MoSET1 (Histone H3K4 Methyltransferase in Magnaporthe oryzae) Regulates Global Gene Expression during Infection-Related Morphogenesis

Here we report the genetic analyses of histone lysine methyltransferase (KMT) genes in the phytopathogenic fungus Magnaporthe oryzae. Eight putative M. oryzae KMT genes were targeted for gene disruption by homologous recombination. Phenotypic assays revealed that the eight KMTs were involved in various infection processes at varying degrees. Moset1 disruptants (Δmoset1) impaired in histone H3 lysine 4 methylation (H3K4me) showed the most severe defects in infection-related morphogenesis, including conidiation and appressorium formation. Consequently, Δmoset1 lost pathogenicity on wheat host plants, thus indicating that H3K4me is an important epigenetic mark for infection-related gene expression in M. oryzae. Interestingly, appressorium formation was greatly restored in the Δmoset1 mutants by exogenous addition of cAMP or of the cutin monomer, 16-hydroxypalmitic acid. The Δmoset1 mutants were still infectious on the super-susceptible barley cultivar Nigrate. These results suggested that MoSET1 plays roles in various aspects of infection, including signal perception and overcoming host-specific resistance. However, since Δmoset1 was also impaired in vegetative growth, the impact of MoSET1 on gene regulation was not infection specific. ChIP-seq analysis of H3K4 di- and tri-methylation (H3K4me2/me3) and MoSET1 protein during infection-related morphogenesis, together with RNA-seq analysis of the Δmoset1 mutant, led to the following conclusions: 1) Approximately 5% of M. oryzae genes showed significant changes in H3K4-me2 or -me3 abundance during infection-related morphogenesis. 2) In general, H3K4-me2 and -me3 abundance was positively associated with active transcription. 3) Lack of MoSET1 methyltransferase, however, resulted in up-regulation of a significant portion of the M. oryzae genes in the vegetative mycelia (1,491 genes), and during infection-related morphogenesis (1,385 genes), indicating that MoSET1 has a role in gene repression either directly or more likely indirectly. 4) Among the 4,077 differentially expressed genes (DEGs) between mycelia and germination tubes, 1,201 and 882 genes were up- and down-regulated, respectively, in a Moset1-dependent manner. 5) The Moset1-dependent DEGs were enriched in several gene categories such as signal transduction, transport, RNA processing, and translation

Effects of water scarcity awareness and climate change belief on recycled water usage willingness: Evidence from New Mexico, United States

The global water crisis is being exacerbated by climate change, even in the United States. Recycled water is a feasible alternative to alleviate the water shortage, but it is constrained by humans’ perceptions. The current study examines how residents’ water scarcity awareness and climate change belief influence their willingness to use recycled water directly and indirectly. Bayesian Mindsponge Framework (BMF) analytics was employed on a dataset of 1831 residents in Albuquerque, New Mexico, an arid inland region in the US. We discovered that residents’ willingness to use direct recycled potable water is positively affected by their awareness of water scarcity, but the effect is conditional on their belief in the impacts of climate change on the water cycle. Meanwhile, the willingness to use indirect recycled potable water is influenced by water scarcity awareness, and the belief in climate change further enhances this effect. These findings implicate that fighting climate change denialism and informing the public of the water scarcity situation in the region can contribute to the effectiveness and sustainability of long-term water conservation and climate change alleviation efforts

Safety and efficacy of fluoxetine on functional outcome after acute stroke (AFFINITY): a randomised, double-blind, placebo-controlled trial

Background Trials of fluoxetine for recovery after stroke report conflicting results. The Assessment oF FluoxetINe In sTroke recoverY (AFFINITY) trial aimed to show if daily oral fluoxetine for 6 months after stroke improves functional outcome in an ethnically diverse population. Methods AFFINITY was a randomised, parallel-group, double-blind, placebo-controlled trial done in 43 hospital stroke units in Australia (n=29), New Zealand (four), and Vietnam (ten). Eligible patients were adults (aged ≥18 years) with a clinical diagnosis of acute stroke in the previous 2–15 days, brain imaging consistent with ischaemic or haemorrhagic stroke, and a persisting neurological deficit that produced a modified Rankin Scale (mRS) score of 1 or more. Patients were randomly assigned 1:1 via a web-based system using a minimisation algorithm to once daily, oral fluoxetine 20 mg capsules or matching placebo for 6 months. Patients, carers, investigators, and outcome assessors were masked to the treatment allocation. The primary outcome was functional status, measured by the mRS, at 6 months. The primary analysis was an ordinal logistic regression of the mRS at 6 months, adjusted for minimisation variables. Primary and safety analyses were done according to the patient's treatment allocation. The trial is registered with the Australian New Zealand Clinical Trials Registry, ACTRN12611000774921. Findings Between Jan 11, 2013, and June 30, 2019, 1280 patients were recruited in Australia (n=532), New Zealand (n=42), and Vietnam (n=706), of whom 642 were randomly assigned to fluoxetine and 638 were randomly assigned to placebo. Mean duration of trial treatment was 167 days (SD 48·1). At 6 months, mRS data were available in 624 (97%) patients in the fluoxetine group and 632 (99%) in the placebo group. The distribution of mRS categories was similar in the fluoxetine and placebo groups (adjusted common odds ratio 0·94, 95% CI 0·76–1·15; p=0·53). Compared with patients in the placebo group, patients in the fluoxetine group had more falls (20 [3%] vs seven [1%]; p=0·018), bone fractures (19 [3%] vs six [1%]; p=0·014), and epileptic seizures (ten [2%] vs two [<1%]; p=0·038) at 6 months. Interpretation Oral fluoxetine 20 mg daily for 6 months after acute stroke did not improve functional outcome and increased the risk of falls, bone fractures, and epileptic seizures. These results do not support the use of fluoxetine to improve functional outcome after stroke

Stress Is Associated with Quality of Life Reduction among Health Professionals in Vietnam: A Multisite Survey

Stress in healthcare workers is increasingly common in recent times. Stress can have negative effects on the mental health and quality of life of healthcare workers. This study is aimed at measuring the quality of life and determining the relationship between quality of life and stress of health professionals in some hospitals in Vietnam. A cross-sectional descriptive study was conducted on 520 health professionals working at Hanoi Medical University Hospital and Thai Binh Medical University Hospital. The World Health Organization Quality-of-Life Scale (WHOQOL-BREF) and the Depression, Anxiety, and Stress Scale-21 Items (DASS-21) scale were used to assess the quality of life and the stress status of healthcare workers. Multivariate regression was performed to measure the relationships between stress and quality of life. Results showed that the level of stress of health workers according to the DASS-21 scale at mild, moderate, severe, and very severe was 10.7%, 8.7%, 5.6%, and 2.9%, respectively. The mean score of overall quality of life was 60.97±11.39. Health workers under stress had a decrease in quality of life scores in physical, mental, social, and environmental domains. Stressed health workers had a reduced quality of life. Attention should be paid to providing appropriate interventions to reduce stress and improve the quality of life in healthcare workers

Correction: MoSET1 (Histone H3K4 Methyltransferase in <i>Magnaporthe oryzae</i>) Regulates Global Gene Expression during Infection-Related Morphogenesis

<p>Correction: MoSET1 (Histone H3K4 Methyltransferase in <i>Magnaporthe oryzae</i>) Regulates Global Gene Expression during Infection-Related Morphogenesis</p

Comparisons of H3K4 methylation dynamics during germination tube formation between MoSET1-dependent and -independent gene sets.

<p>“MoSET1-dependent genes” were defined based on a comparison of FC values (germination tubes [GT]/mycelia [myc]) between wild-type and Δmoset1 strains (see <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005385#sec002" target="_blank">Results</a>). When the rate of FC increase or decrease of a gene in the Δmoset1 strain was less than 50% of the wild-type strain, the gene was categorized as a MoSET1-dependent gene. Genes not meeting the criterion were classified as “MoSET1-independent genes”. MoSET1-dependent and -independent genes were further divided into “up-regulated (UP)” and “down-regulated (DOWN) gene groups. The levels of H3K4 methylation (H3K4me2 or H3K4me3) in each group is represented as a box plot. In the box plots, the central black line indicates the median value, and the top and bottom edges of the box are the 75th and 25th percentiles. The whiskers extend to the most extreme data points which are no more than 1.5 times the interquartile range from the box.</p

Infection types in wheat and barley cultivars inoculated with the wild type and mokmt deletion strains.

<p>- The size of lesion: 0, no visible evidence of infection; 1, pinpoint spots; 2, small lesion (<1.5 mm); 3, lesion with an intermediate size (<3 mm); 4, large and typical lesion; 5, complete blighting of leaf blades.</p><p>- The color of lesion: G: green, B: brown. 0 to 3B: resistant; 3G to 5G: susceptible</p><p>Infection types in wheat and barley cultivars inoculated with the wild type and mokmt deletion strains.</p

Phenotypic characterization of KMT mutants of <i>Magnaporthe oryzae</i>.

<p>(A) Vegetative growth was measured for 14 days on complete agar medium. (B) Conidiation was measured by counting the number of conidia harvested 3 days after BLB induction by suspending them with 20 ml of sterile distilled water per plate. (C) The rate of conidial germination on hydrophobic surfaces was measured under a light microscope. (D) Appressorium formation was measured as the percentage ratio of appressorium-forming mycelium to germinating mycelium on hydrophobic surfaces after 24 h incubation at 25°C. (B-D) Black bars indicate the wild-type strain Br48 (WT) or the original deletion mutants. White bars represent ectopic strains that have an insertion of a disruption construct somewhere in the genome other than the target locus. Grey bars indicate complemented strains that have random insertion of a plasmid carrying the corresponding wild-type locus in a deletion mutant background. All data are presented as means ± SD from three independent experiments. The order of the <i>M</i>. <i>oryzae</i> strains appeared in the figure is the same as that given in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005385#pgen.1005385.s014" target="_blank">S1 Table</a>. ND, not determined; *, significant difference (<i>p</i>< 0.01, two-tailed t-test after angular transformation).</p

The defect of <i>Δmoset1</i> mutants in appressorium formation was restored by exogenous signal molecules.

<p>(A) Effect of <i>Moset1</i> deletion on conidial morphology and appressorium formation. Conidia (a) and appressoria (b) of <i>M</i>. <i>oryzae</i> wild-type strain Br48. Conidia of the <i>Δmoset1</i> mutant showed an elongated shape (c) and seldom formed an appressorium (d). Bar = 10 μm. Exogenous addition of 5mM cAMP (B) and 5μM 1,16-hexadecanediol (C) restored appressorium formation in the <i>Δmoset1</i> mutant. Error bars represent standard deviation.</p