• Research in Plant Disease
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• v.23 no.2
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• pp.168-176
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• 2017

Root-dipping inoculation method has been widely used to determine the resistance of watermelon to Fusarium oxysporum f. sp. niveum causing Fusarium wilt. Although this method leads to the precise results of plant disease responses, more rapid and efficient assay methods have been still required because the root-dipping inoculation method is labor-intensive and time-consuming. In this study, we established a simple and effective bioassay method based on the comparison of various inoculation methods and growth conditions. To develop the system, the occurrence of Fusarium wilt on four resistant and susceptible cultivars was investigated by four different inoculation methods, root-dipping, scalpel, tip and soil-drenching methods. Of these inoculation methods, scalpel method resulted in clear plant disease resistance responses with the simplicity. With the use of scalpel method, we also explored the disease development of the cultivars depending on inoculum concentration, growth stage of seedlings, and incubation temperature after inoculation. Furthermore, we found that the resistance degrees of 23 cultivars derived by scalpel inoculation method were similar to the results by root-dipping method established previously.

• Horticultural Science & Technology
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• v.31 no.1
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• pp.110-116
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• 2013

This study was carried out to establish the simple mass-screening methods for resistant tomato to Fusarium wilt caused by Fusarium oxysporum f. sp. lycopersici (FOL). Root dip inoculation method has been used in many studies on the resistance of tomato to disease. On the other hand, in mass-screening for resistant tomato to Fusarium wilt, the inoculation method is time-consuming and laborious procedure. Disease development of two FOL isolates on two cultivars of tomato according to inoculation method including root dip, tip and scalpel methods were investigated. In compatible interaction, tomato seedlings of each cultivar inoculated by tip method showed the lower and more variable disease severities than by root dip method. Whereas the seedlings by scalpel method represented clear resistant and susceptible responses to Fusarium wilt as root dip method. The resistance degree of each cultivar inoculated with FOL isolates by scalpel method was hardly affected by the tested incubation temperature and inoculum concentration. On the basis of the results, we suggest scalpel inoculation method as an efficient mass-screening method for resistant of tomato cultivars to Fusarium wilt. Roots of tomato seedlings at two-leaf stage grown in plastic cell tray were injured with scalpel and then spore suspension (more than $1{\times}10^7\;conidia{\cdot}mL^{-1}$) of FOL was poured directly on the roots. The infected plants were cultivated in a growth room at $25-30^{\circ}C$ for 4 weeks with 12-hours light a day.

• Research in Plant Disease
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• v.21 no.3
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• pp.201-207
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• 2015

This study was conducted to establish a simple mass-screening method for resistant melon to Fusarium wilt caused by Fusarium oxysporum f. sp. melonis (FOM). Root-dipping inoculation method has been used to investigate resistance of melon plants to Fusarium wilt. However, the inoculation method requires a lot of labor and time because of complicate procedure. To develop a simple screening method on melon Fusarium wilt, occurrence of Fusarium wilt on susceptible and resistant cultivars of melon according to inoculation method including root-dipping, soil-drenching, tip, and scalpel methods was investigated. Scalpel and tip methods showed more clear resistant and susceptible responses in the melon cultivars than root-dipping inoculation method, but tip method represented slightly variable disease severity. In contrast, in the case of soil-drenching inoculation method, disease severity of the susceptible cultivars was very low. Thus we selected scalpel method as inoculation method of a simple screening method for melon Fusarium wilt. By using the scalpel inoculation method, resistance degrees of the cultivars according to incubation temperature after inoculation (25 and $30^{\circ}C$) and inoculum concentration ($1{\times}10^6$ and $1{\times}10^7conidia/ml$) were measured. The resistance or susceptibility of the cultivars was hardly affected by all the tested conditions. To look into the effectiveness of scalpel inoculation methods, resistance of 22 commercial melon cultivars to FOM was compare with root-dipping inoculation method. When the melon cultivars were inoculated by scalpel method, resistance responses of all the tested cultivars were clearly distinguished as by root-dipping method. Taken together, we suggest that an efficient simple mass-screening method for resistant melon plant to Fusarium wilt is to sow the seeds of melon in a pot (70 ml of soil) and to grow the seedlings in a greenhouse ($25{\pm}5^{\circ}C$) for 7 days, to cut the root of seedlings with a scalpel and then pour a 10 ml-aliquot of the spore suspension of $1{\times}10^6conidia/ml$ on soil. The infected plants were cultivated in a growth room at 25 to $30^{\circ}C$ for about 3 weeks with 12-hr light a day.

• Research in Plant Disease
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• v.17 no.1
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• pp.13-18
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• 2011

This study was conducted to establish the efficient screening method for resistant cabbage to Fusarium wilt caused by Fusarium oxysporum f. sp. conglutinans. The resistance degrees of nine commercial cabbage cultivars to the disease were evaluated. Among them, five cultivars (YR-honam, Ogane, Greenhot, Redmat, and Ccoccoma) showing different resistance to the fungus were selected. Then development of Fusarium wilt of the cultivars according to several conditions including root wounding, dipping period of roots in spore suspension, inoculum concentration, and incubation temperature to develop the disease was investigated. Highly resistant cultivars such as 'YR-honam' and 'Ogane' hardly showed change of resistance to the disease by root wounding, dipping period, and inoculum concentration, while disease severity of Fusarium wilt on the cultivars was changed with incubation temperatures ($20^{\circ}C$, $25^{\circ}C$ and $30^{\circ}C$). When the cabbage cultivars were incubated at $25^{\circ}C$, they represented the most difference of resistance and susceptibility to Fusarium wilt. From above results, we suggest that an efficient screening method for resistant cabbage to F. oxysporum f. sp. conglutinans is to dip the non-cut roots of 14-day-old seedlings in spore suspension of $1{\times}10^7$ conidia/ml for 0.5 hr and to transplant the seedlings to plastic pots with a fertilized soil, and then to cultivate the plants in a growth chamber at $25^{\circ}C$ for 3 weeks to develop Fusarium wilt.

• Research in Plant Disease
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• v.18 no.2
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• pp.93-100
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• 2012

Resistance of pepper (Capsicum spp.) to anthracnose (Colletotrichum acutatum) was evaluated during regeneration of Capsicum spp. in National Agrobiodiversity Center. Disease severity of 896 pepper accessions (430 accessions of C. annuum, 219 accessions of C. baccatum, 14 accessions of C. chacoense, 153 accessions of C. chinense, 70 accessions of C. frutescens, 2 accessions of C. pubescens, and unidentified 8 accessions) was investigated at 14 days after inoculation in $28^{\circ}C$ humid chamber. Forty nine accessions of pepper germplasm were resistant to C. acutatum. Among them, nine accessions were highly resistant to C. acutatum without wounding spray inoculation. Four accessions belonged to the species C. baccatum, one accession to C. chacoense, and four accessions to C. frutescens. Forty two resistant candidate accessions were inoculated with pin-prick wounding using a syringe needle. Five accessions were resistant as a less than 3% of disease severity to C. acutatum with wounding inoculation 5 days after inoculation. All resistant accessions were C. baccatum. These five pepper germplasm might be used as breeding resources for the anthracnose resistance breeding program.

• Horticultural Science & Technology
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• v.29 no.1
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• pp.48-52
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• 2011

This study was conducted to establish the efficient screening system for resistant radish to Fusarium oxysporum f. sp. raphani. Five radish cultivars ('Myoungsan', 'Chungdu', 'Jangsaeng', 'Hannongyeorm', and 'Chungsukungjung') showing different degree of resistance to the fungus were selected. And the development of Fusarium wilt of the cultivars according to several conditions such as root wounding, dipping period of roots in spore suspension, inoculum concentration, and incubation temperature to develop the disease was tested. In distinguishing the resistance degree of the radish cultivars to the disease, non-cut roots were more effective than cut roots. And occurrence of Fusarium wilt of the radish plants increased in the proportion to increase of root-dipping period and spore concentration of the fungus. Thus, optimum conditions to differentiate susceptible and resistant cultivars to the disease were root-dipping period of 0.5 hour and spore concentration of $1{\times}10^7\;conidia{\cdot}mL^{-1}$. Disease severity of Fusarium wilt on the cultivars was changed with incubation temperature and the radish seedlings incubated at $25^{\circ}C$ represented the most difference of resistance and susceptibility to Fusarium wilt. From the above results, we suggest that the efficient screening method for resistant radish to Fusarium oxysporum f. sp. raphani would be to dip non-cut roots of fourteen-day-old radish seedlings in spore suspension of $1{\times}10^7\;conidia{\cdot}mL^{-1}$ for 0.5 hour and to transplant the inoculated plants to plastic pots with fertilized soil, and then to incubate the radish plants at a temperature of $25^{\circ}C$ for development of Fusarium wilt.

• Research in Plant Disease
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• v.23 no.4
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• pp.334-347
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• 2017

Bacterial wilt caused by Ralstonia solanacearum is an important disease in cultivation of chili pepper, causing plant death and significant yield losses. Cultivation of disease-resistant varieties is the most suitable measure to control bacterial wilt of chili pepper. To establish an efficient screening method for resistant chili pepper to R. solanacearum, six resistant or susceptible cultivars to the R. solanacearum were selected and the development of bacterial wilt on the cultivars according to several conditions was investigated. Drenching bacterial suspension into the cut roots using a scalpel was more simple and effective to distinguish resistant and susceptible cultivars than inoculation methods of root-dipping or soil-drenching without wounding. A resistant pepper, 'MC4' to R. solanacearum showed high resistance under the developed conditions which were 21- to 28-day-old pepper inoculated with $1{\times}10^8cfu/ml$ of bacterial suspension. On the other hands, the susceptible cultivars represented high disease severity under the conditions. These results indicated that we developed an efficient method to evaluate resistance of chili pepper cultivars against bacterial wilt. In addition, we successfully evaluated resistance degree of 140 commercial chili pepper cultivars to R. solanacearum using the developed method.

• Horticultural Science & Technology
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• v.33 no.1
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• pp.70-82
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• 2015

This study was conducted to establish an efficient screening system to identify melon resistant to Fusarium oxysporum f. sp. melonis. F. oyxsporum f. sp. melonis GR was isolated from infected melon plants collected at Goryeong and identified as F. oxysporum f. sp. melonis based on morphological characteristics, molecular analyses, and host-specificity tests on cucurbits including melon, oriental melon, cucumber, and watermelon. In addition, the GR isolate was determined as race 1 based on resistance responses of melon differentials to the fungus. To select optimized medium for mass production of inoculum of F. oxysporum f. sp. melonis GR, six media were tested. The fungus produced the most spores (microconidia) in V8-juice broth. Resistance degrees to the GR isolate of 22 commercial melon cultivars and 6 rootstocks for melon plants were investigated. All tested rootstocks showed no symptoms of Fusarium wilt. Among the tested melon cultivars, only three cultivars were susceptible and the other cultivars displayed moderate to high resistance to the GR isolate. For further study, six melon cultivars (Redqueen, Summercool, Superseji, Asiapapaya, Eolukpapaya, and Asiahwanggeum) showing different degrees of resistance to the fungus were selected. The development of Fusarium wilt on the cultivars was tested according to several conditions such as plant growth stage, root wounding, dipping period of roots in spore suspension, inoculum concentration, and incubation temperature to develop the disease. On the basis of the test results, we suggest that an efficient screening method for melon plants resistant to F. oxysporum f. sp. melonis is to remove soil from roots of seven-day-old melon seedlings, to dip the seedlings without cutting in s pore s uspension of $3{\times}10^5conidia/mL$ for 30 min, to transplant the inoculated seedlings to plastic pots with horticulture nursery media, and then to cultivate the plants in a growth room at 25 to $28^{\circ}C$ for about 3 weeks with 12-hour light per day.

• The Korean Journal of Mycology
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• v.23 no.2 s.73
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• pp.114-120
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• 1995

Colletotrichum dematium, C. gloeosporioides and Glomerella cingulata were detected in seed samples collected from diseased red pepper (Capsicum annuum) using blotter method. C. gloeosporioides was the predominant species in seed samples tested and followed by C. dematium and G. cingulata. When the seed components were plated C. dematium, C. gloeosporioides and G. cingulata were detected from seed coat, endosperm and cotyledon. The three anthracnose fungi were recorded more frequently from seed coat than that of observed in the endosperm and cotyledon. Seed infection with C. dematium, C. gloeosporioides and G. cingulata caused seed rotting, damping off and seedling blight of red pepper plants. According to the inoculation experiments, it was shown that C. gloeosporioides was the most virulent among three species. C. dematium showed weak virulence when the plants were wounded, and G. cingulata was wound parasite or weakly virulent on red fruits. Benlate T (benomyl+thiram) and Homai (thiophnate-methyl+thiram) were effective to anthracnose fungi when treated to infected seeds.

• Journal of Plant Biotechnology
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• v.44 no.4
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• pp.394-400
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• 2017

This study investigated whether culture filtrates produced by Xylella fastidiosa can be used to determine the varietal susceptibility to Pierce's disease in grapevines (Vitis spp.) as a substitute for pathogen inoculation or field screening. A bioassay of grape leaves with culture filtrates from the pathogen showed that their phytotoxicities were active and host-selective. Ethyl acetate extracts from them also showed toxicities and host selectivity in both bunches of grapes and muscadine grapes. The sensitive range of plants to the culture filtrates and their ethyl acetate extracts was consistent with the host range of the Pierce's disease pathogen. Susceptible cultivars are sensitive to even highly diluted culture filtrates, while resistant cultivars were not affected even at their original culture filtrates. Susceptible cultivars were more sensitive to the undiluted culture filtrate than were highly diluted culture filtrates, and the younger leaves were the most sensitive to the culture filtrates in grapes. Although some European grape cultivars showed moderately susceptibility in this study, the determination of varietal resistance to Pierce's disease by the treatment of culture filtrates of pathogens could provide valuable information for the preliminary selection of genetic resources and seedlings from hybridization in a disease resistant grape breeding program.