Journal: Journal of Fungi

Article Title: Fungal Endophytes for Grass Based Bioremediation: An Endophytic Consortium Isolated from Agrostis stolonifera Stimulates the Growth of Festuca arundinacea in Lead Contaminated Soil

doi: 10.3390/jof6040254

Figure Lengend Snippet: Design of the in vivo experiment. Galena at each concentration was added separately to the soil and well mixed before dividing into 20 pots for each treatment for a total of 80 pots. In each pot, two seeds of Festuca arundinacea were sown. Pots labelled E were treated with the endophyte consortium and those labeled C were treated with sterile water.

Article Snippet: Then, two seeds of Festuca arundinacea (cultivar Jesup, endophytes-free Nil, type continental, accession number: T9904, % seedling emergence of 92%; supplied by Grasslanz Technology Limited, New Zealand) were sown in each pot at a 1 cm depth.

Techniques: In Vivo, Concentration Assay, Labeling, Sterility

Journal: Journal of Fungi

Article Title: Fungal Endophytes for Grass Based Bioremediation: An Endophytic Consortium Isolated from Agrostis stolonifera Stimulates the Growth of Festuca arundinacea in Lead Contaminated Soil

doi: 10.3390/jof6040254

Figure Lengend Snippet: Germination time (here indicated as days-after-sowing, das ) of Festuca arundinacea seeds. Endophytic treatment is indicated by the letter E and control by the letter C at increasing concentration of lead in the soil. The significant statistical difference ( p < 0.05) between endophytic and control set is marked with a (*).

Article Snippet: Then, two seeds of Festuca arundinacea (cultivar Jesup, endophytes-free Nil, type continental, accession number: T9904, % seedling emergence of 92%; supplied by Grasslanz Technology Limited, New Zealand) were sown in each pot at a 1 cm depth.

Techniques: Control, Concentration Assay

Journal: Journal of Fungi

Article Title: Fungal Endophytes for Grass Based Bioremediation: An Endophytic Consortium Isolated from Agrostis stolonifera Stimulates the Growth of Festuca arundinacea in Lead Contaminated Soil

doi: 10.3390/jof6040254

Figure Lengend Snippet: Summary of germination index (GI) results. Number of germinated seeds of Festuca arundinacea with endophyte treatment ( E ) are compared with the control treatment ( C ) at increasing concentrations of lead in the soil. Significant statistical difference ( p < 0.05) between endophytic and control set is marked with a (*).

Article Snippet: Then, two seeds of Festuca arundinacea (cultivar Jesup, endophytes-free Nil, type continental, accession number: T9904, % seedling emergence of 92%; supplied by Grasslanz Technology Limited, New Zealand) were sown in each pot at a 1 cm depth.

Techniques: Control

Journal: Journal of Fungi

Article Title: Fungal Endophytes for Grass Based Bioremediation: An Endophytic Consortium Isolated from Agrostis stolonifera Stimulates the Growth of Festuca arundinacea in Lead Contaminated Soil

doi: 10.3390/jof6040254

Figure Lengend Snippet: Percentage of total tillered plants at three recorded times ( 29 das, 35 das, and 42 das) of Festuca arundinacea ; endophytic treatment ( E ) and control ( C ) grown at increasing concentrations of lead in the soil. Significant statistical difference ( p < 0.05) between endophytic and control set is marked with a (*).

Article Snippet: Then, two seeds of Festuca arundinacea (cultivar Jesup, endophytes-free Nil, type continental, accession number: T9904, % seedling emergence of 92%; supplied by Grasslanz Technology Limited, New Zealand) were sown in each pot at a 1 cm depth.

Techniques: Control

Journal: Journal of Fungi

Article Title: Fungal Endophytes for Grass Based Bioremediation: An Endophytic Consortium Isolated from Agrostis stolonifera Stimulates the Growth of Festuca arundinacea in Lead Contaminated Soil

doi: 10.3390/jof6040254

Figure Lengend Snippet: Foliar biomass measurement of fresh ( a ) and dry weight ( b ) of Festuca arundinacea with endophytes treatment ( E ) were compared with the control treatment ( C ) at increasing concentrations of lead in the soil.

Article Snippet: Then, two seeds of Festuca arundinacea (cultivar Jesup, endophytes-free Nil, type continental, accession number: T9904, % seedling emergence of 92%; supplied by Grasslanz Technology Limited, New Zealand) were sown in each pot at a 1 cm depth.

Techniques: Control

Journal: Journal of Fungi

Article Title: Fungal Endophytes for Grass Based Bioremediation: An Endophytic Consortium Isolated from Agrostis stolonifera Stimulates the Growth of Festuca arundinacea in Lead Contaminated Soil

doi: 10.3390/jof6040254

Figure Lengend Snippet: Average of total tillers per pot produced by Festuca arundinacea with endophyte treatment ( E ) were compared with control treatment ( C ) at increasing concentration of lead in the soil. Bars in the graph represent the standard deviation.

Article Snippet: Then, two seeds of Festuca arundinacea (cultivar Jesup, endophytes-free Nil, type continental, accession number: T9904, % seedling emergence of 92%; supplied by Grasslanz Technology Limited, New Zealand) were sown in each pot at a 1 cm depth.

Techniques: Produced, Control, Concentration Assay, Standard Deviation

Journal: Scientific Reports

Article Title: Intracellular ion concentrations and cation-dependent remodelling of bacterial MreB assemblies

doi: 10.1038/s41598-020-68960-w

Figure Lengend Snippet: Microscopy imaging of MreB polymers isolated under different ion conditions. ( A and B ) PF filtered, Alexa488-labeled Li -MreB polymers form large superstructures ( A ) in polymerizing ionic conditions (300 mM KCl, 2 mM MgCl 2 , 0.1 mM CaCl 2 ) ( B ) in its intracellular ionic conditions determined in this study (296 mM NaCl, 5 mM KCl, 0.38 mM MgCl 2 , 0.99 mM CaCl 2 ). ( C and D ) PF filtered, Alexa488-labeled Rr- MreB polymers form sheets or assemblies in an ionic strength dependent manner ( C ) under high ionic conditions (300 mM KCl, 2 mM MgCl 2 , 0.1 mM CaCl 2 ), ( D ) and under lower KCl conditions (200 mM KCl, 2 mM MgCl 2 , 0.1 mM CaCl 2 ). ( E and F ) PF filtered, Alexa488-labeled Ec-MreB polymers form large superstructures ( E ) under high ionic conditions (300 mM KCl, 2 mM MgCl 2 , 0.1 mM CaCl 2 ) (Ec-MreB polymers form similar assemblies to Li-MreB), ( F ) in its intracellular ionic conditions determined in this study (219 mM NaCl, 38 mM KCl, 2.38 mM MgCl 2 , 0.24 mM CaCl 2 ).

Article Snippet: The Leptospira interrogans MreB gene (gene bank accession number: AAS69864.1), the Rickettsia rickettsii MreB gene (gene bank accession number: ABY73136.1) and the Escherichia coli MreB gene (gene bank accession number: AJF45056.1) sequences were codon optimized for expression in E. coli , synthesized (GenScript) and engineered into the pSY5 plasmid which encodes an 8-histidine tag, followed by a PreScission protease (GE Healthcare Life Sciences) cleavage site prior to the protein sequence.

Techniques: Microscopy, Imaging, Isolation, Labeling

Journal: Scientific Reports

Article Title: Intracellular ion concentrations and cation-dependent remodelling of bacterial MreB assemblies

doi: 10.1038/s41598-020-68960-w

Figure Lengend Snippet: Effect of divalent cations on MreB monomers and polymers. ( A and B ) Relative light scattering change of 50 µM ( A ) Li -MreB or ( B ) Rr- MreB solutions on adding 2 mM calcium to monomers (purified under denaturing conditions) in the absence of high salt (2 mM TRIS.HCl, 0.1 mM CaCl 2 , 0.2 ATP, 1 mM DTT, pH 8.0) (gray line) or to polymers in the presence of high ionic conditions (300 mM KCl, 2 mM MgCl 2 , 0.1 mM CaCl 2 ) (dashed line). ( C ) Microscopy time-lapse snapshots from the entropic motion of Alexa488 labeled Li -MreB superstructures under high salt conditions (300 mM KCl, 2 mM MgCl 2 , 0.1 mM CaCl 2 ). The state of polymer before (gray), 5 min after (cyan) and 10 min after (green) the 2 mM calcium addition. On the overlay image, white arrows indicate that the length change of assemblies contracted by calcium addition (lower panel). ( D and E ) Microscopy images of Alexa488-labeled ( D ) Li -MreB or ( E ) Rr- MreB polymers after 6 mM EDTA treatment, in the absence of Ca 2+ and Mg 2+ . ( F and G ) Microscopy images of Alexa488 labeled ( F ) Li -MreB or ( G ) Rr- MreB polymers after 6 mM EGTA treatment, in the absence of Ca 2+ and presence of Mg 2+ .

Article Snippet: The Leptospira interrogans MreB gene (gene bank accession number: AAS69864.1), the Rickettsia rickettsii MreB gene (gene bank accession number: ABY73136.1) and the Escherichia coli MreB gene (gene bank accession number: AJF45056.1) sequences were codon optimized for expression in E. coli , synthesized (GenScript) and engineered into the pSY5 plasmid which encodes an 8-histidine tag, followed by a PreScission protease (GE Healthcare Life Sciences) cleavage site prior to the protein sequence.

Techniques: Purification, Microscopy, Labeling, Polymer

Journal: Scientific Reports

Article Title: Intracellular ion concentrations and cation-dependent remodelling of bacterial MreB assemblies

doi: 10.1038/s41598-020-68960-w

Figure Lengend Snippet: Rapid changes in calcium levels caused structural reorganization of MreB polymers. ( A ) Under high ionic conditions (300 mM KCl, 2 mM MgCl , 0.1 mM CaCl ), 6 mM EGTA was added to polymerized Li -MreB (50 µM) (black line) then a 2 mM calcium addition caused a jump in light scattering. EGTA and Ca 2+ were added in the opposite order (orange line). ( B ) Under high ionic conditions (300 mM KCl, 2 mM MgCl , 0.1 mM CaCl ), 6 mM EGTA was added to Rr- MreB polymers (50 µM) (black line) then 2 mM Ca 2+ treatment caused a similar response in light scattering as in case of Li -MreB (arrows indicate the addition of Ca 2+ or EGTA). The initial state of MreB at the beginning of the experiment was similar to that shown in Fig. . ( C and D ) Confocal fluorescent microscope images of Alexa488-labeled Li -MreB polymers ( C ) after the consecutive application of 2 mM Ca 2+ then 6 mM EGTA or ( D ) first 6 mM EGTA then 2 mM Ca 2+ . ( E and F ) Images of Alexa488-labeled ( E ) Rr- MreB, ( F ) Ec -MreB polymers, after consecutive application of 6 mM EGTA then 2 mM Ca 2+ . ( G and H ) Microscopy images of ( G ) Li -MreB or ( H ) Rr- MreB overexpressing E. coli cell lysates (diluted two times with distilled water and spread on glass slides) after 6 mM EGTA then 2 mM calcium addition. All cysteines of the sample were labeled with Alexa488-maleimide.

Article Snippet: The Leptospira interrogans MreB gene (gene bank accession number: AAS69864.1), the Rickettsia rickettsii MreB gene (gene bank accession number: ABY73136.1) and the Escherichia coli MreB gene (gene bank accession number: AJF45056.1) sequences were codon optimized for expression in E. coli , synthesized (GenScript) and engineered into the pSY5 plasmid which encodes an 8-histidine tag, followed by a PreScission protease (GE Healthcare Life Sciences) cleavage site prior to the protein sequence.

Techniques: Microscopy, Labeling

Journal: Scientific Reports

Article Title: Intracellular ion concentrations and cation-dependent remodelling of bacterial MreB assemblies

doi: 10.1038/s41598-020-68960-w

Figure Lengend Snippet: Persistence lengths of different MreB assemblies. ( A ) Analysis of microscopy images fitting with the WLC model in the Easyworm software. Alexa488-labeled Rr- MreB polymers were grown under different KCl conditions (50–400 mM), which reveals the longest persistence length at its polymerization optimum at 200 mM of KCl (blue circles). The persistence length of Alexa488 labeled Li -MreB is shorter at its polymerization optimum at 300 mM KCl (red circle). ( B ) The persistence length of Alexa488-labeled Li -MreB assemblies (red columns) changed significantly in the presence of 2 mM calcium, and in the absence of calcium by the treatment with 6 mM EGTA, and Rr- MreB assemblies (blue columns) showed a non-significant change in the presence of 2 mM calcium, but a significant change in the presence of 6 mM EGTA. Both assemblies show non-significant differences to the pretreated samples on subsequent addition of 2 mM calcium after EGTA treatment. The persistence length of Alexa488-labeled Ec -MreB assemblies (green columns) did not show a large difference on the addition of 6 mM EGTA followed by 2 mM calcium. Error bars refer to mean ± SD of five independent measurements. Double asterisks indicate statistically significant difference relative to the initial conditions. The analysis was based on a two-sample t -test, p < 0.005.

Article Snippet: The Leptospira interrogans MreB gene (gene bank accession number: AAS69864.1), the Rickettsia rickettsii MreB gene (gene bank accession number: ABY73136.1) and the Escherichia coli MreB gene (gene bank accession number: AJF45056.1) sequences were codon optimized for expression in E. coli , synthesized (GenScript) and engineered into the pSY5 plasmid which encodes an 8-histidine tag, followed by a PreScission protease (GE Healthcare Life Sciences) cleavage site prior to the protein sequence.

Techniques: Microscopy, Software, Labeling

Journal: Scientific Reports

Article Title: Intracellular ion concentrations and cation-dependent remodelling of bacterial MreB assemblies

doi: 10.1038/s41598-020-68960-w

Figure Lengend Snippet: A hypothetical model to interpret the dynamics of MreB polymers.

Article Snippet: The Leptospira interrogans MreB gene (gene bank accession number: AAS69864.1), the Rickettsia rickettsii MreB gene (gene bank accession number: ABY73136.1) and the Escherichia coli MreB gene (gene bank accession number: AJF45056.1) sequences were codon optimized for expression in E. coli , synthesized (GenScript) and engineered into the pSY5 plasmid which encodes an 8-histidine tag, followed by a PreScission protease (GE Healthcare Life Sciences) cleavage site prior to the protein sequence.

Techniques: