• The Plant Pathology Journal
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• v.40 no.4
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• pp.346-357
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• 2024

This study was carried out to screen the antifungal activity against Colletotrichum acutatum, Colletotrichum dematium, and Colletotrichum coccodes. Bacterial isolate GP-P8 from pepper soil was found to be effective against the tested pathogens with an average inhibition rate of 70.7% in in vitro dual culture assays. 16S rRNA gene sequencing analysis result showed that the effective bacterial isolate as Bacillus siamensis. Biochemical characterization of GP-P8 was also performed. According to the results, protease and cellulose, siderophore production, phosphate solubilization, starch hydrolysis, and indole-3-acetic acid production were shown by the GP-P8. Using specific primers, genes involved in the production of antibiotics, such as iturin, fengycin, difficidin, bacilysin, bacillibactin, surfactin, macrolactin, and bacillaene were also detected in B. siamensis GP-P8. Identification and analysis of volatile organic compounds through solid phase microextraction/gas chromatography-mass spectrometry (SPME/GC-MS) revealed that acetoin and 2,3-butanediol were produced by isolate GP-P8. In vivo tests showed that GP-P8 significantly reduced the anthracnose disease caused by C. acutatum, and enhanced the growth of pepper plant. Reverse transcription polymerase chain reaction analysis of pepper fruits revealed that GP-P8 treated pepper plants showed increased expression of immune genes such as CaPR1, CaPR4, CaNPR1, CaMAPK4, CaJA2, and CaERF53. These results strongly suggest that GP-P8 could be a promising biocontrol agent against pepper anthracnose disease and possibly a pepper plant growth-promoting agent.

• Korean Journal of Environmental Agriculture
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• v.37 no.1
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• pp.34-40
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• 2018

BACKGROUND: In order to select a fungicide that can effectively control anthracnose disease in Japanese plum fruit, mycelial growth inhibition effect and spore germination inhibition effect of six fungicides were tested in vitro against six isolates of Colletotrichum acutatum and five isolates of C. gloeosporioides that were isolated from diseased Japanese plum fruit. METHODS AND RESULTS: Inhibitory effects of fungicides on mycelial growth were investigated after inoculating each isolate on potato dextrose agar amended with four discriminatory concentrations of each fungicide for 7 days at $25^{\circ}C$. For spore germination inhibitory effect, each isolate of the Colletotrichum spp. was cultured in potato dextrose agar for 7-14 days at $25^{\circ}C$. After adjusting the concentration of spores of each isolate to $1{\times}10^6mL^{-1}$ by diluting with 0.025% PDB, the spore suspension was mixed with each fungicide (1:4, v/v), and $60{\mu}L$ aliquots were dispensed to sterile hole slide glass. Hole slide glasses were placed in a humidified box and incubated for 15 hours at $25^{\circ}C$. Then, spore germination was observed under an optical microscope. At recommended concentration of fungicide prochloraz manganese showed the highest mycelial growth inhibitory effect and dithianon showed the lowest mycelial growth inhibition. The $EC_{50}$ values for the inhibition of spore germination by dithianon and pyraclostrobin were $0.069-0.126{\mu}g/mL$ and $0.37-1.59{\mu}g/mL$, respectively. Although benomyl, prochloraz manganese, azoxystrobin, and tebuconazole did not inhibit the spore germination, they appeared to restrain mycelial growth by abnormal growth of germ tube and mycelium after germination. CONCLUSION: Dithianon seemed to have preventive effect. Prochloraz manganese, azoxystrobin, and tebuconazole were likely to have control effect. Pyraclostrobin is considered to have both preventive and control effect against anthracnose disease of Japanese plum fruit.

• The Plant Pathology Journal
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• v.26 no.2
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• pp.178-184
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• 2010

Four major diseases of chili pepper including two fungal diseases, anthracnose (Colletotrichum acutatum) and Phytophthora blight (Phytophthora capsici), and two bacterial diseases, bacterial wilt (Ralstonia solanacearum) and bacterial spot (Xanthomonas campestris pv. vesicatoria), were investigated under future climate-change condition treatments in growth chambers. Treatments with elevated $CO_2$ and temperature were maintained at $720ppm{\pm}20ppm$ $CO_2$ and $30^{\circ}C{\pm}0.5^{\circ}C$, whereas ambient conditions were maintained at $420ppm{\pm}20ppm$ $CO_2$ and $25^{\circ}C{\pm}0.5^{\circ}C$. Pepper seedlings or fruits were infected with each pathogen, and then the disease progress was evaluated in the growth chambers. According to paired t-test analyses, bacterial wilt and spot diseases significantly increased by 24% (p=0.008) and 25% (p=0.016), respectively, with elevated $CO_2$ and temperature conditions. On the other hand, neither Phytophthora blight (p=0.906) nor anthracnose (p=0.125) was statistically significant. The elevated $CO_2$ and temperature accelerated the progress of bacterial wilt by two days and bacterial spot by one day compared to the ambient treatment. Temperature regime studies of the diseases without changes in $CO_2$ confirmed that the accelerated bacterial disease progress was mainly due to the increased temperature rather than the elevated $CO_2$ conditions.

• The Plant Pathology Journal
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• v.37 no.3
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• pp.307-314
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• 2021

Pepper (Capsicum annuum L.) is an important agricultural crop worldwide. Recently, Colletotrichum scovillei, a member of the C. acutatum species complex, was reported to be the dominant pathogen causing pepper anthracnose disease in South Korea. In the present study, we isolated bacterial strains from rhizosphere soil in a pepper field in Gangwon Province, Korea, and assessed their antifungal ability against C. scovillei strain KC05. Among these strains, a strain named BS1 significantly inhibited mycelial growth, appressorium formation, and disease development of C. scovillei. By combined sequence analysis using 16S rRNA and partial gyrA sequences, strain BS1 was identified as Bacillus velezensis, a member of the B. subtilis species complex. BS1 produced hydrolytic enzymes (cellulase and protease) and iron-chelating siderophores. It also promoted chili pepper (cv. Nockwang) seedling growth compared with untreated plants. The study concluded that B. velezensis BS1 has good potential as a biocontrol agent of anthracnose disease in chili pepper caused by C. scovillei.

• The Korean Journal of Mycology
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• v.39 no.2
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• pp.91-98
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• 2011

Oudemansiella mucida, an edible and medicinal mushrooms belonging to Tricholomataceae of Basidiomycota, has been known to produce antifungal substances to inhibit the mycelial growth and spore germination of the plant pathogenic fungi. To produce good amount of antifungal substances from culture media, the optimal culture conditions of O. mucida were investigated. The most favorable conditions for the mycelial growth were $25^{\circ}C$ and pH 5 in potato dextrose agar. The most favorable carbon and nitrogen sources promoting mycelial growth were maltose and calcium nitrate, respectively. The optimum C/N ratio was about 20 : 1 in case that 3% glucose was supplemented to the basal medium as a carbon source. The optimal mycelial growth of O. mucida was found in the Hennerberg medium. The crude extract from submerged culture of potato dextrose broth exhibited inhibition of mycelial growth of Colletotrichum acutatum, Botrytis cinerea and Pyricularia oryzae but, fungicidal activity is not good enough to compared with commercially available fungicides tested. Therefore, the antifungal substances extracted from submerged culture of O. mucida might have a potential to be used for biocontrol agent of fungal diseases of plants.

• Horticultural Science & Technology
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• v.28 no.6
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• pp.1014-1024
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• 2010

Pepper ($Capsicum$ spp.) anthracnose caused by $Colletotrichum$ $acutatum$ is a destructive disease susceptible to areas where chili peppers are grown. $Capsicum$ $baccatum$ var. $pendulum$ (Cbp) is resistant to anthracnose and has actively been used for interspecific hybridization for the introgression of resistance gene(s) into cultivated chili peppers. The goals of this study were to determine the inheritance of resistance to anthracnose within $Capsicum$ $baccatum$ and to map quantitative trait loci (QTLs) for the anthracnose resistance. A genetic mapping population consisting of 126 $F_2$ plants derived from a cross between $Capsicum$ $baccatum$ var. $pendulum$ (resistant) and $Capsicum$ $baccatum$ 'Golden-aji' (susceptible) was used for linkage mapping. The linkage map was constructed with 52 SSRs, 175 AFLPs, and 100 SRAPs covering 1,896cM, with an average interval marker distance of 4.0cM. Based on this map, the number, location, and effect of QTLs for anthracnose resistance were studied using plants inoculated in the laboratory and field. A total of 19 quantitative trait loci (2 major QTLs and 16 minor QTLs) were detected. Two QTLs ($An8.1$, $An9.1$) showed 16.4% phenotypic variations for anthracnose resistance after wounding inoculation. In addition, five minor QTL loci ($An7.3$, $An7.4$, $An4.1$, $An3.1$, $An3.2$) showed a total of 60.73% phenotypic variations of anthracnose resistance in the field test. Several significant QTLs were also detected and their reproducibility was confirmed under different inoculation conditions. These QTLs are now being confirmed with different breeding populations. Markers tightly linked to the QTLs that are reliable under different environmental conditions will help to determine the success of marker-assisted selection for anthracnose -resistant breeding programs in chili pepper.

• Mycobiology
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• v.46 no.2
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• pp.92-100
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• 2018

The filamentous Ascomycota Colletotrichum gloeosporioides sensu lato is a fungus that has been reported worldwide as a causal agent of anthracnose disease in avocado and other crops. In Mexico, this species affects fruits from an early stage of development in the orchard until the post-harvest stage. Although fungicides are continuously applied to control Colletotrichum species, pericarp cankers and soft rot mesocarp in fruits are still frequently observed. Considering the lack of a precise description of the causative agent, the aim of the current study was to determine the pathogens involved in this symptomatology. Twenty-four isolates were consistently obtained from the pericarp of avocado fruits cv. "Hass" collected in the central avocado-producing area of Mexico. Morphological features such as colony growth, conidia size, and mycelial appressorium were assessed. Bayesian multilocus phylogenetic analyses were performed using amplified sequences of the internal transcribed spacer region of the nuclear ribosomal DNA; actin, chitin synthase, glyceraldehyde-3-phosphate dehydrogenase partial genes; and APn2-Mat1-2 intergenic spacer and mating type Mat1-2 partial gene from the nine selected isolates. In addition, fruits were inoculated with a conidial suspension and reproducible symptoms confirmed the presence of Colletotrichum fructicola in this area. This pathogenic species can now be added to those previously reported in the country, such as C. acutatum, C. boninense, C. godetiae, C. gloeosporioides, and C. karstii. Disease management programs to reduce the incidence of anthracnose should include C. fructicola to determine its response to fungicides that are routinely applied, considering that the appearance of new species is affecting the commercial quality of the fruits and shifting the original population structure.

• The Korean Journal of Pesticide Science
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• v.8 no.4
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• pp.338-346
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• 2004

The methanol extract of Chloranthus japonicus roots effectively controlled the development of rice blast(Magnaporthe grisea), rice sheath blight(Corticium sasaki), tomato pay mold(Btrytis cinerea), tomato late blight(Phytophthora infestans), and wheat leaf rust(Puccinia recondita). From the methanol extract of C. japonicus roots, three antifungal substances were isolated. Their chemical structures were determined to be shizukanols B, C, and D mainly by mass and NMR spectral data. Among the three substances, shizukanol C showed the strongest inhibitory activity against mycelial growth of the plant pathogenic fungi tested; it completely inhibited mycelial growth of M. grisea. Colletotrichum gloeosporioides, and C. acutatum at concentrations of more than $12.5{\mu}g$/ and P. infestans at concentrations of more than $3.13{\mu}g/m\ell$. They also controlled effectively the development of rice blast and wheat leaf rust. On the other hand, they caused phytotoxic symptoms on barley leaves and inhibited the growth of duckweed (Lemna paucicostata) with $EC_{50}$ values of $30.0{\mu}g/m\ell$ for shizukanol B, $49.9{\mu}g/m\ell$ for shizukanol C, and $154{\mu}g/m\ell$ for shizukanol D. In addition, shizukanol C showed an insecticidal activity against brown planthopper (Nilaparavata lugens), peen peach aphid (Myzus persicae), diamond-back moth (Plutella xylostella), and tobacco cutworm (Spodoptera litura) of the 5 arthropod pests tested with mortality values of more than 60% at a concentration of $1,000{\mu}g/m\ell$.

• The Korean Journal of Mycology
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• v.37 no.2
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• pp.155-160
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• 2009

The production of dihydroxynaphthalene (DHN) melanin is known to be essential factor for pathogenicity in Colletotrichum lagenarium. However, the genetic diversity of melanin genes was not much known among Colletotrichum spp. To investigate the variability of melanin gene in Colletotrichum spp. that cause anthracnose on diverse crops including tomato, we cloned and sequenced partial sd, one of DHN melanin genes encoding for scytalone dehydratase, from eight strains of C. coccodes, C. acutatum, C. truncatum C. caricae, and C. musae. The size of PCR-amplified sd ranged 437 bp to 545 bp. The nucleotide sequence identity of sd among the Colletotrichum strains tested varied from 49% to 99%. All of the PCR-amplified sd from eight strains contain an intron and have two exons coding for 122 amino acids. Overall, the size and nucleotide sequence of sd varied among the five Colletotrichum spp. Sequence identity of the predicted scytalone dehydratase protein of 122 amino acids ranged 50 to 99%. Phylogentic analysis based on the sd nucleotide sequences revealed that the five Colletotrichum spp. could be genetically divided.

• The Korean Journal of Pesticide Science
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• v.19 no.4
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• pp.428-439
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• 2015

Pepper anthracnose caused by Collectotrichum acutaum has been known as one of the most damaging diseases of pepper, which has reduced not only yield but also quality of pepper produce almost every year. This study was conducted to develop control strategy against pepper anthracnose by alternate application of agricultural organic materials and chemical fungicides. The alternate application effect of agricultural organic materials and chemical fungicides for controlling pepper anthracnose were examined in vitro and in the field. First, thirteen microbial agents and twenty two agricultural organic materials were screened for antifungal activity against C. acutatum through the dual culture method and bioassay. As a result, one microbial agent (Bacillus subtilis QST-713) and three agricultural organic materials (sulfur, bordeaux mixture, marine algae extracts) were found to show high inhibition effect against C. acutatum. In the field test, when Iminoctadine tris+thiram, a chemical fungicide for controlling pepper anthracnose, was sprayed, it reduced disease incidence by 89.5%. Meanwhile Sulfur, bordeaux mixture, copper, marine algae extracts and Bacillus subtilis QST-713 showed low disease incidence at the range of 33.1~81.0%. However, when Iminoctadine tris+thiram and agricultural organic materials(bordeaux mixture, marine algae extracts) were applied to pepper fruits alternately two times at 7 days interval, there was a 81.7 and 87.1% reduction in disease, respectively. Consequently, the alternate spray of chemical fungicide (Iminoctadine tris+thiram) and agricultural organic materials (bordeaux mixture, marine algae extracts) could be recommended as a control method to reduce the using amount of chemical fungicide.