Image_1_Overexpression of SSBXoc, a Single-Stranded DNA-Binding Protein From Xanthomonas oryzae pv. oryzicola, Enhances Plant Growth and Disease and Salt Stress Tolerance in Transgenic Nicotiana benthamiana.PDF

<p>We previously reported that SSB_Xoc, a highly conserved single-stranded DNA-binding protein from Xanthomonas spp., was secreted through the type III secretion system (T3SS) and functioned as a harpin-like protein to elicit the hypersensitive response (HR) in the non-host plant, tobacco. In this study, we cloned Ssb_Xoc gene from X. oryzae pv. oryzicola (Xoc), the causal agent of bacterial leaf streak in rice, and transferred it into Nicotiana benthamiana via Agrobacterium-mediated transformation. The expression of Ssb_Xoc in transgenic N. benthamiana enhanced growth of both seedling and adult plants. When inoculated with the harpin Hpa1 or the pathogen Pseudomonas syringae pv. tomato DC3000 (Pst DC3000), the accumulation of reactive oxygen species (ROS) was increased more in Ssb_Xoc transgenic lines than that in wild-type (WT) plants. The expression of pathogenesis-related protein genes (PR1a and SGT1), HR marker genes (HIN1 and HSR203J) and the mitogen-activated protein kinase pathway gene, MPK3, was significantly higher in transgenic lines than in WT after inoculation with Pst DC3000. In addition, Ssb_Xoc transgenic lines showed the enhanced resistance to the pathogenic bacteria P. s. tabaci and the improved tolerance to salt stress, accompanied by the elevated transcription levels of the defense- and stress-related genes. Taken together, these results indicate that overexpression of the Ssb_Xoc gene in N. benthamiana significantly enhanced plant growth and increased tolerance to disease and salt stress via modulating the expression of the related genes, thus providing an alternative approach for development of plants with improved tolerance against biotic and abiotic stresses.</p

Administration of α-PGG inhibited <i>ex vivo</i> platelet aggregation induced by ADP or collagen.

<p>A, ADP or B, collagen was added to platelet-rich plasma, prepared from murine blood drawn at 30 min after oral administration of α-PGG (20 mg kg⁻¹) or vehicle, to induce aggregation. The aggregation tracings are representative of three experiments.</p

Insulin or α-PGG induced phosphorylation of insulin receptors and IRS-1.

<p>Washed human platelets were incubated with insulin (100 nM) or α-PGG (10 µM) for five and ten minutes. The reactions were stopped by adding lysis buffer and the total and phosphorylated insulin receptors (A) and total IRS-1 and phosphorylated IRS-1 (B) were visualized after immuno-precipitation and Western blotting. The phosphorylation of insulin receptors and IRS-1 was quantified by densitometry.</p

α-PGG inhibited collagen-induced phosphorylation of Akt.

<p>Washed human platelets were stimulated with collagen (1.0 µg mL⁻¹) in the presence or absence of α-PGG (10 µM). Lysis buffer was added to samples at 6 min to terminate reactions. Total Akt and p-Akt were visualized after PAGE and Western blotting as described in the Experimental Procedures. The β-actin was used as a loading control.</p

α-PGG inhibited thrombin induced rise in cytosolic calcium.

<p>Changes in cytosolic calcium were quantified in Fura2/AM loaded platelets. Platelets were incubated with α-PGG (3 or 10 µM) prior to stimulation with thrombin (0.1 U mL⁻¹) and changes in calcium levels were recorded by fluorescence spectrometry as described in Experimental Procedures. The results are reported as means ± SE (n = 4).</p

α-PGG inhibited ADP- or thrombin-induced lowering of cyclic AMP.

<p>ADP (10 µM), thrombin (0.1 U ml⁻¹) or PGE₁ (1 µM) induced changes in platelet cyclic AMP (pmoles/10⁸ platelets) levels were quantified in the presence or absence of α-PGG (10 µM) using enzyme-linked assay kits as described in Experimental Procedures. ADP and thrombin decreased basal cyclic AMP levels by 24% (*p<0.03) and 22% (*p<0.02) respectively. α-PGG blocked ADP and thrombin induced decrease in cyclic AMP levels.</p

α-PGG inhibited thrombin induced secretion from the α- and dense-granules and platelet aggregation.

<p>Washed human platelets were stimulated with thrombin (0.1 U mL⁻¹) in the presence or absence of α-PGG and expression of P-selectin (A) and secretion of ATP (B) and platelet aggregation (C) was monitored as detailed in Experimental Procedures. The results are reported as means ± SD for P-selectin expression (n = 3). ATP secretion and aggregation tracings are representative of three experiments.</p

Polymorphisms associations with coronary artery disease in our case–control study.

<p>Abbreviations: MAF, minor allele frequency; CI, confidence interval; ORs, odds ratios; ORs (95%CI) and <i>P</i> value were computed with multivariate logistic regression analysis by adjusting for age, sex, BMI, hypertension, diabetes, hyperlipidemia, smoking status, glucose, family history of cardiovascular diseases and renal function.</p

Association between <i>ROCK2</i> polymorphisms and blood pressure values in 927 health control subjects.

<p>Abbreviations: SBP, systolic blood pressure; DBP, Diastolic blood pressure; <i>P</i> value of the blood pressure levels was calculated by multiple linear regression model adjusted for age, sex, BMI and smoking status; A recessive model was used for rs56304104 (MAF <5%), and an additive model was used for rs978906 and rs2230774 (MAF >5%).</p

Frequency of <i>ROCK2</i> functional polymorphisms.

<p>Abbreviations: MAF, minor allele frequency; 3′-UTR, 3′-untranslated region.</p>a<p>Position is base-pair location in NCBI build 37.1 (GRCh37).</p