the left chest location for the heart sounds, the S1 and S2 segments related to cardiac systole and diastole conditions, respectively, are successfully extracted and analyzed with good consistency from those of a commercial medical device. At the upper arm location, recorded Korotkoff sounds are used to characterize the systolic and diastolic blood pressure without a doctor or prior calibration. An Omron blood pressure monitor is used to validate these results. The breath sound detections from the lung/ trachea region are achieved a signal-to-noise ration comparable to those of a medical recorder, BIOPAC, with pattern classification capabilities for the diagnosis of viable respiratory diseases. Finally, a 6×6 sensor array is used to record heart sounds at different locations of the chest area simultaneously, including the Aortic, Pulmonic, Erbs point, Tricuspid, and Mitral regions in the form of mixed data resulting from the physiological activities of four heart valves. These signals are then separated by the independent component analysis algorithm and individual heart sound components from specific heart valves can reveal their instantaneous behaviors for the accurate diagnosis of heart diseases. The combination of these demonstrations illustrate a new class of wearable healthcare detection system for potentially advanced diagnostic schemes.

Adolescent idiopathic scoliosis (AIS), a sideways curvature of the spine, is sexually dimorphic, with increased incidence in females. A genome-wide association study identified a female-specific AIS susceptibility locus near the PAX1 gene. Here, we use mouse enhancer assays, three mouse enhancer knockouts, and subsequent phenotypic analyses to characterize this region. Using mouse enhancer assays, we characterize a sequence, PEC7, which overlaps the AIS-associated variant, and find it to be active in the tail tip and intervertebral disc. Removal of PEC7 or Xe1, a known sclerotome enhancer nearby, or deletion of both sequences lead to a kinky tail phenotype only in the Xe1 and combined (Xe1+PEC7) knockouts, with only the latter showing a female sex dimorphic phenotype. Extensive phenotypic characterization of these mouse lines implicates several differentially expressed genes and estrogen signaling in the sex dimorphic bias. In summary, our work functionally characterizes an AIS-associated locus and dissects the mechanism for its sexual dimorphism.

Adolescent idiopathic scoliosis (AIS), a sideways curvature of the spine, is the most common pediatric musculoskeletal disorder, affecting ~3% of the population worldwide. However, its genetic bases and tissues of origin remain largely unknown. Several genome-wide association studies (GWAS) have implicated nucleotide variants in non-coding sequences that control genes with important roles in cartilage, muscle, bone, connective tissue and intervertebral disks (IVDs) as drivers of AIS susceptibility. Here, we set out to define the expression of AIS-associated genes and active regulatory elements by performing RNA-seq and chromatin immunoprecipitation-sequencing against H3 lysine 27 acetylation in these tissues in mouse and human. Our study highlights genetic pathways involving AIS-associated loci that regulate chondrogenesis, IVD development and connective tissue maintenance and homeostasis. In addition, we identify thousands of putative AIS-associated regulatory elements which may orchestrate tissue-specific expression in musculoskeletal tissues of the spine. Quantification of enhancer activity of several candidate regulatory elements from our study identifies three functional enhancers carrying AIS-associated GWAS SNPs at the ADGRG6 and BNC2 loci. Our findings provide a novel genome-wide catalog of AIS-relevant genes and regulatory elements and aid in the identification of novel targets for AIS causality and treatment.

Degenerative changes of the intervertebral disc (IVD) are a leading cause of disability affecting humans worldwide and has been attributed primarily to trauma and the accumulation of pathology during aging. While genetic defects have also been associated with disc degeneration, the precise mechanisms driving the initiation and progression of disease have remained elusive due to a paucity of genetic animal models. Here, we discuss a novel conditional mouse genetic model of endplate-oriented disc herniations in adult mice. Using conditional mouse genetics, we show increased mechanical stiffness and reveal dysregulation of typical gene expression profiles of the IVD in adhesion G-protein coupled receptor G6 (Adgrg6) mutant mice prior to the onset of endplate-oriented disc herniations in adult mice. We observed increased STAT3 activation prior to IVD defects and go on to demonstrate that treatment of Adgrg6 conditional mutant mice with a small molecule inhibitor of STAT3 activation ameliorates endplate-oriented herniations. These findings establish ADGRG6 and STAT3 as novel regulators of IVD endplate and growth plate integrity in the mouse, and implicate ADGRG6/STAT3 signaling as promising therapeutic targets for endplate-oriented disc degeneration.

Nuclear compartments are thought to play a role in three-dimensional genome organization and gene expression. In mammalian brain, the architecture and dynamics of nuclear compartment-associated genome organization is not known. In this study, we developed Genome Organization using CUT and RUN Technology (GO-CaRT) to map genomic interactions with two nuclear compartments-the nuclear lamina and nuclear speckles-from different regions of the developing mouse, macaque and human brain. Lamina-associated domain (LAD) architecture in cells in vivo is distinct from that of cultured cells, including major differences in LADs previously considered to be cell type invariant. In the mouse and human forebrain, dorsal and ventral neural precursor cells have differences in LAD architecture that correspond to their regional identity. LADs in the human and mouse cortex contain transcriptionally highly active sub-domains characterized by broad depletion of histone-3-lysine-9 dimethylation. Evolutionarily conserved LADs in human, macaque and mouse brain are enriched for transcriptionally active neural genes associated with synapse function. By integrating GO-CaRT maps with genome-wide association study data, we found speckle-associated domains to be enriched for schizophrenia risk loci, indicating a physical relationship between these disease-associated genetic variants and a specific nuclear structure. Our work provides a framework for understanding the relationship between distinct nuclear compartments and genome function in brain development and disease.