Overexpression of GmPAP4 Enhances Symbiotic Nitrogen Fixation and Seed Yield in Soybean under Phosphorus-Deficient Condition.

Legume crops establish symbiosis with nitrogen-fixing rhizobia for biological nitrogen fixation (BNF), a process that provides a prominent natural nitrogen source in agroecosystems; and efficient nodulation and nitrogen fixation processes require a large amount of phosphorus (P). Here, a role of GmPAP4, a nodule-localized purple acid phosphatase, in BNF and seed yield was functionally characterized in whole transgenic soybean (Glycine max) plants under a P-limited condition. GmPAP4 was specifically expressed in the infection zones of soybean nodules and its expression was greatly induced in low P stress. Altered expression of GmPAP4 significantly affected soybean nodulation, BNF, and yield under the P-deficient condition. Nodule number, nodule fresh weight, nodule nitrogenase, APase activities, and nodule total P content were significantly increased in GmPAP4 overexpression (OE) lines. Structural characteristics revealed by toluidine blue staining showed that overexpression of GmPAP4 resulted in a larger infection area than wild-type (WT) control. Moreover, the plant biomass and N and P content of shoot and root in GmPAP4 OE lines were also greatly improved, resulting in increased soybean yield in the P-deficient condition. Taken together, our results demonstrated that GmPAP4, a purple acid phosphatase, increased P utilization efficiency in nodules under a P-deficient condition and, subsequently, enhanced symbiotic BNF and seed yield of soybean.

Fast Discrimination and Quantification Analysis of Atractylodis rhizoma Using NIR Spectroscopy Coupled with Chemometrics Tools.

In this study, near-infrared (NIR) spectroscopy and high-performance liquid chromatography (HPLC) combined with chemometrics tools were applied for quick discrimination and quantitative analysis of different varieties and origins of Atractylodis rhizoma samples. Based on NIR data, orthogonal partial least squares discriminant analysis (OPLS-DA) and K-nearest neighbor (KNN) models achieved greater than 90% discriminant accuracy of the three species and two origins of Atractylodis rhizoma. Moreover, the contents of three active ingredients (atractyloxin, atractylone, and ß-eudesmol) in Atractylodis rhizoma were simultaneously determined by HPLC. There are significant differences in the content of the three components in the samples of Atractylodis rhizoma from different varieties and origins. Then, partial least squares regression (PLSR) models for the prediction of atractyloxin, atractylone, and ß-eudesmol content were successfully established. The complete Atractylodis rhizoma spectra gave rise to good predictions of atractyloxin, atractylone, and ß-eudesmol content with R2 values of 0.9642, 0.9588, and 0.9812, respectively. Based on the results of this present research, it can be concluded that NIR is a great nondestructive alternative to be applied as a rapid classification system by the drug industry.

Preparation and structural characterization of epoxidized soybean oils-based pressure sensitive adhesive grafted with tea polyphenol palmitate.

Vegetable oils-based pressure sensitive adhesives (PSAs) are green and sustainable but face unsatisfactory adhesion strengths and are prone to aging during storage and application due to the existence of residual double bonds and massive ester bonds. Nine common antioxidants (tea polyphenol palmitate (TPP), caffeic acid, ferulic acid, gallic acid, butylated hydroxytoluene, tertiary butylhydroquinone, butylated hydroxyanisole, propyl gallate, and tea polyphenols) were grafted into epoxidized soybean oils-PSA (ESO-PSA) system to enhance antiaging properties and adhesion strengths. Results showed ESO-PSAs grafted with caffeic acid, tertiary butylhydroquinone, butylated hydroxyanisole, propyl gallate, tea polyphenols, or TPP didn't occur failure with TPP having best performance. The optimal conditions were ESO reacted with 0.9 % TPP, 70 % rosin ester, and 7.0 % phosphoric acid at 50 °C for 5 min, under which peel strength and loop tack increased to 2.460 N/cm and 1.66 N, respectively, but peel strength residue reduced to 138.09 %, compared with control (0.407 N/cm, 0.43 N, and 1669.99 %). Differential scanning calorimetry and thermogravimetric results showed TPP grafting increased the glass transition temperature of ESO-PSA slightly but improved its thermal stability significantly. Fourier transform infrared spectroscopy and 1H nuclear magnetic resonance results showed TPP, phosphoric acid, and rosin ester all partially participated in the covalently crosslinking polymerization of ESO-PSAs and the rest existed in the network structures in the free form.

Parameter Optimization of Ultrasonic-Microwave Synergistic Extraction of Taxanes from Taxus cuspidata Needles.

Taxanes are the best-known compounds in Taxus cuspidata owing to their strong anticancer effects. However, the traditional taxanes extraction method is the solid-liquid extraction method, which is limited by a large energy consumption and low yield. Therefore, it is urgent to find an efficient method for taxanes extraction. The ultrasonic microwave synergistic extraction (UME) method integrates the cavitation effect of ultrasound and the intensifying heat transfer (ionic conduction and dipole rotation of molecules) effect of microwave to accelerate the release of intracellular compounds and is used in active ingredient extractions. This study aimed to evaluate the performance of UME in extracting taxanes from T. cuspidata needles (dichloromethane-ethanol as extractant). A single-factor experiment, Plackett-Burman design, and the response surface method showed that the optimal UME parameters for taxanes extraction were an ultrasonic power of 300 W, a microwave power of 215 W, and 130 sieve meshes. Under these conditions, the taxanes yield was 570.32 µg/g, which increased by 13.41% and 41.63% compared with the ultrasound (US) and microwave (MW) treatments, respectively. The reasons for the differences in the taxanes yield were revealed by comparing the physicochemical properties of T. cuspidata residues after the UME, US, and MW treatments. The cell structures were significantly damaged after the UME treatment, and numerous tiny holes were observed on the surface. The absorption peaks of cellulose, hemicellulose, and lignin increased significantly in intensity, and the lowest peak temperature (307.40 °C), with a melting enthalpy of -5.19 J/g, was found after the UME treatment compared with the US and MW treatments. These results demonstrate that UME is an effective method (570.32 µg/g) to extract taxanes from T. cuspidata needles by destroying cellular structures.

Molecular regulatory mechanism of key LncRNAs in subclinical mastitic cows with folic acid supplementation.

BACKGROUND: Folic acid is a water-soluble B vitamin (B9), which is closely related to the body's immune and other metabolic pathways. The folic acid synthesized by rumen microbes has been unable to meet the needs of high-yielding dairy cows. The incidence rate of subclinical mastitis in dairy herds worldwide ranged between 25%~65% with no obvious symptoms, but it significantly causes a decrease in lactation and milk quality. Therefore, this study aims at exploring the effects of folic acid supplementation on the expression profile of lncRNAs, exploring the molecular mechanism by which lncRNAs regulate immunity in subclinical mastitic dairy cows. RESULTS: The analysis identified a total of 4384 lncRNA transcripts. Subsequently, differentially expressed lncRNAs in the comparison of two groups (SF vs. SC, HF vs. HC) were identified to be 84 and 55 respectively. Furthermore, the weighted gene co-expression network analysis (WGCNA) and the KEGG enrichment analysis result showed that folic acid supplementation affects inflammation and immune response-related pathways. The two groups have few pathways in common. One important lncRNA MSTRG.11108.1 and its target genes (ICAM1, CCL3, CCL4, etc.) were involved in immune-related pathways. Finally, through integrated analysis of lncRNAs with GWAS data and animal QTL database, we found that differential lncRNA and its target genes could be significantly enriched in SNPs and QTLs related to somatic cell count (SCC) and mastitis, such as MSTRG.11108.1 and its target gene ICAM1, CXCL3, GRO1. CONCLUSIONS: For subclinical mastitic cows, folic acid supplementation can significantly affect the expression of immune-related pathway genes such as ICAM1 by regulating lncRNAs MSTRG.11108.1, thereby affecting related immune phenotypes. Our findings laid a ground foundation for theoretical and practical application for feeding folic acid supplementation in subclinical mastitic cows.

Effects of a combination of lauric acid monoglyceride and cinnamaldehyde on growth performance, gut morphology, and gut microbiota of yellow-feathered broilers.

A total of 480 one-day-old male yellow-feathered broilers were randomly divided into 4 groups with 6 replicates of 20 chicks per replicate. A basal diet was administered to the control group (CON), whereas CML350, CML500, and CML1000 groups were fed with basal diet supplemented with 350, 500, and 1,000 mg/kg of lauric acid monoglyceride and cinnamaldehyde complex, respectively. However, adding 500 mg/kg of lauric acid monoglyceride and cinnamaldehyde complex improved weight gain (P < 0.01), enhanced intestinal morphology, increased serum total protein and albumin content, and total antioxidant capacity (P < 0.01), and significantly increased the Chao1 and Ace indices (P < 0.01), indicating an increase in the richness of the gut microbiota. At the phylum level, CML500 group reduced the abundance of Fusobacteriota at 21 d and Proteobacteria at 42 d (P < 0.01). At the genus level, CML500 group increased the abundance of Faecalibacterium and Alistipes at 42 d (P < 0.01) and decreased the abundance of Escherichia-Shigella (P < 0.01). At the species level, CML500 group reduced the abundance of Escherichia coli at 42 d (P < 0.01) and increased the abundance of Alistipes_sp_CHKCI003 at 42 d (P < 0.01). According to these results, adding 500 mg/kg of lauric acid monoglyceride and cinnamaldehyde complex in feed can improve the growth performance, intestinal morphology, and gut microbiota of yellow-feathered broilers.

Optimization of dropping process of Xuesaitong dropping pills based on quality by design concept and machine vision.

OBJECTIVE: The drooping process of the Xuesaitong dropping pills (XDPs) was optimized based on quality by design concept. Meanwhile, a machine vision (MV) technology was creatively introduced in this study to predict the critical quality attributes (CQAs) rapidly and accurately. SIGNIFICANCE: This study improves the understanding of dropping process, and has reference value for the guidance of pharmaceutical process research and industrial production. METHODS: The study mainly consisted of three stages: the first stage involved the prediction model to establish and evaluate the CQAs, and the second stage involved assessing the quantitative relationships between critical process parameters (CPPs) and CQAs by the mathematical models that were established through the Box-Behnken experimental design. Finally, a probability-based design space for the dropping process was calculated and verified according to the qualification criteria of each quality attribute. RESULTS: The results show that the prediction accuracy of the random forest (RF) model was high and met the analysis requirements, and the CQAs of dropping pills can meet the standard by running in the design space. CONCLUSION: The MV technology developed in this study can be applied to the optimization process of the XDPs. In addition, the operation in the design space can not only ensure the quality of XDPs to meet the criteria, but also help to improve the consistency of XDPs.

Allelic Variation in GmPAP14 Alters Gene Expression to Affect Acid Phosphatase Activity in Soybean.

Improvement in acid phosphatase (APase) activity is considered as an important approach to enhance phosphorus (P) utilization in crops. Here, GmPAP14 was significantly induced by low P (LP), and its transcription level in ZH15 (P efficient soybean) was higher than in NMH (P inefficient soybean) under LP conditions. Further analyses demonstrated that there were several variations in gDNA (G-GmPAP14Z and G-GmPAP14N) and the promoters (P-GmPAP14Z and P-GmPAP14N) of GmPAP14, which might bring about differential transcriptional levels of GmPAP14 in ZH15 and NMH. Histochemical staining measurements revealed that a stronger GUS signal was present in transgenic Arabidopsis with P-GmPAP14Z under LP and normal P (NP) conditions compared with the P-GmPAP14N plant. Functional research demonstrated that transgenic Arabidopsis with G-GmPAP14Z had a higher level of GmPAP14 expression than the G-GmPAP14N plant. Meanwhile, higher APase activity was also observed in the G-GmPAP14Z plant, which led to increases in shoot weight and P content. Additionally, validation of variation in 68 soybean accessions showed that varieties with Del36 displayed higher APase activities than the del36 plant. Thus, these results uncovered that allelic variation in GmPAP14 predominantly altered gene expression to influence APase activity, which provided a possible direction for research of this gene in plants.

Quality by design-based capillary electrophoresis method development for the quantitative analysis of four iridoid compounds in Gentiana macrophylla Radix.

In this study, the capillary electrophoresis-photodiode array detector was employed for the analysis of four iridoid compounds in Gentiana macrophylla Radix (RGM), and the method was optimized based on the concept of analytical quality by design (AQbD). The peak areas relative standard deviation (n = 3) and resolution of the four analytes were selected as critical method attributes. Fractional factorial design test combined with Pareto analysis were employed to screen critical method parameters (buffer concentration, pH, sodium dodecyl sulfate [SDS] micelle concentration, temperature, and voltage). Subsequently, three main factors (buffer concentration, buffer pH, and SDS concentration) were selected by central composite design test for constructing the design space. The optimal separation conditions as follows: capillary column (50.2 cm × 50 µm, detection length 40 cm). Working background electrolyte consisted of 51 mmol/L borax solution (pH = 9.47) and 40 mmol/L SDS. The samples were injected by pressure (5 s at 0.5 psi) and the detection was performed at 254 nm. Applied voltage was 20 kV and column temperature was 23°C. The developed method is rapid and reliable for the quantitative analysis of four iridoid compounds in RGM, providing a reference for the application of AQbD concept in the analysis of natural products.

[Dissolution test and evaluation of Guizhi Fuling Capsules].

Because of the complex components, simple content determination can hardly reflect the overall quality of Guizhi Fuling Capsules. Therefore, it is necessary to carry out a multi-component dissolution test. The variability of quality among different batches of products from different manufacturers is a common problem of Chinese medicine solid preparations. To comprehensively control the quality of Guizhi Fuling Capsules, we studied the dissolution behaviors of 7 index components in the capsules under different conditions, and investigated the consistency of dissolution behaviors among different batches of products from the same manufacturer. The basket method of general rule 0931 in Chinese Pharmacopoeia was adopted, and the rotating speeds were set at 50, 75, and 100 r·min~(-1), respectively. The hydrochloric acid solution(pH 1.2), acetate buffer solution(pH 4.0), pure water, and phosphate buffer solution(pH 6.8) were used as the dissolution media. Automatic sampling was carried out at the time points of 5, 10, 20, 30, 45, and 60 min, respectively. The cumulative dissolution of 7 index components was measured through ultra-performance liquid chromatography(UPLC). The difference factor f_1 and similarity factor f_2 were calculated to comprehensively evaluate the similarity of the dissolution curves among 8 batches of Guizhi Fuling Capsules, and a variety of dissolution and release equations were fitted. The results showed that multiple components had faster dissolution rates at higher rotating speed and in hydrochloric acid medium. The 8 batches of Guizhi Fuling capsules showed the average f_1 value lower than 15 and the average f_2 value higher than 50, which indicated that different batches of products had similar dissolution behaviors. Most components had synchronous dissolution behaviors and similar release cha-racteristics. This study provides a reference for the quality consistency evaluation among batches, processing optimization, and dosage form improvement of Guizhi Fuling Capsules.

Migration and Deposition Law of Pollutants in Urban Sewage Confluence Pipe Network from the Perspective of Ecology.

Aiming at the problem of pollutant migration and deposition in urban sewage confluence pipe, an experimental simulation system of sewage confluence pipe was established. The confluence conditions of three flow patterns (velocity ratio Vaccess/Vtrunk = 0.1/0.2, Vaccess/Vtrunk = 0.1/0.3, and Vaccess/Vtrunk = 0.2/0.3) were simulated. The changes of sediment thickness, carbon pollutants, nitrogen pollutants, and phosphorus pollutants in different confluence areas were analyzed, and the migration and deposition laws of various pollutants in urban sewage confluence pipe network under different flow patterns were revealed. The results show that when the flow velocity of trunk and branch roads changes, the deposition of various pollutants and the carrying capacity of water flow in the pipeline change, resulting in the change of sediment layer thickness and pollutant content. With the increase of trunk velocity, the sediment thickness in the area before and after confluence decreases, while the increase of branch velocity only reduces the sediment thickness in the area at the back of confluence. Under any flow pattern, the sediment thickness in the retention area (G3 and G4) shows an increasing trend, which is the key area of pollution removal. Under the three flow patterns, the content of carbon pollutants reaches the peak at the TCOD and SCOD values of G4 monitoring point. Increasing the trunk velocity can effectively reduce the content of carbon pollutants. The content of nitrogen pollutants in each flow pattern also reaches the maximum at G4 point, which are 213.6 mg/g, 205.2 mg/g, and 212.8 mg/g, respectively. Increasing the trunk velocity can effectively reduce the nitrogen content at points G1-G4, while increasing the flow velocity of the branch road can reduce the nitrogen content at points G5-G7. The distribution of phosphorus pollutants is complex, and the flow pattern needs to be adjusted according to different monitoring points.

Intestinal microbiomics and liver metabolomics insights into the preventive effects of chromium (III)-enriched yeast on hyperlipidemia and hyperglycemia induced by high-fat and high-fructose diet.

In recent years, organic chromium (III) supplements have received increasing attentions for their low toxicity, high bioavailability and wide range of health-promoting benefits. This study aimed to investigate the preventive effects of chromium (III)-enriched yeast (YCr) on high-fat and high-fructose diet (HFHFD)-induced hyperlipidemia and hyperglycemia in mice, and further clarify its mechanism of action from the perspective of intestinal microbiomics and liver metabolomics. The results indicated that oral administration of YCr remarkably inhibited the aberrant elevations of body weight, blood glucose and lipid levels, hepatic cholesterol (TC) and triglyceride (TG) levels caused by HFHFD. Liver histological examination showed that oral YCr intervention inhibited HFHFD induced liver lipid accumulation. Besides, 16S rDNA amplicon sequencing showed that YCr intervention was beneficial to ameliorating intestinal microbiota dysbiosis by altering the proportion of some intestinal microbial phylotypes. Correlation-based network analysis indicated that the key intestinal microbial phylotypes intervened by YCr were closely related to some biochemical parameters associated with glucose and lipid metabolism. Liver metabolomics analysis revealed that dietary YCr intervention significantly regulated the levels of some biomarkers involved in purine metabolism, glycerophospholipid metabolism, citrate cycle, pyrimidine metabolism, glycerophospholipid metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, and so on. Moreover, dietary YCr intervention regulated the mRNA levels of key genes associated with glucose, cholesterol, fatty acids and bile acids metabolism in liver. These findings suggest that dietary YCr intervention has beneficial effects on glucose and lipid metabolism by regulating intestinal microbiota and liver metabolic pathway, and thus can be served as a functional component to prevent hyperlipidemia and hyperglycemia.

Identification and quality control strategy of impurities in Zhengqing Fengtongning injection.

Zhengqing Fengtongning injection is the sterile aqueous solution of Sinomenine Hydrochloride extracted from the root and stem of Sinomenium acutum, and is widely used to treat rheumatoid arthritis. Due to the processes of extraction, separation, purification, preparation and storage, some related impurities might be formed, which may cause side effects on patients. It is important to rapidly separate and identify the related impurities to ensure the safe use of Zhengqing Fengtongning injection. However, there are few literatures about the impurity in Zhengqing Fengtongning injection. In this work, ultra-high performance liquid chromatography- quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS) was developed to analyze impurities in both Zhengqing Fengtongning injection and its drug substance, with Sinomenine Hydrochloride as its active pharmaceutical ingredient (API). Six impurities of the Zhengqing Fengtongning injection were found. Structures of impurities 1 and 6 were confirmed by NMR and other impurities were identified from the fragmentation pattern of Sinomenine, the similarity of molecular weight and fragment ions in references. Finally, the HPLC analytical technique was developed to achieve the quantification of impurities 1 and 6. In addition, some reasonable suggestions are put forward on the quality control of Zhengqing Fengtongning injection and its drug substance based on the processes and structural characteristics of the related substances. The technical system established in this paper is helpful to strengthen the quality control of Zhengqing Fengtongning injection and improve production, and can also provide references for the production and quality control of similar drugs.

On-line screening of natural antioxidants and the antioxidant activity prediction for the extracts from flowers of Chrysanthemum morifolium ramat.

ETHNOPHARMACOLOGY RELEVANCE: Chrysanthemum morifolium Ramat. (Flos Chrysanthemi, FC) the most economically significant "food and drug dual-use" plants, with positive effects on relieving eye fatigue, and reduce internal heat, shows significant activities, such as anti-inflammatory, antioxidant, and neuroprotective, as well as alleviating diabetes effects. AIM OF THE STUDY: This study was undertaken to a screening of natural antioxidants in five kinds of medicinal FC and development of an integrated quality control method based on the antioxidant activity. MATERIALS AND METHODS: A novel quality control method for FC was established by combining the on-line HPLC-DPPH, ESI-MS, and NIR spectra analysis. Firstly, the on-line HPLC-DPPH-MS system was employed to identify the antioxidants in FC extracts. Then, the relationship between the NIR spectra and antioxidant activities of FC samples was calibrated to evaluate the total antioxidant capacity of FC rapidly. RESULTS: The established antioxidant activity-fingerprints contain both chemical information and antioxidant activity characteristics of FC. A total of 16 antioxidants were identified by on-line HPLC-ESI-MS analysis. The results of heat map analysis and cluster analysis showed that the classification method based on antioxidants in FC can be used to identify different cultivars of FC. The optimal pretreatment of the NIR spectra was determined to be row center (RC) 1st der + multiple-scatter correction (MSC) with an optimal LV value of 11. The developed spectral-antioxidant activity model had the excellent predictive ability and was successfully used to evaluate new batches of FC samples, where Rcal = 0.9445 and Rval = 0.8821. CONCLUSIONS: This comprehensive strategy may prove to be a powerful technique for the rapid screening, identification, and activity prediction of antioxidants, which could be used for the quality control of FC, and can serve as reference for design of quality control of other herbs and foods samples.

Two high hierarchical regulators, PuMYB40 and PuWRKY75, control the low phosphorus driven adventitious root formation in Populus ussuriensis.

Adventitious rooting is an essential biological process in the vegetative propagation of economically important horticultural and forest tree species. It enables utilization of the elite genotypes in breeding programmes and production. Promotion of adventitious root (AR) formation has been associated with starvation of inorganic phosphate and some factors involved in low phosphorus (LP) signalling. However, the regulatory mechanism underlying LP-mediated AR formation remains largely elusive. We established an efficient experimental system that guaranteed AR formation through short-term LP treatment in Populus ussuriensis. We then generated a time-course RNA-seq data set to recognize key regulatory genes and regulatory cascades positively regulating AR formation through data analysis and gene network construction, which were followed by experimental validation and characterization. We constructed a multilayered hierarchical gene regulatory network, from which PuMYB40, a typical R2R3-type MYB transcription factor (TF), and its interactive partner, PuWRKY75, as well as their direct targets, PuLRP1 and PuERF003, were identified to function upstream of the known adventitious rooting genes. These regulatory genes were functionally characterized and proved their roles in promoting AR formation in P. ussuriensis. In conclusion, our study unveiled a new hierarchical regulatory network that promoted AR formation in P. ussuriensis, which was activated by short-term LP stimulus and primarily governed by PuMYB40 and PuWRKY75.

An integration of UPLC-Q-TOF/MS and chemometrics analysis for the holistic quality evaluation of different geographical Paederia scandens.

RATIONALE: Due to the special geographical location and climate of China, there are large differences in the chemical composition and content of Paederia scandens (PS) from different origins, which will have a large impact on its efficacy. METHODS: An efficient quality control method for PS was established by combining ultra-high-performance liquid chromatography (UPLC) and electrospray ionization mass spectrometry analyses. First, a UPLC-quadrupole time-of-flight mass spectrometry system was employed to identify the chemical composition of PS from seven different origins. Then, the chemical variation in 73 batches of PS samples was subsequently investigated by quantitation of four marker compounds. RESULTS: A total of 15 common compounds were identified in the samples of PS from seven origins. And four of the marker compounds were chosen based on VIP values to characterize the differences between PS samples of different origins. The linearity ranged between 0.005 and 2.500 mg/mL; the correlation coefficients (r2 ) ranged from 0.999 to 1; the limits of detection ranged from 0.013 to 0.033 µg/mL; and the relative standard deviations for repeatability, precisions, and stabilities were below 0.2%, 1.6%, and 0.6%, respectively. CONCLUSION: The results show that the method can be effective and comprehensive in evaluating the quality of PS from different origins. And this comprehensive strategy proved to be a powerful technique used to differentiate between different geographical herbs.

GmSPX8, a nodule-localized regulator confers nodule development and nitrogen fixation under phosphorus starvation in soybean.

BACKGROUND: Biological nitrogen fixation (BNF) is an important nitrogen source for legume plants, and highly efficient nitrogen fixation requires sufficient phosphorus (P). However, the mechanism of maintaining nitrogen fixation of the legume nodules under low P concentration remains largely unknown. RESULTS: A nodule-localized SPX protein, GmSPX8, was discovered by transcriptome and functional analysis of its role in N2 fixation was characterized in soybean nodules. GmSPX8 was preferentially expressed in nodules and its expression was gradually increased during nodule development. And also the expression pattern was investigated using reporter gene ß-glucuronidase (GUS) driven by the promoter of GmSPX8. GmSPX8 was greatly induced and the GUS activity was increased by 12.2% under P deficiency. Overexpression of GmSPX8 in transgenic plants resulted in increased nodule number, nodule fresh weight and nitrogenase activity by 15.0%, 16.0%, 42.5%, subsequently leading to increased N and P content by 17.0% and 19.0%, while suppression of GmSPX8 showed significantly impaired nodule development and nitrogen fixation efficiency under low P stress. These data indicated that GmSPX8 conferred nodule development and nitrogen fixation under low P condition. By yeast two-hybrid screening, GmPTF1 was identified as a potential interacting protein of GmSPX8, which was further confirmed by BiFC, Y2H and pull down assay. Transcript accumulation of GmPTF1 and its downstream genes such as GmEXLB1 and EXPB2 were increased in GmSPX8 overexpressed transgenic nodules, and in the presence of GmSPX8, the transcriptional activity of GmPTF1 in yeast cells and tobacco leaves was greatly enhanced. CONCLUSIONS: In summary, these findings contribute novel insights towards the role of GmSPX8 in nodule development and nitrogen fixation partly through interacting with GmPTF1 in soybean under low P condition.

[Determination of five saponins in Xuesaitong Dropping Pills by micellar electrokinetic chromatography and evaluation method of between-batch consistency].

The present study determined five saponins in Xuesaitong Dropping Pills(XDP) by micellar electrokinetic chromatography(MEKC), and evaluated between-batch consistency by MEKC fingerprints and similarity analysis. A background buffer was composed of 20 mmol·L~(-1) sodium tetraborate-20 mmol·L~(-1) boric acid solution(pH 8.5), 55 mmol·L~(-1) sodium dodecyl sulfate(SDS), 23 mmol·L~(-1) ß-cyclodextrin, and 13% isopropyl alcohol. All separations were performed at 25 ℃,20 kV and the detection wavelength was set at 203 nm. The separation channel was a fused silica capillary with a dimension of 75 µm I.D. and a total length of 50.2 cm(effective length of 40.0 cm). The contents of notoginsenoside R_1, and ginsenosides Rg_1, Re, Rb_1, Rd were determined with their quality control ranges set. The fingerprints of XDP were established and the between-batch consistency was evaluated by similarity analysis. The contents of five saponins from the 19 batches of XDP were stable in the fixed ranges. Statistical analysis was carried out on the results of multiple batches of samples, and the specific quality control ranges were recommended as follows: notoginsenoside R_1 21.92-34.16 mg·g~(-1), ginsenosides Rg_1 83.54-131.78 mg·g~(-1), ginsenosides Re 13.58-19.82 mg·g~(-1), ginsenosides Rb_1 89.40-129.90 mg·g~(-1), and ginsenosides Rd 22.34-35.67 mg·g~(-1). Eleven characteristic peaks were identified in the fingerprints. Five peaks, notoginsenoside R_1 and ginsenosides Rg_1, Re, Rb_1, Rd, were identified with reference standards. The similarities of the 19 batches of samples were all above 0.988, indicating good between-batch consistency. This method is green and simple, and can be used for the quantitative determination and quality evaluation of XDP. It can also provide references for the quality control of other Chinese medicinal dripping pills.

Quality assessment of Succus Bambusae oral liquids based on gas chromatography/mass spectrometry fingerprints and chemometrics.

RATIONALE: Succus Bambusae is consumed as a kind of herbal medicine and natural beverage in China. However, the current quality standards for Succus Bambusae are low and lack safety indicators, which makes it difficult to effectively guarantee its quality. Therefore, it is of great significance to study the identification and quality control technology for the product. METHODS: We have developed a set of qualitative and quantitative methods based on gas chromatography/mass spectrometry (GC/MS) for the analysis of volatile components in Succus Bambusae oral liquid (SBOL). Combining GC/MS fingerprint analysis and related chemometrics algorithms, with similarity evaluation, Hotelling T2 and distance to Model X (DModX) as criteria, the quality consistency of different batches was evaluated, and SBOL samples from different manufacturers were differentiated. RESULTS: Twenty-nine volatile components were preliminarily identified from 40 batches of SBOL samples from six manufacturers, and six Q-markers (Quality Markers) for the SBOLs were discussed and determined using GC/MS. The products from different manufacturers were distinguished using chemometrics. CONCLUSIONS: The results showed that the quality of the SBOL samples from different batches and different manufacturers fluctuated greatly, which suggested that research into the raw materials and manufacturing techniques should be strengthened to improve the quality of SBOL and ensure its quality consistency.