• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.125.1-125
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• 2003

Pepper anthracnose is one of the major limiting factors in pepper production. Boring last over 10 years, Colletotrichum gloeosporioides has been known as the most prevalent species among five Colletotrichum spp. involved as anthracnose causing agents. Recently, however, the change of major species with pepper anthracnose has been proposed. Identification study was peformed on 12 test isolates collected from anthracnose disease symptoms on pepper during 2001-2002 and 25 reference isolates obtained from several other host plants. The identification of the isolates with morphological observation and IfS region sequence comparison resulted that 11 ones from 12 test isolates colleted from pepper anthracnose during 2001-2002 were identified as C. acutatum. PCR using species-specific primers designed from ITS region sequence suggested a rapid diagnosis method in identifying C. acutatum from C. gloeosporioides.

• The Plant Pathology Journal
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• v.34 no.6
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• pp.480-489
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• 2018

Bitter rot caused by Colletotrichum species is a common fruit rotting disease of apple and one of the economically important disease in worldwide. In 2015 and 2016, distinct symptoms of bitter rot disease were observed in apple orchards in five regions of South Korea. In the present study, infected apples from these regions were utilized to obtain eighteen isolates of Colletotrichum spp. These isolates were identified and characterized according to their morphological characteristics and nucleotide sequence data of internal transcribed spacer regions and glyceraldehyde-3-phosphate-dehydrogenase. Molecular analyses suggested that the isolates of Colletotrichum causing the bitter rot disease in South Korea belong to 4 species: C. siamense; C. fructicola; C. fioriniae and C. nymphaeae. C. siamense and C. fructicola belonged to Musae Clade of C. gloeosporioides complex species while C. fioriniae and C. nymphaeae belonged to the Clade 3 and Clade 2 of C. acutatum complex species, respectively. Additionally, we also found that the isolates of C. gloeosporioides species-complex were more aggressive than those in the C. acutatum species complex via pathogenicity tests. Taken together, our results suggest that accurate identification of Colletotrichum spp. within each species complex is required for management of bitter rot disease on apple fruit in South Korea.

• The Plant Pathology Journal
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• v.26 no.1
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• pp.49-56
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• 2010

Thirty-six isolates of Colletotrichum spp. were obtained from infected grapes in two different locations of Korea; 18 isolates from Cheonahn, where carbendazim (MBC) and the mixture of MBC and diethofencarb (NPC) had been applied to control grape ripe rot, and 18 isolates from Cheongju, where no fungicides had been used. Sequences analysis of the internal transcribed spacer (ITS) and the $\beta$-tubulin gene identified 34 of the 36 isolates as Colletotrichum gloeosporioides. The remaining two isolates from Cheongju were identified as C. acutatum. Of the 18 isolates from Cheonahn, 12 were resistant to both MBC and the mixture (MBC+NPC), and six were sensitive to them. All C. gloeosporioides isolates from Cheongju, but not the two C. acutatum isolates, were sensitive to these fungicides. Sequence analysis of the $\beta$-tubulin gene in all isolates revealed that C. gloeosporioides resistant to MBC and MBC+NPC had a tyrosine instead of phenylalanine at the amino acid position 200. The appearance of resistance to MBC and the mixture in C. gloeosporioides correlated with the history of fungicide application in Korea.

• The Plant Pathology Journal
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• v.31 no.2
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• pp.165-175
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• 2015

Anthracnose is a fungal disease caused by Colletotrichum species that is detrimental to numerous plant species. Anthracnose control with fungicides has both human health and environmental safety implications. Despite increasing public concerns, fungicide use will continue in the absence of viable alternatives. There have been relatively less efforts to search antagonistic bacteria from mudflats harboring microbial diversity. A total of 420 bacterial strains were isolated from mudflats near the western sea of South Korea. Five bacterial strains, LB01, LB14, HM03, HM17, and LB15, were characterized as having antifungal properties in the presence of C. acutatum and C. gloeosporioides. The three Bacillus atrophaeus strains, LB14, HM03, and HM17, produced large quantities of chitinase and protease enzymes, whereas the B. amyloliquefaciens strain LB01 produced protease and cellulase enzymes. Two important antagonistic traits, siderophore production and solubilization of insoluble phosphate, were observed in the three B. atrophaeus strains. Analyses of disease suppression revealed that LB14 was most effective for suppressing the incidence of anthracnose symptoms on pepper fruits. LB14 produced antagonistic compounds and suppressed conidial germination of C. acutatum and C. gloeosporioides. The results from the present study will provide a basis for developing a reliable alternative to fungicides for anthracnose control.

• The Plant Pathology Journal
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• v.27 no.2
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• pp.156-163
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• 2011

Antifungal activity of three Myxococcus spp., KYC 1126, 1136, and 2001, was tested in vitro against three phytopathogenic fungi (Botrytis cinerea, Colletotrichum acutatum, and Pyricularia grisea). Spore germination and mycelial growth of the three pathogenic fungi were completely inhibited by bioactive substances from a myxobacterium KYC 1126. In addition, the activity of KYC 1126 was fungicidal, but liquid culture filtrate of KYC 1126 did not affect protoplast reversion in C. acutatum. A bioassay of KYC 1126 filtrate against anthracnose in hot pepper was conducted in the greenhouse and field at 2009 and 2010. The incidence of anthracnose in control seedlings was 74%, but was reduced to 29% after KYC 1126 treatment. The control value with KYC 1126 was 60% while that with the fungicide dithianon was 42%. In the greenhouse, disease incidence with KYC 1126 was consistentely 10-35% lower than with fungicide as a positive control. The control value with KYC 1126 was 13.4% and 41.0%, whereas that with the fungicide was 52.3% and 63% in 2009 and 2010, respectively. Although anti-anthracnose activity of KYC 1126 was not maintained for long time in the field, the bacteriolytic myxobacterium KYC 1126 could be a prospective biocontrol agent.

• The Plant Pathology Journal
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• v.30 no.3
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• pp.279-287
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• 2014

KYC 3262 was selected as a biocontrol agent against anthracnose on hot pepper from 813 extracts of myxobacterial isolates. Dual culture with Colletotrichum acutatum and 813 myxobacterial extracts was conducted, and 19 extracts were selected that inhibited germination and mycelial growth of C. acutatum. All selections were Sorangium cellulosum, which are cellulolytic myxobacteria from soil. With the infection bioassay on detached fruits in airtight containers, KYC 3262, KYC 3512, KYC 3279, and KYC 3584 were selected. The listed four myxobacteria were cultured in CSG/1 liquid media, and harvested filtrates were sprayed on the infected fruits. KYC 3262 was selected from the studies of attached fruit in a greenhouse study. KYC 3262 filtrate was applied for 3 years (from 2011 to 2013) in a field study in Asan, Republic of Korea. Control values of the KYC 3262 in the field were 31%, 89%, and 82% in 2011, 2012, and 2013, whereas values of the fungicide spray treatment were 19%, 97%, and 91%, respectively. Yields (kg/20 plants) of the KYC 3262 were 2.66 kg and 18.6 kg in 2011 and 2013, respectively, and those of the fungicide treatment were 2.0 kg and 20.2 kg, in 2011 and 2013, respectively.

• Research in Plant Disease
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• v.16 no.2
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• pp.170-175
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• 2010

A monoterpenoid benzylideneacetone (BZA) is a bacterial metabolite isolated from culture broth of an entomopathogenic bacterium, Xenorhabdus nematophila K1. It was tested in this study the control efficacy of the metabolite against two major fungal diseases occurring in red-pepper plants. BZA exhibited significant antifungal activities against Phytophthora capsici and Colletotrichum acutatum. Under natural light conditions, the antifungal activity of BZA was maintained for more than sixty days. The antifungal activity of BZA was not lost even in soil because the incidence of Phytophthora blight against red-pepper plants was significantly reduced when the suspensions of P. capsici were poured to the rhizosphere soils mixed with BZA. Application of the BZA suspension spray to the fruit surface infected with C. acutatum significantly suppressed the disease occurrence of anthracnose on the red-pepper plants. These results suggest that BZA can be used to develop a promising agrochemical to control phytophthora blight and anthracnose of redpepper plants.

• Journal of Life Science
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• v.20 no.8
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• pp.1254-1260
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• 2010

Microorganisms near the plant rhizosphere usually inhabit the surface or the inside of the plant roots and have a direct effect on plant growth by secreting plant growth promoters or antagonistic materials which protect the root zone system from various pathogens. This study was carried out to identify and isolate the antagonistic materials after isolation of microorganisms showing high antagonistic activities, in hopes of contributing to the development of sustainable agriculture and the preservation of agricultural environments. A number of antagonistic bacteria were isolated from paddy soil. Among isolates, RRj 228 showed plant growth promotion and antagonistic activity. RRj 228 was identified as Pseudomonas sp. according to the results of physiological properties and genetic methods. On the basis of the results of anti-fungal spectrum against several pathogens by RRj 228, the antagonistic effect of the isolate against Botrytis cinerea, Pythium ultimum, Phytopthola capsici, and Rhizoctonia solani, especially against red-pepper anthracnose caused by Colletotrichum acutatum, was remarkable. The experiment evaluating the biological control effect by RRj 228 revealed that the $ED_{50}$ value by the RRj 228 culture against C. acutatum, R. solani and P. ultimum were 0.14 mg/ml, 0.16 mg/ml and 0.29 mg/ml, respectively. An antagonistic substance was isolated and purified by several chromatographies from the RRj 228 culture. The $^1H$ and $^{13}C$ assignment of the antagonistic substance was achieved from two-dimensional $^1H-^1H$ COSY, HMQC, and HMBC. Finally, the antagonistic substance was identified as Phenazine-1-carboxylic acid ($C_{13}H_8N_2O_2$, M.W.=224).

• Research in Plant Disease
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• v.24 no.3
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• pp.213-220
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• 2018

We classified the previously reported antagonistic strains of Sorangium cellulosum into 5 subspecies (A-E). Four strains were antagonistic to Botrytis cinerea (AB group) and two strains were antagonistic to Colletotrichum acutatum (AC group). According to the genetic and sequential analyses with standard genes, xynB1, bglA2, groEL1 for grouping, all strains of AB group were belonged to subspecies C and all strains of AC group were belonged to subspecies D. In addition, high pressure liquid chromatography with the culture filtrates confirmed the genetic results, because AB group had peaks with retention time at 20-22.5 minutes, whereas AC group had no peak. There was positive relationship ($R^2=0.9652$) between the control values of infecting B. cinerea on cherry tomatoes and the main peak areas of chromatograms among the four isolates of AB group. From the subspecies results of AB group, the main peak of KYC 3270 was expected to be epothilone D. However the retention times of the standard of commercial epothilone D and the main peak of KYC 3270 culture filtrate were different as 9.9 and 11.581 min., respectively. Finally, the antagonistic metabolite of AB group was inferred as 7-ketone epothilone D.

• Mycobiology
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• v.47 no.3
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• pp.329-334
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• 2019

Streptomyces blastmyceticus strain 12-6 was isolated from a forest soil sample of Cheonan area on the basis of strong antifungal activities against plant pathogenic fungi. Butanol extracts of the cultural filtrates were active against C. acutatum, C. coccodes, C. gloeosporioides, F. oxysporum, and T. roseum. Active fractions were prepared by thin layer chromatography using silica gel plate; 12-6-2 ($R_f$ 0.36), 12-6-3 ($R_f$ 0.44). Scanning electron microscopy showed that the active fractions caused a change in surface texture of fungal spores from smooth surface to wrinkled surface. The lethal effect on the spores of the active fractions varied from 56% to 100%. It was shown that the spores of C. acutatum were more sensitive to the antifungal fractions than the spores of F. oxysporum. Fluorescence staining using TOTO-1 indicated that the antifungal fractions could make the spores more sensitive to the fluorescence dye. Thus, it was suggested that antifungal agents prepared in this study exhibited the antifungal activity by damaging the plasma membrane of both fungal spores and hyphae. Identification of antifungal agents in the active fraction using GC-MS analysis revealed the presence of cyclo-(Leu-Pro) and 9-octadecenamide as major components that have already been known as antifungal substances.