• The Plant Pathology Journal
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• v.25 no.4
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• pp.322-327
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• 2009

A total of 23 isolates of Botrytis cinerea causing the grey mold were collected from infected ginseng in several fields of Korea. The sensitivity to carbendazim and the mixture of carbendazim plus diethofencarb was determined through a mycelial inhibition test on PDA amended with or without fungicides. B. cinerea isolates were classified as 3 phenotypes, which were the first phenotype resistant to both of carbendazim and the mixture ($Car^RMix^R$), the second one resistant to carbendazim and sensitive to the mixture ($Car^RMix^S$), and the last one sensitive to both of them ($Car^RMix^S$). Carbendazim resistance correlated with a single mutation $\beta$-tubulin gene of B. cinerea amplified with primer pair tubkjhL and tubkjhR causing a change of glutamate to alanine at amino acid position 198. Furthermore, the substitution of valine for glutamate led the resistance to carbendazim and the mixture at the same position of amino acid. PCR-restriction fragment length polymorphism (PCR-RFLP) analysis using the restriction endonuclease, Tsp451 and BstUI allowed differentiation of the PCR fragment of $\beta$-tubulin gene of $Car^SMix^S$ isolates from that of $Car^RMix^R$ and $Car^RMix^S$ isolates. This method will aid in a fast detection of resistance of carbendazim and the mixture of carbendazim plus diethofencarb in B. cinerea in ginseng field.

• The Korean Journal of Pesticide Science
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• v.11 no.1
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• pp.18-26
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• 2007

Fungicidal screening was performed to control soybean black root rot caused by Calonectria ilicicola through in vitro and greenhouse assays. In in vitro assay, 25 fungicides were assessed by an agar dilution method and a 96-well microtiter plate method. While protective fungicides including dithianon, dichlofluanid, mancozeb, and captan showed a very low activity against the mycelial growth C. ilicicola SC03-15 in the agar dilution method, they displayed potent inhibitory activity against spore germination in a 96-well microtiter plate method with $EC_{50}$ values of 4.65, 0.61, 4.64, and $0.29{\mu}g\;mL^{-1}$, respectively. Ergosterol biosynthesis-inhibiting (EBI) fungicides showed different antifungal activity against mycelial growth and spore germination according to molecules. Difenconazole displayed higher antifungal activity against spore germination rather than mycelial growth, and prochloraz inhibited potently both mycelial growth and spore germination with EC50 values less than $1.8{\mu}g\;ml^{-1}$. In contrast, the other EBI fungicides inhibited more highly mycelial growth than spore germination. Carbendazim+diethofencarb and dazomet also inhibited both mycelial growth and spore germination of C. ilicicola SC03-15 at very low concentrations. In greenhouse assay, carbendazim+diethofencarb effectively controlled a soybean black root rot by drenching 2 days before or after inoculation. In addition, tebuconazole showed potent curative activity against soybean black root rot.

• The Plant Pathology Journal
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• v.26 no.1
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• pp.49-56
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• 2010

Thirty-six isolates of Colletotrichum spp. were obtained from infected grapes in two different locations of Korea; 18 isolates from Cheonahn, where carbendazim (MBC) and the mixture of MBC and diethofencarb (NPC) had been applied to control grape ripe rot, and 18 isolates from Cheongju, where no fungicides had been used. Sequences analysis of the internal transcribed spacer (ITS) and the $\beta$-tubulin gene identified 34 of the 36 isolates as Colletotrichum gloeosporioides. The remaining two isolates from Cheongju were identified as C. acutatum. Of the 18 isolates from Cheonahn, 12 were resistant to both MBC and the mixture (MBC+NPC), and six were sensitive to them. All C. gloeosporioides isolates from Cheongju, but not the two C. acutatum isolates, were sensitive to these fungicides. Sequence analysis of the $\beta$-tubulin gene in all isolates revealed that C. gloeosporioides resistant to MBC and MBC+NPC had a tyrosine instead of phenylalanine at the amino acid position 200. The appearance of resistance to MBC and the mixture in C. gloeosporioides correlated with the history of fungicide application in Korea.

• The Korean Journal of Pesticide Science
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• v.19 no.3
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• pp.335-344
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• 2015

It was conducted to identify all 47 isolates obtained from infected fruits of pepper and boxthorn, and to investigate the mycological characteristics and the response to fungicides. All of 11 isolates from pepper were identified as Colletotrichum acutatum included into A2 group. Among 36 isolates from boxthorn, 14 isolates were identified as C. gloeosporioides, and the others were done as C. acutatum, which were composed as A1 group with 15 isolates and A3 with 7 isolates. After incubating the isolates on PDA at $25^{\circ}C$ for 10 days, the colony color of C. acutatum was greyish white, while that of C. gloeosporioides was orange at center of colony and was gradually turned into an greyish white to the periphery. The rate of conidia showing ellongated ellipsoidal shape with round ends was over 95% in C. acutatum isolated from pepper. However, C. acutatum isolated from boxthorn produced ellongated ellipsoidal conidia with the rate of 75%, and the others were pointed at one or both ends. Regardless of species of Colletotrichum, all isolated used in this study was showed an optimal temperature at $25^{\circ}C$. $EC_{50}$ values of all isolates of Colletotrichum spp. to 2 fungicides as carbendazim and the mixture of carbendazim and diethofencarb was investigated by an agar dilution method. With C. acutatum isolates from pepper belonged to A2 group, the mean of $EC_{50}$ value to carbendazim and the mixture of carbendazim and diethofencarb was 0.68 and $3.16{\mu}g/ml$, respectively. In the case of C. acutatum isolates from boxthorn, which were divided into 2 groups as A1 and A3 group, that to carbendazim was 0.21 at A1 and $0.24{\mu}g/ml$ at A3, while that to the mixture was 1.52 and $3.35{\mu}g/ml$. Isolates of C. gloeosporioides showed the mean of $EC_{50}$ value was $0.12{\mu}g/ml$ to carbendazim and $0.92{\mu}g/ml$ to the mixture. The value of resistant factor was higher in the isolates of C. acutatum obtained in boxthorn than those from pepper.

• Research in Plant Disease
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• v.13 no.3
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• pp.170-176
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• 2007

Effects of fungicides on the mycelial growth of Botrytis cinerea isolated from ginseng leaves were investigated by an agar dilution method. By using a agar dilution method, it was investigated the effect of fungicides, procymidone, carbendazim and the mixture with both of carbendazim and diethofencarb, on the mycelial growth of Botrytis cinerea isolates, which were isolated from infected leaves of ginseng in 2005 and 2006. With MIC (minimum inhibiton concentration) of procymidone against B. cinerea, pathogens were divided into two groups. While one showed the low MIC between 0.8 and $4.0{\mu}g/ml$, the other showed higher MIC above $20{\mu}g/ml$. In terms of the inhibition ratio of mycelial growth at the indicated concentration of procymidone, isolates of B. cinerea were divided into three groups; the sensitive, the intermediate resistant, and the resistant group. Each group was differentiated by $EC_{50}$; the sensitive group showed below $2.0{\mu}g/ml$, the intermediate resistant group between 2.0 to $5.0{\mu}g/ml$, and resistant group above $5.0{\mu}g/ml$. Compared with the ratio of resistant isolates of B. cinerea in 2005, the ratio in 2006 increased from 19.3% to 27.5%. Furthermore, the average $EC_{50}$ value of them increased from $10.0{\mu}g/ml$ in 2005 to $237.3{\mu}g/ml$ in 2006. The ratio of isolates showing the multiple resistance between procymidone and carbendazim was 40.2%, whereas the ratio was 4.0% showing the multiple resistance in the mixture.