T-2 Toxin Caused Mice Testicular Inflammation Injury via ROS-Mediated NLRP3 Inflammasome Activation.

T-2 toxin treatment causes male reproduction system dysfunction, although the exact mechanism remains unclear. In this research, male Kunming mice and TM4 cells were treated with varying concentrations of the T-2 toxin for evaluating the adverse effect of T-2 toxin on male reproductive function. MCC950 or NAC was used to block NLRP3 inflammasome activation and eliminate reactive oxygen species (ROS) accumulation in the TM4 cell, respectively. The results showed that: (1) T-2 toxin caused testicular atrophy, destroyed the microstructure and ultrastructure of the testis, and caused sperm deformities; (2) T-2 toxin increased the content and gene expressions of TNF-α and IL-6 and decreased the IL-10 content and gene expression, causing testis and TM4 cell inflammatory injury; (3) T-2 toxin activated NLRP3 inflammasome in the testis and TM4 cells and caused ROS accumulation in the testis; (4) suppressing NLRP3 inflammasome activation using 20 nM MCC950 alleviated the TM4 cell inflammatory damage caused via the T-2 toxin; nevertheless, 20 nM MCC950 did not reduce ROS accumulation in TM4 cells; and (5) NAC relieved the inflammatory damage in TM4 cells by inhibiting NLRP3 inflammasome activation. Taken together, T-2 toxin caused testicular inflammation injury through ROS-mediated NLRP3 inflammasome activation, resulting in male reproductive dysfunction.

PINK1/Parkin-mediated mitophagy is activated to protect against testicular damage caused by aluminum.

Oxidative stress is the main mechanism of aluminum (Al) reproductive toxicity and can also lead to mitochondrial damage. Damaged mitochondria can trigger apoptosis, leading to testicular damage. PINK1 (phosphatase and tensin homolog (PTEN)-induced putative kinase1)/Parkin (E3 ubiquitin ligase PARK2)-mediated mitophagy can remove damaged mitochondria to maintain cellular homeostasis. However, its role in testicular damage caused by Al is unclear. In this study, first, 24 male wild type (WT) C57BL/6 N mice were divided into 4 groups and orally administered with 0, 44.825, 89.65, and 179.3 mg/kg body weight AlCl3 for 90 days, respectively. We demonstrated that apoptosis and PINK1/Parkin-mediated mitophagy were activated in a dose-effect relationship in the damaged testis caused by AlCl3, and were most significant at 179.3 mg/kg body weight AlCl3. Then, 40 male WT C57BL/6 N mice and 40 male Parkin knockout (Parkin-/-) C57BL/6 N mice were divided into 4 groups and orally administered with 0, 179.3, 0 or 179.3 mg/kg body weight AlCl3 for 90 days, respectively. Parkin-/- inhibited mitophagy, exacerbated the growth inhibition, testicular damage, mitochondrial damage, oxidative stress, and apoptosis under AlCl3 exposure. In general, the results reveal that AlCl3 exposure can activate PINK1/Parkin-mediated mitophagy, and the PINK1/Parkin-mediated mitophagy plays a protective role in the testicular damage caused by AlCl3.

Activation of PINK1/Parkin-mediated mitophagy protects against apoptosis in kidney damage caused by aluminum.

Aluminum (Al) induces apoptosis via oxidative stress and/or mitochondrial damage. Kidney is the main organ of Al excretion, but whether Al causes apoptosis in kidney of mice remains unclear. Mitophagy maintains cell homeostasis via clearing damaged mitochondria and reducing oxidative stress, but the role in kidney damage caused by Al has also not been investigated. In this study, firstly, forty wild type (WT) male C57 mice were randomly exposed to AlCl3 at 0, 44.825, 89.65 or 179.3 mg/kg body weight in drinking water for 90 days, respectively. Our results confirmed that Al induced apoptosis, and activated PINK1 (phosphatase and tensin homolog (PTEN)-induced putative kinase1)/Parkin (E3 ubiquitin ligase PARK2)-mediated mitophagy with the dose increased. And secondly, to further assess the role of PINK1/Parkin-mediated mitophagy in Al-induced kidney damage, twenty Parkin knockout (Parkin-/-) mice and twenty WT mice were divided into WT group, WT + Al group, Parkin-/- group, and Parkin-/- + Al group, and they were provided with AlCl3 at a dose of 0 or 179.3 mg/kg body weight in drinking water for 90 days, respectively. The results showed that Parkin-/- induced more severe kidney injury caused by Al. Besides, Parkin-/- aggravated oxidative stress and apoptosis caused by Al. Overall, our findings indicate that the activation of PINK1/Parkin-mediated mitophagy protects against apoptosis in kidney damage caused by Al.

ROS-mediated mitophagy and apoptosis are involved in aluminum-induced femoral impairment in mice.

The problem of excessive aluminum (Al) content in food is widespread. After Al enters the body, it can cause mineral metabolism imbalance and reactive oxygen species (ROS) overproduction, which ultimately leads to bone impairment. ROS is mainly produced in mitochondria and acts on mitochondria. Mitochondrial damage is closely related to mitophagy and apoptosis. In order to clarify whether ROS-mediated mitophagy and apoptosis are involved in Al-induced femoral impairment, forty-eight male C57BL/6 N mice were exposed to AlCl3 (179.3 mg/kg) and/or NAC (100 mg/kg) for 90 days. Our results showed that NAC inhibited the mitophagy and apoptosis, and alleviated growth inhibition, mineral metabolism imbalance, structural damage, decreased bone mineral density and decreased bone formation factor expressions in the femora of Al-treated mice. These results suggest that ROS-mediated mitophagy and apoptosis are involved in Al-induced femoral impairment in mice, exogenous ROS clearance is a potential strategy for the treatment of Al-induced bone impairment.

T-2 toxin causes dysfunction of Sertoli cells by inducing oxidative stress.

T-2 toxin is an inevitable mycotoxin in food products and feeds. It is a proven toxicant impairing the male reproductive system. However, previous studies have concentrated on the toxic effect of T-2 toxin on Leydig cells, with little attention on the Sertoli cell cytotoxicity. Therefore, this study aimed to establish the toxic mechanism of T-2 toxin on Sertoli cells. The Sertoli cell line (TM4 cell) was cultured and exposed to different concentrations of T-2 toxin with/without N-acetyl-L-cysteine (NAC) for 24 h. A CCK-8 assay then measured the cell viability. In addition, the expression of TM4 cell biomarkers (FSHR and ABP) and functional factors (occludin (Ocln), zonula occluden-1 (ZO-1), Connexin 43 (Cx-43), and N-Cadherin (N-cad)) were measured by qRT-PCR and Western blotting. The oxidative stress status (ROS, MDA, CAT, and SOD) and apoptosis rate, including the caspase-9, 8, and 3 activities in TM4 cells, were analyzed. We established that (1): T-2 toxin decreased TM4 cells viability and the half-maximal inhibitory concentration was 8.10 nM. (2): T-2 toxin-induced oxidative stress, evidenced by increased ROS and MDA contents, and inhibited CAT and SOD activities. (3): T-2 toxin inhibited FSHR, ABP, ocln, ZO-1, Cx-43, and N-Cad expressions. (4): T-2 toxin promoted TM4 cell apoptosis and caspase-9, 8, and 3 activities. (5): N-acetyl-L-cysteine relieved oxidative stress, functional impairment, and apoptosis in TM4 cells treated with T-2 toxin. Thus, T-2 toxin induced TM4 cell dysfunction through ROS-induced apoptosis.

Parkin-mediated mitochondrial quality control protects against aluminum-induced liver damage in mice.

Aluminum (Al) is known to be hepatotoxic. Oxidative stress is the main mechanism of liver injury caused by Al, and can also lead to mitochondrial damage. Mitochondrial damage is a prerequisite for mitochondrial quality control (MQC) dysregulation. Parkin can activate MQC and maintain mitochondrial homeostasis. However, the role of Parkin-mediated MQC in Al-induced liver damage has not been elucidated. In this study, forty male wild type (WT) C57BL/6N mice were treated with 0, 44.825, 89.65 or 179.3 mg/kg body weight AlCl3 in drinking water for 90 days, respectively. We found that Al induced mitophagy and disrupted mitochondrial dynamics and mitochondrial biogenesis. Then, twenty male WT C57BL/6N mice and twenty male Parkin knockout (Parkin-/-) C57BL/6N mice were divided into four groups and treated with 0, 89.65, 0, 89.65 mg/kg body weight AlCl3 in drinking water for 90 days, respectively. We found that Parkin-/- inhibited mitophagy and further disrupted mitochondrial dynamics and mitochondrial biogenesis. These results indicated that Parkin-mediated MQC could be disrupted by Al and protected against Al-induced liver damage.

Mitophagy and apoptosis mediated by ROS participate in AlCl3-induced MC3T3-E1 cell dysfunction.

Aluminum (Al), as a common environmental pollutant, causes osteoblast (OB) dysfunction and then leads to Al-related bone diseases (ARBD). One of the mechanisms of ARBD is oxidative stress, which leads to an increase in the production of reactive oxygen species (ROS). ROS can induce mitochondrial damage, thereby inducing mitophagy and apoptosis. But whether mitophagy and apoptosis mediated by ROS, and the role of ROS in AlCl3-induced MC3T3-E1 cell dysfunction is still unclear. In this study, MC3T3-E1 cells used 0 mM Al (control group), 2 mM Al (Al group), 5 mM N-acetyl cysteine (NAC) (NAC group), 2 mM Al and 5 mM NAC (Al + NAC group) for 24 h. We found AlCl3-induced MC3T3-E1 cell dysfunction accompanied by oxidative stress, apoptosis, and mitophagy. While NAC, a ROS scavenger treatment, restored cell function and alleviated the mitophagy and apoptosis. These results suggested that mitophagy and apoptosis mediated by ROS participate in AlCl3-induced MC3T3-E1 cell dysfunction.

Protective effect of mitophagy against aluminum-induced MC3T3-E1 cells dysfunction.

Aluminum (Al) is a ubiquitous environmental metal toxicant that causes osteoblast (OB) damage which leads to Al-related bone diseases. Mitochondrial damage plays a key role in Al-related bone diseases, and while mitophagy can clear damaged mitochondria and improve OB function, the relationship between mitophagy and Al-induced OB dysfunction is unknown. To explore the role of mitophagy in Al-induced OB dysfunction in vitro, we used 2 µM carbonyl cyanide m-chlorophenylhydrazone (CCCP) and 0.4 µM Cyclosporin A (CsA) to activate and inhibit mitophagy, respectively. MC3T3-E1 cells were treated with 0 mM AlCl3 (control group); 2 mM AlCl3 (Al group); 2 µM CCCP (CCCP group); 2 µM CCCP and 2 mM AlCl3 (CCCP + Al group); 0.4 µM CsA (CsA group); 0.4 µM CsA and 2 mM AlCl3 (CsA + Al group). The results showed that Al induced ultrastructural and functional impairment of MC3T3-E1 cells. Compared to the Al group, mitophagy activation caused mitochondrial membrane potentials to collapse, up-regulated PINK1, Parkin, and LC3 expression, down-regulated p62 expression, and increased mitophagosome numbers. Mitophagy activation also reduced Al-induced oxidative stress and MC3T3-E1 cell functional damage, as seen in improvement in cell viability, cellular calcium and phosphorus contents, and collagen I, osteocalcin, and bone alkaline phosphatase gene expression. Mitophagy inhibition had the opposite effects on activation. Overall, these results show that mitophagy can protect against Al-induced OB dysfunction.

Effect of different combined mechanical and thermal treatments on the quality characteristics of garlic paste.

The aim of this study was to investigate the effects of the sequence of different thermal and mechanical treatments on the physicochemical parameters and microstructure of garlic paste, in order to improve the quality of the product. The total organosulfur compounds (OSCs) decreased steadily in blended-heated garlic, whereas OSCs decreased sharply after 2 min at 75 °C or 5 min at 85 and 95 °C in heated-blended garlic. After blanching for 5 min, allicin could maintain over 4.0 mg/g only at 75 °C; and OSCs of heated-blended garlic paste were found to drop by 29.56%, 90.63% and 94.79% at 75, 85 and 95 °C, respectively. In blended-heated garlic, the color values of L* (lightness) and a* (redness) decreased (P < 0.05), while the b* (yellowness) and C* (chroma) increased (P < 0.05), obtaining green discoloration garlic paste. The total color differences of blended-heated samples were greater than 12.08, which were 2-6 folds higher compared with heated-blended garlic. Total phenolic content and antioxidant activity decreased (P < 0.05) in all thermal treatments, thermal treatment of heated-blended garlic less than 5 min maintained over 30% of antiradical activity. The sequence of unit operations determined the pattern of garlic microstructure disruption, resulting in various enzymic and non-enzymic reactions. Our results indicated that use of heat treatment prior to blend processing is an effective and feasible method to inhibit garlic discoloration and retain high content of bioactive OSCs. It is recommended that garlic paste be prepared using heated-blended processing, with thermal processing limited to 75 °C for less than 5 min.

Review of the Reproductive Toxicity of T-2 Toxin.

T-2 toxin, an inevitable environmental pollutant, is the most toxic type A trichothecene mycotoxin. Reproductive disruption is a key adverse effect of T-2 toxin. Herein, this paper reviews the reproductive toxicity of T-2 toxin and its mechanisms in male and female members of different species. The reproductive toxicity of T-2 toxin is evidenced by decreased fertility, disrupted structures and functions of reproductive organs, and loss of gametogenesis in males and females. T-2 toxin disrupts the reproductive endocrine axis and inhibits reproductive hormone synthesis. Furthermore, exposure to T-2 toxin during pregnancy results in embryotoxicity and the abnormal development of offspring. We also summarize the research progress in counteracting the reproductive toxicity of T-2 toxin. This review provides information toward a comprehensive understanding of the reproductive toxicity mechanisms of T-2 toxin.

Polymer-Based Device Fabrication and Applications Using Direct Laser Writing Technology.

Polymer materials exhibit unique properties in the fabrication of optical waveguide devices, electromagnetic devices, and bio-devices. Direct laser writing (DLW) technology is widely used for micro-structure fabrication due to its high processing precision, low cost, and no need for mask exposure. This paper reviews the latest research progresses of polymer-based micro/nano-devices fabricated using the DLW technique as well as their applications. In order to realize various device structures and functions, different manufacture parameters of DLW systems are adopted, which are also investigated in this work. The flexible use of the DLW process in various polymer-based microstructures, including optical, electronic, magnetic, and biomedical devices are reviewed together with their applications. In addition, polymer materials which are developed with unique properties for the use of DLW technology are also discussed.

Effects and mechanism of free amino acids on browning in the processing of black garlic.

BACKGROUND: Black garlic is produced by heating raw garlic at a high temperature for a long time without any additives. The thermal processing induces many chemical reactions, such as the Maillard reaction, which causes the color change from white to dark brown. Garlic contains a variety of amino acids, and the effect of each amino acid on browning is not fully understood. This work investigated the effect and mechanism of free amino acids on the browning of black garlic using model solutions containing garlic neutral polysaccharide, hydrolyzed garlic neutral polysaccharide, fructose, and free amino acids. RESULTS: A significant increase in reducing sugar was detected when garlic neutral polysaccharide was heated with glycine. The browning intensity of garlic neutral polysaccharide-glycine model solution was obviously higher after heating at 80 °C compared with that of garlic neutral polysaccharide solution. The model solution containing histidine had the greatest browning degree. The histidine model has a stable pH value, and almost no 5-hydroxymethylfurfural (5-HMF) was detected. CONCLUSION: Amino acid can promote the breaking of the garlic neutral polysaccharide chain and can react with the fructose generated to form browning. Histidine has the greatest effect on the browning, because histidine could eliminate the inhibiting effect of organic acid on Maillard reaction due to the buffer ability, and histidine had high reactivity in the late stage of Maillard reaction. © 2019 Society of Chemical Industry.

Characterization, Variables, and Antioxidant Activity of the Maillard Reaction in a Fructose⁻Histidine Model System.

Fructose and its polysaccharides are widely found in fruits and vegetables, with the Maillard reaction of fructose affecting food quality. This study aimed to investigate the Maillard reaction of fructose using a fructose⁻histidine model system. The reaction process was characterized using fluorescence spectroscopy and ultraviolet spectroscopy. The effects of temperature, initial reactant concentration, initial fructose concentration, initial histidine concentration, and initial pH value on the different stages of the Maillard reaction were studied. Reactant reduction, ultraviolet and fluorescence spectra, acetic acid content, 5-hydroxymethylfurfural (5-HMF) content, and browning intensity were evaluated. The results showed that increasing the temperature and reactant concentration promoted the condensation reaction of fructose and amino acid in the early stage, the formation of intermediate products with ultraviolet absorption and fluorescence in the intermediate stage, and the formation of pigment in the final stage. The 5-HMF concentration decreased with increasing histidine concentration and initial pH value. Changes in the shape of ultraviolet and fluorescence spectra showed that the initial pH value affected not only the reaction rate, but also the intermediate product types. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging rate of the Maillard reaction products increased with increasing temperature, reactant concentration, and initial pH value.

Natural Variation in OsLG3 Increases Drought Tolerance in Rice by Inducing ROS Scavenging.

Improving the performance of rice (Oryza sativa) under drought stress has the potential to significantly affect rice productivity. Here, we report that the ERF family transcription factor OsLG3 positively regulates drought tolerance in rice. In our previous work, we found that OsLG3 has a positive effect on rice grain length without affecting grain quality. In this study, we found that OsLG3 was more strongly expressed in upland rice than in lowland rice under drought stress conditions. By performing candidate gene association analysis, we found that natural variation in the promoter of OsLG3 is associated with tolerance to osmotic stress in germinating rice seeds. Overexpression of OsLG3 significantly improved the tolerance of rice plants to simulated drought, whereas suppression of OsLG3 resulted in greater susceptibility. Phylogenetic analysis indicated that the tolerant allele of OsLG3 may improve drought tolerance in cultivated japonica rice. Introgression lines and complementation transgenic lines containing the elite allele of OsLG3IRAT109 showed increased drought tolerance, demonstrating that natural variation in OsLG3 contributes to drought tolerance in rice. Further investigation suggested that OsLG3 plays a positive role in drought stress tolerance in rice by inducing reactive oxygen species scavenging. Collectively, our findings reveal that natural variation in OsLG3 contributes to rice drought tolerance and that the elite allele of OsLG3 is a promising genetic resource for the development of drought-tolerant rice varieties.

High-Density Genetic Map Construction and Stem Total Polysaccharide Content-Related QTL Exploration for Chinese Endemic Dendrobium (Orchidaceae).

Plants of the Dendrobium genus are orchids with not only ornamental value but also high medicinal value. To understand the genetic basis of variations in active ingredients of the stem total polysaccharide contents (STPCs) among different Dendrobium species, it is of paramount importance to understand the mechanism of STPC formation and identify genes affecting its process at the whole genome level. Here, we report the first high-density single-nucleotide polymorphism (SNP) integrated genetic map with a good genome coverage of Dendrobium. The specific-locus amplified fragment sequencing (SLAF-seq) technology led to identification of 7,013,400 SNPs from 1,503,626 high-quality SLAF markers from two parents (Dendrobium moniliforme ♀ × Dendrobium officinale ♂) and their interspecific F1 hybrid population. The final genetic map contained 8, 573 SLAF markers, covering 19 linkage groups (LGs). This genetic map spanned a length of 2,737.49 cM, where the average distance between markers is 0.32 cM. In total, 5 quantitative trait loci (QTL) related to STPC were identified, 3 of which have candidate genes within the confidence intervals of these stable QTLs based on the D. officinale genome sequence. This study will build a foundation up for the mapping of other medicinal-related traits and provide an important reference for the molecular breeding of these Chinese herb.

Semi-Preparative Separation of 10 Caffeoylquinic Acid Derivatives Using High Speed Counter-Current Chromatogaphy Combined with Semi-Preparative HPLC from the Roots of Burdock (Arctium lappa L.).

Burdock roots are healthy dietary supplements and a kind of famous traditional Chinese medicine, which contains large amounts of caffeoylquinic acid derivatives. However, little research has been reported on the preparative separation of these compounds from burdock roots. In the present study, a combinative method of HSCCC and semi-preparative HPLC was developed for the semi-preparative separation of caffeoylquinic acid derivatives from the burdock roots. The ethyl acetate extract of burdock roots was first fractionated by MCI macroporous resin chromatography and give three fractions (Fr. 1-3) from the elution of 40% methanol. Then, these three fractions (120 mg) were separately subjected to HSCCC for purification with the solvent system composed of petroleum ether-ethyl acetate-methanol-water at different volume ratios, and the mixtures were further purified by semi-preparative HPLC. As a result, a total of eight known caffeoylquinic acid derivatives including 3-O-caffeoylquinic acid (32.7 mg, 95.7%), 1,5-O- dicaffeoylquinic acid (4.3 mg, 97.2%), 3-O-caffeoylquinic acid methyl ester (12.1 mg, 93.2%), 1,3-O-dicaffeoylquinic acid (42.9 mg, 91.1%), 1,5-O-dicaffeoyl-3-O-(4-maloyl)-quinic acid (4.3 mg, 84.5%), 4,5-O-dicaffeoylquinic acid (5.3 mg, 95.5%), 1,5-O-dicaffeoyl-3-O-succinylquinic acid (8.7 mg, 93.4%), and 1,5-O-dicaffeoyl-4-O-succinylquinic acid (1.7 mg, 91.8%), and two new compounds were obtained. The new compounds were 1,4-O-dicaffeoyl-3-succinyl methyl ester quinic acid (14.6 mg, 96.1%) and 1,5-O-dicaffeoyl-3-O-succinyl methyl ester quinic acid (3.1 mg, 92.6%), respectively. The research indicated that the combination of HSCCC and semi-preparative HPLC is a highly efficient approach for preparative separation of the instability and bioactive caffeoylquinic acid derivatives from natural products.

Composition analysis and antioxidant properties of black garlic extract.

Black garlic produced from fresh garlic under controlled high temperature and humidity has strong antioxidant properties. To determine these compounds, five fractions (from F1 to F5) were separated and purified by elution with chloroform:methanol at different ratios (8:1, 6:1, 4:1, 2:1, and 0:1; v/v). The antioxidant activity of each fraction was analyzed. The results showed that F3 and F4 had higher phenolic contents and stronger 2,2-diphenyl-2-picrylhydrazyl radical scavenging activity than the others. Seven purified individual components were further separated using semipreparation high-performance liquid chromatography from these two intensely antioxidant fractions (F3 and F4), their structures were elucidated by high-performance liquid chromatography coupled to diode array detection, electrospray ionization, mass spectrometry, 1H nuclear magnetic resonance, and 13C nuclear magnetic resonance spectrometry. Three compounds including adenosine, uridine, and 2-acetylpyrrole were first identified in black garlic, except for 5-hydroxymethylfurfural, (1S, 3S)-1-methyl-1,2,3,4-tetrahydro-ß-carboline-3-carboxylic acid, and (1R, 3S)-1-methyl-1,2,3,4-tetrahydro-ß-carboline-3-carboxylic acid. The cellular antioxidant activities of uridine, adenosine, carboline alkaloids, 5-hydroxymethylfurfural, and ethyl acetate extracts were consistent with the results of in vitro experimental antioxidant properties. The results provide useful information for understanding the health benefits of black garlic products.

Effects of temperature on the quality of black garlic.

BACKGROUND: Black garlic is a type of garlic product that is generally produced by heating raw garlic at high temperature with controlled humidity for more than 30 days. Black garlic has appeared on the market for many years. It is crucial to investigate the characteristics of quality formation of black garlic during processing at various temperatures. RESULTS: In this study, fresh garlic was processed to black garlic at temperatures of 60, 70, 80 and 90 °C. Moisture, amino acid nitrogen and allicin contents decreased gradually during thermal processing of various temperatures. Reducing sugar, 5-hydroxymethylfurfural, total phenols, total acids contents and browning increased. The changing rate of quality indicators and flavour of black garlic varied at different temperatures. Browning intensity reached about 74 when black garlic aged. The sensory score was significantly higher in black garlic aged at 70 °C (39.95 ± 0.31) compared with that at other temperatures, suggesting that 70 °C might facilitate formation of good quality and flavour of black garlic during processing. CONCLUSION: Temperature had a remarkable impact on the quality and flavour of black garlic. © 2015 Society of Chemical Industry.

Overexpression of OsMYB48-1, a novel MYB-related transcription factor, enhances drought and salinity tolerance in rice.

MYB-type transcription factors (TFs) play essential roles in plant growth, development and respond to environmental stresses. Role of MYB-related TFs of rice in drought stress tolerance is not well documented. Here, we report the isolation and characterization of a novel MYB-related TF, OsMYB48-1, of rice. Expression of OsMYB48-1 was strongly induced by polyethylene glycol (PEG), abscisic acid (ABA), H2O2, and dehydration, while being slightly induced by high salinity and cold treatment. The OsMYB48-1 protein was localized in the nucleus with transactivation activity at the C terminus. Overexpression of OsMYB48-1 in rice significantly improved tolerance to simulated drought and salinity stresses caused by mannitol, PEG, and NaCl, respectively, and drought stress was caused by drying the soil. In contrast to wild type plants, the overexpression lines exhibited reduced rate of water loss, lower malondialdehyde (MDA) content and higher proline content under stress conditions. Moreover, overexpression plants were hypersensitive to ABA at both germination and post-germination stages and accumulated more endogenous ABA under drought stress conditions. Further studies demonstrated that overexpression of OsMYB48-1 could regulate the expression of some ABA biosynthesis genes (OsNCED4, OsNCED5), early signaling genes (OsPP2C68, OSRK1) and late responsive genes (RAB21, OsLEA3, RAB16C and RAB16D) under drought stress conditions. Collectively, these results suggested that OsMYB48-1 functions as a novel MYB-related TF which plays a positive role in drought and salinity tolerance by regulating stress-induced ABA synthesis.

OsMIOX, a myo-inositol oxygenase gene, improves drought tolerance through scavenging of reactive oxygen species in rice (Oryza sativa L.).

Myo-inositol oxygenase (MIOX), a unique monooxygenase, catalyzes the oxidation of myo-inositol to d-glucuronic acid. However, the protective role of MIOX in plants against oxidative stress or drought stress remains unknown. In this study, the functional characterization of MIOX obtained from the cDNA library of upland rice (Oryza sativa L. cv. IRAT109), was performed. OsMIOX was expressed predominantly in the roots and induced by drought, H2O2, salt, cold and abscisic acid. The transgenic rice lines overexpressing OsMIOX showed obviously improved growth performance in the medium containing 200 mM mannitol. Further, the survival rate of leaves from the transgenic rice lines was significantly higher than that of the wild type plants under polyethylene glycol treatment. It was discovered that the activity of ROS-scavenging enzymes and proline content, as well as the transcript levels of many ROS scavenging genes were significantly increased in transgenic plants compared to the wild type plants under drought stress conditions. Together, these data suggest that OsMIOX has a specific function in drought stress tolerance by decreasing oxidative damage.