• Research in Plant Disease
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• v.9 no.3
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• pp.174-178
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• 2003

Pseudomonas sp. antagonistic to Colletotrichum truncatum and C. gloesporioies was selected as a biological control agent for soybean anthracnose. Pseudomonas sp. inhibited the mycelial growth of pathogens effectively as the funhicides such as benomyl and fluazinam in vitro tests without any adverse effects on soybean. Seed treatment with Pseudomonas sp. increased emergence rate of soybean seeds significantly after inoculaton with C. truncatum. When the suspension of Pseudomonas sp. was sprayed on soybean plants, the control efficacy was not different from that of fungicides, benomyl and fluazinam two weeks after treatment, however the efficacy did not last long enough after three weeks.

• International Journal of Industrial Entomology and Biomaterials
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• v.11 no.1
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• pp.37-42
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• 2005

We cloned the $\beta$-tubulin genes from the wild type strain and two benomyl-resistant mutants of Metahizium anisopliae and determined their nucleotide sequences. A $\beta$-tubulin encoding 448-residue protein from wild type M. anisopliae shows strong homology to other $\beta$-tubulins. The coding region is interrupted by four introns. Comparisons of intron position between the M. anisopliae gene and other fungal $\beta$-tubulin genes show considerable positional conservation. The mutations responsible for benomyl resistance were determined in two spontaneous mutants, 8-18 and 8­19. One mutant 8-18 substituted glutamate for aspar­agine at position 33 and lysine for glutamine at position 134. The other mutant 8-19 showed alterations at three positions of $\beta$-tubulin arginine for tryptophan at position 21, lysine for asparagine at position 33, and phenylalanine for leucine at position 240. These data suggest that regions of $\beta$-tubulin containing amino acids 21, 33,134, and 240 interact to form the binding site of benomyl.

• Applied Biological Chemistry
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• v.41 no.3
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• pp.219-223
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• 1998

We have established a transformation system for entomopathogenic fungus, Metarhizium anisopliae, in order to develop mycoinsecticide by recombinant DNA techniques. Protoplasts of M. anisopliae would be transformed to a benomyl-resistant by introducing pBRG-4 plasmid DNA, which contains a ${\beta}-tubulin$ gene of Aspergillus flavus conferring resistance to benomyl and a pyr4 gene of Neurospora crassa, in the presence of 5% polyethylene glycol and 10 mM calcium chloride. Transformants occuring at a frequency of 10 colonies per $50\;{\mu}g$ pBRG-4 DNA grew on the $5\;{\mu}g/ml$ concentrations of benamyl, while the wild type was inhibited by $2.5\;{\mu}g/ml$. From the Southern analysis using genomic DNAs isolated from M. anisopliae transformants, the positive signals suggested that the ${\beta}-tubulin$ gene had integrated in the M. anisopliae genome by homologous recombination.

• Journal of Bio-Environment Control
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• v.12 no.4
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• pp.180-183
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• 2003

P. wasabiae was isolated from discolored seeds of wasabi(Wasabia japonical Mtsum.) and inoculated to fresh seeds, then the effect of fungicides on suppression of diseases were determined. Emergence rate of wasabi seeds where suppressed to 52.5% by the inoculation and it reached up to 92.7% by dipping treatment of inoculated seeds ito benomyl solution. The incidence rate of black leg disease and damping off were 32.0 and 22.0%, respectively, in control treatment that sown in the soil inoculated with P. wasabiae. But dipping treatment of inoculated seeds into benomyl solution resulted in 12.0% and 10.7% in incidence rate of those two diseases, respectively.

• The Korean Journal of Mycology
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• v.22 no.3
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• pp.260-265
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• 1994

Since around 1980 apple rot caused by Botryosphaeria dothidea has become prevalent throughout the growing areas in Korea, during which period chemical controls have been executed with no notable improvement. Results of investigations on resistance of the causal fungus to its control chemical are as followings; The susceptible fungal isolates showed no mycelial growth at $150\;{\mu}g/ml$ of benomyl whereas the resistant isolates showed 7-13 mm growth at $300\;{\mu}g/ml$ and 6-8 mm at $2,400\;{\mu}g/ml$ of this fungicide. At the latter high concentration, spore germination of the resistant isolates was 5-9% while that of the susceptible isolates was 0%. Within the range of $20-2,400\;{\mu}g/ml$ tested, the susceptible isolates were unable to form pycnidia, but the resistant isolates formed abundant pycnidia at the lower concentration with decreasing pycnidia along with the higher concentration.

• Proceedings of the Korea Society of Environmental Biology Conference
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• 2005.12a
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• pp.38-45
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• 2005

The egg bags of Korean salamander(Hynobius leechii) were collected from farmlands in Gyeongsangnam-do area. The assumed breeding time, numerical variation of embryos in each egg bag, mortality and the rates of abnormalities were investigated. The toxicity of benomyl, the metabolite carbendazim and BIC which were frequently spread in agricultural area and caused spontaneous embryonic malformation was investigated. The assumed breeding time between the end of February and the end of March has the difference about a month because of a habitat and it takes about 2 or 3 weeks from laying eggs to hatching. The length of each egg bag and the number of embryos were very varied in each area. It is due to geographical variation. Among egg bags in total study area, only 406 of egg bags(17.70% of total egg bags) developed all of embryos to normal larvae, and 78.49% of total embryos were normally developed. The patterns of spontaneous embryonic malformation were 26 species from A to Z and the abnormal patterns in individual were 8 species and above. the geographical differences about the abnormal pattern were identified and 11 habitats categorized 4 groups. The most frequent abnormality in Gyeongsangnam-do area is the dysplasia of external gill. The caudal dysplasia, abdominal blister and dysplasia of fin were also frequently observed. Individuals showing severe external defect were histologically studied and they showed retinal hypo-pigmentation, thyroid carcinoma, somatic muscular dysplasia, degeneration of cephalic neuron and various organ dysplasia. Benomyl and carbendazim were treated by 10pM$^{\sim}$10uM and BIC was treated by 1$^{\sim}$40ppm to know the effect of toxicity about toxic substance of salamander. After benomyl was treated, a survival rate was sharply dropped from 2 to 8 days. $LC_{100}$ identified in $1{\mu}M$, $LC_{50}$ identified between 100nM and $1{\mu}M$. $EC_{50}$ was assumed between 10nM and 100nM. The prevalent external malformation was abdomen swelled abnormally and histo-pathological effects were abdomen, neural tube and lens hernia. This suggests that benomyl is the toxicitic substance which inhibits the development of digestive system and nervous system. The result of treated carbendazim was similar to that of the treated benomyl. The survival rate is sharply dropped between 2 and 6 days. $LC_{100}$ was identified $1{\mu}M$ and $LC_{50}$ was identified between 10nM and 100nM. This shows that cabendazim has stronger lethal toxicity than benomyl. Ventral blister, eye dysplasia and cephalic dysplasia in the individual of external malformation mean that cabendazim affected nervous system much more than benomyl. Because the toxicity of BIC affected less in the beginning but affected more in the near hatching period, the period causing toxicity is somewhat different. $LC_{100}$ identified near 40ppm and $LC_{50}$ identified near 25ppm. The external defect shows mainly ventral blister and histo-pathological results show intestinal deformities. This result suggests the BIC inhibited strongly the development of digestive system. These abnormal developments may be caused by antimitotic action, inhibition of tubulin complex, destruction of microtubule, inhibitions of neurulation and closing of neural fold, and by the inhibition of movement of neural crest cells of benomyl. These abnormal developments may be caused by the rupture of epithelium, the loss of microtubule, the reduction of spindle size, the inhibition of spindle assembly formation, the destruction of spindle poles of carbendazim. These abnormal developments may be caused cytotoxicity by inhibition of the synthesis of a number of macromolecules and similar reaction the inhibition of benomyl.

• 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 Korean Journal of Mycology
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• v.19 no.1
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• pp.85-87
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• 1991

Prochloraz of fungicide was applied on Mycogone perniciosa causing wet bubble in cultivated mushroom, Agaricus bisporus. In vitro, Prochloraz was an excellent fungicide on two strains of Mycogone, tolerant and non-tolerant to Benomyl, respectively. At the low dosage, Prochloraz more inhibited mycelial growth of mushrooms than Benomyl. At the higher dosage, Benomyl more inhibited the mycelial growth than prochloraz. The higher yield of sporophore of the mushroom with low inferction rate was abtained from several trial of Prochloraz. Prochloroz was concluded to be effective fungicide on Mycogone perniciosa on Agaricus cultivation.

• Proceedings of the Korean Society of Toxicology Conference
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• 2002.11b
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• pp.165-165
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• 2002

These experiments tested whether exposure to benomyl (BN) or lindane (LD) during late pregnancy affect the development of reproductive tract in adult offspring. Timed pregnant Sprague Dawley rats (N=7∼8/treatment) were gavaged with LD (1, 5, or 10 mg/kg), BN (50, 250, or 500 mg/kg), or com oil on gestation days 15∼19.(omitted)

• The Korean Journal of Pesticide Science
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• v.7 no.3
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• pp.198-206
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• 2003

To assess potential risk of the benzimidazole fungicides, their cytotoxicities were evaluated. Activities of LDH(Lactic dehydrogenase) in the culture fluid of CHL(chinese hamster lung) fiberoblast cell treated with 4.0, 16.0 or $32.0{\mu}g/mL$ of carbendazim for 24 hours were elevated 2.16, 2.94 and 2.64 folds compared to the control, respectively. DNA synthesis was inhibited by 45% at $2.0{\mu}g/mL$ of carbendazim. Benzimidazole fungicides showed high toxicity to cell and mitochondria of CHL cell by Giemsa and MTT (3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide) assay. $IC_{50}$ by the Giemsa assay of thiophanate-methyl, benomyl, carbendazim and captafol were over 125, 1.2, 30.0 and $0.3{\mu}g/mL$, respectively. $IC_{50}$ by the MTT assay of thiophanate-methyl, benomyl, carbendazim and captafol were over 125, 18.7, 20.4 and $2.6{\mu}g/mL$, respectively. Inhibitory concentration of cell median proliferation by SRB (sulforhodamin B) assay for thiophanate-methyl, carbendazim, benomyl, and captafol were 17.4, 5.3, 1.5 and $0.5{\mu}g/mL$, respectively. Accordingly, benzimidazole fungicides inhibited DNA synthesis, mitochondrial function, cell proliferation and induced cell necrosis.