• Research in Plant Disease
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• v.29 no.1
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• pp.11-22
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• 2023

Resazurin-based microtiter assay was used to evaluate the inhibitory effect of fungicides on the respiration of Colletotrichum acutatum s. lat. 20JDS8 sensitive and 20CDJ6 resistant to strobilurin fungicides. The spores of C. acutatum s. lat. 20JDS8 were inoculated into potato dextrose broth (PDB) at densities of 1x10⁴, 1x10⁵ and 1x10⁶ spores/ml, respectively. The relative fluorescence unit (RFU) of all treatments inoculated at each spore density started to rise after 12 hr of incubation, and were 1,965.5, 5,412.5, and 10,061.0, respectively, after 24 hr of incubation. To evaluate the inhibitory effect of fungicide on the respiration of the pathogen, the spores of the pathogen were inoculated into the PDB and treated with the fungicides 0, 6, 12, and 24 hr after incubation, respectively. After keeping the pathogen culturing for another 24 hr, PrestoBlue reagent was treated into the PDB culturing the pathogen. The RFU of each treatment was examined 1 hr after the reagent was treated. When dithianon, isopyrazam, pyraclostrobin, and fluazinam were treated at high concentrations in the stages of spores (immediately after inoculation [0 hr]), spore germination (after incubation for 6 hr), and hyphal growth (after incubation for 12 hr), the respiration of pathogens was inhibited by 90-100%. When the fungicides were treated after culturing the pathogen for 24 hr, the respiratory inhibitory effects were greatly reduced. With pyraclostrobin-resistant C. acutatum s. lat. 20CDJ6, azxoystrobin, trifloxystrobin and kresoxim-methyl, which have the same mode of action, had very little or no respiratory inhibitory effect in all growth stages of pathogens. Based on the above results, it was thought that the resazurin-based microtiter assay could quickly and accurately evaluate the inhibitory efficacy of the fungicides that inhibited respiration.

• Journal of Pharmacopuncture
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• v.15 no.4
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• pp.42-51
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• 2012

Objectives: This study used the basic principle of Oriental medicine, the sovereign, minister, assistant and courier principle (君臣佐使論) to investigate the effects of the component of ONGABO, which is composed of Ginseng Radix (Red Ginseng), Angelica Gigantis Radix, Schisandrae Fructus, Cuscuta Semen and Curcumae tuber on the viability of HepG2 cells. Methods: Single and mixed extracts of the component of ONGABO were prepared by lypohilizing powder of Red Ginseng (6-year root from Kanghwa), Angelica Gigantis Radix, Schisandrae Fructus, Cuscuta Semen, Curcumae Tuber (from Omniherb Co., Ltd., Korea) at the laboratory of herbal medicine in Woosuk University and were eluted after being macerated with 100% ethanol for three days. The cell viability of HepG2 was determined by using an absorptiometric analysis with PrestoBlue (Invitrogen) reagent after the plate had been incubated for 48 hours. All of the experiments were repeated three times to obtain the average value and standard deviation. The statistical analysis was done and the correlation factor was obtained by using Microsoft Office Excel 2007 and Origin 6.0 software. Results: Although Ginseng Radix (Red Ginseng) and Schisandrae Fructus did not enhance the viability of HepG2 cells, they were shown to provide protection of those cells. On the other hand, Angelica Gigantis Radix decreased the viability of HepG2 cells significantly, Cuscuta Semen and Curcumae Tuber had a small or no effect on the viability of HepG2 cells. Conclusions: In the sovereign, minister, assistant and courier principle (君臣佐使論), Ginseng Radix (Red Ginseng) corresponds to the sovereign component because it provides cell protection effects, Angelica Gigantis Radix corresponds to minister medicinal because it kills cells, Schisandrae Fructus corresponds to the assistant medicinal to help red ginseng having cell protect effects. Cuscuta Semen and Curcumae Tuber correspond to the courier medicinal having no effect in cell viability in HepG2. We hope this study provides motivation for advanced research on the sovereign, minister, assistant and courier principle.