• Microbiology and Biotechnology Letters
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• v.33 no.1
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• pp.16-23
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• 2005

In order to develop a new microbial fungicide using endophytic bacteria for the control of anthracnoses occurring on various crops, a total of 260 bacterial strains were isolated from fresh tissues of 5 plant species. After they were cultured in broth medium, their antifungal activities were tested for in vivo antifungal activity against cucumber anthracnose caused by Colletotrichum orbiculare. As the results, liquid cultures of 28 strains showed potent antifungal activities more than $90\%$ against cucumber anthracnose. At 3-fold dilutions of liquid cultures, 18 strains inhibited the development of cucumber anthracnose of more than $70\%$. They were further tested for in vivo antifungal activity against red pepper anthracnose caused by C. coccodes and in vitro antifungal activity against C. acutatum, a fungal agent causing red pepper anthracnose. Among 18 strains, a bacterial strain EB215 isolated from cucumber roots displayed the most potent antifungal activity against Colletotrichum species. It was identified as Burkholderia cepacia based on its physiological and biochemical characteristics, Biolog test and 16S rDNA gene sequence. It also controlled effectively the development of rice blast (Magnaporthe grisea), rice sheath blight (Corticium sasaki), tomato gray mold (Botrytis cinerea), and tomato late blight (Phytophthora infestans). Studies on the characterization of antifungal substances produced by B. cepacia EB215 are in progress.

• Journal of Korea Foresty Energy
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• v.22 no.3
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• pp.49-56
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• 2003

As a part of agricultural utilization of charcoal and pyroligneous acid, the effect of wood carbonization products on the growth of red pepper and soil microorganisms was investigated. The treatment of charcoal and pyroligneous acid provided good growth conditions to microorganisms through neutralizing soil acidity and improving the physicochemical properties of soil. Therefore the density of useful microorganism in the soil has been increased. In the growth of red pepper, the length, diameter, and the fruit numbers of red pepper have been increased by treating with wood carbonization products. It was especially shown that yield has increased about 50% in the fruit number, by treating charcoal 1kg, 1000 time-diluted solution of pyroligneous acid and bacteria, compared with the control. It was estimated that increasing the length of seedling and the diameter of red pepper stem contributed to the resistance against the prerequisites of various environmental changes in open field. Therefore, the final yield would be increased. In the antagonism experiment of red pepper mold (Colletotrichum gloeosporioides), the mold became extinct in the 2- and 10-time diluted solution of pyroligneous acid, compared with the control. On the other hand, their growth speed was delayed in the 100- and 1000 time-diluted solution.

• Korean Journal of Environmental Biology
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• v.39 no.4
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• pp.479-485
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• 2021

A semi-selective agar medium was developed for detecting and enumerating Colletotrichum scovillei in chili (Capsicum annum) plant material. Potato-dextrose-agar(PDA) was used as the basic medium. The composition of the semi-selective agar medium was established after several attempts to favor the development of C. scovillei and inhibit the growth of other fungi and bacteria. The semi-selective agar medium contained PDA amended with pyribencarb and pydiflumetofen at 40 ㎍ mL^-1 each and streptomycin at 100㎍ mL^-1 for preventing bacterial growth. The pH was adjusted to 4.8 with 85% lactic acid. The inhibition of the mycelial growth of C. scovillei was significantly less than that of most other fungi including Fusarium species when grown on the semi-selective medium. C. scovillei was detected from naturally infected chili plants by plating fruit and stem tissue suspensions on the semi-selective medium, which was found to be reliable and quantifiable. This was the first report of a semi-selective agar medium to detect the presence of C. scovillei in naturally infected chili tissue.

• Research in Plant Disease
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• v.30 no.3
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• pp.219-228
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• 2024

Pepper anthracnose, caused by Colletotrichum spp., leads to a decrease in the quantity of pepper fruit production. Molecular diagnosis is crucial for rapid identification of pathogens and determination of fungicide resistance. However, the traditional process of isolating the pathogen, extracting genomic DNA, and analyzing the gene sequence is time-consuming, which delays rapid diagnosis. In this study, we introduced a method using conidia of Colletotrichum spp. instead of genomic DNA, eliminating the need for DNA extraction or special processing for diagnosis. To elucidate this method, sensitivity was assessed through polymerase chain reaction (PCR) and quantitative real-time PCR (qPCR) tests using internal transcribed spacer-based primer pairs. Both PCR and qPCR tests showed that detection is feasible with just one conidia, with over 1,000 conidia yielding results comparable to approximately 1 pg of genomic DNA. For amplifying the cytochrome b gene for quinone-outside inhibitor fungicide susceptibility testing, detection from a single conidium is achievable, but a stable PCR product is obtained by increasing the number of cycles to 35. Additionally, the addition of 10% grinding fresh chili pepper paste to V8-Juicea gar medium, which is known for inducing conidia rapidly from the isolates, resulted in 3.2 to 6.0 times more conidia compared to the commonly used potato dextrose agar medium, enhancing the potential for swift testing. Taken together, this study presents a direct utilization of pepper anthracnose conidia through PCR or qPCR, offering a valuable technique for amplifying target genes, such as the minimum conidial amount and barcode genes, for molecular identification of anthracnose disease in pepper through PCR and qPCR analysis.

• Korean Journal of Plant Resources
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• v.30 no.4
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• pp.343-351
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• 2017

Plant extracts were screened for antifungal activity against major plant pathogens, Botrytis sp., Collectotrichum sp., Alternaria sp. and Cylindrocarpon sp. using 96-well microdilution method. Among the 662 methanol extracts from 401 plant species, 36 extracts showed complete inhibition of spore germination against at least one of four pathogenic fungi. Extracts of Morus alba twig and Sophora flavescens root showed minimum inhibition concentration (MIC) at $1,250{\mu}g/ml$ against Botrytis sp.. Extracts of Chloranthus japonicus root showed MIC at $1,250{\mu}g/ml$ against Collectotrichum sp.. Extracts of Glycyrrhiza uralensis aerial part, Inula helenium root and Menispermum dauricum root showed MIC between 625 and $1,250{\mu}g/ml$ against Alternaria sp.. G. uralensis aerial part and I. helenium root showed MIC at $1,250{\mu}g/ml$ against Cylindrocarpon sp.. Specifically, the extracts of Agrimonia pilosa root, Angelica tenuissima root, Asarum sieboldii root, Campsis grandifolia leaf and twig, Cnidium officinale root, Dictamnus dasycarpus root, G. uralensis aerial part, I. helenium root and M. alba twig completely inhibited spore germination at lower than $5,000{\mu}g/ml$ against all of four pathogenic fungi. Two methanol extracts from G. uralensis aerial part and M. alba twig may used as a candidate to develop into effective disease management materials in plant cultivation.

• Research in Plant Disease
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• v.13 no.2
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• pp.93-97
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• 2007

The microorganisms with the antifungal activity against Phytophthora capsici and Colletotrichum acutatum and the plant growth promotion activity were screened from forest soils of Moon-gyeong (Juheul Mountain), Gyeongsangbuk-do. One of the isolates, strain MG 121 showed antifungal activity against P. capsici and C. acutatum and possessed phosphate solubilization activity was selected to development biocontrol agent. The strain MG 121 was identified as Streptomyces sp. by analysis of 16S rDNA. On the test with pepper fruits, the strain inhibited disease incidences of late blight and anthracnose over 80%. In greenhouse test, plant height, the number of leaf, fresh weight and roots length of pepper plants upon treatment of culture suspension of Streptomyces sp. MG 121 were significantly higher than those without the bacterial cells. In addition, strain MG 121 was capable to solublize rock-phosphate after incubation for 144 hours in potato dextrose broth. The concentration of soluble phosphate in PDB amended with 0.5% rock-phosphate was increased up to $765{\mu}g/ml$.

• Journal of Life Science
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• v.16 no.3 s.76
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• pp.420-426
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• 2006

Bacillus sp. KMU-991 was isolated from Oslo city soils at Norway and shown a strong antifungal activity on red-pepper anthracnose, Colletotrichum gloeosporioides. Bacillus sp. KMU-991 produced a maximum level of antifungal substrate under aerobic incubation at $30^{\circ}C$, 180 rpm for 48 hours in TSB medium(initial pH 7.0) containing 1.0% mannitol and 1.0% ammonium chloride. Precipitate of culture broth by $30{\sim}60%$ ammonium sulfate precipitation exhibited strong antifungal activity against C. gloeosporioides KACC 40804. Butanol extract of cultured broth also shown fungal growth inhibitory activity against Fusarium oxysporum f. sp. radicus-lycopersici KACC 40537, Rhizoctonia solani AG-4 KACC 40142, Botrytis cinerea KACC 40573, Colletotrichum orbiculare KACC 40808, and Phytophthora cambivora KACC 40160 by agar diffusion method.

• 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.

• The Korean Journal of Mycology
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• v.37 no.1
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• pp.96-103
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• 2009

For the control of pathogenic microorganisms, Bacillus spp. were isolated from diseased pepper fruits in Korea. Among them, Bacillus sp. IUB158-03 showed high inhibitory effect on mycelial growth and spore germination of C. gloeosporioides and Botrytis cinerea. The strain was identified as B. amyloliquefaciens IUB158-03 based on its physiological, biochemical characteristics and Microlog analysis. The highest level of antifungal substances by B. amyloliquefaciens IUB158-03 were obtained when the bacterium was cultured in medium containing 2% soluble starch, 3% yeast extract, 0.5% tryptone, 0.5% $NH_4H_2PO_4$, and 1% NaCl (pH 6.0) at $25^{\circ}C$ for 72 hrs. The antifungal substances were purified by butanol extraction, silica gel column chromatography, preparative thin layer chromatography, and high performance liquid chromatography. The purified antifungal substance was confirmed $R_f$ 0.27 by TLC. This substance exhibited antifungal activity against Fusarium solani, Rhizoctonia solani, Botrytis cineria, Alternata alternaria of plant pathogenic fungi and Trichophyton mentagrophytes, Epidermophyton floccosum, Cryptococcus neoformans of human pathogenic fungi.

• The Korean Journal of Pesticide Science
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• v.10 no.1
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• pp.56-65
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• 2006

An antagonistic bacterial isolate, that inhibits the growth of plant pathogens, was selected and identified from 5,000 isolates screened from the rhizosphere of various crop plants. An isolate Bacillus sp. N32, tested against Colletotrichum gloeosporioides causing anthracnose disease in hot pepper, produced both a heat resistant antifungal protein and a heat sensitive antifungal protein. The heat resistant protein was partially purified by Ammonium sulfate fractionation and gel filtration chromatography. The bioautography showed that the proteins possessed high antifungal activity. The biosynthetic gene cluster responsible for the heat resistant antifungal protein was cloned from cosmid library using DNA probe obtained from PCR product with the primers targeting the conserved nucleotide sequence of the synthetic genes reported earlier, Most of the clones obtained showed higher homology to fengycin antibiotic synthetic gene family reported earlier. On the other hand, the heat sensitive protein was isolated from SDS-PAGE and electroblotting to determine the N-terminal amino acid sequences. The heat sensitive antifungal protein gene was cloned from the ${\lambda}-ZAP$ libraries using a DNA probe based on the N-terminal amino acid sequences of the heat sensitive protein. We are contemplating to clone and sequence the whole gene cluster encoding the heat sensitive protein for further analysis.