Journal: bioRxiv

Article Title: A proteostasis clock underlies the timing of bacterial dormancy and antibiotic tolerance

doi: 10.64898/2026.03.09.710435

Figure Lengend Snippet: a , Representative brightfield and fluorescence images of WT and pheT D1 bacteria stained with Peroxy Orange 1 (PO-1). Cells from the stationary phase were re-cultured in LB or LB supplemented with 0.1 mg/mL catalase and sampled at the indicated time points. Arrows indicate PO-1-positive aggregates. Scale bars, 2 μm. b , PO-1 fluorescence intensity distribution for WT and pheT D1 cells at the indicated time points during recovery. c , Representative fluorescence images of pheT D1 bacteria stained with Proteostat after re-culturing in LB or LB supplemented with 0.1 mg/mL catalase. Scale bars, 2 μm. d , Quantification of the fraction of pheT D1 cells containing Proteostat-positive aggregates upon re-culturing in LB or LB supplemented with 0.1 mg/mL catalase. Data are presented as mean ± SD (n = 4). e , Growth curves of pheT D1 bacteria were monitored by measuring optical density at 600 nm (OD 600 ) over time in LB or LB supplemented with 0.1 mg/mL catalase. Data are presented as mean ± SD (n = 3).

Article Snippet: For Peroxy Orange 1 (PO-1) staining, bacteria were stained with PO-1 (MCE, HY-103469) at a concentration of 10 μM in PBS for 1 hour at 37 °C.

Techniques: Fluorescence, Bacteria, Staining, Cell Culture

Journal: bioRxiv

Article Title: A proteostasis clock underlies the timing of bacterial dormancy and antibiotic tolerance

doi: 10.64898/2026.03.09.710435

Figure Lengend Snippet: a , Representative brightfield and fluorescence images of WT and pheT D1 bacteria stained with Peroxy Orange 1 (PO-1). Cells from the stationary phase were re-cultured in LB or LB supplemented with 0.1 mg/mL catalase and sampled at the indicated time points. Arrows indicate PO-1-positive aggregates. Scale bars, 2 μm. b , PO-1 fluorescence intensity distribution for WT and pheT D1 cells at the indicated time points during recovery. c , Representative fluorescence images of pheT D1 bacteria stained with Proteostat after re-culturing in LB or LB supplemented with 0.1 mg/mL catalase. Scale bars, 2 μm. d , Quantification of the fraction of pheT D1 cells containing Proteostat-positive aggregates upon re-culturing in LB or LB supplemented with 0.1 mg/mL catalase. Data are presented as mean ± SD (n = 4). e , Growth curves of pheT D1 bacteria were monitored by measuring optical density at 600 nm (OD 600 ) over time in LB or LB supplemented with 0.1 mg/mL catalase. Data are presented as mean ± SD (n = 3).

Article Snippet: For Peroxy Orange 1 (PO-1) staining, bacteria were stained with PO-1 (MCE, HY-103469) at a concentration of 10 μM in PBS for 1 hour at 37 °C.

Techniques: Fluorescence, Bacteria, Staining, Cell Culture

Journal: Plant Biotechnology Journal

Article Title: MsWRKY49 Modulates Pollen Tube Elongation in Response to Boron Deficiency by Regulating Reactive Oxygen Species Homeostasis in Alfalfa

doi: 10.1111/pbi.70366

Figure Lengend Snippet: Low boron inhibits pollen tube elongation by increasing ROS levels in alfalfa. (a) Fluorescence microscopy of pollen tubes stained with the ROS sensor CM‐H 2 DCFDA under low boron conditions. BF refers to bright field. Scale bars = 10 μm. (b) Quantification of DCF fluorescence intensity in pollen tubes. Statistical significance is denoted by **, corresponding to p < 0.01. (c) DCF fluorescence intensity along the length of the pollen tube under low boron conditions. (d) Fluorescence microscopy of pollen tubes stained with the H 2 O 2 ‐specific dye PO1 under low boron conditions. BF refers to bright field. Scale bars = 10 μm. (e) Quantification of PO1 fluorescence intensity in pollen tubes. Mean ± SE values were determined from three biological replicates. n = 20–25. Statistical significance is denoted by **, corresponding to p < 0.01. (f) PO1 fluorescence intensity along the length of the pollen tube under low boron conditions. n = 15, and the shaded area represents the SE value from three independent experiments. (g) The effect of 1 mM H 2 O 2 treatment on pollen tube elongation under low boron conditions. Scale bars = 500 μm. (h) Statistical analysis of pollen tube length after 1 mM H 2 O 2 treatment. n = 300, mean ± SD from three independent experiments. Different letters represent significant differences at p < 0.05 (one‐way ANOVA). (i) Effect of 40 μM AsA treatment on pollen tube elongation under low boron conditions. Scale bars = 500 μm. (j) Statistical analysis of pollen tube length after 40 μM AsA treatment. n = 200, mean ± SD values were determined from three independent experiments. Different letters represent significant differences at p < 0.05 (one‐way ANOVA).

Article Snippet: Subsequently, the pollen tubes were stained with 10 μM 5‐(and 6)‐chloromethyl‐2′,7′‐dichlorodihydrofluorescein diacetate (CM‐H 2 DCFDA) (Thermo Fisher Scientific, USA) or 5 μM Peroxy Orange 1 (PO1) (Tocris Bioscience, UK) for 10 min at room temperature in the dark.

Techniques: Fluorescence, Microscopy, Staining

Journal: Plant Biotechnology Journal

Article Title: MsWRKY49 Modulates Pollen Tube Elongation in Response to Boron Deficiency by Regulating Reactive Oxygen Species Homeostasis in Alfalfa

doi: 10.1111/pbi.70366

Figure Lengend Snippet: The mswrky49 mutant promotes pollen tube elongation under low‐boron conditions by reducing ROS accumulation. (a) Pollen tube phenotypes of WT and mswrky49 mutants under different boron concentrations. Scale bars = 500 μm. (b) Length measurements of WT and mswrky49 mutant pollen tubes after 1 h of different boron concentrations treatment. The data represent mean ± SD of three independent experiments ( n = 200). Different letters represent significant differences at p < 0.05 (one‐way ANOVA). (c) Observation of H 2 O 2 levels in WT and mswrky49 mutant pollen tubes under different boron concentrations using PO1 fluorescence staining. Scale bars = 10 μm. (d) Quantification of fluorescence intensity of PO1 in WT and mswrky49 mutant pollen tubes under different boron concentrations. The data represent mean ± SE of three independent experiments ( n = 25–20). Different letters represent significant differences at p < 0.05 (one‐way ANOVA).

Article Snippet: Subsequently, the pollen tubes were stained with 10 μM 5‐(and 6)‐chloromethyl‐2′,7′‐dichlorodihydrofluorescein diacetate (CM‐H 2 DCFDA) (Thermo Fisher Scientific, USA) or 5 μM Peroxy Orange 1 (PO1) (Tocris Bioscience, UK) for 10 min at room temperature in the dark.

Techniques: Mutagenesis, Fluorescence, Staining

Journal: Plant Biotechnology Journal

Article Title: MsWRKY49 Modulates Pollen Tube Elongation in Response to Boron Deficiency by Regulating Reactive Oxygen Species Homeostasis in Alfalfa

doi: 10.1111/pbi.70366

Figure Lengend Snippet: Overexpression of MsWRKY49 in alfalfa reduces length of pollen tubes by increasing ROS accumulation. (a) Pollen tube phenotypes of WT and MsWRKY49‐OE under different boron concentrations. Scale bars = 500 μm. (b) Length measurements of WT and MsWRKY49‐OE pollen tubes after 1 h of different boron concentrations treatment. The data represent mean ± SD of three independent experiments ( n = 200). Different letters represent significant differences at p < 0.05 (one‐way ANOVA). (c) Observation of H 2 O 2 levels in WT and MsWRKY49‐OE pollen tubes under different boron concentrations using PO1 fluorescence staining. Scale bars = 10 μm. (d) Quantification of fluorescence intensity of PO1 in WT and MsWRKY49‐OE pollen tubes under different boron concentrations. The data represent mean ± SE of three independent experiments ( n = 15–20). Different letters represent significant differences at p < 0.05 (one‐way ANOVA).

Article Snippet: Subsequently, the pollen tubes were stained with 10 μM 5‐(and 6)‐chloromethyl‐2′,7′‐dichlorodihydrofluorescein diacetate (CM‐H 2 DCFDA) (Thermo Fisher Scientific, USA) or 5 μM Peroxy Orange 1 (PO1) (Tocris Bioscience, UK) for 10 min at room temperature in the dark.

Techniques: Over Expression, Fluorescence, Staining