OsDWD1 E3 ligase-mediated OsNPR1 degradation suppresses basal defense in rice.

Many ubiquitin E3 ligases function in plant immunity. Here, we show that Oryza sativa (rice) DDB1 binding WD (OsDWD1) suppresses immune responses by targeting O. sativa non-expresser of pathogenesis-related gene 1 (OsNPR1) for degradation. Knock-down and overexpression experiments in rice plants showed that OsDWD1 is a negative regulator of the immune response and that OsNPR1 is a substrate of OsDWD1 and a substrate receptor of OsCRL4. After constructing the loss-of-function mutant OsDWD1R239A , we showed that the downregulation of OsNPR1 seen in rice lines overexpressing wild-type (WT) OsDWD1 (OsDWD1WT -ox) was compromised in OsDWD1R239A -ox lines, and that OsNPR1 upregulation enhanced resistance to pathogen infection, confirming that OsCRL4OsDWD1 regulates OsNPR1 protein levels. The enhanced disease resistance seen in OsDWD1 knock-down (OsDWD1-kd) lines contrasted with the reduced disease resistance in double knock-down (OsDWD1/OsNPR1-kd) lines, indicating that the enhanced disease resistance of OsDWD1-kd resulted from the accumulation of OsNPR1. Moreover, an in vivo heterologous protein degradation assay in Arabidopsis thaliana ddb1 mutants confirmed that the CUL4-based E3 ligase system can also influence OsNPR1 protein levels in Arabidopsis. Although OsNPR1 was degraded by the OsCRL4OsDWD1 -mediated ubiquitination system, the phosphodegron-motif-mutated NPR1 was partially degraded in the DWD1-ox protoplasts. This suggests that there might be another degradation process for OsNPR1. Taken together, these results indicate that OsDWD1 regulates OsNPR1 protein levels in rice to suppress the untimely activation of immune responses.

Comparison of Floaters after Cataract Surgery with Different Viscoelastics.

Purpose: To investigate whether there is a difference between symptoms of floaters according to the type of ophthalmic viscosurgical devices(OVDs) used during phacoemulsification. Methods: A total of 112 eyes had undergone standard phacosurgery with the dispersive OVDs(Group1). Group2 comprised 117 eyes that underwent phacosurgery with the dispersive OVDs, but between continuous curvilinear capsulorhexis and hydrodissection, some OVDs had been removed. Group3 included 120 eyes that had undergone phacosurgery with the cohesive OVDs. Results: 14 eyes (12.5%) of Group1 had new-onset floater after surgery whereas 6 eyes (5.13%) in Group2, and 7 eyes (5.83%) in Group3 at the day after and a week after surgery. This was significantly higher in Group1 than Group2 and Group3, respectively (p=0.047,0.049). Conclusion: Cataract surgery with dispersive OVD can predispose the eye to an increased floater symptom. Therefore, surgeons should consider release some OVDs during hydrodissection with dispersive viscoelastics and keep trying to avoid IOP surge during surgery.

OsWRKY51, a rice transcription factor, functions as a positive regulator in defense response against Xanthomonas oryzae pv. oryzae.

KEY MESSAGE: OsWRKY51 functions as a positive transcriptional regulator in defense signaling against Xanthomonas oryzae pv. oryzae by direct DNA binding to the promoter of defense related gene, OsPR10a. OsWRKY51 in rice (Oryza sativa L.) is induced by exogenous salicylic acid (SA) and inoculation with Xanthomonas oryzae pv. oryzae (Xoo). To examine the role of OsWRKY51 in the defense response of rice, we generated OsWRKY51 overexpressing and underexpressing transgenic rice plants. OsWRKY51-overexpressing transgenic rice lines were more resistant to Xoo and showed greater expression of defense-related genes than wild-type (WT) plants, while OsWRKY51-underexpressing lines were more susceptible to Xoo and showed less expression of defense-associated genes than WT plants. Transgenic lines overexpressing OsWRKY51 showed growth retardation compared to WT plants. In contrast, transgenic lines underexpressing OsWRKY51 by RNA interference showed similar plant height with WT plants. Transient expression of OsWRKY51-green fluorescent protein fusion protein in rice protoplasts revealed that OsWRKY51 was localized in the nucleus. OsWRKY51 bound to the W-box and WLE1 elements of the OsPR10a promoter. Based on these results, we suggest that OsWRKY51 is a positive transcriptional regulator of defense signaling and has direct DNA binding ability to the promoter of OsPR10a, although it is reported to be a negative regulator in GA signaling.

Molecular characterization of Oryza sativa WRKY6, which binds to W-box-like element 1 of the Oryza sativa pathogenesis-related (PR) 10a promoter and confers reduced susceptibility to pathogens.

WRKY proteins are transcription factors (TFs) that regulate the expression of defense-related genes. The salicylic acid (SA)-inducible Oryza sativa WRKY6 (OsWRKY6) was identified as a positive regulator of Oryza sativa pathogenesis-related 10a (OsPR10a) by transient expression assays. A physical interaction between OsWRKY6 and W-box-like element 1 (WLE1), which positively regulates OsPR10a/probenazole induced protein 1 expression, was verified in vitro. Several pathogenesis-related (PR) genes were constitutively activated, including OsPR10a, and transgenic rice (Oryza sativa) plants overexpressing (ox) OsWRKY6 exhibited enhanced disease resistance to pathogens. By contrast, PR gene induction was compromised in transgenic OsWRKY6-RNAi lines, suggesting that OsWRKY6 is a positive regulator of defense responses. OsWRKY6-ox lines displayed leaf lesions, and increased OsWRKY6 levels caused cell death. Salicylic acid (SA) concentrations were higher in OsWRKY6-ox lines than in wild-type (WT) plants, and transcript levels of Oryza sativa isochorismate synthase 1 (OsICS1), which encodes a major enzyme involved in SA biosynthesis, were higher in OsWRKY6-ox lines than in WT. OsWRKY6 directly bound to the OsICS1 promoter in vivo. This indicates that OsWRKY6 can directly regulate OsICS1 expression and thereby increase SA concentrations. OsWRKY6 autoregulates its own expression. OsWRKY6 protein degradation is possibly regulated by ubiquitination. Our results suggest that OsWRKY6 positively regulates defense responses through activation of OsICS1 expression and OsWRKY6 stabilization.

The Rab GTPase RabG3b positively regulates autophagy and immunity-associated hypersensitive cell death in Arabidopsis.

A central component of the plant defense response to pathogens is the hypersensitive response (HR), a form of programmed cell death (PCD). Rapid and localized induction of HR PCD ensures that pathogen invasion is prevented. Autophagy has been implicated in the regulation of HR cell death, but the functional relationship between autophagy and HR PCD and the regulation of these processes during the plant immune response remain controversial. Here, we show that a small GTP-binding protein, RabG3b, plays a positive role in autophagy and promotes HR cell death in response to avirulent bacterial pathogens in Arabidopsis (Arabidopsis thaliana). Transgenic plants overexpressing a constitutively active RabG3b (RabG3bCA) displayed accelerated, unrestricted HR PCD within 1 d of infection, in contrast to the autophagy-defective atg5-1 mutant, which gradually developed chlorotic cell death through uninfected sites over several days. Microscopic analyses showed the accumulation of autophagic structures during HR cell death in RabG3bCA cells. Our results suggest that RabG3b contributes to HR cell death via the activation of autophagy, which plays a positive role in plant immunity-triggered HR PCD.

Comparison of natural course, intravitreal triamcinolone and macular laser photocoagulation for treatment of mild diabetic macular edema.

PURPOSE: To assess the natural course of the mild diabetic macular edema(DME) and to compare the visual outcomes with the patients with receiving either macular laser photocoagulation or intravitreal injection of triamcinolone acetonide(IVTA). METHODS: 28 eyes with central macular thickness (CMT) of between 250 to 300µm were followed without treatment and 48 eyes with CMT between 300 to 500µm had been divided into 3 subgroups according to treatment. We evaluated the best corrected visual acuity (BCVA) and CMT of natural course group and compared the BCVA and CMT of the patients who had been treated with IVTA or macular laser treatment. RESULTS: The eyes with DME between 250 to 300µm showed no significant change in BCVA and CMT at 6 month. Among the eyes with DME between 300 to 500µm, all 3 subgroups showed no statistically significant change of BCVA at any follow up period and no significant difference was revealed among the subgroups. All subgroups showed significant reduction of CMT after 1 month and maintained until final follow-up and there was no significant difference among subgroups. CONCLUSIONS: Mild DME between 250 to 500µm did not show significant worsening of BCVA or macular edema without any specific treatment.

Passivation of precipitation-hardened UNS N07718 in a shallow sour aqueous solution.

Passivation of precipitation-hardened UNS N07718 in 5 wt% NaCl + 0.5 wt% CH3COOH was investigated. Cyclic potentiodynamic polarisation revealed that the alloy surface was passivated without active-passive transition behaviour. The alloy surface was in a stable passive state during potentiostatic polarisation at 0.5 VSSE for 12 h. Bode and Mott-Schottky plots showed that the passive film became electrically resistive and less defective with n-type semiconductive properties during the polarisation. X-ray photoelectron spectra revealed that Cr- and Fe-enriched hydro/oxide layers were formed on the outer and inner layers of the passive film, respectively. The thickness of the film was almost constant with the increase of the polarisation time. The outer Cr-hydroxide layer changed into a Cr-oxide layer during the polarisation, resulting in a decreased donor density in the passive film. The film's composition change during the polarisation should be related to the corrosion resistance of the alloy in the shallow sour conditions.

The Rab GTPase RabG3b functions in autophagy and contributes to tracheary element differentiation in Arabidopsis.

The tracheary elements (TEs) of the xylem serve as the water-conducting vessels of the plant vascular system. To achieve this, TEs undergo secondary cell wall thickening and cell death, during which the cell contents are completely removed. Cell death of TEs is a typical example of developmental programmed cell death that has been suggested to be autophagic. However, little evidence of autophagy in TE differentiation has been provided. The present study demonstrates that the small GTP binding protein RabG3b plays a role in TE differentiation through its function in autophagy. Differentiating wild type TE cells were found to undergo autophagy in an Arabidopsis culture system. Both autophagy and TE formation were significantly stimulated by overexpression of a constitutively active mutant (RabG3bCA), and were inhibited in transgenic plants overexpressing a dominant negative mutant (RabG3bDN) or RabG3b RNAi (RabG3bRNAi), a brassinosteroid insensitive mutant bri1-301, and an autophagy mutant atg5-1. Taken together, our results suggest that autophagy occurs during TE differentiation, and that RabG3b, as a component of autophagy, regulates TE differentiation.

Cysteamine suppresses human peripheral blood mononuclear cells--human corneal endothelial cell reaction via reactive oxygen species reduction.

PURPOSE: To investigate the effect of cysteamine (CYS) on mixed peripheral blood mononuclear cells (PBMCs)--human corneal endothelial cell (HCEC) reaction (MLER). METHODS: PBMC stimulation assay was performed using cultured HCEC. MLERs were treated with various concentrations of CYS (0-20 mM). The proliferation rate and secretion profiles of transforming growth factor-ß1 (TGF-ß1) and interleukin-6 (IL-6) of PBMCs stimulated by cultured HCEC were determined using bromodeoxyuridine proliferation assay and enzyme-linked immunosorbent assay, respectively. RESULTS: CYS suppressed PBMC proliferation in a dose-dependent manner (p<0.001). The intracellular reactive oxygen species (ROS) levels decreased with an increase in CYS concentration (p<0.001). The levels of TGF-ß1 and IL-6 decreased in a dose-dependent manner as well (p=0.011 and 0.003, respectively). CONCLUSIONS: This study showed that CYS decreased PBMC proliferation, IL-6 and TGF-ß1 levels via ROS formation. Our results suggest that CYS could suppress inflammation associated with PBMCs to corneal endothelial cells.

Overexpression of constitutively active Arabidopsis RabG3b promotes xylem development in transgenic poplars.

An Arabidopsis small GTPase, RabG3b, was previously characterized as a component of autophagy and as a positive regulator for xylem development in Arabidopsis. In this work, we assessed whether RabG3b modulates xylem-associated traits in poplar in a similar way as in Arabidopsis. We generated transgenic poplars (Populus alba × Populus tremula var. glandulosa) overexpressing a constitutively active form of RabG3b (RabG3bCA) and performed a range of morphological, histochemical and molecular analyses to examine xylogenesis. RabG3bCA transgenic poplars showed increased stem growth due to enhanced xylem development. Autophagic structures were observed in differentiating xyelm cells undergoing programmed cell death (PCD) in wild-type poplar, and were more abundant in RabG3bCA transgenic poplar plants and cultured cells. Xylogenic activation was also accompanied by the expression of secondary wall-, PCD- and autophagy-related genes. Collectively, our results suggest that Arabidopsis RabG3b functions to regulate xylem growth through the activation of autophagy during wood formation in Populus, as does the same in Arabidopsis.

SRFR1, a suppressor of effector-triggered immunity, encodes a conserved tetratricopeptide repeat protein with similarity to transcriptional repressors.

Effector-triggered immunity provides plants with strong protection from pathogens. However, this response has the potential to be highly deleterious to the host and needs to be tightly controlled. The molecular mechanisms in the plant that regulate the balance between activation and suppression of resistance are not fully understood. Previously, we identified Arabidopsis suppressor of rps4-RLD 1 (srfr1) mutants with enhanced resistance to the bacterial effector AvrRps4. These mutants were recessive and retained full susceptibility to virulent bacteria, suggesting that SRFR1 functions as a negative regulator and that AvrRps4-triggered immunity was specifically enhanced in the mutants. Consistent with this, we show here that the response to flagellin, an elicitor of basal resistance, is unaltered in srfr1-1. In contrast, resistance to AvrRps4 in srfr1-1 requires EDS1, a central regulator of effector-triggered immunity via multiple resistance genes. SRFR1 is a single-copy gene encoding a pioneer tetratricopeptide repeat protein conserved between plants and animals. The SRFR1 tetratricopeptide repeat domain shows sequence similarity to those of transcriptional repressors in Saccharomyces cerevisiae and Caenorhabditis elegans. Indeed, a sub-pool of SRFR1 transiently expressed in Nicotiana benthamiana leaf cells localizes to the nucleus. Identification of SRFR1 may therefore provide insight into the regulation of the transcriptional reprogramming that is activated by effector-triggered immunity.

GDSL lipase-like 1 regulates systemic resistance associated with ethylene signaling in Arabidopsis.

Systemic resistance is induced by necrotizing pathogenic microbes and non-pathogenic rhizobacteria and confers protection against a broad range of pathogens. Here we show that Arabidopsis GDSL LIPASE-LIKE 1 (GLIP1) plays an important role in plant immunity, eliciting both local and systemic resistance in plants. GLIP1 functions independently of salicylic acid but requires ethylene signaling. Enhancement of GLIP1 expression in plants increases resistance to pathogens including Alternaria brassicicola, Erwinia carotovora and Pseudomonas syringae, and limits their growth at the infection site. Furthermore, local treatment with GLIP1 proteins is sufficient for the activation of systemic resistance, inducing both resistance gene expression and pathogen resistance in systemic leaves. The PDF1.2-inducing activity accumulates in petiole exudates in a GLIP1-dependent manner and is fractionated in the size range of less than 10 kDa as determined by size exclusion chromatography. Our results demonstrate that GLIP1-elicited systemic resistance is dependent on ethylene signaling and provide evidence that GLIP1 may mediate the production of a systemic signaling molecule(s).

Metabolite Profiling and Antioxidant Activity of 10 New Early- to Mid-Season Apple Cultivars and 14 Traditional Cultivars.

Early- to mid-season apple cultivars have recently been developed in response to global warming; however, their metabolite compositions remain unclear. Herein, metabolites, such as free sugars, and organic acids and antioxidant activity were determined in 10 new and 14 traditional apple cultivars. Additionally, the phenolic profiles of the apple pulp and peel were characterized by high-resolution mass spectrometry. Major phenolic compounds in apples varied depending on the cultivar and tissue (i.e., peel or pulp). Among the new apple cultivars, Decobell and Tinkerbell, showed high antioxidant activity and contained higher phenolic compound content than other cultivars in the peel and pulp, respectively. Honggeum showed high phenolic content with similar sugar to acid ratio compared to popular traditional cultivars. In addition to antioxidant phenolic contents, metabolite profile information can be used to select apple cultivars for various purposes. For example, Indo can be selected for sweet apple taste because of its higher sugar to acid ratio. This information can be used to select apple cultivars for various purposes. For example, Decobell peel could be used as sources of food supplements and food additives, and Tinkerbell pulp can be utilized for apple juice making according to its metabolite profile.

Characterization of Fruit Quality Attributes and Cell Wall Metabolism in 1-Methylcyclopropene (1-MCP)-Treated 'Summer King' and 'Green Ball' Apples During Cold Storage.

This study aimed to elucidate whether 1-methylcyclopropene (1-MCP) treatment delays the fruit softening mechanism associated with the fruit quality of the newly released apple cultivars "Summer King" and "Green Ball" during cold storage. For both cultivars, the fruit treated with 1-MCP exhibited lower internal ethylene concentration, higher firmness, and higher titratable acidity relative to the control fruit, in association with less fruit softening. In addition, the treated fruit significantly delayed fresh weight loss and reduction of soluble solids content, especially in "Green Ball." Moreover, slower degradation of cell wall components (water-soluble pectin, sodium carbonate-soluble pectin, hemicellulose, and cellulose) was also observed in the treated fruit in comparison to the control fruit. Similarly, the enzymatic activities (of polygalacturonase, pectin methylesterase, cellulase, ß-galactosidase, and α-L-arabinofuranosidase) that cause cell wall degradation were relatively lower in the treated fruit than in the control fruit for both cultivars, which was further proved by transcriptional analysis of the genes encoding the enzymes. Overall, the results suggested that the usage of 1-MCP is useful to delay fruit softening of the two cultivars during cold storage by delaying the degradation of cell wall components and enzymatic activities of cell wall hydrolysis.

Structural, optical, and electrical properties of semiconducting ZnO nanosheets.

ZnO nanosheets were fabricated by an oxygen-assisted carbothermal reduction process and their properties were evaluated. In particular, the FET characteristics and photoluminescence properties of ZnO nanosheets were evaluated. The conduction type of ZnO nanosheets was determined as an n-type and the mobility was 20-40 cm2/ V-s, which is fairly high compared to ZnO nanowires. This might be attributed to the wide conduction area of ZnO nanosheet compared to nanowire structures and their better crystallinity.

Optic disc edema as a possible complication of Imatinib mesylate (Gleevec).

BACKGROUND: Imatinib is effective for a variety of hematologic and nonhematologic malignancies. Well-known ocular side effects are periorbital edema, epiphora, extraocular muscle palsy, and blepharoconjunctivitis. However, optic disc edema has not been reported as a complication of imatinib mesylate. Herein, we describe a patient with chronic myeloid leukemia (CML) who developed optic disc edema during treatment with imatinib. CASE: A 14-year-old Korean girl was referred to ophthalmology for a 3-week history of photopsia in both eyes. She had started taking imatinib daily 2 months previously for CML. At the initial exam, slit lamp showed optic disc edema in both eyes, even though visual acuity was 20/20 and other optic nerve function evaluations were within normal limits. We recommended to the oncologist discontinuation of the imatinib treatment. OBSERVATIONS: When the patient was followed for 1 week after stopping imatinib treatment, the frequency of photopsia decreased and the optic disc edema improved. Because a second examination 3 weeks after discontinuation of imatinib revealed much improved optic disc edema, she restarted the imatinib treatment. No ocular side effects have been noted so far. CONCLUSIONS: Optic disc edema should be considered one of the complications associated with imatinib. We propose discontinuation of the treatment for a short period when optic disc edema occurs.

Upper eyelid retraction after periorbital trauma.

We report four unusual cases of upper eyelid retraction following periorbital trauma. Four previously healthy patients were evaluated for unilateral upper eyelid retraction following periorbital trauma. A 31-year-old man (Case 1) and a 24-year-old man (Case 2) presented with left upper eyelid retraction which developed after blow-out fractures, a 44-year-old woman (Case 3) presented with left upper eyelid retraction secondary to a periorbital contusion that occurred one week prior, and a 56-year-old man (Case 4) presented with left upper eyelid retraction that developed 1 month after a lower canalicular laceration was sustained during a traffic accident. The authors performed a thyroid function test and orbital computed tomography (CT) in all cases. Thyroid function was normal in all patients, CT showed an adhesion of the superior rectus muscle and superior oblique muscle in the first case and diffuse thickening of the superior rectus muscle and levator complex in the third case. CT showed no specific findings in the second or fourth cases. Upper eyelid retraction due to superior complex adhesion can be considered one of the complications of periorbital trauma.

Differential expression of three catalase genes in the small radish (Rhaphanus sativus L. var. sativus).

Three catalase cDNA clones were isolated from the small radish (Raphanus sativus L.). Their nucleotide and deduced amino acid sequences showed the greatest homology to those of Arabidopsis. Genomic Southern blot analysis, using RsCat1 cDNA as a probe, showed that catalases are encoded by small multigene family in the small radish. Nondenaturing polyacrylamide gels revealed the presence of several catalase isozymes, the levels of which varied among the organs examined. The isozyme activities were assigned the individual catalase genes by Northern analysis using total RNA from different organs. The three catalase genes were differentially expressed in response to treatments such as white light, xenobiotics, osmoticum, and UV. Their expression in seedlings was controlled by the circadian clock under a light/dark cycle and/or in constant light. Interestingly, RsCat1 transcripts peaked in the morning, while those of RsCat2 and RsCat3 peaked in the early evening. Our results suggest that the RsCat enzymes are involved in defense against the oxidative stress induced by environmental changes.

Overexpression in Arabidopsis of a plasma membrane-targeting glutamate receptor from small radish increases glutamate-mediated Ca2+ influx and delays fungal infection.

Ionotropic glutamate receptors (iGluRs) are ligand-gated nonselective cation channels that mediate fast excitatory neurotransmission. Although homologues of the iGluRs have been identified in higher plants, their roles are largely unknown. In this work we isolated a full-length cDNA clone (RsGluR) encoding a putative glutamate receptor from small radish. An RsGluR: mGFP fusion protein was localized to the plasma membrane. In Arabidopsis thaliana overexpressing the full-length cDNA, glutamate treatment triggered greater Ca2+ influx in the root cells of transgenic seedlings than in those of the wild type. Transgenic plants exhibited multiple morphological changes such as necrosis at their tips and the margins of developing leaves, dwarf stature with multiple secondary inflorescences, and retarded growth, as previously observed in transgenic Arabidopsis overexpressing AtGluR3.2 [Kim et al. (2001)]. Microarray analysis showed that jasmonic acid (JA)-responsive genes including defensins and JA-biosynthetic genes were up-regulated. RsGluR overexpression also inhibited growth of a necrotic fungal pathogen Botrytis cinerea possibly due to up-regulation of the defensins. Based on these results, we suggest that RsGluR is a glutamate-gated Ca2+ channel located in the plasma membrane of higher plants and plays a direct or indirect role in defense against pathogen infection by triggering JA biosynthesis.

Erratum to: The relationship between quality of sleep and night shift rotation interval.

[This corrects the article DOI: 10.1186/s40557-015-0084-x.].