Transcription factor OsWRKY72 controls rice leaf angle by regulating LAZY1-mediated shoot gravitropism.

Leaf angle is a major trait of ideal architecture, which is considered to influence rice (Oryza sativa) cultivation and grain yield. Although a few mutants with altered rice leaf inclination angle have been reported, the underlying molecular mechanism remains unclear. In this study, we showed that a WRKY transcription factor gene, OsWRKY72, was highly expressed in the leaf sheath and lamina joint. Phenotypic analyses showed that oswrky72 mutants have smaller leaf angles than the wild type (WT), while OsWRKY72 overexpression lines exhibited increased leaf angle. This observation suggests that OsWRKY72 functions as a positive regulator, promoting the enlargement of the leaf angle. Our bioinformatics analysis identified LAZY1 as the downstream gene of OsWRKY72. Electrophoretic mobility shift assays (EMSA) and dual luciferase analysis revealed that OsWRKY72 directly inhibits LAZY1 by binding to its promoter. Moreover, knocking out OsWRKY72 enhanced shoot gravitropism, which is in contrast to the phenotype of lazy1 plants. These results imply that OsWRKY72 regulates leaf angle through gravitropism by reducing the expression of LAZY1. In addition, OsWRKY72 could directly regulate the expression of other leaf angle-related genes, such as FLOWERING LOCUS T-LIKE 12 (OsFTL12) and WALL-ASSOCIATED KINASE 11 (OsWAK11). Our study indicates that OsWRKY72 contributes positively to the expansion of the leaf angle by interfering with shoot gravitropism in rice.

Longitudinal relationship between harsh parenting and adolescent non-suicidal self-injury: The roles of basic psychological needs frustration and self-concept clarity.

BACKGROUND: Harsh parenting has been shown to be associated with adolescents' non-suicidal self-injury (NSSI) behavior. However, less is known about the mechanism underlying the association. OBJECTIVE: The aim of this study was to use a one-year longitudinal design to investigate the mediating role of basic psychological needs frustration and the moderating role of self-concept clarity in the association between harsh parenting and NSSI. METHODS: Three waves of data (called T1, T2 and T3) were collected 6 months apart, between April 2021 and 2022, in a sample of 786 Chinese adolescents (52.0 % girls; Mage at Wave 1 = 13.27 years). Structural equation model was used to test a longitudinal moderated mediation model, with gender, age and baseline NSSI as covariates. RESULTS: The results showed process of mediation in which T1 harsh parenting was longitudinally and positively associated with T3 NSSI through T2 basic psychological needs frustration. Furthermore, moderated mediation analyses revealed that T2 self-concept clarity buffered the adverse impact of T2 needs frustration on T3 NSSI, thereby mitigating the mediation process. CONCLUSIONS: The findings support the self-determination theory, and suggest that fostering adolescents' self-concept clarity and satisfying their psychological needs may be useful in programs designed to lower the risk of NSSI.

Functions of WRKYs in plant growth and development.

As sessile organisms, plants must overcome various stresses. Accordingly, they have evolved several plant-specific growth and developmental processes. These plant processes may be related to the evolution of plant-specific protein families. The WRKY transcription factors originated in eukaryotes and expanded in plants, but are not present in animals. Over the past two decades, there have been many studies on WRKYs in plants, with much of the research concentrated on their roles in stress responses. Nevertheless, recent findings have revealed that WRKYs are also required for seed dormancy and germination, postembryonic morphogenesis, flowering, gametophyte development, and seed production. Thus, WRKYs may be important for plant adaptations to a sessile lifestyle because they simultaneously regulate stress resistance and plant-specific growth and development.

Preparation and identification of an anti-nicarbazin monoclonal antibody and its application in the agriculture and food industries.

Background: As a broad-spectrum drug against chicken coccidiosis, nicarbazine is widely used. The international community has made regulations and requirements on the residue limits of nicarbazine metabolites in chicken. The research reports on the detection methods of nicarbazine residues are mainly based on large-scale instruments such as high-performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry-mass spectrometry (LC/MS/MS) and so on. However, in the practical monitoring and detection application, the rapid, sensitive, efficient and accurate detection of nicarbazine residues is becoming more and more urgent. Methods: This study aimed to establish an enzyme-linked immunosorbent assay (ELISA)-based method to detect nicarbazin drug residues with high sensitivity and specificity, and wide applicability. Artificial immunogens were prepared by molecular modification synthesis. Nuclear magnetic resonance (NMR) and ultraviolet analyses were conducted to confirm that the correct product was obtained. Monoclonal antibodies were acquired by immunizing mice and preparing hybridoma cells. Results: In this study, 4,4'-dinitrocarbanilide (DNC), a metabolite of nicarbazine, was synthesized and modified to make it have immunogenicity. Fifteen healthy female mice of 6-8 weeks old were immunized in three groups. The successfully immunized mice were screened by serum titer. One mouse with the highest titer was fused and cloned three times, and four positive cell lines were obtained. Nine monoclonal antibodies were obtained from mouse ascites. The best matched antigens and antibodies were screened by an ELISA chessboard method. A detection method of nicarbazine ELISA kit was developed. Our prepared anti-nicarbazin monoclonal antibody had a half-maximal inhibitory concentration (IC50) of 0.825 ng/mL, and the curve range was 0.3-24.3 ng/mL. There was no cross reaction to other six common anti-coccidiosis drugs. The recovery results showed that the fortified recovery of the chicken and duck samples ranged from 74.4-111.7%, the test results of which all met the requirements for veterinary drug residue detection. Conclusions: This method, which uses a specific antibody against the nicarbazin metabolic product DNC, enables rapid quantitative detection. Our new ELISA-based method should facilitate the development of assays to monitor and detect agricultural and veterinary drug residues.

Generation and Characterization of an Anti-diclazuril Monoclonal Antibody and Development of a Diagnostic Enzyme-Linked Immunosorbent Assay for Poultry.

An anti-diclazuril monoclonal antibody (mAb) was developed for use in enzyme-linked immunosorbent assay (ELISA)-based detection of diclazuril with high sensitivity and specificity, which can be used to measure anti-coccidial drug residues. The anti-diclazuril mAb had a half-maximal inhibitory concentration of 0.449-0.517 ng/mL. The mAb cross-reactivity with toltrazuril, toltrazuril 18 sulfone, clozaril, monesin, madurmycin, and salinomycin was very minimal (< 0.1%). The detection limit of the ELISA using this mAb was 0.10 ng/mL and the sensitivity was 0.05 ng/mL. A standard curve generated in the range of 0.05-16.2 ng/mL had a linear correlation coefficient value of ≥ 0.99. The average recoveries of diclazuril from chicken and duck samples ranged from 85.0 to 102.5%.Intra- and inter-assay coefficients of variation ranged from 5.9 to 8.5% and 9.2 to 12.6%, respectively. Using the International Immunogenetics Information System®, the VH domain of the mAb was found to be encoded by an IGHV3 family gene and had the following complementarity determining region (CDR) sequences: GFTFSRY (CDR1), SRGGS (CDR2), and GDDNYAFAY (CDR3). The VL domain was encoded by an IGKV1 family gene and had the following CDR sequences: KSSQSLLNSRTRKNYLA (CDR1), WASTRES (CDR2), and KQSYNLHT (CDR3). This study provides a method to generate anti-diclazuril mAbs and determine their variable region sequences. The diagnostic ELISA developed using this mAb may drive additional studies on the monitoring and detection of food and veterinary drug residues.

Construction of α-Fe2O3/Sulfur-Doped Polyimide Direct Z-Scheme Photocatalyst with Enhanced Solar Light Photocatalytic Activity.

A novel two-dimensional α-Fe2O3/sulfur-doped polyimide (FO/SPI) direct Z-scheme photocatalyst was successfully constructed by a facile thermal treatment method. The effects of α-Fe2O3 nanosheets on the morphology, chemical structure, and photoelectronic properties of FO/SPI composites were systematically characterized by different spectroscopic means. These methods include X-ray diffraction, scanning electron microscopy, X-ray photoelectron spectroscopy, transient fluorescence spectra, and so forth. It was confirmed that the small amounts of α-Fe2O3 can availably facilitate exfoliation of bulk SPI, resulting in a transformation of SPI from bulk to 2D layered composite that illustrates tight interface through the coordination Fe-N bond and an all-solid-state direct Z-scheme junction. Thus, the transfer and separation efficiency of photogenerated electron/hole pairs were significantly enhanced, which greatly promoted improvement of the photocatalytic activity of the FO/SPI composite for methyl orange degradation under solar light. This work provides a new approach to constructing efficient inorganic-organic Z-scheme photocatalyst based on strong interface interaction.

Combined analysis of the transcriptome and metabolome provides insights into the fleshy stem expansion mechanism in stem lettuce.

As a stem variety of lettuce, the fleshy stem is the main product organ of stem lettuce. The molecular mechanism of fleshy stem expansion in stem lettuce is a complex biological process. In the study, the material accumulation, gene expression, and morphogenesis during fleshy stem expansion process were analyzed by the comparative analysis of metabolome, transcriptome and the anatomical studies. The anatomical studies showed that the occurrence and activity of vascular cambium mainly led to the development of fleshy stems; and the volume of pith cells gradually increased and arranged tightly during the expansion process. A total of 822 differential metabolites and 9,383 differentially expressed genes (DEGs) were identified by the metabolomics and transcriptomics analyses, respectively. These changes significantly enriched in sugar synthesis, glycolysis, and plant hormone anabolism. The expression profiles of genes in the sugar metabolic pathway gradually increased in fleshy stem expansion process. But the sucrose content was the highest in the early stage of fleshy stem expansion, other sugars such as fructose and glucose content increased during fleshy stem expansion process. Plant hormones, including IAA, GA, CTK, and JA, depicted important roles at different stem expansion stages. A total of 1,805 DEGs were identified as transcription factors, such as MYB, bHLH, and bZIP, indicating that these transcription factor families might regulate the fleshy stems expansion in lettuce. In addition, the expression patterns identified by qRT-PCR were consistent with the expression abundance identified by the transcriptome data. The important genes and metabolites identified in the lettuce fleshy stem expansion process will provide important information for the further molecular mechanism study of lettuce fleshy stem growth and development.

The emerging role of jasmonate in the control of flowering time.

Plants dynamically synchronize their flowering time with changes in the internal and external environments through a variety of signaling pathways to maximize fitness. In the last two decades, the major pathways associated with flowering, including the photoperiod, vernalization, age, autonomous, gibberellin, and ambient temperature pathways, have been extensively analyzed. In recent years, an increasing number of signals, such as sugar, thermosensory, stress, and certain hormones, have been shown to be involved in fine-tuning flowering time. Among these signals, the jasmonate signaling pathway has a function in the determination of flowering time that has not been systematically summarized. In this review, we present an overview of current knowledge of jasmonate control of flowering and discuss jasmonate crosstalk with other signals (such as gibberellin, defense, and touch) during floral transition.

Phytomelatonin inhibits seed germination by regulating germination-related hormone signaling in Arabidopsis.

Seed germination is a vital initial stage in the life cycle of a plant, which determines subsequent vegetative growth and reproduction. Melatonin acts as a plant's master regulator and is also involved in the process of seed germination. In a recent study, we show that the high concentration melatonin inhibited seed germination in Arabidopsis. Transcriptome and phenotype analysis implied that melatonin-mediated seed germination interacted with phytohormones abscisic acid (ABA), gibberellin (GA), and auxin. In this short communication, we discuss the mechanism of phytomelatonin that inhibits seed germination through ABA, GA, and IAA in Arabidopsis.

Iron deficiency-induced transcription factors bHLH38/100/101 negatively modulate flowering time in Arabidopsis thaliana.

The mechanisms regulating flowering have been extensively studied and the roles of many environmental signals in this process have been reported. However, little is known on the relationship between iron deficiency and flowering regulation, although the response mechanism to iron deficiency has been studied for decades. In this study, we observed that the flowering time of wild-type Arabidopsis thaliana was significantly repressed by iron deficiency under long days. Phenotype analysis showed that iron deficiency delayed flowering of Arabidopsis through the iron deficiency-induced transcription factors bHLH38, bHLH100, and bHLH101 (bHLH38/100/101), which redundantly regulated flowering time and expression of FLOWERING LOCUS T (FT) specifically under long days. Genetic analysis indicated that disruption of FT expression suppressed the early-flowering phenotype of bhlh38/100/101 triple-mutant plants, indicating that bHLH38/100/101 are dependent on functional FT. Furthermore, bHLH38/100/101 interacted with CONSTANS (CO), thereby interfering with the transcriptional activation of CO to regulate FT expression. Therefore, the results indicated that iron deficiency affects flowering of Arabidopsis under long days through bHLH38/100/101-CO-FT signaling.

Rapid determination of sulfonamides in chicken using two-dimensional online cleanup mode with three columns coupled to liquid chromatography-tandem mass spectrometry.

A novel method based on online cleanup mode combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) was established. After an automated sample cleanup system with aqueous gel column, sulfonamides in chicken were detected in multiple reaction monitoring (MRM) mode. The total run time of the system, which included automated extraction, analytical chromatography and re-equilibration, was within 30 min. Different experimental processes containing extraction, purification, separation, and detection have been evaluated respectively to obtain optimized parameters. The developed method was fully validated and the efficient and superior performance of the developed method was demonstrated. The method produced linear results for all sulfonamides from 1 to 10 ng g-1 with a linearity >0.99. The intra-day precision of the method was <8.45% while the inter-day precision was <9.11%. The matrix effect was 77.5% to 105.1%. The recovery was in the range of 72.66% to 116.7% for all sulfonamides. The limit of quantitation in the chicken was 0.6 ng g-1 and the limit of detection was 0.2 ng g-1. Compared with traditional procedures, the automated sample clean-up strategy could significantly shorten the analysis time and offer higher detectability, with the advantage of sufficient sensitivity. Also, the use of gel chromatography column employed the water phase and reduced the organic reagent to achieve the level of green chemistry.

Simultaneous determination of 131 pesticides in tea by on-line GPC-GC-MS/MS using graphitized multi-walled carbon nanotubes as dispersive solid phase extraction sorbent.

A rapid and sensitive method was developed to determine 131 pesticides in tea by on-line gel permeation chromatography-gas chromatography tandem mass spectrometer (on-line GPC-GC-MS/MS) using graphitized multi-walled carbon nanotubes (MWCNTs) as dispersive solid phase extraction (DSPE) sorbent in sample pretreatment. Sample clean-up performance of various functionalized MWCNTs and their influences to recoveries were investigated. In the method validation, good linearity correlation coefficients (R2) were obtained, which were higher than 0.99 for all pesticides. The percent recoveries were 78.2%-113.9% at the spiked concentration levels of 20, 50 and 200 µg/kg with relative standard deviations (RSDs, n = 6) lower than 15.8%. Limits of detections (LODs) were in the range of 0.5-5.0 µg/kg. This method is simple and fast with relatively high sensitivity and can achieve the requirements of pesticides analysis.

Serum bisphenol A concentration and premature thelarche in female infants aged 4-month to 2-year.

OBJECTIVES: To estimate the association between serum bisphenol A and premature thelarche in female infants aged 4-mo to 2-y. METHODS: A total of 251 female infants (aged 4 mo to 2 y) with premature thelarche and 33 healthy age-matched control subjects were analyzed. All participants underwent physical examination and serum bisphenol A was measured by ultra-performance liquid chromatography tandem mass spectrometry. RESULTS: Serum bisphenol A concentration in the premature thelarche group (3.48 ng/ml, 95%CI: 0.09-140.26) was significantly higher than that in the control group (1.70 ng/ml, 95%CI: 0.06-51.78) (p = 0.039). There was no correlation between age and serum bisphenol A (BPA) level. Univariate logistic regression analysis showed that serum BPA concentration positively associated with premature thelarche, and the effect of BPA fell down as the age grew. CONCLUSIONS: This hospital-based study implied that there is an association between serum BPA concentrations and premature thelarche. Additionally, serum BPA levels were markedly higher in infants aged 4-mo to 2-y-old, raising a concern for public health authorities.

[Determination of dicyandiamide in dairy products by high performance liquid chromatography].

A high performance liquid chromatographic (HPLC) method for the determination of dicyandiamide in dairy products was developed. The sample was extracted by acetonitrile, separated on an XBridge Amide column (250 mm x 4.6 mm, 3.5 microm) using acetonitrile-water (90/10, v/v) containing 0.2% (v/v) formic acid as the mobile phase at a flow rate of 1.0 mL/min, and determined by a photodiode array detector at the wavelength of 218 nm. The calibration curve was linear in the range of 0.5 - 50 mg/L with the correlation coefficient (r2) of 0.999 9. The recoveries were from 96.7% to 101.0% with the relative standard deviations (RSDs) in the range of 4.5% to 4.9% (n = 6). The limit of detection was 0.2 mg/kg, and the limit of quantification was 0.5 mg/kg. The method is simple, sensitive, accurate and precise for the determination of dicyandiamide in dairy products.