Powerpac basic

A solution of 0.5 μM CaCl₂.2H₂O and 0.5 mM MgCl₂.6H₂O [31 ] (CaMg solution) was autoclaved, 200 mL was decanted into each of the two glass beakers, and platinum electrodes were immersed in the solutions: the anode in one beaker and the cathode in another. The circuit was completed with an agar-based potassium chloride salt bridge (30% KCl), and the solution was electrolysed at 60V potential difference using a Bio-Rad PowerPacTM Basic (Bio-Rad, USA) power pack for 1 h at room temperature, yielding anodic (oxidizing) water at pH 2.4 and cathodic (reducing) water at pH 11.2 [27 (link)]. The anodic water was discarded, and the cathodic water was used for invigoration of seeds within 1 h.
The priming solutions used were cathodic water, un-electrolysed CaMg solution and deionized water. Seeds (50 seeds per treatment) were hydrated by placing them between 20 layers of single-ply paper towel (Twinsaver Wiper Roll), which was placed on aluminium foil. To prime the seeds, 50 mL of the solutions was poured onto these paper towels. The aluminium foil containing the seeds was placed in plastic pouches, and after 24 h, just before radicle emergence, the seeds were dried back to their original masses under ambient laboratory conditions for 7 days and kept at 4 °C in air-tight bottles until required.

The proteins (40 μg / well) were subjected to 10–12% SDS polyacrylamide gel or 4–20% ExpressPlus PAGE Gel (Genscript, Nanjing, China) for electrophoresis and transferred onto PVDF membranes by PowerPac^TM Basic (Bio-Rad, Hercules, USA). The PVDF membranes were incubated in blocking buffer (5% skim milk in TBS-T buffer) at room temperature (RT) for 1 h and then incubated with the antibodies to C5a, C5aR, His-tag, p38 MAPK, p-p38 MAPK, ERK1/2, p-ERK1/2, JNK, p-JNK and β-actin at 4°C overnight. β-actin expression in each sample was identified as the internal standard. The phosphorylation levels of p38 MAPK, ERK1/2 and JNK were normalized to the total levels of p38 MAPK, ERK1/2 and JNK respectively. After 5 times of washing with TBST-T, the PVDF membranes were further incubated with IRDye 800CW-conjugated anti-mouse IgG, HRP-conjugated anti-rabbit IgG and HRP-conjugated anti-mouse IgG at RT for 1 h. The bands were visualized by Odyssey system (for the recombinant rat C5a identification) or regular X-ray film through ECL detection system after washing the PVDF membranes 5 times. Finally, the density of radiographic band onto PVDF membranes was analyzed by using the software of Quantity One (Bio-Rad).

Our plasma corona discharge device (3DT, MULTIDYNE 1000, Germantown, WI, USA) consisted of a treating head that contained two hook-shaped wire electrodes. The plasma was generated under high voltage at electrode 2 × 12 kV and a frequency of 50 Hz at atmospheric pressure conditions, using ambient air as a carrier gas. A rotating table (15 × 15 cm) made of steel covered by a thick layer of PVC was placed under the treating head. A power supply (PowerPac^tm basic, Bio-Rad, Hercules, CA, USA) was attached to the table, allowing the upper part of the table to rotate at a desired number of rounds per min. A Petri dish containing the soil sample (3–5 replicates) or the isolated bacteria eluted from the soil (3–5 replicates) was placed in the center of the rotating table. The distance between the sample and the treating head was adjusted to 2 cm. The corona discharge device scheme is illustrated in Figure 1.

The plasma corona discharge device (3DT, MULTIDYNE 1000, Germantown, WI, USA) (Figure 1) used in this study was comprised of a treating head containing two electrodes. To generate the plasma, the device was operated under a high voltage of 2 × 12 kV and a frequency of 50 Hz under atmospheric pressure conditions, using ambient air as a carrier gas. A rotating table (Figure 1) covered with a layer of PVC was positioned beneath the treating head. The upper part of the table was rotated at controlled rounds per min (rpm) using a power supply (PowerPac^tm Basic, Bio-Rad, Hercules, CA, USA). A plastic Petri dish containing the S. cerevisiae sample was placed in the center of the rotating table.

The plasma corona discharge device (3DT, Multidyne 1000, Germantown Wisconsin USA) consisted of two hook-shaped wire electrodes that served as a treating head. The physical conditions for generating plasma are high voltage at the electrode (2 × 12 kV and 50 Hz; output voltage: 24VDC maximum; and output current: 0.3 AMPS maximum) with ambient air as a carrier gas at atmospheric pressure (measurements of the voltage between the anode of the plasma device and the cathode of the sensor, and the UV intensity generated by the plasma corona device, are shown at Supplementary Materials Figures S1–S3). A rotating table (15 × 15 cm), made of steel coated by polyvinyl chloride (PVC), was adjusted under the treating head at a distance of two cm. The table was connected to a power supply (PowerPacTM Basic, Bio-Rad) to enable a rotation rate of 11 rpm. As described in our previous study, this is the optimal rotation rate for soil bacterial eradication [13 (link)]. The bacterial sample was spread in a Petri dish that was placed at the center of the rotating table (Figure 1).