Phytochrome B mediates dim-light-reduced insect resistance by promoting the ethylene pathway in rice.

Increasing planting density is one of the most effective ways to improve crop yield. However, one major factor that limits crop planting density is the weakened immunity of plants to pathogens and insects caused by dim light (DL) under shade conditions. The molecular mechanism underlying how DL compromises plant immunity remains unclear. Here, we report that DL reduces rice (Oryza sativa) resistance against brown planthopper (BPH; Nilaparvata lugens) by elevating ethylene (ET) biosynthesis and signaling in a Phytochrome B (OsPHYB)-dependent manner. The DL-reduced BPH resistance is relieved in osphyB mutants, but aggravated in OsPHYB overexpressing plants. Further, we found that DL reduces the nuclear accumulation of OsphyB, thus alleviating Phytochrome Interacting Factor Like14 (OsPIL14) degradation, consequently leading to the up-regulation of 1-Aminocyclopropane-1-Carboxylate Oxidase1 (OsACO1) and an increase in ET levels. In addition, we found that nuclear OsphyB stabilizes Ethylene Insensitive Like2 (OsEIL2) by competitively interacting with EIN3 Binding F-Box Protein (OsEBF1) to enhance ET signaling in rice, which contrasts with previous findings that phyB blocks ET signaling by facilitating Ethylene Insensitive3 (EIN3) degradation in other plant species. Thus, enhanced ET biosynthesis and signaling reduces BPH resistance under DL conditions. Our findings provide insights into the molecular mechanism of the light-regulated ET pathway and host-insect interactions and potential strategies for sustainable insect management.

Development of a sensitive direct competitive chemiluminescent enzyme immunoassay for gentamicin based on the construction of a specific single-chain variable fragment-alkaline phosphatase fusion protein.

A sensitive chemiluminescent enzyme immunoassay (CLEIA) was established for the determination of gentamicin (GEN) residue levels in animal tissue. This assay is based on a fusion protein of single-chain variable fragment (scFv) and alkaline phosphatase (AP). Initially, VL and VH derived from anti-gentamicin monoclonal antibody were linked by a short peptide to construct a scFv. Subsequently, the constructed scFv sequence was accessed into the pLIP6/GN vector, and a soluble scFv-AP fusion protein was generated. The scFv-AP fusion protein was used to develop a direct competitive CLEIA (dcCLEIA) for the determination of gentamicin. In the dcCLEIA, the half inhibitory concentration (IC50) and limit of detection (LOD) were 1.073 ng/mL and 0.380 ng/mL, respectively. The average recoveries of gentamicin spiked in animal tissue samples ranged from 78% to 96%. These results showed a strong correlation with ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). The above results suggest that the anti-GEN scFv-AP fusion protein is suitable for detecting gentamicin residues in edible animal tissues.

The phosphoproteomic and interactomic landscape of qGL3/OsPPKL1-mediated brassinosteroid signaling in rice.

Oryza sativa L. (rice) is one of the most important crops in the world, and grain size is a major component determining rice yield. Recent studies have identified a number of grain size regulators, which are involved in phytohormone signaling, G protein signaling, the mitogen-activated protein kinase signaling pathway, the ubiquitin-proteasome pathway or transcriptional regulation. In a previous study, we cloned qGL3/OsPPKL1 encoding a rice protein phosphatase that negatively modulates brassinosteroid (BR) signaling and grain length. Here, to further explore the 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 INSENSITIVE 3-LIKE 1 (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.

Analysis of the Properties of 44 ABC Transporter Genes from Biocontrol Agent Trichoderma asperellum ACCC30536 and Their Responses to Pathogenic Alternaria alternata Toxin Stress.

ATP-binding cassette (ABC) transporters are involved in transporting multiple substrates, such as toxins, and may be important for the survival of Trichoderma when encountering biotic toxins. In this study, genome searching revealed that there are 44 ABC transporters encoded in the genome of Trichoderma asperellum. These ABC transporters were divided into six types based on three-dimensional (3D) structure prediction, of which four, represented by 39 ABCs, are involved in transport and the remaining two, represented by 5 ABCs, are involved in regulating translation. The characteristics of nucleotide-binding domain (NBD) are important in the identification of ABC proteins. Even though the 3D structures of the 79 NBDs in the 44 ABCs are similar, multiple sequence alignment showed they can be divided into three classes. In total, 794 motifs were found in the promoter regions of the 44 ABC genes, of which 541 were cis-regulators related to stress responses. To characterize how their ABCs respond when T. asperellum interact with fungi or plants, T. asperellum was cultivated in either minimal media (MM) control, C-hungry, N-hungry, or poplar medium (PdPap) to simulate normal conditions, competition with pathogens, interaction with pathogens, and interaction with plants, respectively. The results show that 17 of 39 transport ABCs are highly expressed in at least one condition, whereas four of the five translation-regulating ABCs are highly expressed in at least one condition. Of these 21 highly expressed ABCs, 6 were chosen for RT-qPCR expression under the toxin stress of phytopathogen Alternaria alternata, and the results show ABC01, ABC04, ABC05, and ABC31 were highly expressed and may be involved in pathogen interaction and detoxifying toxins from A. alternata.

qGL3/OsPPKL1 induces phosphorylation of 14-3-3 protein OsGF14b to inhibit OsBZR1 function in brassinosteroid signaling.

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.

OsBAK2/OsSERK2 expression is repressed by OsBZR1 to modulate brassinosteroid response and grain length in rice.

Brassinosteroids (BRs) are a class of polyhydroxylated steroidal phytohormones that are essential for plant growth and development. Rice BRASSINOSTEROID-INSENSITIVE1 (BRI1)-ASSOCIATED RECEPTOR KINASES (OsBAKs) are plasma membrane-localized receptor kinases belonging to the subfamily of leucine-rich repeat receptor kinases. It has been found that in Arabidopsis, BRs induce the formation of a BRI1-BAK1 heterodimer complex and transmit the cascade signal to BRASSINAZOLE RESISTANT1/bri1-EMS-SUPPRESSOR1 (BZR1/BES1) to regulate BR signaling. Here, in rice (Oryza sativa ssp. japonica), we found that OsBZR1 binds directly to the promoter of OsBAK2, but not OsBAK1, and represses the expression of OsBAK2 to form a BR feedback inhibition loop. Additionally, the phosphorylation of OsBZR1 by OsGSK3 reduced its binding to the OsBAK2 promoter. The osbak2 mutant displays a typical BR-deficiency phenotype and negative modulates the accumulation of OsBZR1. Interestingly, the grain length of the osbak2 mutant was increased whereas in the cr-osbak2/cr-osbzr1 double mutant, the reduced grain length of the cr-osbzr1 mutant was restored, implying that the increased grain length of osbak2 may be due to the rice somatic embryogenesis receptor kinase-dependent pathway. Our study reveals a novel mechanism by which OsBAK2 and OsBZR1 engage in a negative feedback loop to maintain rice BR homeostasis, facilitating a deeper understanding of the BR signaling network and grain length regulation in rice.

Egg white protein-based delivery system for bioactive substances: a review.

Some bioactive substances in food have problems such as poor solubility, unstable chemical properties and low bioavailability, which limits their application in functional food. Recently, many egg white protein-based delivery carriers have been developed to improve the chemical stability, biological activity and bioavailability of bioactive substances. This article reviewed the structure and properties of several major egg white proteins commonly used to construct bioactive substance delivery systems. Several common carrier types based on egg white proteins, including hydrogels, emulsions, micro/nanoparticles, aerogels and electrospinning were then introduced. The biological functions of common bioactive substances, the limitations, and the role of egg white protein-based delivery systems were also discussed. At present, whole egg white protein, ovalbumin and lysozyme are most widely used in delivery systems, while ovotransferrin, ovomucoid and ovomucin are less developed and applied. Egg white protein-based nanoparticles are currently the most commonly used delivery carriers. Egg white protein-based hydrogels, emulsions, and microparticles are also widely used. Future research on the application of various egg white proteins in developed new delivery systems will provide more choices for the delivery of various bioactive substances.

Cytokinin Confers Brown Planthopper Resistance by Elevating Jasmonic Acid Pathway in Rice.

Plants have evolved a sophisticated defense system that employs various hormone pathways to defend against attacks by insect pests. Cytokinin (CK) plays an important role in plant growth and stress tolerance, but the role of CKs in plant-insect interaction remains largely unclear. Here, we report that CKs act as a positive regulator in rice resistance against brown planthopper (BPH), a devastating insect pest of rice. We found that BPH feeding promotes CK biosynthesis and signaling in rice. Exogenous application of CKs significantly increased the rice resistance to BPH. Increasing endogenous CKs by knocking out cytokinin oxidase/dehydrogenase (OsCKXs) led to enhanced resistance to BPH. Moreover, the levels of the plant hormone jasmonic acid (JA) and the expression of JA-responsive genes were elevated by CK treatment and in OsCKXs knockout plants. Furthermore, JA-deficient mutant og1 was more susceptible to BPH, and CK-induced BPH resistance was suppressed in og1. These results indicate that CK-mediated BPH resistance is JA-dependent. Our findings provide the direct evidence for the novel role of CK in promoting insect resistance, and demonstrate that CK-induced insect resistance is JA-dependent. These results provide important guidance for effective pest management strategies in the future.

Effects of salt and heat treatment on the physicochemical properties, microstructure, secondary structure, and simulated in vitro gastrointestinal digestion of duck egg white.

BACKGROUND: The texture and structure of the duck egg white (DEW) gel under salt and heat treatment are crucial to its digestibility. Specifically, the structural changes of food protein gels have been recognized for their potential to regulate in vitro digestion. In this study, the effects of gel characteristics and simulated in vitro gastrointestinal digestion of DEW under combined salt and heat treatment were investigated. RESULTS: With the increase in salting time and temperature, a porous opaque gel with large particles was formed, the moisture content of DEW showed a downward trend, and the same was true for hardness changes. The microstructure suggested that, with the penetration of NaCl, DEW proteins were denatured, and the protein molecules gradually unfolded and then aggregated after 7 days. The secondary structure revealed that, as the salting time and temperature increased, the proportion of intermolecular ß-sheets and α-helices decreased. In terms of in vitro digestion, the highest digestibility was obtained at 14 days of salting combined with 100 °C heat treatment, and the digestibility was the lowest when marinated for 7 days at 121 °C. Liquid chromatography and tandem mass spectrometry (LC-MS/MS) indicated that the number of different types of peptides and specific peptides was positively correlated with the salting time and temperature of the DEW at the end of gastric digestion. CONCLUSIONS: Heat treatment at 100 °C has a higher in vitro digestibility than at 121 °C. Gels with low hardness, large pores, and rough textures are easier to digest by pepsin and release more peptides. © 2021 Society of Chemical Industry.

Transcriptome analysis reveals delaying of the ripening and cell-wall degradation of kiwifruit by hydrogen sulfide.

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 < 0.05). Transmission electron microscopy (TEM) showed that degradation of the cell wall was inhibited after H2 S treatment. Based on transcriptome data analysis and quantitative real-time polymerase chain reaction (qRT-PCR), expression levels of endo-1,4-ß-glucanase (ß-glu), ß-galactosidase (ß-gal) and pectinesterase (PME) decreased whereas pectinesterase inhibitor (PMEI) significantly increased in response to H2 S. The members of the signal transduction pathway involved in ethylene were also identified. Hydrogen sulfide inhibited the expression of ethylene receptor 2 (ETR2), ERF003, ERF5, and ERF016, and increased the expression of ethylene-responsive transcription factor 4 (ERF4) and ERF113. CONCLUSION: Hydrogen sulfide could delay the ripening and senescence of kiwifruit by regulating the cell-wall degrading enzyme genes and affecting ethylene signal transduction pathway genes. Our results revealed the effect of H2 S treatment on the softening of kiwifruit at the transcription level, laying a foundation for further research. © 2020 Society of Chemical Industry.

Physicochemical and nutritional characteristics of preserved duck egg white.

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.

Effects of alkaline concentration, temperature, and additives on the strength of alkaline-induced egg white gel.

Egg whites can undergo gelation at extreme pH. In this paper, the effects of NaOH concentration (1.5, 2, 2.5, and 3%), temperature (10, 20, 30, and 40°C), and additives (metallic compounds, carbohydrates, stabilizers, and coagulants) on the strength of alkaline-induced egg white gel were investigated. Results showed that NaOH concentration and induced temperature significantly affected the rate of formation and peak strength of the egg white gel. Of the 6 metallic compounds used in this experiment, CuSO4exhibited the optimal effect on the strength of alkaline-induced egg white gel, followed by MgCl2, ZnSO4, PbO, and CaCl2. When CuSO4concentration was 0.2%, the gel strength increased by 31.92%. The effect of Fe2(SO4)3was negligible. Of the 5 carbohydrate additives, xanthan gum (0.2%) caused the highest increase (54.31%) in the strength of alkaline-induced egg white gel, followed by sodium alginate, glucose, starch, and sucrose. Meanwhile, propylene glycol (0.25%) caused the highest improvement (15.78%) in the strength of alkaline-induced egg white gel among the 3 stabilizing agents and coagulants used, followed by Na2HPO4and glucono-δ-lactone.

Hydrogen sulfide extends the postharvest life and enhances antioxidant activity of kiwifruit during storage.

BACKGROUND: Exogenous hydrogen sulfide (H2S) treatment can prolong the postharvest life of cut flowers and strawberries. Little work has been done to explore the effects of H2S on respiratory climacteric fruits such as kiwifruits during storage. Therefore the aim of the present study was to evaluate the effects of H2S treatment at concentrations of 15­1000 µmol L⁻¹ on the postharvest life of kiwifruit during 25 °C storage and the role of H2S in regulating the antioxidant defensive system of kiwifruit. RESULTS: Treatments with 45 and 90 µmol L⁻¹ H2S significantly inhibited the increase in soluble sugar content and the decrease in vitamin C (Vit C), chlorophyll content and firmness, inhibited ethylene production and both superoxide production rate (O(·2)⁻) and hydrogen peroxide content. Kiwifruits with 45 and 90 µmol L⁻¹ H2S exhibited significantly higher activities of superoxide dismutase, catalase and peroxidase. Treatment with 180 µmol L⁻¹ H2S promoted the ripening of kiwifruits. CONCLUSION: Treatments with 45 and 90 µmol L⁻¹ H2S could delay the maturation and senescence of kiwifruits and maintain higher titratable acid (TA) and Vit C during eating-ripe storage by inhibiting ethylene production, improving protective enzyme activities and decreasing the accumulation of reactive oxygen species to protect the cell membrane during storage.

Egg Yolk Lipids Alleviated Dextran Sulfate Sodium-Induced Colitis by Inhibiting NLRP3 Inflammasome and Regulating Gut Microbiota.

The increasing incidence of inflammatory bowel disease (IBD) has become a global phenomenon. Egg yolk lipids are one of the essential dietary foods, but its effects on intestinal immunity remain unclear. Here, egg yolk lipids are obtained using ethanol extraction and a total of 601 kinds of lipids are detected via lipidomics, including 251 kinds of triglycerides, 133 kinds of phosphatidylcholines, 44 kinds of phosphatidylethanolamines. Then, the study finds that egg yolk lipids significantly alleviate dextran sulfate sodium-induced colitis and reduce the production of inflammatory factors. Meanwhile, egg yolk lipids also maintain intestinal barrier integrity and decrease lipopolysaccharide translocation by alleviating intestinal structure damage and increasing the numbers of goblet cells and mucin 2. Mechanistically, egg yolk lipids attenuate colitis by inhibiting the assembly and activation of NLRP3 inflammasome. Moreover, the study also finds that egg yolk lipids reverse gut microbiota dysbiosis referring to increased relative abundance of Bacteroides acidifaciens and decrease relative abundance of Akkermansia muciniphila, as well as increased short chain fatty acids concentration in the gut. Together, the study elucidates the anti-colitis effect of egg yolk lipids and provides positive evidences for egg yolk lipids involving in dietary strategy and IBD therapy.

Hydrogen sulfide enhances kiwifruit resistance to soft rot by regulating jasmonic acid signaling pathway.

As the third active gas signal molecule in plants, hydrogen sulfide (H2S) plays important roles in physiological metabolisms and biological process of fruits and vegetables during postharvest storage. In the present study, the effects of H2S on enhancing resistance against soft rot caused by Botryosphaeria dothidea and the involvement of jasmonic acid (JA) signaling pathway in kiwifruit during the storage were investigated. The results showed that 20 µL L-1 H2S fumigation restrained the disease incidence of B. dothidea-inoculated kiwifruit during storage, and delayed the decrease of firmness and the increase of soluble solids (SSC) content. H2S treatment increased the transcription levels of genes related to JA biosynthesis (AcLOX3, AcAOS, AcAOC2, and AcOPR) and signaling pathway (AcCOI1, AcJAZ5, AcMYC2, and AcERF1), as well as the JA accumulation. Meanwhile, H2S promoted the expression of defense-related genes (AcPPO, AcSOD, AcGLU, AcCHI, AcAPX, and AcCAT). Correlation analysis revealed that JA content was positively correlated with the expression levels of JA biosynthesis and defense-related genes. Overall, the results indicated that H2S could promote the increase of endogenous JA content and expression of defense-related genes by regulating the transcription levels of JA pathway-related genes, which contributed to the inhibition on the soft rot occurrence of kiwifruit.

Protective Effect of Egg Yolk Lipids against Dextran Sulfate Sodium-Induced Colitis: The Key Role of Gut Microbiota and Short-Chain Fatty Acids.

Egg yolk lipids significantly alleviate dextran sulfate sodium (DSS)-induced colitis by inhibiting NLRP3 inflammasome, reversing gut microbiota dysbiosis, and increasing short chain fatty acids (SCFAs) concentrations. However, the role of gut microbiota and the relationship between SCFAs and NLRP3 inflammasome are still unknown. Here, this study confirms that antibiotic treatment abolishes the protective effect of egg yolk lipids on DSS-induced colonic inflammation, intestinal barrier damage, and lipopolysaccharide translocation. Fecal microbiota transplantation also supports that egg yolk lipids alleviate colitis in a gut microbiota-dependent manner. Then, the study investigates the relationship between SCFAs and NLRP3 inflammasome, and finds that SCFAs significantly suppress colitis via inhibiting colonic NLRP3 inflammasome activation and proinflammatory cytokines secretions (interleukin, IL)-1ß and IL-18, and combined treatment of SCFAs and MCC950 (NLRP3 inhibitor) shows a better activity against colitis and NLRP3 inflammasome activation. Together, these findings provide positive evidence for gut microbiorta-SCFAs-NLRP3 axis as a novel target involving in the therapy of inflammatory bowel disease.

Ovotransferrin alleviated acute gastric mucosal injury in BALB/c mice caused by ethanol.

Acute gastric mucosal injury is a common gastrointestinal disorder, which influences patients' life quality. It was found that ovotransferrin (OVT) reduces the abundance of Helicobacter pylori associated with gastric disease in the intestine of immunosuppressed mice. To clarify its gastric protective function, the present study investigated the effect of OVT on BALB/c mice with ethanol-induced gastric mucosal injury. Results showed that OVT attenuated the ethanol-induced gastric mucosal injury. Furthermore, OVT effectively downregulated the expression of inflammatory markers (tumor necrosis factor-α, interleukin (IL)-1ß and IL-6) but enhanced the secretion of IL-4, IL-10 and prostaglandin E2. And OVT pretreatment significantly inhibited the activation of the MAPK/NF-κB pathway. Additionally, OVT improved gastric antioxidant ability by increasing superoxide dismutase and glutathione levels and decreasing malondialdehyde and myeloperoxidase content. Pretreatment with OVT modulated the equilibrium between B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X. The above results indicated that OVT alleviated inflammatory responses, oxidative stress and apoptosis in gastric mucosal injury mice caused by ethanol.

Ovotransferrin Inhibits TNF-α Induced Inflammatory Response in Gastric Epithelial Cells via MAPK and NF-κB Pathway.

Ovotransferrin (OVT) has been confirmed to have anti-inflammatory activity. However, its effect and mechanism on gastric inflammation are unclear. In this study, the effect and mechanism of the OVT on the tumor necrosis factor-α (TNF-α) induced inflammatory response in gastric epithelial cells (GES-1) were investigated. The enzyme linked immunosorbent assay (ELISA) was used to determine the levels of inflammation cytokines, followed by RNA sequencing to explore the potential pathways of its anti-inflammatory effect, and then it was validated by Western blotting and pathways inhibitors. Results showed that the OVT at concentrations of 50-400 µg/mL displayed nontoxicity against GES-1 cells. Additionally, 100 µg/mL of OVT obviously reduced the secretion of interleukin (IL)-8, IL-6, and TNF-α by 63.02% (630.09/1703.98), 35.53% (935.81/1451.43), and 36.19% (964.60/1511.63), respectively. The results of RNA sequencing exhibited that the OVT significantly influences the activation of mitogen-activated protein kinase (MAPK) and the nuclear factor kappa-light-chain enhancer of activated B cell (NF-κB) pathways, which was verified by the levels of p-IKK, p-IκB, p-P65, p-ERK, p-JNK, and p-P38 protein. IL-8 contents released by GES-1 cells after incubation with inhibitors of NF-κB and MAPK pathways further confirmed that OVT hindered activation of these two pathways. Collectively, these results suggested that OVT was a natural protein with the potential to treat gastric inflammation.

Production of high affinity monoclonal antibody and development of indirect competitive chemiluminescence enzyme immunoassay for gentamicin residue in animal tissues.

To determine gentamicin residues in animal tissues, a monoclonal antibody (Mab) was produced and a sensitive indirect competitive chemiluminescent enzyme immunoassay (icCLEIA) was developed. At first, gentamicin was conjugated with bovine serum albumin as immunogens which were used to immunize BALB/c mice. Then, an anti-gentamicin Mab was prepared by hybridoma technology. Finally, a sensitive icCLEIA was developed with an 50% inhibition concentration (IC50) of 0.067 ng/mL for gentamicin. The limit of detection of the icCLEIA was 0.002 ng/mL. The cross reactivity of the Mab with structural analogues were<0.01%. The recoveries of gentamicin ranged from 80 to 101% and coefficient of variation was <6.4% in pork and fish samples. Samples were detected by UPLC-MS/MS for evaluating reliability of the icCLEIA. The results suggested that the prepared anti-gentamicin Mab can be used for rapid and convenient immunoassays to detect gentamicin residues in animal tissues.

Beneficial and biocontrol effects of Trichoderma atroviride, a dominant species in white birch rhizosphere soil.

White birch (Betula platyphylla Suk.) is a typical pioneer tree species that is important in forest restoration in northern China, Japan, and Korea. In the present study, 37 isolates were obtained from B. platyphylla rhizosphere soils in Heilongjiang Province; they were identified as T. pleuroticola (3 isolates), T. virens (2 isolates), T. hamatum (8 isolates), T. atroviride (21 isolates, dominant species) and T. asperelloides (3 isolates). Stress tolerance tests (salt, alkali, and nutritional stress that simulated saline alkali or barren soil) and confrontation assays (with four pathogens) were performed to determine which isolates had good biocontrol ability in barren soil; the results show that T. atroviride was outstanding. Then, in order to determine the effect of T. atroviride on plants and soil, Gynura cusimbua seeds were sown and treated with a T. atroviride spore suspension, as was unsown soil. The seedlings treated using T. atroviride had significantly greater height, stem diameter, soluble protein content, soluble sugar content, and malonaldehyde (MDA) content and their catalase (CAT) activity was also significantly increased. In addition, when the plants were inoculated with Alternaria alternata, the plants treated using T. atroviride had stronger CAT activity, significantly higher soluble protein content and soluble sugar content, and significantly lower MDA content, which indicates stronger resistance and less injury caused by the pathogen. In addition, T. atroviride not only increased the content of available nitrogen and available phosphorus in the soil, but also promoted G. cusimbua seedlings' absorption of available nitrogen and available phosphorus. Thus, the characteristics of T. atroviride may make it the main factor that helps B. platyphylla colonise cut-over lands. T. atroviride, a promising biocontrol candidate, can be used in agriculture and forestry.