• The Korean Journal of Mycology
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• v.38 no.1
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• pp.21-24
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• 2010

This study was carried out to select cell wall degrading enzymes for maximizing protoplast yield from Basidiomycetes. The protoplasts were released from spore suspension, mycelia cultured on cellophane membrane, and homogenized mycelia of Flammulina velutipes using commercial cell wall degrading enzymes. The highest yield of protoplasts was obtained from the homogenized mycelia treated with the enzyme combination of $Glucanex^R$ 200G and cellulase onozuka R-10. The protocol was also available for Pleurotus ostreatus, P. eryngii, and Hypsizygus marmoreus.

• The Plant Pathology Journal
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• v.26 no.4
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• pp.353-359
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• 2010

In order to control postharvest spoilage of satsuma mandarin fruits, rhizobacteria were isolated from soil samples. The Pantoea agglomerans strain 59-4 (Pa 59-4) which suppresses the decay of mandarin fruit by green and blue mold, was tested for the control efficacy and its mode of action was investigated. Pa 59-4 inhibited infection by green and blue mold on wounded mandarins, which were artificially inoculated with a spore suspension of Penicillium digitatum and P. italicum with control efficacies of 85-90% and 75-80%, respectively. The biocontrol efficacy was increased by raising the concentration of cells to between $10^8$ and $10^9\;cfu/ml$, and pretreatment with the antagonist prevented subsequent infection by green mold. The population of Pa 59-4 was increased more than 10 fold during the 24 hr incubation at $20^{\circ}C$, indicating that colonization of the wound site might prevent the infection by green mold. Despite poor antifungal activity, the Pa 59-4 isolate completely inhibited the germination and growth of P. digitatum spores at $1{\times}10^8\;cfu/ml$. We argue that the control efficacy was mediated by nutrient competition. Overall, the effective rhizobacterium, Pa 59-4, was shown to be a promising biocontrol agent for the postharvest spoilage of mandarin fruits by green and blue mold.

• Research in Plant Disease
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• v.28 no.4
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• pp.204-208
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• 2022

Hydrogen peroxide is an eco-friendly oxidizing agent, which has exhibited a broad spectrum of antimicrobial activity without adverse environmental impact. This study was conducted to investigate the antifungal effect of hydrogen peroxide treatment against Cylindrocarpon destructans, and consequently to evaluate its control efficacy against root rot disease of 2-year-old ginseng plants. Hydrogen peroxide treatment strongly inhibited the viability of C. destructans conidia in vitro. The hydrogen peroxide at a concentration of 300 mg/l significantly reduced disease infection of the ginseng root when treated to spore suspension (10⁷ conidia/ml). Spraying with 300 mg/l of hydrogen peroxide reduced the root rot disease of the ginseng sprouts by 15% compared to the untreated control at 14 days after the inoculation. However, 300 mg/l of hydrogen peroxide delayed the emergence of ginseng plants during sprouting under aeroponic conditions. Further works need to be done to provide an acceptable control efficacy of hydrogen peroxide against the disease and its good safety to ginseng plants.

• The Plant Pathology Journal
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• v.40 no.5
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• pp.512-524
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• 2024

Transcription factors (TFs) regulate gene expression by binding to DNA. The NAC gene family in plants consists of crucial TFs that influence plant development and stress responses. The whole genome of Capsicum annuum shows over 100 NAC genes (CaNAC). Functional characteristics of the most CaNAC TFs are unknown. In this study, we identified CaNAC4, a novel NAC TF in C. annuum. CaNAC4 expression increased after inoculation with the pathogens, Xanthomonas axonopodis pv. vesicatoria race 3 and X. axonopodis pv. glycines 8ra, and following treatment with the plant hormones, salicylic acid and abscisic acid. We investigated the functional characteristics of the CaNAC4 gene and its roles in salt tolerance and anti-pathogen defense in transgenic Nicotiana benthamiana. For salt stress analysis, the leaf discs of wild-type and CaNAC4-transgenic N. benthamiana plants were exposed to different concentrations of sodium chloride. Chlorophyll loss was more severe in salt stress-treated wild-type plants than in CaNAC4-transgenic plants. To analyze the role of CaNAC4 in anti-pathogen defense, a spore suspension of Botrytis cinerea was used to infect the leaves. The disease caused by B. cinerea gradually increased in severity, and the symptoms were clearer in the CaNAC4-transgenic lines. We also investigated hypersensitive response (HR) in CaNAC4-transgenic plants. The results showed a stronger HR in wild-type plants after infiltration with the apoptosis regulator, BAX. In conclusion, our results suggest that CaNAC4 may enhance salt tolerance and act as a negative regulator of biotic stress in plants.

• Research in Plant Disease
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• v.20 no.4
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• pp.235-244
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• 2014

Clubroot and Fusarium wilt of cole crops (Brassica oleracea L.) are destructive diseases which for many years has brought a decline in quality and large losses in yields all over the world. The breeding of resistant cultivars is an effective approach to reduce the use of chemical fungicides and minimize crop losses. This study was conducted to evaluate the resistance of 60 cabbage (B. oleracea var. capitata) and 6 broccoli (B. oleracea var. italica) lines provided by The RDA-Genebank Information Center to clubroot and Fusarium wilt. To investigate resistance to clubroot, seedlings of the genetic resources were inoculated with Plasmodiophora brassicae by drenching the roots with a mixed spore suspension (1 : 1) of two isolates. Of the tested genetic resources, four cabbage lines were moderately resistant and 'K166220' represented the highest resistance to P. brassicae. The others were susceptible to clubroot. On the other hand, to select resistant plants to Fusarium wilt, the genetic resources were inoculated with Fusarium oxysporum f. sp. conglutinans by dipping the roots in spore suspension of the fungus. Among them, 17 cabbage and 5 broccoli lines were resistant, 16 cabbage lines were moderately resistant, and the others were susceptible to Fusarium wilt. Especially, three cabbage ('IT227115', 'K161791', 'K173350') and two broccoli ('IT227100', 'IT227099') lines were highly resistant to the fungus. We suggest that the resistant genetic resources can be used as a basic material for resistant B. oleracea breeding system against clubroot and Fusarium wilt.

• Horticultural Science & Technology
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• v.35 no.2
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• pp.210-219
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• 2017

This study was conducted to establish an efficient screening method for radish (Raphanus sativus) cultivars that are resistant to black spot, which is caused by Alternaria brassicicola. Seven A. brassicicola isolates were selected and investigated for their ability to produce spores and pathogenicity. Of these isolates, A. brassicicola KACC 40036 and 43923 produced abundant spores in V-8 juice agar medium and showed pathogenicity and strong virulence on radish seedlings. We examined the resistance of 61 commercial cultivars of radish to A. brassicicola KACC40036, and found that there are no highly resistant radish cultivars; however, some cultivars, such as 'Geumbong' and 'Searom', showed weak resistance to A. brassicicola. For further study, we selected four radish cultivars that showed different disease responses to A. brassicicola KACC40036. According to the growth stage of the radish seedlings, inoculum concentration, and incubation temperature of radish, development of black spot on four cultivars has been investigated. The results showed that younger seedlings were more sensitive to A. brassicicola than older seedlings, and the disease severity depended on the concentration of the spore suspension. The disease severity of plants incubated in humidity chamber at $25^{\circ}C$ was greater than that of plants grown at $20^{\circ}C$ or $30^{\circ}C$. Taken together, we suggest the following method for screening for radish plants that are resistant to A. brassicicola: 1) inoculate 16-day-old radish seedlings with an A. brassicicola spore suspension ($2.0{\times}10^5spores{\cdot}mL^{-1}$) using the spray method, 2) incubate the inoculated plants in a humidity chamber at $25^{\circ}C$ for 24 h and then transfer the plants to a growth chamber at $25^{\circ}C$ with 80% relative humidity under a 12 h light/dark cycle, and 3) assess the disease severity of the plants two days after inoculation.

• The Korean Journal of Pesticide Science
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• v.14 no.2
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• pp.148-156
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• 2010

S59-4 isolate was evaluated as a potential biocontrol agent using in vivo wounded garlic bulb assay. When the spore suspension ($10^5$ spores/$m\ell$) of Penicillium hirsutum was co-inoculated with cell suspension of S59-4 isolate on wounded garlics, the isolate showed high suppressive effect to disease development. The isolate was identified as Pantoea agglomerans S59-4(Pa59-4) through Biolog system. Furthermore, soaking garlic bulbs in the suspension of Pa59-4 significantly reduced garlic decay caused by P. hirsutum. The optimal concentration of Pa59-4 for controlling garlic blue mold was $10^7\sim10^8$ cfu/$m\ell$. And suppressive effect of Pa59-4 on garlic storage decay reduced as inoculation concentration of Penicillium hirsutum increased. In addition in order to investigate population dynamics of Pa59-4 on application site of garlic cloves, two antibiotic markers, pimaricin and vancomycin were selected. Bacterial density of Pa59-4 on the wounded garlic cloves increased continuously both under room temperature condition and low temperature condition until 30days after application of Pa59-4, meanwhile that of Pa59-4 on intact garlic cloves increased until 15days after application of Pa59-4 and thereafter decreased continuously. Two culture media for mass-production of Pa59-4, LB medium and TSB medium, were selected. By-product of bio-fungicide formulated by mixing white carbon and bacterial suspension of Pa59-4 suppressed by 40 to 50% garlic blue mold. Above results suggest that Pa59-4 be a promising control agent against garlic blue mold.

• Research in Plant Disease
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• v.23 no.2
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• pp.159-167
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• 2017

This study was conducted to evaluate the control efficacy of the organic farming materials on cucumber scab caused by Cladosporium cucumerinum PT1 (KACC 48094). The antifungal activities in vitro as well as the suppressive effect of 43 organic farming materials on the spore germination and germ tube growth by inoculating spore suspension on cucumber seedlings in vivo were investigated. Thirteen organic farming materials inhibited the mycelial growth of C. cucumerinum and nine of these were microbial agents. In the screening using cucumber seedlings, six organic farming materials were very effective with control efficacy value of 90%. Among them, Bacillus amyloliquefaciens M27 provided suppressive effect on both mycelial growth and spore germination against cucumber scab. Finally, nine organic farming materials were selected to test the protective and curative effects, and all chosen organic farming materials significantly suppressed disease incidence when applied in the preventive action, in comparison with the curative action. Especially, Bordeaux mixture I and III gave excellent protective control efficacy with control values of 96.7% and 73.3%, respectively, whereas its curative control effect was significant low. Among these, only Thymus quinquecostatus+Sophora extract showed curative activity, although the control value was as low as 50%. This study suggests that cucumber scab can be controlled by some organic farming materials in the farmhouses under comparatively cold and wet condition and protective treatment is more important and efficient.

• Journal of Food Hygiene and Safety
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• v.36 no.5
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• pp.447-453
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• 2021

In this study, the effectiveness of physical control technology, a combined light sterilization (LED, UV) and hot water treatment in reducing Aspergillus ochraceus for food production environment was investigated. In brief, 1 mL aliquot of A. ochraceus spore suspension (10^7-8 spore/mL) was inoculated onto stainless steel chips, which was then dried at 37℃, and each was subjected to different physical treatment. Treatments were performed for 0.5, 1, 2, 5, 8, and 11 hours to reduce the strains using a light-emitting diode, but no significant difference was confirmed among the treatments. However, a significant reduction was observed on the chips treated with UV-C exposure and hot water immersion. After being treated solely with 360 kJ/m² of UV-C on stainless steel chip, the fungi were significantly reduced to 1.27 log CFU/cm². Concerning the hot water treatment, the initial inoculum amount of 6.49 log CFU/cm² was entirely killed by immersion in 83℃ water for 5 minutes. Maintaining a high temperature for 5 minutes at the site is difficult. Thus, considering economic feasibility and usability, we attempted to confirm the appropriate A. ochraceus reduction conditions by combining a relatively low temperature of 60℃ and UV rays. With the combined treatments, even in lukewarm water, A. ochraceus decreased significantly through the increases in the immersion time and the amount of UV-C irradiation, and the yield was below the detection limit. Based on these results, if work tools are immersed in 60℃ lukewarm water for 3 minutes and then placed in a UV sterilization device for more than 10 minutes, the possibility of A. ochraceus cross-contamination during work is expected to be reduced.

• Research in Plant Disease
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• v.22 no.2
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• pp.72-80
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• 2016

Gummy stem blight, caused by the fungus Didymella bryoniae, is major disease of watermelons worldwide. The objective of the present study was to establish an efficient screening system to identify watermelon resistant to D. bryoniae. An GSB3 isolate was prepared from a watermelon plant showing typical symptoms of gummy stem blight in Haman-gun and identified as D. bryoniae based on molecular analysis of internal transcribed spacer sequence. A simple mass-production technique of inoculum was developed based on spore production of D. bryoniae GSB3 under several incubation conditions and their virulence on watermelon plants. Resistance degrees of 22 commercial watermelon cultivars to the GSB3 isolate were evaluated. Among them, four watermelon cultivars showing different degree of resistance response were selected for further study. Development of disease on the cultivars according to various conditions including inoculum concentrations, incubation periods in dew chamber, and incubation temperatures was investigated. From the results, we suggest an efficient screening method for resistant watermelon cultivars to gummy stem blight. Seeds of watermelon cultivar are sown and grown in a greenhouse until plant stage of 2-fully expanded leaves. Seedlings are inoculated with D. bryoniae by spraying spore suspension of the fungus at a concentration of $5.0{\times}10^5spores/ml$. The infected plants are incubated in humidity chamber at $25^{\circ}C$ for 48 hours and then transferred to a growth chamber at $25^{\circ}C$ and 80% relative humidity with 12-hour light a day. Three to four days after inoculation, disease severity of the plant are measured using percentage of infected leaf area.