• The Plant Pathology Journal
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• v.39 no.6
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• pp.614-624
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• 2023

Botrytis cinerea is a major fungal plant pathogen that causes gray mold disease in strawberries, leading to a decrease in strawberry yield. While benzimidazole is widely used as a fungicide for controlling this disease, the increasing prevalence of resistant populations to this fungicide undermines its effectiveness. To investigate benzimidazole resistant B. cinerea in South Korea, 78 strains were isolated from strawberries grown in 78 different farms in 2022, and their EC₅₀ values for benzimidazole were examined. As a result, 64 strains exhibited resistance to benzimidazole, and experimental tests using detached strawberry leaves and the plants in a greenhouse confirmed the reduced efficacy of benzimidazole to control these strains. The benzimidazole resistant strains identified in this study possessed two types of mutations, E198A or E198V, in the TUB2 gene. To detect these mutations, TaqMan probes were designed, enabling rapid identification of benzimidazole resistant B. cinerea in strawberry and tomato farms. This study utilizes TaqMan real-time polymerase chain reaction analysis to swiftly identify benzimidazole resistant B. cinerea, thereby offering the possibility of effective disease management by identifying optimum locations and time of application.

• Korean Journal Plant Pathology
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• v.14 no.4
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• pp.367-370
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• 1998

Monilina fructicola, the brown rot fungus of stone fruits, was isolated from overwintering mummies and peduncles on peach trees from February to March, 1998. The resistant population of these isolates to benzimidazole (benomyl, carbendazim and thiophanate-methyl) and dicarboximide (iprodione, vinclozolin and procymidone) was examined. Among 417 isolates, the incidence of isolates resistant to benomyl, carbendazim, and thiophanate-methyl were 45 (10.8%), 47 (11.3%), and 46 (11.0%), respectively. Forty two (10.0%) isolates showed cross-resistance to benzimidazole fungicides. On the other hand, the resistant isolates against iprodione, vinclozolin and procymidone were 186 (44.6%), 1 (0.2%) and 150 (36.0%), respectively. Among the isolates, 116 (27.8%) showed cross-resistance to iprodione and procymidone. Moreover, 27 (6.5%) of 417 isolates showed double-resistance to both benzimidazole (benomyl) and dicarboximide (iprodione).

• The Plant Pathology Journal
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• v.22 no.4
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• pp.369-374
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• 2006

To characterize benzimidazole-resistant and -sensitive Monilinia fructicola populations, the fungal isolates were obtained from peach plants showing brown rot and bloosom blight. Benzimidazole-sensitive isolates did not grow on potato dextrose agar(PDA) amended with $\geq1.0{\mu}g$ active ingredient(a.i.)/ml of the fungicides. However, benzimidazole-resistant isolates grew on PDA regardless of the tested concentrations of fungicides. Benzimidazole-resistant isolates did not grow on diethofencarb-PDA, but sensitive isolates grew on the same PDA. In the nucleotide sequences of $\beta$-tubulin gene, only codon 198(GAG: glutamic acid), a target site for benzimidazole, was replaced with GCG(alanine) in all of the resistant isolates, and this substitution seems to play an important role in the development of resistance. Other interesting codons such as 165(GCT), 200(TTC), and 241(GCT) were not changed among the isolates. Benzimidazole-resistant and -sensitive isolates were clustered clearly in random amplified polymerphic DNA analyses and the results revealed that low levels of genetic diversity between benzimidazole-sensitive and -resistant isolates of M. fructicola in the investigated regions.

• Korean Journal Plant Pathology
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• v.14 no.6
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• pp.682-688
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• 1998

Fungicide resistance of 48 isolates of Botrytis cinerea collected from citrus in Cheju was investigated and genetic diversity was analyzed with random amplified polymorphic DNA(RAPD). High levels of resistance to benzimidazole fungicides benomyl and thiophanate-methyl and N-phenylcarbamate fungicide diethofencarb were observed. Negative cross resistance was clear between benzimidazole and N-phenylcarbamate fungicides, and multiple resistance to the fungicides was also observed. There was cross resistance among the dicarboximide fungicides procymidione, vinclozolin and iprodione as it was observed between the benzimidazole fungicides benomyl and thiophanate-methyl. The lowest levels of resistance were to the dicarboximide fungicides, but no sensitive isolate to polyoxin B was observed. The isolates showed genetically diverse RAPD profiles according to the geographic origin collected, but there was no significant correaltion between RAPD profiles of genomic DNA and the levels of fungicide resistance of the isolates. The isolates showed genetically diverse RAPD profiles, indicating that genetic differentiation had already occurred in the populations of B. cinerea distributed in Cheju.

• The Plant Pathology Journal
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• v.22 no.1
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• pp.1-6
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• 2006

The population shifts of Monilinia fructicola isolates which were resistant to the fungicide benzimidazoles were investigated in four regions of Korea from 1998 to 2000. The isolation frequency of benzimidazole-resistant isolates ranged from 18.8% to 29.6% in Chongdo and from $22.0\%$ to $26.8\%$ in Gyeongsan during the same period. However, the frequency of benzimidazoleresistant isolates was less than $4.0\%$ in Chochiwon and Youngduk during the same period. Benzimidazoleresistant isolates showed cross-resistance among benzimidazoles. On the other hand, none of the isolates showed cross-resistance to diethofencarb and carbendazim. Regardless of the year, the benzimidazole-resistant isolates of $EC_{50}$ higher than 500 $\mug%$ a.i./ml were isolated more frequently in mid and late season than in early season. In an orchard of Gyeongsan that had not been exposed to any fungicides for several years, the population of benzimidazole-resistant isolate had persisted without much fluctuation for three years. These results suggest that benzimidazole resistance of M. fructicola is becoming a problem in controlling brown rot and blossom blight of peach in regions like Chongdo and Gyeongsan.

• The Plant Pathology Journal
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• v.23 no.3
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• pp.187-192
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• 2007

The anthracnose disease on pepper fruits in Korea was caused by Colletotrichum acutatum as well as C. gloeosporioides. Since C. acutatum showed less sensitivity to benomyl, it was analyzed whether the less sensitivity was given by the same mechanism for the fungicide resistance of C. gloeosporioides. The isolates of C. acutatum were less sensitive to the three benzimidazole fungicides tested, benomyl, carbendazim, and thiophanate-methyl. However, the of C. acutatum isolates were different from the resistant isolates of C. gloeosporioides in their response to diethofencarb, one of N-phenyl-carbamates; the former was still less sensitive to diethofencarb than the latter. The differences in the resistance mechanisms in two species were conspicuous in sequence analysis of the tub2 genes. The genes from C. acutatum did not show any non-synonymous base substitutions at the regions known to be correlated with the benzimidazole-resistance. All of these data may indicate that the less sensitivity of C. acutatum to benomyl is based on different mechanism(s) from that of C. gloeosporioides.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.1
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• pp.7-11
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• 2007

The study was conducted on 162 adult male Haemonchus contortus of sheep collected from Avikanagar, Jaipur and Bikaner regions to diagnose the benzimidazole (BZ) resistance in H. contortus. The BZ resistance is primarily linked with the mutation in ${\beta}$-tubulin isotype 1 gene which substitute phenylalanine (Phe) into tyrosine (Tyr) at the 200 codon of the gene. An allele specific polymerase chain reaction (AS-PCR) technique was used for diagnosis of BZ resistance in H. contortus. In AS-PCR, one reverse primer (TGG 312) was used in two separate reactions with each of 2 forward primers (resistant TGG 331 and susceptible CAW 106 primer) that differed only at 3' nucleotide position. Therefore, the amplified products from resistant and susceptible parasites were produced 267 and 266 bp, respectively. A total of 162 parasites were genotyped, of which 130 parasites found homozygous resistant 'rr', 22 heterozygous 'rS' and 10 homozygous susceptible 'SS' type. The prevalence of 'rr' individuals was higher in Jaipur (98%) followed by Avikanagar (93%) and Bikaner (50%) regions. Overall, the prevalence of BZ resistant allele (r) was higher (87%) as compared to 13% of BZ susceptible allele (S).

• Korean Journal Plant Pathology
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• v.11 no.2
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• pp.146-150
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• 1995

1994년, 1995년에 주요 채소 작물에서 분리한 잿빛곰팡이병균(Botrytis cinerea)의 방제 살균제에 대한 저항성을 조사한 결과, 1994년 분리된 713균주 중 610균주(85.6%)가 benzimidazole계에 저항성(Ben\ulcorner)이었고, 249균주(34.9%)는 dicarboximide계 살균제에 저항성(Pro\ulcorner)이었으며, benzimidazole계 및 N-phenyl carbamate계 두약제 모두에 저항성인 균주는 분리되지 않았다. 1995년에는 현재까지 520균주를 분리하였는데, 그중 Ben\ulcorner이 317균주(61%)이며 Pro\ulcorner은 214균주(41.2%)이었고, Ben\ulcorner이며 NPC\ulcorner인 균주(Ben\ulcorner+NPC\ulcorner)는 15균주(2.9%)로 나타났다. 이러한 저항성 균주의 최소 억제 농도는 carbendazim 1,000 $\mu\textrm{g}$/ml 이상, diethofencarb 1,000 $\mu\textrm{g}$/ml 이상이었다. 또 Ben\ulcorner+NPC\ulcorner인 15균주 중 3균주는 dicarboximide계 살균제인 procymidone에도 저항성이었으며, 한 균주를 제외한 모든 Pro\ulcorner 균주는 Ben\ulcorner로 나타났다.

• The Korean Journal of Physiology and Pharmacology
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• v.26 no.5
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• pp.377-387
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• 2022

Benzimidazole anthelmintic agents have been recently repurposed to overcome cancers resistant to conventional therapies. To evaluate the anti-cancer effects of benzimidazole on resistant cells, various cell death pathways were investigated in 5-fluorouracil-resistant colorectal cancer cells. The viability of wild-type and 5-fluorouracil-resistant SNU-C5 colorectal cancer cells was assayed, followed by Western blotting. Flow cytometry assays for cell death and cell cycle was also performed to analyze the anti-cancer effects of benzimidazole. When compared with albendazole, fenbendazole showed higher susceptibility to 5-fluorouracil-resistant SNU-C5 cells and was used in subsequent experiments. Flow cytometry revealed that fenbendazole significantly induces apoptosis as well as cell cycle arrest at G2/M phase on both cells. When compared with wild-type SNU-C5 cells, 5-fluorouracil-resistant SNU-C5 cells showed reduced autophagy, increased ferroptosis and ferroptosis-augmented apoptosis, and less activation of caspase-8 and p53. These results suggest that fenbendazole may be a potential alternative treatment in 5-fluorouracil-resistant cancer cells, and the anticancer activity of fenbendazole does not require p53 in 5-fluorouracil-resistant SNU-C5 cells.

• The Plant Pathology Journal
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• v.38 no.6
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• pp.629-636
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• 2022

Colletotrichum species is known as the major causal pathogen of red pepper anthracnose in Korea and various groups of fungicides are registered for the management of the disease. However, the consistent use of fungicides has resulted in the development of resistance in many red pepper-growing areas of Korea. Effective management of the occurrence of fungicide resistance depends on constant monitoring and early detection. Thus, in this study, various methods such as agar dilution method (ADM), gene sequencing, allele-specific polymerase chain reaction (PCR), and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were applied for the detection of benzimidazole resistance among 24 isolates of Colletotrichum acutatum s. lat. and Colletotrichum gloeosporioides s. lat. The result of the ADM showed that C. gloeosporioides s. lat. was classified into sensitive and resistant isolates to benomyl while C. acutatum s. lat. was insensitive at ≥1 ㎍/ml of benomyl. The sequence analysis of the β-tubulin gene showed the presence of a single nucleotide mutation at the 198th amino acid position of five isolates (16CACY14, 16CAYY19, 15HN5, 15KJ1, and 16CAYY7) of C. gloeosporioides s. lat. Allele-specific PCR and PCR-RFLP were used to detect point mutation at 198th amino acid position and this was done within a day unlike ADM which usually takes more than one week and thus saving time and resources that are essential in the fungicide resistance management in the field. Therefore, the molecular techniques established in this study can warrant early detection of benzimidazole fungicide resistance for the adoption of management strategies that can prevent yield losses among farmers.