• The Plant Pathology Journal
• /
• v.24 no.4
• /
• pp.392-399
• /
• 2008

Detached chili pepper fruits were inoculated with the conidial suspension of Colletotrichum acutatum JC-24 simultaneously (simultaneous inoculation, SI) and at delayed time (delayed inoculation, DI) after wounding with (delayed wound inoculation, DWI) or without additional wounding (delayed non-wound inoculation, DNI) at the inoculation time. Disease severity was significantly lowered by DNI, compared to SI. By DNI, the disease reduction rates were proportional with the length of delayed time, and greater at the high temperature range (18, 23 and $28^{\circ}$) than at the low temperature ($13^{\circ}$) tested. DWI was also effective in reducing the disease severity especially at 18oC; however, its effectiveness was lower than for DNI. In light microscopy, parenchyma cells at the wounding sites were modified structurally, initially forming new cell walls crossing cytoplasm, enlarged with multiple periclinal cell divisions, and finally layered like wound periderms. In DWI, the above structural modifications occurred, showing the restriction of the fungal invasion by the cell walls in enlarged modified cells, while no definite cellular modifications were found with proliferation of fungal hyphae in SI. Sclerenchyma-like cells with thickened cell walls were proliferated around the wounding sites, which were partially dissolved by DWI, probably leading to some disease development. All of these results suggest that the decline of the anthracnose disease in pepper fruit by the delayed inoculations may be derived from the structural modifications related to the healing processes of the previous wound inflicted on the tissues.

• Journal of Ginseng Research
• /
• v.45 no.1
• /
• pp.156-162
• /
• 2021

Background: It is estimated that 20-30% of ginseng crops in Canada are lost to root rot each harvest. This disease is commonly caused by fungal infection with Ilyonectria, previously known as Cylindrocarpon. Previous reports have linked the virulence of fungal disease to the production of siderophores, a class of small-molecule iron chelators. However, these siderophores have not been identified in Ilyonectria. Methods: High-resolution LC-MS/MS was used to screen Ilyonectria and Cylindrocarpon strain extracts for secondary metabolite production. These strains were also tested for their ability to cause root rot in American ginseng and categorized as virulent or avirulent. The differences in detected metabolites between the virulent and avirulent strains were compared with a focus on siderophores. Results: For the first time, a siderophore N,N',N"-triacetylfusarinine C (TAFC) has been identified in Ilyonectria, and it appears to be linked to disease virulence. Siderophore production was suppressed as the concentration of iron increased, which is in agreement with previous reports. Conclusion: The identification of the siderophore produced by Ilyonectria gives us further insight into the root rot disease that heavily affects ginseng crop yields. This research identifies a molecular pathway previously unknown for ginseng root rot and could lead to new disease treatment options.

• Microbiology and Biotechnology Letters
• /
• v.45 no.4
• /
• pp.358-364
• /
• 2017

Asymmetric PCR preferentially amplifies one DNA strand for use in DNA hybridization studies. Linear-After-The-Exponential-PCR (LATE-PCR) is an advanced asymmetric PCR method which uses innovatively designed primers at different concentrations. This study aimed to optimise LATE-PCR parameters to produce single-stranded DNA of Candida spp. and Aspergillus spp. for detection via probe hybridisation. The internal transcribed spacer (ITS) region was used to design limiting primer and excess primer for LATE-PCR. Primer annealing and melting temperature, difference of melting temperature between limiting and excess primer and concentration of primers were optimized. In order to confirm the presence of single-stranded DNA, the LATE-PCR product was hybridised with digoxigenin labeled complementary oligonucleotide probe specific for each fungal genus and detected using anti-digoxigenin antibody by dot blotting. Important parameters that determine the production of single-stranded DNA in a LATE-PCR reaction are difference of melting temperature between the limiting and excess primer of at least $5^{\circ}C$ and primer concentration ratio of excess primer to limiting primer at 20:1. LATE-PCR products of Candida albicans, Candida parapsilosis, Candida tropicalis and Aspergillus terreus at up to 1:100 dilution and after 1 h hybridization time, successfully hybridised to respective oligonucleotide probes with no cross reactivity observed between each fungal genus probe and non-target products. For Aspergillus fumigatus, LATE-PCR products were detected at 1:10 dilution and after overnight hybridisation. These results indicate high detection sensitivity for single-stranded DNA produced by LATE-PCR. In conclusion, this advancement of PCR may be utilised to detect fungal pathogens which can aid the diagnosis of invasive fungal disease.

• Journal of Dairy Science and Biotechnology
• /
• v.30 no.2
• /
• pp.75-81
• /
• 2012

Molds cause severe cheese deterioration, even though some white and blue molds are used for the manufacture of Camembert and Blue cheese, respectively. The species of Geotrichum, Moniliella, Aspergillus, Penicillium, Mucor, Fusarium, Phoma, and Cladosporium are the main fungi that affect contamination during cheese ripening. Once deteriorated by fungal spoilage, cheese becomes toxic and inedible. Fungal deterioration of cheese decreases the nutritional value, flavor profiles, physicochemical and organoleptic properties, and increases toxicity and infectious disease. Fungal contamination during cheese ripening is highly damaging to cheese production in Korean farmstead milk processing companies. Therefore, these companies hesitate to develop natural and ripened cheese varieties. This article discusses the recent and ongoing developments in the removal techniques of fungal contamination during cheese ripening. There are 2 categories of antifungal agents: chemical and natural. Major chemical agents are preservatives (propionic acid, sodium propionate, and calcium propionate) and ethanol. Among the natural agents, grapefruit seed extract, phytoncide, essential oils, and garlic have been investigated as natural antifungal agents. Additionally, some studies have shown that antibiotics such as natamycin and Delvocid$^{(R)}$, have antifungal activities for cheese contaminated with fungi. Microbial resources such as probiotic lactic acid bacteria, Propionibacterium, lactic acid bacteria from Kimchi, and bacteriocin are well known as antifungal agents. In addition, ozonization treatment has been reported to inhibit the growth activity of cheese-contaminating fungi.

• The Plant Pathology Journal
• /
• v.15 no.3
• /
• pp.162-167
• /
• 1999

Integration of microbial antagonists with fungicides was tried to control the gray mold caused by Botrytis cinerea on pepper in greenhouse conditions and to reduce fungicide uses. All of the selected bacterial antagonists, Bacillus amyloliquefaciens BL3, Paenibacillus polymyxa BL4, and Pseudomonas putida Cha94, completely inhibited the conidial germination of B. cinerea until 30 days after treatment. However, bacterial colonization of pepper phylloplane was poor in BL4, while the other bacterial isolates and the fungal antagonist Trichoderma harzianum TM colonized well on the phylloplane, maintaining the population density of 104-105 cfu/g until 15 days after microbial treatments. Out of 13 kinds of selected fungicides used for gray mold diseases, polyoxin B and BKF 1995 showed the most discriminatory activity on the fungal growth between B. cinerea and TM. TM grew readily on the media containing those fungicides, while B. cinerea showed poor or no mycelial growth on them. The selected fungicides and antagonists alone reduced incidence of gray mold on pepper, showing disease indices of about 2.4 to 3.0, while its was increased up to 4.2 in the untreated control. Alternate treatments with the antagonists and 2-fold diluted fungicides inhibited the disease incidence as much as the antagonists or fungicides alone, and reduced the secondary inoculum more than the single treatments. This suggests that integration of antagonists and fungicides may be an efficient way to reduce fungicide sprays with reliable control efficacy of the disease. However, there was not much difference in the early and mid-term disease progress among the treatments and the untreated control, probably due to extremely favorable environmental conditions for the disease development in this experiment.

• The Korean Journal of Pesticide Science
• /
• v.12 no.2
• /
• pp.168-176
• /
• 2008

Recently damage of rice bakanae disease disseminated by infected seeds increased in paddy field in Korea. For controlling rice bakanae disease, the efficacy of 17 fungicides was assessed by 5 kinds of bioassay, spore germination test (SGT), mycelial growth test, detection test on Komada's medium (KDT), pouch test (PT) and greenhouse test (GT). Among ergosterol biosynthesis inhibiting fungicides, prochloraz showed a high controlling activity in all the assay systems while the others showed very low activity except for $500\;{\mu}g/ml$ of hexaconazole in GT and $500\;{\mu}g/ml$ of triflumizole in KDT. Although benomyl and the mixture of benomyl and thiram showed a good activity at 100 and $500\;{\mu}g/ml$ in SGT and PT, respectively, in GT they did a middle activity. Trifloxystrobin and kresoxim-methyl included in strobilurins showed a good activity even at $20\;{\mu}g/ml$ in KDT as well as a middle activity in SGT. Also a high activity not only at $10\;{\mu}g/ml$ in SGT but also at $100\;{\mu}g/ml$ KDT was detected in thiram. The activity of fludioxonil was confirmed in SGT, KDT and PT. Based on these results, it is very important to determine a bioassay system, because the fungicidal activity against rice bakanae disease was fluctuated depending on a assay systems as well as the mechanism of fungicide.

• The Plant Pathology Journal
• /
• v.25 no.1
• /
• pp.86-90
• /
• 2009

In November 2008, typical powdery mildew symptoms were observed on leaves of Arabidopsis thaliana ecotype Col-0 plants in a growth room under controlled laboratory conditions at Korea University, Seoul. The disease was characterized by the appearance of white powder-like fungal growth on the surface of infected leaves. As the disease progressed, infected leaves exhibited chlorotic or necrotic brown lesions, and leaf distortion and senescence. Conidiophores of the causal fungus were hyaline, unbranched, 3-4 celled, cylindrical, and $80-115{\times}6-9{\mu}m$ in size. Singly produced conidia (pseudoidium type) were hyaline, oblong to cylindrical or oval in shape, and $26-55{\times}15-20{\mu}m$ in size with a length/width ratio of average 3, angular/rectangular wrinkling of outer wall and no distinct fibrosin bodies. Appressoria on the hyphae were multi-lobed. These structures are typical of the powdery mildew Oidium subgenus Pseudoidium, anamorph of the genus Erysiphe. The measurements of the fungal structures coincided with those of Erysiphe cruciferarum. The phylogenetic analysis using ITS rDNA sequences revealed that the causal fungus Erysiphe sp. KUS-F23994 is identical to E. cruciferarum. The isolated fungus incited powdery mildew symptoms on the inoculated Arabidopsis leaves, which proved Koch's postulates. Taken all data together, we first report the occurrence of powdery mildew disease of A. thaliana caused by Erysiphe cruciferarum in Korea.

• Journal of Forest and Environmental Science
• /
• v.34 no.1
• /
• pp.61-65
• /
• 2018

To know if small changes in rearing water temperature and density affect the number of bacterial and fungal colonies in metamorphosed frogs, Dybowski's frog tadpoles were reared from Gosner 25-26 stages at either low ($1^{\circ}C$ low to ambient water temperature), ambient, or high ($1^{\circ}C$ high) water temperature (each 15 tadpoles in 20 L water) condition and at either low (10 tadpoles/20 L water), medium (20 tadpoles), or high (30 tadpoles) density condition. Immediately after metamorphosis, we sampled bacteria and fungi from skin, liver, and heart of six metamorphosed frogs, randomly selected for each treatment group. After separate incubation of bacteria and fungi on 3M Petrifilm plates, we counted the number of bacterial and fungal colonies appeared on the plates and compared the numbers among the temperature and density treatment groups. For temperature treatment, high-temperature group had fewer bacterial colonies, while low-temperature group had more fungal colonies than the other two groups. For density treatment, low-density group had fewer bacterial colonies than the other two groups, but the number of fungal colonies were not different among the groups. Our results suggest that small increased rearing water temperature and lowered rearing density could potentially reduce pathogens in farming frogs.

• Proceedings of the Botanical Society of Korea Conference
• /
• 1998.07a
• /
• pp.10-18
• /
• 1998

For improvement of plants in disease resistance, it is most important to elucidate the mechanism to perceive and respond to the signal molecules of invaders. A model system with pea and its pathogen, Mycosphaerella pinodes, showed that the fungal elicitor induced defense responses in all plant species tested but that the suppressor of the fungus blocked or delayed the expression of defense responses and induced accessibility only in the host plant. In the world, many researchers believe that the pathogens` signals are recognized only on the receptors in the plasma membranes. Though we found that the ATPase and polyphosphoinositide metabolism in isolated plasma membranes responded to these fungal signals, we failed to detect specific actions of the suppressor in vitro on these plasma membrane functions. Recently, we found that ATPase (NTPases) and superoxide generating system in isolated cell wall were regulated by these fungal signals even in vitro, especially, by the suppressor in a strictly species-specific manner and also that the cell wall alone prepared an original defense system. The effects of both fungal signals on the isolated cell wall functions in vitro coincide perfectly with those on defense responses in vivo. In this treatise, we discuss the key role of the cell wall, which is plant-specific and the most exterior organelle, in determining host-parasite specificity and molecular target for improvement of plants.

• The Plant Pathology Journal
• /
• v.35 no.1
• /
• pp.77-83
• /
• 2019

Chlorine dioxide ($ClO_2$) can be used as an alternative disinfectant for controlling fungal contamination during postharvest storage. In this study, we tested the in vitro and in vivo inhibitory effects of gaseous $ClO_2$ against Diaporthe batatas SP-d1, the causal agent of sweetpotato dry rot. In in vitro tests, spore suspensions of SP-d1 spread on acidified potato dextrose agar were treated with various $ClO_2$ concentrations (1-20 ppm) for 0-60 min. Fungal growth was significantly inhibited at 1 ppm of $ClO_2$ treatment for 30 min, and completely inhibited at 20 ppm. In in vivo tests, spore suspensions were drop-inoculated onto sweetpotato slices, followed by $ClO_2$ treatment with different concentrations and durations. Lesion diameters were not significantly different between the tested $ClO_2$ concentrations; however, lesion diameters significantly decreased upon increasing the exposure time. Similarly, fungal populations decreased at the tested $ClO_2$ concentrations over time. However, the sliced tissue itself hardened after 60-min $ClO_2$ treatments, especially at 20 ppm of $ClO_2$. When sweetpotato roots were dip-inoculated in spore suspensions for 10 min prior to treatment with 20 and 40 ppm of $ClO_2$ for 0-60 min, fungal populations decreased with increasing $ClO_2$ concentrations. Taken together, these results showed that gaseous $ClO_2$ could significantly inhibit D. batatas growth and dry rot development in sweetpotato. Overall, gaseous $ClO_2$ could be used to control this fungal disease during the postharvest storage of sweetpotato.