1.Functional analysis of a nitrate-induced GARP transcription factor AhNIGT1.2 in peanut nodulation.
Xiaoliang LI ; Haitong HE ; Suqin HE ; Luyao WANG ; Wei ZHANG ; Zhaosheng KONG ; Lixiang WANG
Chinese Journal of Biotechnology 2025;41(2):657-669
Peanut, a major economic and oil crop known for the high protein and oil content, is extensively cultivated in China. Peanut plants have the ability to form nodules with rhizobia, where the nitrogenase converts atmospheric nitrogen into ammonia nitrogen that can be utilized by the plants. Analysis of nodule fixation is of positive significance for avoiding overapplication of chemical fertilizer and developing sustainable agriculture. In this study, AhNIGT1.2, a member of the NIGT family predominantly expressed in peanut nodules, was identified by bioinformatics analysis. Subsequent spatiotemporal expression analysis revealed that AhNIGT1.2 was highly expressed in nodules and showed significant responses to high nitrogen, low nitrogen, high phosphorus, low phosphorus, and rhizobia treatments. Histochemical staining indicated that the gene was primarily expressed in developing nodules and at the connection region between mature nodules and peanut roots. The fusion protein AhNIGT1.2-GFP was located in the nucleus of tobacco epidermal cells. The AhNIGT1.2-OE significantly increased the number of peanut nodules, while AhNIGT1.2-RNAi reduced the number of nodules, which suggested a positive regulatory role of AhNIGT1.2 in peanut nodulation. The AhNIGT1.2-OE in roots down-regulated the expression levels of NRT1.2, NRT2.4, NLP1, and NLP7, which indicated that AhNIGT1.2 influenced peanut nodulation by modulating nitrate transport and the expression of NLP genes. The transcriptome analysis of AhNIGT1.2-OE and control roots revealed that overexpressing AhNIGT1.2 significantly enriched the differentially expressed genes associated with nitrate response, nodulation factor pathway, enzymes for triterpene biosynthesis, and carotenoid biosynthesis. These findings suggest that AhNIGT1.2 play a key role in peanut nodulation by regulating nitrate transport and response and other related pathways. This study gives insights into the molecular mechanisms of nitrogen and phosphorus in regulating legume nodulation and nitrogen fixation, and sheds light on the development of legume crops that can efficiently fix nitrogen in high nitrogen environments.
Arachis/physiology*
;
Nitrates/metabolism*
;
Plant Proteins/physiology*
;
Transcription Factors/metabolism*
;
Plant Root Nodulation/physiology*
;
Gene Expression Regulation, Plant
;
Root Nodules, Plant/metabolism*
;
Nitrogen Fixation
2.BnaNRT1.5s mediates nitrate transporter to regulate nitrogen use efficiency in Brassica napus.
Shilong CHEN ; Lei YAO ; Rumeng WANG ; Jian ZENG ; Jianghe LI ; Shiyao CUI ; Xu WANG ; Haixing SONG ; Zhenhua ZHANG ; Pan GONG
Chinese Journal of Biotechnology 2025;41(7):2954-2965
Improving the nitrogen use efficiency (NUE) of Brassica napus is of significant importance for achieving the national goal of zero growth in chemical fertilizer application and ensuring the green development of the rapeseed industry. This study aims to explore the effects of the nitrate transporter gene BnaNRT1.5s on the nitrogen transport and NUE of B. napus, providing excellent genetic resources for the development of nitrogen-efficient B. napus varieties. The spatiotemporal expression of BnaA05.NRT1.5 as a key nitrogen responsive gene was profiled by qRT-PCR at different growth stages and for different tissue samples of B. napus 'Westar'. Subcellular localization was employed to examine its expression pattern in the cells. Additionally, CRISPR/Cas9 was used to create BnaNRT1.5s knockout lines, which were subjected to hydroponic experiments under high nitrogen (12.0 mmol/L) and low nitrogen (0.3 mmol/L) conditions. After the seedlings were cultivated for 21 days, root and shoot samples were collected for weighing, nitrogen content determination, xylem sap nitrate content assessment, and calculation of total nitrogen and NUE. The B. napus nitrate transporter BnaA05.NRT1.5 was localized to the cell membrane. During the seedling and early bolting stages, BnaA05.NRT1.5 was predominantly expressed in roots, while it was highly expressed in old leaves and mature silique skin during the reproductive stage. Compared with the wild type, the mutant BnaNRT1.5s showed significant increases in the dry weight and total nitrogen of seedlings under both high and low nitrogen conditions. Under low nitrogen conditions, NUE in the roots of BnaNRT1.5s significantly improved. Notably, under both high and low nitrogen conditions, the nitrate content in the shoots of BnaNRT1.5s decreased significantly, while that in the roots increased significantly, resulting in a significantly decreased shoot-to-root nitrate content ratio. BnaNRT1.5s is involved in regulating the transport of nitrate from the roots to the shoots, and its mutation enhances nitrogen absorption and utilization in B. napus seedlings, promoting seedling growth. This study not only provides references for understanding the physiological and molecular mechanisms by which BnaNRT1.5s regulates NUE but also offers valuable genetic resources for improving NUE in B. napus.
Brassica napus/genetics*
;
Anion Transport Proteins/metabolism*
;
Nitrogen/metabolism*
;
Nitrate Transporters
;
Plant Proteins/metabolism*
;
Nitrates/metabolism*
;
Gene Expression Regulation, Plant
;
Biological Transport
3.Nitrate reduction capacity of the oral microbiota is impaired in periodontitis: potential implications for systemic nitric oxide availability.
Bob T ROSIER ; William JOHNSTON ; Miguel CARDA-DIÉGUEZ ; Annabel SIMPSON ; Elena CABELLO-YEVES ; Krystyna PIELA ; Robert REILLY ; Alejandro ARTACHO ; Chris EASTON ; Mia BURLEIGH ; Shauna CULSHAW ; Alex MIRA
International Journal of Oral Science 2024;16(1):1-1
The reduction of nitrate to nitrite by the oral microbiota has been proposed to be important for oral health and results in nitric oxide formation that can improve cardiometabolic conditions. Studies of bacterial composition in subgingival plaque suggest that nitrate-reducing bacteria are associated with periodontal health, but the impact of periodontitis on nitrate-reducing capacity (NRC) and, therefore, nitric oxide availability has not been evaluated. The current study aimed to evaluate how periodontitis affects the NRC of the oral microbiota. First, 16S rRNA sequencing data from five different countries were analyzed, revealing that nitrate-reducing bacteria were significantly lower in subgingival plaque of periodontitis patients compared with healthy individuals (P < 0.05 in all five datasets with n = 20-82 samples per dataset). Secondly, subgingival plaque, saliva, and plasma samples were obtained from 42 periodontitis patients before and after periodontal treatment. The oral NRC was determined in vitro by incubating saliva with 8 mmol/L nitrate (a concentration found in saliva after nitrate-rich vegetable intake) and compared with the NRC of 15 healthy individuals. Salivary NRC was found to be diminished in periodontal patients before treatment (P < 0.05) but recovered to healthy levels 90 days post-treatment. Additionally, the subgingival levels of nitrate-reducing bacteria increased after treatment and correlated negatively with periodontitis-associated bacteria (P < 0.01). No significant effect of periodontal treatment on the baseline saliva and plasma nitrate and nitrite levels was found, indicating that differences in the NRC may only be revealed after nitrate intake. Our results suggest that an impaired NRC in periodontitis could limit dietary nitrate-derived nitric oxide levels, and the effect on systemic health should be explored in future studies.
Humans
;
Nitrates
;
Nitric Oxide
;
Nitrites
;
RNA, Ribosomal, 16S/genetics*
;
Periodontitis/microbiology*
;
Bacteria
;
Dental Plaque/microbiology*
;
Saliva/microbiology*
;
Microbiota/genetics*
4.One new galloyl derivative from Isodon henryi.
Yi-Xiao LI ; Jun CHI ; Zhi-Min WANG ; Li-Ping DAI
China Journal of Chinese Materia Medica 2024;49(21):5909-5913
Seven compounds(1-7) were isolated from Isodon henryi through silica gel, Sephadex LH-20, ODS, MCI column chromatography and semi-preparative HPLC. Their structures were identified as isogallicacid(1), caffeic acid(2), syringic acid(3), protocatechuic acid(4), oresbiusin A(5), lophanthoside A(6), and 8-hydroxypinoresinol(7), by spectroscopic techniques including HR-MS, IR, UV, NMR, and ECD. Compound 1 was a new galloyl derivative. Moreover, it demonstrated a significant inhibitory effect on the lipopolysaccharide-induced release of nitric oxide from RAW264.7 cells.
Mice
;
Animals
;
RAW 264.7 Cells
;
Nitric Oxide/metabolism*
;
Isodon/chemistry*
;
Molecular Structure
;
Drugs, Chinese Herbal/chemistry*
;
Macrophages/drug effects*
;
Magnetic Resonance Spectroscopy
;
Gallic Acid/analogs & derivatives*
5.Effect of High-Concentration Uric Acid on Nitric Oxide.
Si-Yu QIN ; Rong-Yu LAN ; Jia ZENG ; Xue BAI ; Jing-Tao WANG ; Xiang-Lin YIN ; Rui-Jie QU ; Ming-Hai QU ; Hao JIANG ; Wen-Long LI ; Si-Ying PEI ; Zhi-Ling HOU ; Bao-Sheng GUAN ; Hong-Bin QIU
Acta Academiae Medicinae Sinicae 2023;45(4):666-671
Uric acid (UA) is the final product of purine metabolism in human body,and its metabolic disorder will induce hyperuricemia (HUA).The occurrence and development of HUA are associated with a variety of pathological mechanisms such as oxidative stress injury,activation of inflammatory cytokines,and activation of renin-angiotensin-aldosterone system.These mechanisms directly or indirectly affect the bioavailability of endogenous nitric oxide (NO).The decrease in NO bioavailability is common in the diseases with high concentration of UA as an independent risk factor.In this review,we summarize the mechanisms by which high concentrations of UA affect the endogenous NO bioavailability,with a focus on the mechanisms of high-concentration UA in decreasing the synthesis and/or increasing the consumption of NO.This review aims to provide references for alleviating the multisystem symptoms and improving the prognosis of HUA,and lay a theoretical foundation for in-depth study of the correlations between HUA and other metabolic diseases.
Humans
;
Nitric Oxide
;
Uric Acid
;
Hyperuricemia
;
Biological Availability
;
Cytokines
6.Near-infrared excited graphene oxide/silver nitrate/chitosan coating for improving antibacterial properties of titanium implants.
Yifan WANG ; Yingde XU ; Xuefeng ZHANG ; Jingyu LIU ; Jintong HAN ; Shengli ZHU ; Yanqin LIANG ; Shuilin WU ; Zhenduo CUI ; Weijia LÜ ; Zhaoyang LI
Chinese Journal of Reparative and Reconstructive Surgery 2023;37(8):937-944
OBJECTIVE:
To design and construct a graphene oxide (GO)/silver nitrate (Ag3PO4)/chitosan (CS) composite coating for rapidly killing bacteria and preventing postoperative infection in implant surgery.
METHODS:
GO/Ag3PO4 composites were prepared by ion exchange method, and CS and GO/Ag3PO4 composites were deposited on medical titanium (Ti) sheets successively. The morphology, physical image, photothermal and photocatalytic ability, antibacterial ability, and adhesion to the matrix of the materials were characterized.
RESULTS:
The GO/Ag3PO4 composites were successfully prepared by ion exchange method and the heterogeneous structure of GO/Ag3PO4 was proved by morphology phase test. The heterogeneous structure formed by Ag3PO4 and GO reduced the band gap from 1.79 eV to 1.39 eV which could be excited by 808 nm near-infrared light. The photothermal and photocatalytic experiments proved that the GO/Ag3PO4/CS coating had excellent photothermal and photodynamic properties. In vitro antibacterial experiments showed that the antibacterial rate of the GO/Ag3PO4/CS composite coating against Staphylococcus aureus reached 99.81% after 20 minutes irradiation with 808 nm near-infrared light. At the same time, the composite coating had excellent light stability, which could provide stable and sustained antibacterial effect.
CONCLUSION
GO/Ag3PO4/CS coating can be excited by 808 nm near infrared light to produce reactive oxygen species, which has excellent antibacterial activity under light.
Chitosan
;
Silver Nitrate
;
Titanium
;
Anti-Bacterial Agents/pharmacology*
;
Coloring Agents
7.Homeostatic medicine: new strategy and concept of health maintenance as well as diagnosis and treatment of diseases.
Li Zheng QIN ; Jian ZHOU ; Lei HU ; Song Ling WANG
Chinese Journal of Stomatology 2023;58(2):109-117
Homeostasis is a dynamic balance process of self-regulating. Biological systems remain stable through adapting to changing external conditions to maintain normal life activities. Homeostatic medicine is the science of studying homeostasis of human molecules, cells, organs and the whole body. It is a comprehensive discipline based on maintaining homeostasis to keep human health and assist for diseases prevention and diagnoses. Homeostatic medicine focuses on the whole body and on the role of homeostasis in health and disease, which is expected to provide new ideas and strategies for maintaining health as well as diagnosing and treating diseases. Nitric oxide (NO) plays an important role in the control of multisystem homeostasis. Nitrate is an important substance in regulating NO homeostasis through the nitrate-nitrite-NO pathway. Sialin, nitrate transporter which is located in the cell membrane and cytoplasm, mediates multiple cellular biological functions. The nitrate-nitrite-NO pathway and sialin-mediated biological functions play an important role in the regulation of body homeostasis.
Humans
;
Nitrates/metabolism*
;
Nitrites/metabolism*
;
Homeostasis
;
Nitric Oxide
8.Environmental exposure to perchlorate, nitrate, and thiocyanate in relation to chronic kidney disease in the general US population, NHANES 2005-2016.
Wei LI ; Hong WU ; Xuewen XU ; Yange ZHANG
Chinese Medical Journal 2023;136(13):1573-1582
BACKGROUND:
Few studies have explored the impact of perchlorate, nitrate, and thiocyanate (PNT) on kidney function. This study aimed to evaluate the association of urinary levels of PNT with renal function as well as the prevalence of chronic kidney disease (CKD) among the general population in the United States.
METHODS:
This analysis included data from 13,373 adults (≥20 years) from the National Health and Nutrition Examination Survey 2005 to 2016. We used multivariable linear and logistic regression, to explore the associations of urinary PNT with kidney function. Restricted cubic splines were used to assess the potentially non-linear relationships between PNT exposure and outcomes.
RESULTS:
After traditional creatinine adjustment, perchlorate (P-traditional) was positively associated with estimated glomerular filtration rate (eGFR) (adjusted β: 2.75; 95% confidence interval [CI]: 2.25 to 3.26; P < 0.001), and negatively associated with urinary albumin-to-creatinine ratio (ACR) (adjusted β: -0.05; 95% CI: -0.07 to -0.02; P = 0.001) in adjusted models. After both traditional and covariate-adjusted creatinine adjustment, urinary nitrate and thiocyanate were positively associated with eGFR (all P values <0.05), and negatively associated with ACR (all P values <0.05); higher nitrate or thiocyanate was associated with a lower risk of CKD (all P values <0.001). Moreover, there were L-shaped non-linear associations between nitrate, thiocyanate, and outcomes. In the adjusted models, for quartiles of PNT, statistically significant dose-response associations were observed in most relationships. Most results were consistent in the stratified and sensitivity analyses.
CONCLUSIONS
Exposures to PNT might be associated with kidney function, indicating a potential beneficial effect of environmental PNT exposure (especially nitrate and thiocyanate) on the human kidney.
Adult
;
Humans
;
United States/epidemiology*
;
Nitrates/adverse effects*
;
Nutrition Surveys
;
Thiocyanates/urine*
;
Perchlorates/urine*
;
Creatinine
;
Environmental Exposure
;
Renal Insufficiency, Chronic/epidemiology*
;
Logistic Models
9.Identification, expression and DNA variation analysis of high affinity nitrate transporter NRT2/3 gene family in Sorghum bicolor.
Shanshan ZHAO ; Zhiqiang GUO ; Lixun ZHU ; Jiali FAN ; Bohui YANG ; Wenting CHAI ; Huiqiong SUN ; Fan FENG ; Yuexiu LIANG ; Chunlei ZOU ; Xiaodong JIANG ; Weijun ZHAO ; Jinhui LÜ ; Chunlai ZHANG
Chinese Journal of Biotechnology 2023;39(7):2743-2761
Nitrate is the main form of inorganic nitrogen that crop absorbs, and nitrate transporter 2 (NRT2) is a high affinity transporter using nitrate as a specific substrate. When the available nitrate is limited, the high affinity transport systems are activated and play an important role in the process of nitrate absorption and transport. Most NRT2 cannot transport nitrates alone and require the assistance of a helper protein belonging to nitrate assimilation related family (NAR2) to complete the absorption or transport of nitrates. Crop nitrogen utilization efficiency is affected by environmental conditions, and there are differences between varieties, so it is of great significance to develop varieties with high nitrogen utilization efficiency. Sorghum bicolor has high stress tolerance and is more efficient in soil nitrogen uptake and utilization. The S. bicolor genome database was scanned to systematically analyze the gene structure, chromosomal localization, physicochemical properties, secondary structure and transmembrane domain, signal peptide and subcellular localization, promoter region cis-acting elements, phylogenetic evolution, single nucleotide polymorphism (SNP) recognition and annotation, and selection pressure of the gene family members. Through bioinformatics analysis, 5 NRT2 gene members (designated as SbNRT2-1a, SbNRT2-1b, SbNRT2-2, SbNRT2-3, and SbNRT2-4) and 2 NAR2 gene members (designated as SbNRT3-1 and SbNRT3-2) were identified, the number of which was less than that of foxtail millet. SbNRT2/3 were distributed on 3 chromosomes, and could be divided into four subfamilies. The genetic structure of the same subfamilies was highly similar. The average value of SbNRT2/3 hydrophilicity was positive, indicating that they were all hydrophobic proteins, whereas α-helix and random coil accounted for more than 70% of the total secondary structure. Subcellular localization occurred on plasma membrane, where SbNRT2 proteins did not contain signal peptides, but SbNRT3 proteins contained signal peptides. Further analysis revealed that the number of transmembrane domains of the SbNRT2s family members was greater than 10, while that of the SbNRT3s were 2. There was a close collinearity between NRT2/3s of S. bicolor and Zea mays. Protein domains analysis showed the presence of MFS_1 and NAR2 protein domains, which supported executing high affinity nitrate transport. Phylogenetic tree analysis showed that SbNRT2/3 were more closely related to those of Z. mays and Setaria italic. Analysis of gene promoter cis-acting elements indicated that the promoter region of SbNRT2/3 had several plant hormones and stress response elements, which might respond to growth and environmental cues. Gene expression heat map showed that SbNRT2-3 and SbNRT3-1 were induced by nitrate in the root and stem, respectively, and SbNRT2-4 and SbNRT2-3 were induced by low nitrogen in the root and stem. Non-synonymous SNP variants were found in SbNRT2-4 and SbNRT2-1a. Selection pressure analysis showed that the SbNRT2/3 were subject to purification and selection during evolution. The expression of SbNRT2/3 gene and the effect of aphid infection were consistent with the expression analysis results of genes in different tissues, and SbNRT2-1b and SbNRT3-1 were significantly expressed in the roots of aphid lines 5-27sug, and the expression levels of SbNRT2-3, SbNRT2-4 and SbNRT3-2 were significantly reduced in sorghum aphid infested leaves. Overall, genome-wide identification, expression and DNA variation analysis of NRT2/3 gene family of Sorghum bicolor provided a basis for elucidating the high efficiency of sorghum in nitrogen utilization.
Nitrate Transporters
;
Nitrates/metabolism*
;
Sorghum/metabolism*
;
Anion Transport Proteins/metabolism*
;
Phylogeny
;
Protein Sorting Signals/genetics*
;
Nitrogen/metabolism*
;
DNA
;
Gene Expression Regulation, Plant
;
Plant Proteins/metabolism*
10.Determination of cobalt and tungsten in human urine by inductively coupled plasma mass spectrometry.
Liu YANG ; Chao Yang WANG ; Yu Dan ZHANG ; Qi GENG ; Wen Hua QIN
Chinese Journal of Industrial Hygiene and Occupational Diseases 2023;41(5):371-374
Objective: To establish a inductively coupled plasma mass spectrometry method for the determination of trace cobalt and tungsten in human urine. Methods: The authors used 1% nitric acid solution as diluent in October-December 2021, the sample dilution factor and internal standard element were optimized by single factor rotation experiment, and the difference between the working curve and the standard curve was compared. Results: The method uses working curve to determine cobalt and tungsten in urine, the linear range of this method was 0.0~10.0 μg/L, the correlation coefficient was 0.999 9, the detection limits respectively were 0.005 μg/L (cobalt) and 0.09 μg/L (tungsten), the recoveries of samples respectively were 87.0%~100.2% (cobalt) and 89.4%~104.8% (tungsten), the relative standard deviations respectively were 0.4%~4.4% (cobalt) and 0.6%~3.8% (tungsten) . Conclusion: A simple and rapid method for determination of cobalt and tungsten in urine has been established. This method has the advantages of simple operation, high sensitivity, low detection limit and good stability. It is suitable for determination of cobalt and tungsten in urine of all kinds of people.
Humans
;
Cobalt/analysis*
;
Tungsten/analysis*
;
Spectrum Analysis
;
Nitric Acid
;
Mass Spectrometry

Result Analysis
Print
Save
E-mail