The antifungal activity of compounds 1 –4 isolated from A. alternata was tested against B. cinerea, as reported in Zdorovenko et al. (2021) [40 (link)], with some modifications. Compounds 1 –4 , and the positive control, the fungicide pentachloronitrobenzene (Sigma-Aldrich), were dissolved in 4% of DMSO at a final concentration of 5 × 10−3 M. The sensitivity of B. cinerea to these compounds was evaluated on PDA as inhibition of mycelial radial growth. In brief, mycelial plugs 4 mm diameter were cut from the margin of actively growing 5-day-old colonies and one plug was placed in the center of the Petri dish with the mycelia in contact with the medium. Then, the different compounds were applied separately on the top of each plug. The negative control was obtained by applying 20 μL of 4% of DMSO. The plates were incubated at 28 °C for 5 days. Inhibition of the fungal growth was observed as a decrease of growth compared to the negative control.
Pentachloronitrobenzene
Manufactured by Merck Group
Sourced in United States
Pentachloronitrobenzene is a chemical compound used in laboratory settings. It is a crystalline solid that is used as a precursor in the synthesis of other organic compounds. The core function of this product is to serve as a building block for further chemical reactions and analyses.
Automatically generated - may contain errors
Lab products found in correlation
Rifampicin, by Merck Group (2 mentions)
Potato dextrose agar (pda), by BD (1 mentions)
Ampicillin, by Merck Group (1 mentions)
Caco3, by Thermo Fisher Scientific (1 mentions)
V8 juice, by Merck Group (1 mentions)
Fluorescein diacetate, by Merck Group (1 mentions)
Toto 3 iodide, by Thermo Fisher Scientific (1 mentions)
Ckx53 inverted light microscope, by Olympus (1 mentions)
Pimaricin, by Merck Group (1 mentions)
Ampicillin, by GoldBio (1 mentions)
3 protocols using pentachloronitrobenzene
1
Antifungal Activity of Alternaria Compounds
2
Isolation and Culturing of Plant Pathogens
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Small sections of root lesions from the infected plant, surface sterilized in 70% ethanol, were cultured on potato dextrose agar (Becton, Dickinson and Company, Sparks, MD), V8 agar, and 10% V8-PARPH. For V8 medium, 1% CaCO 3 (98%; Acros Organics, Geel, Belgium) was added to V8 juice (Campbell Soup Company, Camden, NJ) and centrifuged for 10 min at 10,000 rpm. Then, 50 ml of buffered and clarified V8 juice was added to 450 ml of deionized water (10% V8), along with 8 g of agar (Sigma-Aldrich, St. Louis, MO), and autoclaved at 121°C at 15 psi for 15 min. Similarly, for V8-PARPH selective medium, 500 µl of a fungicide and antibiotic mixture containing pentachloronitrobenzene (99% [GC]; Sigma-Aldrich) (0.63 g/50 ml ethanol), ampicillin (Sigma-Aldrich) (1.25 g/50 ml ethanol), rifampicin (Sigma-Aldrich) (0.05 g/50 ml ethanol), pimaricin (2.5%) (MP Biomedicals, Santa Ana, CA), and hymexazol (Sigma-Aldrich) (250 mg/50 ml sterilized water) was added to the V8 agar medium after autoclaving (Ferguson and Jeffers 1999; (link)Jeffers and Martin 1986) (link). The cultured plates were incubated at room temperature (25°C). Appearance of the colonies, as well as vegetative and reproductive structures, was described after 7 days of incubation.
3
Isolation and Characterization of P. agathidicida
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P. agathidicida isolates NZFS 3770 and NZFS 3772 were obtained from the culture collection held at Scion (Rotorua, New Zealand). P. agathidicida 3770 was originally isolated from Coromandel, New Zealand, while 3772 was isolated near Auckland, New Zealand (Studholme et al. 2016) . Oxathiapiprolin (≥98% purity) was purchased from Carbosynth (Compton, Berskhire, UK). It was stored at 1 mg/ml in 100% dimethyl sulfoxide (DMSO) at -20 °C. To account for possible solvent effects, DMSO concentrations were standardized and DMSO-only controls were performed in all experiments. Pimaricin (2.5%, w/v, aqueous solution), rifampicin, pentachloronitrobenzene, b-sitosterol and fluorescein diacetate were from Sigma Chemical Co. (St. Louis, MO, USA). Ampicillin was from GoldBio (St. Louis, MO, USA). TOTO-3 iodide was from Invitrogen. Brightfield microscopy was performed using an Olympus CKX53 inverted light microscope with Olympus cellSens Standard software for image processing.
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