Millimeter-wave over fiber integrated sensing and communication system using self-coherent OFDM.

Orthogonal frequency-division multiplexing (OFDM) waveform is highly preferred as a dual-function candidate for integrated sensing and communication (ISAC) systems. However, the sensitivity to both carrier frequency offset (CFO) and phase noise greatly impedes its applications in millimeter-wave ISAC systems. Here, we propose and experimentally demonstrate a photonic millimeter-wave ISAC system employing the virtual-carrier-aided self-coherent OFDM technique, wherein a digitally-generated local oscillator is transmitted along with the OFDM signal. Then, a compact CFO-immune and phase noise-immune envelope detection method is implemented for down-converting millimeter-wave communication and radar echo signals. In experiments, a V-band ISAC system is successfully implemented with a simplified remote radio unit, using the remote photonic millimeter-wave heterodyning up-conversion for downlink and the envelope detection-assisted down-conversion for uplink (or radar echoes). In the converged transmission link with a 5-km fiber link and 2-m space link, the Kramers-Kronig (KK) receiver supports a communication data rate up to 16-Gbit/s by mitigating signal-signal beat interference (SSBI). More significantly, the SSBI leads to negligible effects on the sensing performance when classic matched filtering is adopted for target identification. Consequently, a 4.8-cm range resolution and a 4-mm range accuracy are obtained for the radar sensing function.

Alginate Oligosaccharides Enhance Antioxidant Status and Intestinal Health by Modulating the Gut Microbiota in Weaned Piglets.

Alginate oligosaccharides (AOSs), which are an attractive feed additive for animal production, exhibit pleiotropic bioactivities. In the present study, we investigated graded doses of AOS-mediated alterations in the physiological responses of piglets by determining the intestinal architecture, barrier function, and microbiota. A total of 144 weaned piglets were allocated into four dietary treatments in a completely random design, which included a control diet (CON) and three treated diets formulated with 250 mg/kg (AOS250), 500 mg/kg (AOS500), and 1000 mg/kg AOS (AOS1000), respectively. The trial was carried out for 28 days. Our results showed that AOS treatment reinforced the intestinal barrier function by increasing the ileal villus height, density, and fold, as well as the expression of tight junction proteins, especially at the dose of 500 mg/kg AOS. Meanwhile, supplementations with AOSs showed positive effects on enhancing antioxidant capacity and alleviating intestinal inflammation by elevating the levels of antioxidant enzymes and inhibiting excessive inflammatory cytokines. The DESeq2 analysis showed that AOS supplementation inhibited the growth of harmful bacteria Helicobacter and Escherichia_Shigella and enhanced the relative abundance of Faecalibacterium and Veillonella. Collectively, these findings suggested that AOSs have beneficial effects on growth performance, antioxidant capacity, and gut health in piglets.

Experimental demonstration of 201.6-Gbit/s coherent probabilistic shaping QAM transmission with quantum noise stream cipher over a 1200-km standard single mode fiber.

A probabilistic shaping (PS) quadrature amplitude modulation (QAM) based on Y-00 quantum noise stream cipher (QNSC) has been proposed. We experimentally demonstrated this scheme with data rate of 201.6Gbit/s over a 1200-km standard single mode fiber (SSMF) under a 20% SD-FEC threshold. Accounting for the 20% FEC and 6.25% pilot overhead, the achieved net data rate is ∼160Gbit/s. In the proposed scheme, a mathematical cipher (Y-00 protocol) is utilized to convert the original low-order modulation PS-16 (22 × 22) QAM into ultra-dense high-order modulation PS-65536 (28 × 28) QAM. Then, the physical randomness of quantum (shot) noise at photodetection and amplified spontaneous emission (ASE) noise from optical amplifiers are employed to mask the encrypted ultra-dense high-order signal for further improving the security. We further analyze the security performance by two metrics known in the reported QNSC systems, namely the number of masked signals (NMS) of noise and the detection failure probability (DFP). Experimental results show it is difficult or even impossible to extract transmission signals from quantum or ASE noise for an eavesdropper (Eve). We believe that the proposed PS-QAM/QNSC secure transmission scheme has the potential to be compatible with existing high-speed long-distance optical fiber communication systems.

Suppressive role of lovastatin in intracerebral hemorrhage through repression of autophagy.

Statins possess critical function in the brain. Here, we intended to investigate the role of lovastatin in brain damage after intracerebral hemorrhage (ICH). A collagenase-induced ICH rat model was established followed by lovastatin treatment. Then, the effect of lovastatin on ICH-induced brain damage was explored with cognitive function, learning and memory abilities, and neurological damage of rats analyzed. Besides, brain water content, number of degenerate neurons, Nissl's body, and apoptosis of neurons were detected. Oxidative stress levels, inflammation, and autophagy levels in ICH were measured after treatment of lovastatin. Lovastatin improved the cognitive impairment of rats, enhanced their spatial learning and memory abilities, reduced nervous system damage, lesion area, and brain water content after ICH. Lovastatin was capable of reducing the number of degenerated neurons, the apoptosis level, autophagy level, and increasing the number of Nissl's body. Lovastatin inhibited the oxidative stress response and inflammatory factors in the brain tissue after ICH, and increased the expression of anti-inflammatory factor IL-10. Lovastatin inhibited AMPK/mTOR signaling pathway after ICH. Our study highlighted the suppressive role of lovastatin in ICH-induced brain damage.

New Insights of Biological Functions of Natural Polyphenols in Inflammatory Intestinal Diseases.

The intestine is critically crucial for nutrient absorption and host defense against exogenous stimuli. Inflammation-related intestinal diseases, including enteritis, inflammatory bowel disease (IBD), and colorectal cancer (CRC), are heavy burdens for human beings due to their high incidence and devastating clinical symptoms. Current studies have confirmed that inflammatory responses, along with oxidative stress and dysbiosis as critical pathogenesis, are involved in most intestinal diseases. Polyphenols are secondary metabolites derived from plants, which possess convincible anti-oxidative and anti-inflammatory properties, as well as regulation of intestinal microbiome, indicating the potential applications in enterocolitis and CRC. Actually, accumulating studies based on the biological functions of polyphenols have been performed to investigate the functional roles and underlying mechanisms over the last few decades. Based on the mounting evidence of literature, the objective of this review is to outline the current research progress regarding the category, biological functions, and metabolism of polyphenols within the intestine, as well as applications for the prevention and treatment of intestinal diseases, which might provide ever-expanding new insights for the utilization of natural polyphenols.

High-Speed Extraction of Regions of Interest in Optical Camera Communication Enabled by Grid Virtual Division.

Optical camera communication (OCC), enabled by light-emitting diodes (LEDs) and embedded cameras on smartphones, has drawn considerable attention thanks to the pervasive adoption of LED lighting and mobile devices. However, most existing studies do not consider the performance bottleneck of Region of Interest (RoI) extraction during decoding, making it challenging to improve communication capacity further. To this end, we propose a fast grid virtual division scheme based on pixel grayscale values, which extracts RoI quickly without sacrificing computational complexity, thereby reducing the decoding delay and improving the communication capacity of OCC. Essentially, the proposed scheme uses a grid division strategy to divide the received image into blocks and randomly sample several pixels within different blocks to quickly locate the RoI with high grayscale values in the original image. By implementing the lightweight RoI extraction algorithm, we experimentally verify its effectiveness in reducing decoding latency, demonstrating its superior performance in terms of communication capacity. The experimental results clearly show that the decoding delay of the proposed scheme is 70% lower than that provided by the Gaussian blur scheme for the iPhone receiver at a transmission frequency of 5 kHz.

High-Accuracy Height-Independent 3D VLP Based on Received Signal Strength Ratio.

Visible light positioning (VLP) has attracted intensive attention from both academic and industrial communities thanks to its high accuracy, immunity to electromagnetic interference, and low deployment cost. In general, the receiver in a VLP system determines its own position by exploring the received signal strength (RSS) from the transmitter according to a pre-built RSS attenuation model. In such model-based methods, the LED's emission power and the receiver's height are usually required known and constant parameters to obtain reasonable positioning accuracy. However, the LED's emission power is normally time-varying due to the fact that the LED's optical output power is prone to changing with the LED's temperature, and the receiver's height is random in a realistic application scenario. To this end, we propose a height-independent three-dimensional (3D) VLP scheme based on the RSS ratio (RSSR), rather than only using RSS. Unlike existing RSS-based VLP methods, our method is able to independently find the horizontal coordinate, i.e., two-dimensional (2D) position, without a priori height information of the receiver, and also avoids the negative effect caused by fluctuation of the LED's emission power. Moreover, we can further infer the height of the receiver to achieve three-dimensional (3D) positioning by iterating the 2D results back into positioning equations. To quickly verify the proposed scheme, we conduct theoretical analysis with mathematical proof and experimental results with real data, which confirm that the proposed scheme can achieve high position accuracy without known information of the receiver's height and LED's emission power. We also implement a VLP prototype with five LED transmitters, and experimental results show that the proposed scheme can achieve very low average errors of 2.73 cm in 2D and 7.20 cm in 3D.

Active magnetic dipole emission by the Ge2Sb2Te5 nanodisk.

Active light manipulation plays a critical role in nanophotonics. In this Letter, we investigate the modulation properties of magnetic dipole (MD) emission based on the phase change material Ge2Sb2Te5 hollow nanodisk (GST-HND). The results show that the amorphous GST-HND supports a strong MD response with a radiative decay enhancement of 282 times and quantum efficiency of 100%. More importantly, by tuning the crystallization rate of GST, the active manipulation of MD radiation is achieved with a quantum efficiency modulation depth of up to 95% at a specific wavelength. Our work may provide significant instruction for the active tuning of optical nanodevices.

Genome-wide analysis of wheat DNA-binding with one finger (Dof) transcription factor genes: evolutionary characteristics and diverse abiotic stress responses.

BACKGROUND: DNA binding with one finger (Dof) transcription factors play important roles in plant growth and abiotic stress responses. Although genome-wide identification and analysis of the DOF transcription factor family has been reported in other species, no relevant studies have emerged in wheat. The aim of this study was to investigate the evolutionary and functional characteristics associated with plant growth and abiotic stress responses by genome-wide analysis of the wheat Dof transcription factor gene family. RESULTS: Using the recently released wheat genome database (IWGSC RefSeq v1.0), we identified 96 wheat Dof gene family members, which were phylogenetically clustered into five distinct subfamilies. Gene duplication analysis revealed a broad and heterogeneous distribution of TaDofs on the chromosome groups 1 to 7, and obvious tandem duplication genes were present on chromosomes 2 and 3.Members of the same gene subfamily had similar exon-intron structures, while members of different subfamilies had obvious differences. Functional divergence analysis indicated that type-II functional divergence played a major role in the differentiation of the TaDof gene family. Positive selection analysis revealed that the Dof gene family experienced different degrees of positive selection pressure during the process of evolution, and five significant positive selection sites (30A, 31 T, 33A, 102G and 104S) were identified. Additionally, nine groups of coevolving amino acid sites, which may play a key role in maintaining the structural and functional stability of Dof proteins, were identified. The results from the RNA-seq data and qRT-PCR analysis revealed that TaDof genes exhibited obvious expression preference or specificity in different organs and developmental stages, as well as in diverse abiotic stress responses. Most TaDof genes were significantly upregulated by heat, PEG and heavy metal stresses. CONCLUSIONS: The genome-wide analysis and identification of wheat DOF transcription factor family and the discovery of important amino acid sites are expected to provide new insights into the structure, evolution and function of the plant Dof gene family.

Secretory phospholipase A2 group IIA enhances the metabolic rate and increases glucose utilization in response to thyroid hormone.

Secretory phospholipase A2 group IIA (PLA2G2A) is a phospholipase which has a role in inflammation, atherogenesis, and host defense. Previously, we found that PLA2G2A protects mice on high-fat diets from weight gain and insulin resistance. Here, we examined the regulation of PLA2G2A and the metabolic changes that occur in response to variations in thyroid status. In particular, the impact of PLA2G2A on the brown adipose tissue (BAT) thermogenic gene expression was explored. We induced hypothyroidism in C57BL/6 and PLA2G2A-overexpressing (IIA+) mice over a 10 wk period or treated them with thyroid hormone (T3) for 5 wk. There were no significant changes in PLA2G2A abundance in response to thyroid status. The energy expenditure of hypothyroid IIA+ mice did not increase; however, the energy expenditure, substrate utilization, insulin sensitivity, and glucose tolerance were all elevated in the IIA+ mice given T3. Moreover, white adipocytes from IIA+ mice were much more prone to "beiging," including increased expression of brown adipose thermogenic markers such as uncoupling protein 1 (UCP1), PR domain containing 16, and early B cell factor 2. Finally, the BAT of IIA+ mice had increased UCP1 and other proteins indicative of mitochondrial uncoupling and nonshivering adaptive thermogenesis. These data reveal a novel role for PLA2G2A on adipose tissue thermogenesis depending on thyroid status.-Kuefner, M. S., Deng, X., Stephenson, E. J., Pham, K., Park, E. A. Secretory phospholipase A2 group IIA enhances the metabolic rate and increases glucose utilization in response to thyroid hormone.

Genome-wide identification and expression analysis of expansin gene family in common wheat (Triticum aestivum L.).

BACKGROUND: Expansin loosens plant cell walls and involves in cell enlargement and various abiotic stresses. Plant expansin superfamily contains four subfamilies: α-expansin (EXPA), ß-expansin (EXPB), expansin-like A (EXLA), and expansin-like B (EXLB). In this work, we performed a comprehensive study on the molecular characterization, phylogenetic relationship and expression profiling of common wheat (Triticum aestivum) expansin gene family using the recently released wheat genome database (IWGSC RefSeq v1.1 with a coverage rate of 94%). RESULTS: Genome-wide analysis identified 241 expansin genes in the wheat genome, which were grouped into three subfamilies (EXPA, EXPB and EXLA) by phylogenetic tree. Molecular structure analysis showed that wheat expansin gene family showed high evolutionary conservation although some differences were present in different subfamilies. Some key amino acid sites that contribute to functional divergence, positive selection, and coevolution were detected. Evolutionary analysis revealed that wheat expansin gene superfamily underwent strong positive selection. The transcriptome map and qRT-PCR analysis found that wheat expansin genes had tissue/organ expression specificity and preference, and generally highly expressed in the roots. The expression levels of some expansin genes were significantly induced by NaCl and polyethylene glycol stresses, which was consistent with the differential distribution of the cis-elements in the promoter region. CONCLUSIONS: Wheat expansin gene family showed high evolutionary conservation and wide range of functional divergence. Different selection constraints may influence the evolution of the three expansin subfamilies. The different expression patterns demonstrated that expansin genes could play important roles in plant growth and abiotic stress responses. This study provides new insights into the structures, evolution and functions of wheat expansin gene family.

A novel splice-site mutation in the ATP2C1 gene of a Chinese family with Hailey-Hailey disease.

Hailey-Hailey disease (HHD), also known as familial benign chronic pemphigus, is an autosomal dominant genodermatosis. It is characterized by erosions, blisters and erythematous plaques at sites of friction or intertriginous areas. The pathogenic gene of HHD has been revealed as the ATPase secretory pathway Ca2+ transporting 1 gene ( ATP2C1), which encodes the protein, secretory pathway Ca 2+/Mn 2+-ATPase 1 (SPCA1). ATP2C1 gene mutations are responsible for HHD by resulting in abnormal Ca 2+ homeostasis in the skin and giving rise to acantholysis, a characteristic pathology of HHD. In this study, a four-generation family containing three HHD sufferers was recruited. Direct sequencing of the ATP2C1 gene was performed in the proband and other available family members. Reverse-transcriptase polymerase chain reaction analysis was conducted to show the potential variant effect on ATP2C1 splicing. A novel heterozygous c.325-2A>G transition at the splice acceptor site of intron 4 in the ATP2C1 gene was identified, and it co-segregated with the disease in this family. The mutation resulted in exon 5 skipping and an in-frame deletion of 12 amino acids (p.Ala109_Gln120del) in SPCA1. This splice-site mutation may be responsible for HHD in this family. This study would further expand the mutation spectrum of the ATP2C1 gene and may be helpful in the genetic counseling and prenatal diagnosis of HHD.

Integrated physiological and proteomic analysis of embryo and endosperm reveals central salt stress response proteins during seed germination of winter wheat cultivar Zhengmai 366.

BACKGROUND: Salinity is a major abiotic stressor that affects seed germination, plant growth, and crop production. Seed germination represents the beginning of plant growth and is closely linked with subsequent crop development and ultimate yield formation. This study attempted to extend findings regarding the potential proteomic dynamics during wheat seed germination under salt stress and to explore the mechanism of crop salt response. RESULTS: Salt stress significantly affected seed physiological activities during the germination process, resulting in significant decreases in phytohormone and α-amylase activity and significant increases in soluble sugar, starch, and ADP glucose pyrophosphorylase activity. A comparative proteomics approach was applied to analyze the dynamic proteome changes of embryo and endosperm during seed germination in Chinese winter wheat cultivar Zhengmai 366 under salt stress. Two-dimensional electrophoresis identified 92 and 61 differentially accumulated proteins (DAPs) in response to salt stress in embryo and endosperm, respectively. Both organs contained a high proportion of DAPs involved in stress defense, energy metabolism, and protein/amino acid metabolism. The endosperm had more DAPs related to storage proteins and starch metabolism than the embryo, and 2% of DAPs participating in lipid and sterol metabolism were specifically detected in the embryo. CONCLUSIONS: Seed physiological activities were significantly affected during the germination process when subjected to salt stress. The DAPs involved in stress defense and energy metabolism were upregulated whereas those related to reserve substance degradation and protein/amino acid metabolism were significantly downregulated, leading to delayed seed germination under salt stress. Our proteomic results revealed synergistic regulation of the response to salt stress during seed germination.

Retraction Note to: Integrated physiological and proteomic analysis of embryo and endosperm reveals central salt stress response proteins during seed germination of winter wheat cultivar Zhengmai 366.

The editor has retracted this article [1] because parts of Figs. 1 and 4 were duplicated from a previously published paper by the same authors [2] without appropriate disclosure. None of the authors have responded to any correspondence from the editor about publication of this retraction notice.

Low complexity OFDM VLC system enabled by spatial summing modulation.

Commercial-off-the-shelf (COTS) devices enabled visible light communication (VLC) for Internet of things (IoT) applications has attracted extensive attentions from both academic and industrial communities, thanks to the pervasive deployments of light emitting diode (LED) lighting infrastructure. However, due to the limitation of frequency response and non-linearity of the commercial illuminating LED light consisting of multiple LED chips, the achievable data rate is far less than that provided by the experimental VLC system with a single LED with specialized devices, e.g., lens. To this end, we propose a power-of-2 arrangement scheme for LED chips to generate spatial summing modulation with low control complexity, and demonstrate its availability in an orthogonal frequency division multiplexing (OFDM) VLC system purely built upon COTS devices. It significantly differs from a conventional OFDM VLC system relying on digital-to-analog converter (DAC) and analog signal chain, which is complex and confined by LED's non-linearity, thanks to we design a novel digital-to-light converter (DLC) which can output 256 light intensities linearly and be directly controlled by the discrete digital signals generated by the OFDM modulator. An experimental demonstration with employing the QAM-OFDM modulation scheme successfully confirms the effectiveness of the proposed spatial summing VLC system, which can achieve low BERs of below the forward error correct (FEC) threshold of 3.8×10-3 for both QAM8 and QAM16 running transmission frequency of 300 kHz under a communication distance of 0.8 m. It demonstrates the promising potential for delivering a data rate at hundred kbps level with this novel spatial summing based OFDM VLC system, which is sufficient for many IoT applications.

Mutation Analysis of the ATP7B Gene in Seven Chinese Families with Wilson's Disease.

BACKGROUND: Wilson's disease (WD) is an autosomal recessive disease, which is characterized by an excessive copper accumulation in the liver and brain, leading to subsequent hepatic and/or neurological disorders. The causative gene for WD has been identified as the ATPase Cu2+ transporting beta polypeptide gene (ATP7B), which encodes a protein called copper-transporting ATPase 2. ATP7B mutations may lead to reduced biliary excretion of excess copper and disrupted copper homeostasis, resulting in various clinical symptoms of WD. METHODS: Direct sequencing of the ATP7B gene was performed in 7 Han Chinese families with WD, and haplotype analysis was conducted in families having the same mutation. RESULTS: Nine ATP7B gene mutations were identified, including 7 missense mutations (p.Asp765Gly, p.Arg778Leu, p.Thr888Pro, p.Pro992Leu, p.Asp1047Val, p.Ile1148Thr and p.Ala1295Val), 1 duplication mutation (c.525dupA), and 1 nonsense mutation (p.Gly837*). Combined with our previous data, haplotype analysis revealed that the founder effect accounted for 48% of alleles in Han Chinese, constituted by high allele frequency mutations p.Arg778Leu, p.Pro992Leu and p.Ala1295Val. CONCLUSION: This study revealed genetic defects of 7 Han Chinese families with WD, and has implications for their genetic counseling and clinical management.

Effects of water deficit and different nitrogen fertilizer treatments on the quality of wheat for Chinese fresh white noodles and steamed bread and the composition of storage proteins.

BACKGROUND: Noodles and steamed bread are popular wheat products consumed worldwide, particularly in China and other Asian countries. We performed the first comprehensive study of the influence of water deficits and different nitrogen fertilizer applications on two elite Chinese bread wheat cultivars, Zhongmai 175 and Jimai 22, which are distinct in gluten strength. These wheat cultivars were tested to determine the qualities that are optimal for the production of Chinese fresh white noodles (CFWN) and northern-style Chinese steamed bread (NCSB), and storage protein composition. RESULTS: Water deficit and high nitrogen (N) fertilizer application promoted total grain protein content and the accumulation of gliadins and glutenins, while low N resulted in the opposite results. Water deficit and high N fertilizer in Jimai 22, with medium-to-strong gluten strength significantly improved NCSB and CFWN qualities. The quality of CFWN under low N, and that of NCSB under both high and low N conditions, was significantly reduced. However, NCSB and CFWN quality in Zhongmai 175 with weak-to-medium gluten strength was not significantly affected by water deficit and different N fertilizer applications. Grain subproteome analysis revealed that considerable cultivar-specific gliadins and glutenins showed significant accumulation changes in response to water deficit and high / low N fertilizer treatment, which could be responsible for NCSB and CFWN quality changes under different treatments. CONCLUSION: Water deficit and high / low N fertilizer treatments caused changes in cultivar-specific storage protein composition, resulting in differences in the accumulation of gliadins, glutenins, and the quality of NCSB and CFWN. © 2019 Society of Chemical Industry.

Genome-wide identification, molecular evolution, and expression analysis of auxin response factor (ARF) gene family in Brachypodium distachyon L.

BACKGROUND: The auxin response factor (ARF) gene family is involved in plant development and hormone regulation. Although the ARF gene family has been studied in some plant species, its structural features, molecular evolution, and expression profiling in Brachypodium distachyon L. are still not clear. RESULTS: Genome-wide analysis identified 19 ARF genes in B. distachyon. A phylogenetic tree constructed with 182 ARF genes from seven plant species revealed three different clades, and the ARF genes from within a clade exhibited structural conservation, although certain divergences occurred in different clades. The branch-site model identified some sites where positive selection may have occurred, and functional divergence analysis found more Type II divergence sites than Type I. In particular, both positive selection and functional divergence may have occurred in 241H, 243G, 244 L, 310 T, 340G and 355 T. Subcellular localization prediction and experimental verification indicated that BdARF proteins were present in the nucleus. Transcript expression analysis revealed that BdARFs were mainly expressed in the leaf and root tips, stems, and developing seeds. Some BdARF genes exhibited significantly upregulated expression under various abiotic stressors. Particularly, BdARF4 and BdARF8 were significantly upregulated in response to abiotic stress factors such as salicylic acid and heavy metals. CONCLUSION: The ARF gene family in B. distachyon was highly conserved. Several important amino acid sites were identified where positive selection and functional divergence occurred, and they may play important roles in functional differentiation. BdARF genes had clear tissue and organ expression preference and were involved in abiotic stress response, suggesting their roles in plant growth and stress resistance.

Secure and private NOMA VLC using OFDM with two-level chaotic encryption.

In this paper, we propose and demonstrate a secure and private non-orthogonal multiple access (NOMA) based visible light communication (VLC) system. Orthogonal frequency division multiplexing (OFDM) modulation is applied in the system and a two-level chaotic encryption scheme is further implemented, which can guarantee both the security of legitimate users against eavesdroppers and the privacy among all the legitimate users. An experimental demonstration with two legitimate users and one eavesdropper successfully verifies the feasibility of the proposed secure and private NOMA VLC system. To the best of our knowledge, it is the first time that simultaneous security and privacy improvement is considered for NOMA VLC systems.

2D-DIGE comparative proteomic analysis of developing wheat grains under high-nitrogen fertilization revealed key differentially accumulated proteins that promote storage protein and starch biosyntheses.

Nitrogen (N) serves as a macronutrient that is essential to plant growth and development, and significantly influences storage protein and starch biosyntheses and, ultimately, grain yield and quality. In this study, we performed the first comparative proteomic analysis of developing wheat grains under high-N conditions using 2D-DIGE and tandem mass spectrometry. High-N fertilizer application caused significant increases in ear number, ear grain number, and grain yield. 2D-DIGE identified 142 differentially accumulated proteins (DAPs) during grain development in the elite Chinese bread wheat cultivar Zhongmai 175, of which 132 (93%) were identified by MALDI-TOF/TOF-MS, representing 92 unique proteins. These proteins are involved mainly in energy, N and protein metabolism, carbon metabolism, and starch biosynthesis. Subcellular localization prediction and fluorescence confocal microscopic analysis showed that the DAPs identified were localized mainly in the cytosol and chloroplast. Principal component analysis (PCA) revealed a greater proteomic difference among grain developmental periods than between the high-N and control groups. Protein-protein interaction analysis highlighted a complex network centered around enzymes involved in energy, N and protein metabolism, and starch biosynthesis. Six key DAP genes showed expression patterns consistent with their protein accumulation trends during grain development. A putative metabolic pathway was proposed, with synergistic regulatory networks of grain storage protein and starch biosyntheses in response to high-N application.