The antagonistic activity of all Pseudomonas isolates against phytopathogenic Fusarium and Alternaria strains and the saprophytic Ulocladium spec. 219 (U219) was tested by dual culturing of the bacteria and fungi on Potato Dextrose Agar (ROTH, Karlsruhe, Germany) adjusted to pH 6.5. On each agar plate, four bacterial isolates pre-cultivated overnight in Standard I Nutrient Broth (MERCK, Darmstadt, Germany) were spread in straight lines of 30 mm and at a distance of 35 mm from the center of a 90 mm Petri dish. After 2 days of incubation at 25°C, an agar plug of 8 mm diameter with growing mycelium of one of the fungal indicator strains was placed in the center of the agar plate. The plates were incubated at 25°C until the growing fungal colony on a control plate without bacterial inoculation reached a radius of 35 mm. Criterion for antagonistic activity was a distance between the edge of the fungal colony and the bacterial smear (the inhibition zone) of at least 3 mm. Antagonistic activity was quantified as none (0–3 mm inhibition zone), low (3–5 mm), moderate (5–10 mm), and strong (>10 mm) (Supplementary Figure S1 ). The tests were repeated twice.
Potato dextrose agar (pda)
Manufactured by Carl Roth
Sourced in Germany
The PDA (Personal Digital Assistant) is a compact, handheld device designed for various laboratory tasks. It serves as a digital tool for data collection, analysis, and record-keeping. The core function of the PDA is to provide a portable and versatile platform for laboratory professionals to manage their work efficiently.
Automatically generated - may contain errors
Lab products found in correlation
Potato dextrose broth (pdb), by Carl Roth (3 mentions)
Standard 1 nutrient broth, by Merck Group (1 mentions)
Magnesium sulfate, by Merck Group (1 mentions)
α zel, by Merck Group (1 mentions)
Deuterated dimethyl sulfoxide, by Merck Group (1 mentions)
Ammonium acetate, by Mallinckrodt (1 mentions)
Tetracycline, by Carl Roth (1 mentions)
Lb medium, by Carl Roth (1 mentions)
Fast real time pcr system, by Thermo Fisher Scientific (1 mentions)
Malt extract, by Carl Roth (1 mentions)
Potato dextrose agar (pda), by BD (1 mentions)
Stereodiscovery v8 dissection microscope, by Zeiss (1 mentions)
20 protocols using potato dextrose agar (pda)
1
Antagonistic Activity of Pseudomonas Isolates
2
Characterization of Chitosan Batches
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In this study, a set of CS batches, representing various physicochemical parameters (viscosity, molecular weight, degree of deacetylation) and of different origins from different providers were used (Table 1 ). Acetic acid (99.5–99.9%) was supplied by POCH, Poland. Sodium hydroxide (NaOH) was supplied by POCH, (Warsaw, Poland), and Trypan blue was supplied by Pol-Aura, (Warsaw, Poland). Potato dextrose agar (PDA) and Potato dextrose broth (PDB) were supplied by ROTH, (Karlsruhe, Germany). Milli-Q water was used to prepare dilutions of the CS samples.
3
Quantification of Mycotoxins in Potato Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Potato dextrose agar (PDA) and potato dextrose broth (PDB) were purchased from Carl Roth GmbH + Co. KG (Karlsruhe, Germany). Rice flour was purchased from Biokorn GmbH + Co. KG (Aalen, Germany). ZEN was acquired from Tocris Bioscience (Bristol, England). A stock (1 mg mL−1) and working (5 µg mL−1) solution of ZEN was prepared as methanolic solution and stored at −20 °C. α-ZEL was purchased from Sigma-Aldrich Chemie GmbH (Steinheim, Germany). ZEN-14-S, ZEN-14-G und ZEN-16-G as reference standard were kindly provided by Prof. Franz Berthiller (University of Natural Resources and Life Sciences, Vienna, Austria). Acetonitrile and methanol were of HPLC-grade and obtained from Th. Geyer (Renningen, Germany). Ethyl acetate p. a. and sodium chloride p. a. were also purchased from Th. Geyer (Renningen, Germany). Magnesium sulfate was acquired from Sigma Aldrich (Steinheim, Germany). Ammonium acetate was purchased from Mallinckrodt Baker Inc. (Griesheim, Germany). Ultrapure water was obtained from a Seralpur PRO 90 CN purification system by Seral (Ransbach-Baumbach, Germany). Deuterated dimethyl sulfoxide (99.8 atom-% D) was acquired from Merck Switzerland. Trimesic acid trimethyl ester were purchased from OrganoSpezialChemie GmbH Bitterfeld. It’s purity has been traced back to that of NIST standard MRM 350b by the inhouse 1H qNMR method.
4
Fungal Inoculation of Grains
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The fungal inoculum was added 2 days after incubation of the grains inoculated with the bacteria. The strains F. culmorum 13 and F. graminearum 23 were pre-cultivated on Synthetic Nutrient Agar (Nirenberg, 1976 ). A. tenuissima 220 was grown on Potato Dextrose Agar (ROTH, Karlsruhe/Germany). After pre-incubation of the fungi for 7 days at 25°C in the dark, an incubation followed under mixed black light (near UV, emission 310–360 nm) and artificial daylight with a 12 h:12 h light:dark cycle for 5 days. Fungal spores were harvested by washing the culture surface with sterile 1/4 strength Ringer’s solution. A Thoma counting chamber of 0.1 mm depth (Poly-Optik GmbH, Bad Blankenburg, Germany) was used to adjust a spore density of 6 × 104 cells mL-1 in 1/4 strength Ringer’s solution. By adding 250 μL of this suspension, an inoculation titre of approximately 1 × 103 spores g-1 of grains (FM) was achieved. To prepare untreated controls, 250 μL of sterile 1/4 strength Ringer’s solution were added twice to the grains.
5
Isolation and Cultivation of Colletotrichum from Lupin Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Symptomatic (dried) lupin stem or pod tissue (Fig. 1 ) of 1–3 cm was surface sterilized (after rehydration in sterile ddH2O for dried samples) for 5 s with 0.25% sodium hypochlorite solution and rinsed thrice for 5 s in sterile ddH2O. Thin slices of 1 mm were cut and placed on PDA (potato dextrose agar, Carl Roth, Karlsruhe, Germany) amended with Tetracycline (0.02 g/l, Carl Roth) for 3 to 4 days at 22 °C in the dark. Single cultures were selected and grown on fresh PDA plates amended with Tetracycline for 4 to 6 days at 22 °C in the dark and suspected Colletotrichum species were sub-cultured. Single spore cultures were obtained and transferred to PDA and maintained at 22 °C in the dark as working cultures and stored at − 80 °C in 25% glycerol for long-term storage.
6
Cultivation and Preparation of Entomopathogenic Microbes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All of the parasites were cultivated from their respective glycerol stocks (50% glycerol, Carl-Roth) stored at −80 °C. B. thuringiensis was cultured strictly as per the protocol in Milutinovic et al. [35 (link)], allowing for the production of spores which in turn produce Cry toxins. P. entomophila was grown overnight in LB medium (Carl-Roth) in a 250 mL culture flask at 30 °C and under shaking conditions of 200 RPM. The overnight culture of P. entomophila was centrifuged at 3200 G-force to obtain bacterial pellets while the culture supernatant was discarded. The pellets thus obtained were suspended in Phosphate Saline Buffer (pH = 7) prior to use in survival assay. Non-evolved B. bassiana was plated on Potato-Dextrose agar (Carl-Roth) and stored at room temperature for 2–3 weeks prior to spore collection.
7
Fungal Cultivation and Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The fungi studied included the saprophyte Aspergillus niger AvT 14.203, the mycoparasite Gliocladium roseum DSM 62726 (syn. Clonostachys rosea; obtained from Deutsche Sammlung von Mikroorganismen und Zellkulturen, Braunschweig, Germany), and the phytopathogen Fusarium verticillioides FRC M-8114 (obtained from the Fusarium Research Center, PA, USA). The fungi were maintained on Potato dextrose agar plates. Potato dextrose agar was purchased from Carl Roth GmbH (Karlsruhe, Germany) and prepared according to the manufacturer’s instruction. For the experiments, the fungi were grown on agar plates containing GM7 medium which contains asparagin as nitrogen source [10 (link)], and spore suspensions were made in sterile-distilled water. For liquid cultures, GM7 medium was prepared in the same way except that glucose was autoclaved separately and no agar for solidification was used.
8
GLA and Beta-Carotene Production via Umbelopsis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The GLA and beta-carotene-producing strain Umbelopsis isabellina CCF2412 (Culture Collection of Fungi, Charles University, Prague, Czech Republic) was used for the preparation of prefermented cereals. The strain was kept on potato-dextrose agar (Carl Roth, Germany) at 4°C in dark conditions. Spore suspension was prepared by cultivation of the strain on rye for 7 d (Jeennor et al., 2008 (link)). The spores were then washed with distilled water with 0.05% Tween 80, filtered and adjusted to 2.105 spores/ml. A total of 100 g of wheat bran (substrate) in HDPE sacks (30 × 40 cm) were moistened with 75 ml of water and soaked for 2 h at laboratory temperature. Subsequently, substrates were autoclaved for 60 min (110 kPa, 110°C). After chilling, substrates were inoculated with 20 ml of spore suspension and cultivated for 7 d at 28±1°C with additional cultivating medium (D-glucose 15 g/l, yeast extract 5 g/l) containing mycelium on the 3rd day of cultivation. Mycelium was grown in mentioned cultivating medium for 3 d at 28±1°C on rotary shaker (165 rpm; Innova 40R, New Brunswick, Canada) under constant illumination with blue light (λmax = 440 nm). The final prefermented product contained 1.2 mg/g of GLA and 0.14 μg/g of beta-carotene.
9
Cultivation and Isolation of Fusarium Spp.
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Fusarium spp. isolates used in this study are listed in Table 1 . The isolates were provided either on synthetic nutrient-poor agar [52 (link)] or on potato-dextrose agar (PDA; Potato Dextrose Agar, Carl Roth GmbH, Karlsruhe, Germany) plates. Single-spore isolates were made of each isolate and cultivated on PDA in the dark at 25 °C for 10 days for the studies.
10
Isolation and Antimicrobial Profiling of Enterobacteria
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the isolation of enterobacteria, homogenized cell suspensions from the second sampling were plated on growth media. In total, 180 aerobic bacteria were isolated from the arugula phyllosphere and rhizosphere as well as bulk soil. Different media including PDA, MacConkey, KingsB, R2A (all obtained from Carl Roth GmbH, Karlsruhe, Germany), MIS (prepared according to [61 (link)]), and NAII (Sifin, Berlin, Germany) allowed a broad-spectrum coverage to create a representative strain library. None of the media used for the initial isolation was supplemented with antibiotics, in order to avoid an isolation bias caused by selective conditions. A disk diffusion antimicrobial susceptibility test method [35 ] was used to test for susceptibility towards a defined set of antibiotics. Instead of MH agar, NAII agar was used for the test plates. Unique morphotypes were tested for their sensitivity against Ampicillin (10 μg), Chloramphenicol (30 μg), Erythromycin (15 μg), Gentamicin (10 μg), Penicillin G (10 Units; 6 μg), Polymyxin B (300 Units; 30 μg), Streptomycin (10 μg), and Tetracycline (30 μg).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!