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Brassinosteroids (BRs) play essential roles in regulating plant growth and development, however, gaps still remain in our understanding of the BR signaling network. We previously cloned a grain length quantitative trait locus qGL3, encoding a rice (Oryza sativa L.) protein phosphatase with Kelch-like repeat domain (OsPPKL1), that negatively regulates grain length and BR signaling. To further explore the BR signaling network, we performed phosphoproteomic analysis to screen qGL3-regulated downstream components. We selected a 14-3-3 protein OsGF14b from the phosphoproteomic data for further analysis. qGL3 promoted the phosphorylation of OsGF14b and induced the interaction intensity between OsGF14b and OsBZR1. In addition, phosphorylation of OsGF14b played an important role in regulating nucleocytoplasmic shuttling of OsBZR1. The serine acids (Ser258Ser259) residues of OsGF14b play an essential role in BR-mediated responses and plant development. Genetic and molecular analyses indicated that OsGF14b functions as a negative regulator in BR signaling and represses the transcriptional activation activity of OsBZR1. Collectively, these results demonstrate that qGL3 induces the phosphorylation of OsGF14b, which modulates nucleocytoplasmic shuttling and transcriptional activation activity of OsBZR1, to eventually negatively regulate BR signaling and grain length in rice.

The immune regulation function of ovotransferrin (OVT) explored using the RAW264.7 was induced by lipopolysaccharide (LPS) as vitro model in this study. The results showed that RAW264.7 cultured with OVT (200 μg/ml) alone not only enhanced the phagocytic activity and the production and expression of inflammatory factors, but also expression of toll‐like receptor 4 (TLR4) gene was significantly promoted by OVT. OVT (50 μg/ml) significantly inhibited the secretion and expression of inflammatory factors in LPS‐stimulated RAW264.7, but CD14 and TLR4 genes expressions were no obvious effects. Inflammatory cytokines and NO secreted by OVT‐induced macrophages pretreated with inhibitors of TLR4 were down‐regulated. We further verified the effects of OVT on inflammatory signaling pathway‐related proteins through immunofluorescence and western blotting, MyD88, TLR4 and the phosphorylation of IκBα and p65 were significantly promoted by OVT, but there was no significant effects on the phosphorylation of IRF3. OVT promoted the phosphorylation of ERK and p38 in RAW264.7 and inhibited the phosphorylated expression of MAPK in LPS‐mediated inflammation. These results indicated that OVT had the bidirectional immunoregulatory function through TLR4‐mediated NF‐κB/MAPK signaling pathway, that is, anti‐inflammatory effect of low concentration and immune‐enhancing activity of high concentration were showed. That provides a theoretical utilization for the development and utilization of OVT., Ovotransferrin had the bidirectional immunoregulatory function through TLR4‐mediated NF‐κB/MAPK signaling pathway, that is, anti‐inflammatory effect of low concentration and immune‐enhancing activity of high concentration were showed.

To determine furaltadone metabolites 5-methylmorpholino-3-amino-2-oxazolidinone (AMOZ) residual levels in ctenopharyngodon idellus, direct competitive enzyme-linked immunosorbent assay (dcELISA) was developed based on a single-chain variable fragment (scFv)-alkaline phosphatase (AP) fusion protein. Firstly, VH and VL gene sequences were cloned from hybridoma cell lines secreting monoclonal antibody against AMOZ derivative, and then the productive VH and VL genes were assembled to a scFv gene and engineered to produce scFv-AP fusion protein. The fusion protein was further identified as a bifunctional reagent for immunoassay, then a sensitive one-step dcELISA against AMOZ derivative was developed with a half-maximal inhibitory concentration (IC50) and limit of detection (LOD) of 10.62 and 4.83 ng/mL, respectively. The recovery values of AMOZ from spiked fish samples determined by dcELISA were in 71.31%∼82.88%. The results suggested that the prepared scFv-AP fusion protein in this work can be used for rapid and convenient immunoassays to detect AMOZ residues in fish samples.

BACKGROUND Hydrogen sulfide (H2 S) is a known signaling molecule in plants, which has the ability to delay fruit ripening. Our previous studies have shown that H2 S treatment could delay the maturation of kiwifruits by inhibiting ethylene production, improving protective enzyme activities, and decreasing the accumulation of reactive oxygen species to protect the cell membrane during storage. The mechanism related to the way in which H2 S affected kiwifruit maturation was still unclear. We performed transcriptome sequencing to explore the influences of H2 S on the softening of kiwifruit. RESULTS The firmness and the soluble solids content (SSC) of the kiwifruit were significantly better maintained with H2 S treatment compared to the control during the storage period (P

Rice (Oryza sativa L.) is one of the most important crops in the world, and grain size is a major component determining rice yield. Recent studies identified a number of grain size regulators, which are involved in the phytohormone signaling, G protein signaling, mitogen-activated protein kinase signaling pathway, ubiquitin-proteasome pathway, or transcriptional regulation. In previous study, we cloned qGL3/OsPPKL1 encoding a rice protein phosphatase that negatively modulates brassinosteroid (BR) signaling and grain length. Here, to further explore qGL3-mediated BR signaling network, we performed phosphoproteomic screenings using two pairs of rice materials: the indica rice cultivar 9311 and its near-isogenic line NILqgl3 and the japonica rice cultivar Dongjin and its qGL3 knockout mutant m-qgl3. Together with qGL3-interacting proteins, we constructed the qGL3-mediated network, which reveals the relationships between BR signaling and other critical signaling pathways. Transgenic plants of these network components showed BR-related alterations in plant architecture. From this network, we validated a qGL3-interacting protein, O. sativa VERNALIZATION INSENSITIVE3-LIKE1 (OsVIL1), and demonstrated that qGL3 dephosphorylates OsVIL1 to modulate BR signaling. The qGL3-dependent network uncovered in this study increases our understanding of BR signaling and provides a profound foundation for addressing how BR modulates plant architecture in rice.

Current research aimed at studying the effect of benzothiazole (BTH) on the fruit quality and resistance against Penicillium italicum (P. italicum). Recently, a synthetically prepared novel BTH was introduced that elicits the induction of resistance against various diseases of fruits. However, little was reported on the effect of BTH on the disease resistance and fruit quality of postharvest navel orange fruit. In this study, 50 mg·L−1 BTH significantly reduced the decay rate of fruits during 36 days of storage at 20 ± 0.5°C ( P < 0.05 ). BTH markedly inhibited the weight loss rate in fruits ( P < 0.05 ) and effectively maintained higher soluble solid content (SSC), titratable acid (TA), and vitamin C (VC) content compared with control navel orange fruits. Further, BTH significantly suppressed the increase of disease incidence and lesion area of orange fruits challenged with P. italicum ( P < 0.05 ). BTH treatment significantly enhanced antioxidant capacity (DPPH, ABTS radical scavenging activity, and reducing power), and superoxide dismutase (SOD) and peroxidase (POD) activities were significantly increased, while the activity of catalase (CAT) was opposite to the former ( P < 0.05 ). The activities of β-1,3-glucanase (GLU), phenylalanine ammonia-lyase (PAL), and chalcone isomerase (CHT) were significantly higher in BTH-treated navel orange fruits ( P < 0.05 ). Our results suggested that BTH treatment may be a promising treatment for maintaining the quality and inhibiting blue mold of postharvest navel orange in the future.

Scope Inflammation is the pathological basis of many chronic diseases, and persistent intestinal inflammation is a key factor in the further development of colon cancer. Egg-derived peptides have been proven to have anti-intestinal inflammation activity. Egg white treated with salt contains a lot of rich protein, whether its peptides have anti-inflammatory activity and how their mechanism of action is still unclear. Methods and results In this study, ELISA was used to determine the anti-inflammatory activity of the peptides (VF-4 and DR-8 from salted egg white), and then RNA-seq was used to explore the mechanism of their anti-inflammatory activity, and then verified by western blotting and inhibitors. The results showed that VF-4 and DR-8 significantly inhibited TNF-α-induced IL-8 secretion in HT-29 cells in a concentration-dependent manner, and VF-4 showed a more significant anti-inflammatory effect than DR-8. The anti-inflammatory mechanism of VF-4 and DR-8 was through inhibiting the activation of NF-κB, MAPK, and PI3K-Akt pathways, reducing the production of inflammatory mediators. Conclusion VF-4 and DR-8 have obvious anti-inflammatory activity, which can reduce intestinal inflammation and inhibit its further development into colon cancer. This article is protected by copyright. All rights reserved.

Ethnopharmacological relevance Egg yolk oils (EYO) is a traditional Chinese medicine obtained from Gallus gallus domesticus Brisson, which has been used to treat inflammatory related diseases such as cheilitis, ulceration and acute anal fissure. However, the detailed anti-inflammatory mechanism of EYO is still unknown. Aim of the study The anti-inflammatory activity and mechanism of EYO were investigated in tumor necrosis factor (TNF)-α induced Caco-2 cells. Materials and methods EYO was obtained by direct-heat extraction (HE), ethanol extraction (EE) and petroleum ether extraction (PE), respectively. Fatty acid compositions of three EYO were measured by gas chromatography (GC). Cell viability, enzyme-linked immunosorbent assay (ELISA), transcriptome, RT-PCR and Western blotting were also performed. Results Fatty acid compositions of three EYO were different with varied extraction methods. EYO significantly reduced interleukin (IL)-8 secretion. EYO exerted anti-inflammatory effect via coordinating regulation of Nrf2/NF-κB pathways based on the results of transcriptome, Q-PCR and Western blotting. In detail, PE and HE inhibited the NF-κB pathway, whereas EE exerted anti-inflammatory activity via the Nrf2/NF-κB pathways. Conclusions The aforementioned results showed the anti-inflammatory mechanism of EYO. These findings might be beneficial to clinical applications of EYO.

Changes in the gelation behavior and mechanism were investigated in low-density lipoprotein (LDL) and high-density lipoprotein (HDL) under the treatment of alkali. Three textural parameters of two kinds of alkali-induced lipoprotein increased, especially the springiness of LDL and the hardness of HDL. Alkaline treatment contributes to the sulfhydryl (SH) oxidation and SH-disulfide bonds (SS) exchange reactions to produce SS, promoting the aggregation of proteins and the formation of lipoprotein gels. For LDL gels, the increased hydrophobic interaction improves its thermal stability. HDL underwent deep unfolding and reassemble to form a network structure with high thermal stability and uniform density. Continuously alkaline treatment destroyed less SS and exposed more buried SH. The decrease of β-structures in both lipoproteins was observed by Fourier Transform Infrared (FTIR). These results suggested that alkali-induced LDL and HDL gelation behavior highly related to the protein structure, SS and hydrophobic interactions.

Nitric oxide (NO) is a signal molecule that can regulate fruit physiology. Several studies have indicated that NO can inhibit the ripening of kiwifruit, but the mechanism underlying this process is unresolved. This study used transcriptome analysis to identify the essential genes related to NO regulation during kiwifruit softening. NO gas fumigation (15 μl L-1) significantly delayed kiwifruit softening. There were 736 differentially expressed genes (DEGs) between the NO treatment and the control. The expression levels of polygalacturonase (PG), pectate lyase (PL), β-galactosidase (β-GAL), pectinesterase (PE), and the beta-amylases-related genes decreased in response to the NO treatment, while those of four genes encoding cellulose synthase increased. The expression of genes related to ethylene biosynthesis and signal transduction also differed; the expression levels of 1-aminocyclopropane carboxylic acid oxidase (ACO), the ethylene receptors (ERS1, ETR2), and the ethylene-responsive transcription factors (ERF016, ERF7, ERF010, ERF062, ERF110, ERF037, ERF008, ERF113, ERF12, ERF095) were lower in the NO-treated kiwifruit. Expression of the calcium ion (Ca2+) signal-related genes (CNGC1, CPK1, CIPK2, CML31, CML48, ZIFL1) significantly differed and may be involved in the regulation of the NO softening response. These findings add to our understanding of the molecular mechanisms of the NO-delayed softening response in kiwifruit.

Current study aimed to evaluate the effects of fumigation with 15 μl L−1 nitric oxide (NO) on the quality of ‘Newhall’ navel orange fruit during storage at 5 °C. Fruits quality parameters, decay rate, antioxidant capacity, antioxidant enzymes activities and disease resistance related enzymes activities were evaluated. NO treatment significantly inhibited the increasing weight loss, maintained soluble solid content (SSC), titratable acid (TA) and vitamin C (VC) contents (P

In this paper, the physicochemical and nutritional characteristics of preserved duck egg white were analyzed and compared with fresh egg and hard-cooked egg white (n = 3). The data obtained showed that the preserved egg white was rich in essential amino acids and minerals, such as Ca, Mg, Fe, Zn, Cu, K, and Na. After fresh duck eggs were processed into preserved eggs, contents of moisture, CP, amino acid, and water-soluble vitamin of egg white significantly decreased (P < 0.05); however, pH, free amino acid content, and most inorganic elemental contents of egg white significantly increased (P < 0.05). The preserved egg white had higher a* (redness/greenness) and b* values (yellowness/blueness; P < 0.05) and lower L* value (lightness; P < 0.05) than hard-cooked egg white. The gel hardness of preserved egg white was approximately 50% of hard-cooked egg white; however, its springiness and cohesiveness were approximately 1.5 times of hard-cooked egg white. The results indicated that pickling with alkaline and other additives can significantly change physical properties and chemical composition of duck egg white, which make preserved egg white with characteristics of rich elements, brown color, and high springiness, but low vitamin.

The effects of fumigation with NO, H2S and combination of both on the quality, ethylene biosynthesis and softening of postharvest ‘Dahong’ peach fruit were investigated in current study. Results showed that NO or H2S significantly inhibited the increase of rot index, soluble solid contents, decrease of firmness along with titratable acid contents (P