• 한국식물병리학회지
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• 제14권6호
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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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• 제22권4호
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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.

• 농약과학회지
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• 제7권3호
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• pp.198-206
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• 2003

Benzimidazole계 살균제의 세포독성을 평가하기 위하여 CHL(chinese hamster lung) 세포를 이용한 Giemsa stain assay, Lactic dehydrogenase(LDH) activity, MTT assay, DNA synthesis inhibition test 및 Sulforhodamin B(SRB) assay를 수행하여 benzimidazole 계 살균제에 대한 약제별 세포독성을 평가하고 위해도 평가를 위한 자료로 사용하고자 하였다. 시험결과 LDH 활성은 carbendazim 4.0, 16.0 및 $32.0{\mu}g/mL$을 24 시간 처리하였을 때 음성대조군에 비해 농도별로 세포독성이 각각 2.16, 2.94 및 2.64배 강하게 나타났다. DNA 합성시험에서는 carbendazim $2.0{\mu}g/mL$ 농도에서 약 45% DNA 합성을 저해하였으며, Giemsa assay와 MTT assay에서도 세포와 mitochondria에 독성을 일으키는 것으로 나타났다. Giemsa assay의 $IC_{50}$은 thiophanate-methyl, benomyl, carbendazim 및 양성대조 약제인 captafol에서 각각 >125, 1.2, 30.0 및 $0.3{\mu}g/mL$ 이었고, MTT assay의 $IC_{50}$은 thiophanate-methyl, benomyl, carbendazim 및 captafol에서 각각 >125, 18.7, 20.4 및 $2.6{\mu}g/mL$이었다. 그리고 세포 증식에 대한 영향을 조사하기 위하여 수행된 SRB assay 의 $IP_{50}$(반수세포증 식억제농도)에서도 thiophanate-methyl, carbendazim, benomyl 및 captafol이 각각 17.4, 5.3, 1.5, 및 $0.5{\mu}g/mL$으로 관찰되었다. 이상의 결과는 급성독성이 낮은 benzimidazole계 살균제가 유전독성 등 특수독성이 강하게 나타나는 원인일 것으로 판단되었다.

• Bulletin of the Korean Chemical Society
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• 제30권12호
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• pp.2930-2934
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• 2009

Surface-enhanced Raman Scattering (SERS) spectroscopy is applied to the study of the adsorption of benzoimidazolic fungicides benzimidazole (BIZ) and thiabendazole (TBZ) on silver mirrors. The influence of pH on the adsorption mechanism was investigated. In case of BIZ, two different adsorption mechanisms are deduced depending on the experimental conditions: via the $\pi$ electrons of the ring in neutral conditions and through an ionic pairing of protonated nitrogen atom with the chloride adsorbed on the metal surface. The SERS spectra of TBZ revealed that most molecules were adsorbed on silver surface by the ${\pi}$ electrons in neutral and acidic conditions but in acid conditions, some molecules were adsorbed via the sulfur and nitrogen atoms tilted slightly to the surface.

• 한국식물병리학회지
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• 제14권4호
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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).

• 한국식물병리학회지
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• 제14권6호
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• pp.693-699
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• 1998

Three three hundred and ninety seven isolates of Botrytis cinerea were isolated from infected plants of strawberry, tomato and cucumber from several areas in Korea during 1994∼1996 and the resistance of these isolates against some fungicides were examined. The isolation frequency of phenotypes resistant to carbendazim, procymidone, and diethofencarb were found to be 69.9 43.7, and 31.8%, respectively. The isolates were divided into six phenotypic groups; SSR, SRR, RSS, RRS, RSR and RRR, representing sensitive (S) or resistant (R) to benzimidazole, dicarboximide, and N-phenylcarbamate fungicides in order. The percentage of six phenotypes were 28.2, 2.0, 27.2, 41.0, 0.9 and 0.8%, respectively. On the basis of the mycelial growth inhibition (%) B. cinerea isolates were divided into three classes (class 1; 0∼50%, class 2; 51-99%, class 3; 100% inhibition) on carbendazim and three classes (class 1; 0∼75%, class 2; 76∼99%, class 3; 100% inhibition) on procymidone and the mixture of carbendazim+diethofencarb, respectively. Changes in sensitivity levles to carbendazim and carbendazim+diethofencarb were affected by introduction and increasing ratio of the use of diethofencarb.

• The Plant Pathology Journal
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• 제22권1호
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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.

• 한국식물병리학회지
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• 제11권2호
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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 Plant Pathology Journal
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• 제23권3호
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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.

• 농약과학회지
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• 제5권1호
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• pp.61-67
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• 2001

강원도 고랭지의 백합 재배 농가로부터 1998년부터 2000년에 걸쳐 516균주의 Botrytis elliptica를 분리하여 주요 방제 살균제에 대한 저항성 실태를 조사하였다. 분리한 균주들 가운데 benomyl, procymidone, diethofencarb에 대하여 저항성을 나타내는 균주의 출현율은 각각 90.1, 32.3, 40.9%로 높게 나타나 약제 저항성 문제가 심각한 것으로 나타났다. 분리된 균주의 benzimidazole계, dicarboximide계, N-phenylcarbamate계에 대한 감수성(S) 및 저항성(R) 반응은 6그룹(RSS, RRS, SSR, SRR, RSR, RRR)으로 나누어졌다. 6개 반응형 각각의 분리 빈도는 40.7, 8.5, 7.2, 2.7, 19.8, 21.1%로 나타났다. 이 중 carbendazim에만 저항성인 RSS형이 40.7%로 가장 놀게 나타났고, Procymidone과 diethofencarb에 저항성인 SRR형은 2.7%로 가장 낮게 나타났다. 이러한 약제 반응형은 월동기를 지나면서 상대적으로 증가하는 반응형과 감소하는 반응형이 있는데 이는 이들 균 자체 적응력 차이 때문인 것으로 보여진다. 적응력이 높은 약제 반응형은 RSS, SSR형으로 나타났으며, 적응력이 낮은 약제 반응형은 SRR, RSR, RRR형으로 나타났고 RRS형은 큰 변동이 없었다. Benzimidazole계 살균제에 저항성균의 방제를 위하여 1998년 봄부터 benzimidazole계와 N-phenylcarbamate계의 합제 사용으로 효과는 얻고 있으나, 이 두 계열의 약제에 대한 다중 저항성균이 점차 증가하고 있어 점차 약효가 감소하고있는 실정이다. 이 연구는 강원도 고랭지에서 백합 잎마름병균(Botrytis elliptica)의 방제 위하여 살균제 사용에 철저한 관리가 필요함을 시사한다.