• Research in Plant Disease
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• v.24 no.4
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• pp.302-307
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• 2018

From 2014 to 2016, approximately 5% of okra fruit were observed displaying gray mold symptoms at the research field of Jeollabuk-do Agricultural Research and Extension Services, Korea. The symptoms observed were water-soaked, brown or gray spots, and abundant mycelial with conidia appearing on the infected fruit. Initial infection commenced from the base of fruit and gradually moved to the pod, where it finally resulted in collapse. Colonies on potato dextrose agar were gray to grayish brown, felted and cottony expanding 65-80 mm after one week. The fungus formed several black sclerotia ranging $1.0-3.5{\times}0.5-3.0mm$ on the Petri dish after two weeks. The conidia were one-celled, ellipsoidal or ovoid, colorless or pale brown, and $6.2-15.4{\times}5.0-10.4{\mu}m$. Conidiophores arose solitary or in groups, straight or flexuous, septate, with an inflated basal cell brown to light brown, and measured $85-450{\times}10.0-40.0{\mu}m$. On the basis of the morphological characteristics and phylogenetic analyses of internal transcribed spacer rDNA, the fungus was identified as Botrytis cinerea Pers. Pathogenicity of a representative isolate was proved by artificial inoculation, fulfilling Koch's postulates. To our knowledge, this is the first report on the occurrence of B. cinerea on okra in Korea.

• The Korean Journal of Mycology
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• v.47 no.1
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• pp.1-12
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• 2019

This study was conducted to investigate the optimum culture conditions, for mass production of Tremella fuciformis in M9 basic medium. The strain KMCC04674, used in this study was identified T. fuciformis by internal transcribed spacer (ITS). To define the optimum conditions for the mass production of T. fuciformis, we investigated the effects of different culture conditions and various nutrient sources on the fungal growth. The optimum initial pH and temperature for the fungal growth were 5.0 and $25^{\circ}C$, respectively. The optimal composition of the growth medium was 4.0% mannitol, 3.0% $NH_4H_2PO_4$, 1.0% malt extract, 1.0% glutamic acid, 5mM $CaCl_2$, and 0.5% glucanic acid.

• Journal of Microbiology and Biotechnology
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• v.32 no.3
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• pp.294-301
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• 2022

In our greenhouse experiment, soil heat treatment groups (50, 80, and 121℃) significantly promoted growth and disease suppression of Panax notoginseng in consecutively cultivated soil (CCS) samples (p < 0.01), and 80℃ worked better than 50℃ and 121℃ (p < 0.01). Furthermore, we found that heat treatment at 80℃ changes the microbial diversity in CCS, and the inhibition ratios of culturable microorganisms, such as fungi and actinomycetes, were nearly 100%. However, the heat-tolerant bacterial community was preserved. The 16S rRNA gene and internal transcribed spacer (ITS) sequencing analyses indicated that the soil heat treatment had a greater effect on the Chao1 index and Shannon's diversity index of bacteria than fungi, and the relative abundances of Firmicutes and Proteobacteria were significantly higher than without heating (80 and 121℃, p < 0.05). Soil probiotic bacteria, such as Bacillus (67%), Sporosarcina (9%), Paenibacillus (6%), Paenisporosarcina (6%), and Cohnella (4%), remained in the soil after the 80℃ and 121℃ heat treatments. Although steam increased the relative abundances of most of the heat-tolerant microbes before sowing, richness and diversity gradually recovered to the level of CCS, regardless of fungi or bacteria, after replanting. Thus, we added heat-tolerant microbes (such as Bacillus) after steaming, which reduced the relative abundance of pathogens, recruited antagonistic bacteria, and provided a long-term protective effect compared to the steaming and Bacillus alone (p < 0.05). Taken together, the current study provides novel insight into sustainable agriculture in a consecutively cultivated system.

• Journal of Environmental Health Sciences
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• v.48 no.5
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• pp.282-289
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• 2022

Background: Airborne fungi are ubiquitous in the air and exposure to an airborne fungus can be a significant risk factor. The composition of fungi has been potentially important for human health, especially for respiratory diseases like asthma and atopic dermatitis. Therefore, we attempted to ascertain what kind of airborne fungi affect human health at a nationwide level. Objectives: This study was carried out to provide information on indoor fungi distribution at multi-use facilities throughout South Korea. Methods: We classified our data by region and public facility after collection, cultivation, and identification via the sequencing of the ITS (internal transcribed spacer) region. We investigated whether or not the proliferation of HaCaT cells was affected by the identified airborne fungi. Results: In our data, the most isolated airborne fungi by region were Penicillium spp (Seoul, Daegu), Periconia sp (Gyeonggi-do), Iprex sp (Gangwon-do), Phanerochaete sp (Busan), Bjerkandera sp (Gwangju), and Aspergillus sp (Jeju-do). In the public facilities, the most detected fungi were Cladosporium sp (public transport), Penicillium sp (apartment house, retail market, financial institution, karaoke room), Bjerokandera sp (underground parking lot, public toilet, medical institution), Periconia sp (retail store), and Fusarium sp (general restaurant). Next, we selected twenty airborne fungi to examine their cytotoxicity and proliferation of human skin cells. In this experiment, the proliferation of the cells was influenced by most of the identified fungi. In case of the cytotoxicity test, most genera except for Rhodotorula sp and Moesziomyces sp showed cytotoxicity in HaCaT cells. Conclusions: The distribution of mold in the indoor air in multi-use facilities in South Korea differs from region to region, and this is an indicator that should be considered in future health impact studies. In addition, as a result of culturing about 20 types of bacteria dominant in indoor air, it was found that most (90%) inhibit the growth of skin cells, which can be harmful to health. An in-depth study of the health effects of floating fungi is needed.

• The Korean Journal of Mycology
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• v.24 no.2 s.77
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• pp.155-165
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• 1996

This study was carried out to identify the phylogenetic relationship among several isolates of Pleurotus species by comparing ITS II region of ribosomal DNA(rDNA) repeat unit. Two primers from ribosomal DNA sequences were chosen to amplify the specific internal transcribed spacer (ITS) II region of Pleurotus spp. The exact ITS II region with an unique band from six species of Pleurotus genus could be amplified using the two primers taken from at the 3'-end of 5.8S rDNA and 5'-end of 28S rDNA. Six representative species of the Pleurotus genus were easily characterized according to the length differences of ITS II region. Furthermore, within P. ostreatus species, different sizes of ITS II region could be observed in the isolates of ASI 2025 and ASI 2095 although they were classified as P. ostreatus by the conventional observation. The nucleotide sequence analyses of PCR-amplified ITS II region indicated that the isolates ASI 2025 and ASI 2095 were different from other Pleurotus spp. When the nucleotide sequences of six Pleurotus species were compared, three typical ITS II regions were highly variable especially at both ends of this region. The phylogenetic tree obtained by the Neighbor program of Felsenstein PHYLIP package with all the nucleotide sequence of Pleurotus spp. indicated that P. ostreatus, P. florida, P. sajor-caju and P. eryngii were closely related to one phylogenetic branch and P. cystidious was related to other branch with P. cornucopiae. The isolates ASI 2025 and 2038, however, were not closely related to any other Pleurotus spp. and formed their own individual branches.

• Horticultural Science & Technology
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• v.34 no.2
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• pp.305-313
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• 2016

Brassica juncea (2n = 4x = 36, AABB genome, 1,068 Mb) is a U's triangle species and an amphidiploid derivative of B. rapa and B. nigra. Fifteen varieties were used to study the ITS (internal transcribed spacer) regions of ribosomal DNA and MITEs (miniature inverted-repeat transposable elements) with a view of developing specific molecular markers. ITSs and MITEs are an excellent resource for developing DNA markers for genomics and evolutionary studies because most of them are stably inherited and present in high copy numbers. The ITS (ITS1 and ITS2) sequence was compared with the consensus sequence of B. rapa and B. nigra. Variation in ITS1 created two separate groups among 15 varieties, with 10 varieties in one group and 5 in the other. Phylogenetic analysis revealed two major clusters for those 10 and 5 varieties. Among the 160 different MITE primers used to evaluate the selected 15 varieties of B. juncea, 70 were related to the Stowaway, 79 to the Tourist, 6 to the hAT, and 5 to the Mutator super-families of MITEs. Of 160 markers examined, 32 were found to be polymorphic when fifteen different varieties of B. juncea were evaluated. The variety 'Blackgat' was different from the other mustard varieties with respect to both phenotype and genotype. The diversity of 47 additional accessions could be verified using eight selected molecular markers derived from MITE family sequences. The polymorphic markers identified in this study can be used for varietal classification, variety protection, and other breeding purposes.

• Korean Journal of Breeding Science
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• v.41 no.4
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• pp.420-428
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• 2009

In this study, the variation of 20 Pinus thunbergii isolates of F. circinatum from 2 damaged sites in Jeju-Island were compared with a known Fusarium circinatum using molecular biological techniques. Two- and four-year-old seedlings of Pinus rigida${\times}$Pinus x rigitaeda and two-, three- and six-year-old seedlings of P. thunbergii were inoculated with one of the most virulent isolates, FT-7, to determine differences in susceptibility. In site 1 (FT), 13 isolates of F. circinatum were isolated from 14 individuals and in site 2 (FS), 7 isolates of F. circinatum were isolated from 9 individuals. No difference was found in the sequences of the internal transcribed spacer (ITS) region of the ribosomal RNA genes in the FS and FT isolates, and also even in the known isolate of F. circinatum, FE 1-1. However, the ITS sequences of the FS and FT isolates differed from those of a fungus, Botrytis cinerea. Two-year-old seedlings of P. rigida${\times}$P. x rigitaeda showed higher susceptibility (93.3% of mortality) than four-year-old ones. Three-year-old seedlings of P. thunbergii showed the highest susceptibility (66.7% of mortality) compared to those at other ages in the same species. We found a positive correlation between basal diameter and lesion length in the seedlings of P. rigida${\times}$P. x rigitaeda ($R^2=0.66$) and P. thunbergii (p < 0.0001), respectively. There were significant differences in susceptibility by the age of seedlings in each of P. rigida${\times}$P. x rigitaeda (p < 0.0001) and P. thunbergii (p < 0.0001) based on lesion length.

• Korean journal of applied entomology
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• v.57 no.4
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• pp.393-399
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• 2018

A loop-mediated isothermal amplification (LAMP) primer set (WBPH-65) was designed for the species-specific detection of white-backed planthopper (WBPH) Sogatella furcifera based on the full-length sequence of the internal transcribed spacer 2 (ITS2) (KC417469.1). The WBPH-65 primer set consists of six primers (total 165 bp), F3 (18 bp), B3 (18 bp), FIP (43 bp), BIP (40 bp), LF (21 bp), and LB (25 bp). After the LAMP reaction of three rice planthoppers, S. furcifera, Nilaparvata lugens, and Laodelphax striatellus, with the WBPH-65 primer set for 60 min at $65^{\circ}C$, the LAMP products were observed in the genomic DNA of S. furcifera only. According to the DNA amount of S. furcifera and incubation duration at $65^{\circ}C$, the difference of fluorescence relative to the negative control (0 ng) was clearly observed in a 40-min incubation with 10 and 100 ng or in case of 60-min incubation with 0.01, 0.1, 1, 10, and 100 ng. There was little difference in fluorescence between the negative control and all the other DNAs tested in 20- and 30-min incubations. On the other hand, the WBPH-65 primer set without LF and LB primers showed little amplification in the genomic DNAs of the three rice planthoppers, S. furcifera, N. lugens, and L. striatellus in a 60-min incubation. These results suggest that all six primers (F3, B3, FIP, BIP, LF, and BF) are necessary for the WBPH-65 primer set to detect S. furcifera within a 60-min incubation, and is able to discriminate S. furcifera from at least N. lugens and L. striatellus.

• Horticultural Science & Technology
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• v.33 no.3
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• pp.409-419
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• 2015

This study was conducted to establish an efficient screening method for watermelon plants resistant to Fusarium wilt (FW), which is caused by Fusarium oxysporum f. sp. niveum (Fon). An HA isolate was prepared from a wilted watermelon plant in Haman-gun and identified as F. oxysporum f. sp. niveum based on morphological characteristics, molecular analyses of ITS (internal transcribed spacer) and TEF (translation elongation factor $1{\alpha}$) sequences, and host specificity on cucurbits including watermelon, melon, oriental melon, and cucumber. The assay for disease response of watermelon differentials indicated that the HA isolate was race 0. Among seven liquid media tested, the highest amount of Fon spores was produced from V8-juice broth, which was selected as a medium for mass production of Fon. The disease assay for 21 watermelon and 11 watermelon-rootstock cultivars demonstrated that 20 watermelon cultivars except for 'Soknoranggul' were susceptible; 'Soknoranggul' was moderately resistant. All the tested rootstock cultivars were highly resistant to the HA isolate. The evaluation of disease development depending on various conditions suggested that an efficient screening method for FW resistance in watermelon plants is to dip the roots of 10-day-old seedlings in spore suspension of $1.0{\times}10^5-1.0{\times}10^6conidia{\cdot}mL^{-1}$ for 30 min., to transplant the seedlings to plastic pots with a fertilized soil, and then to cultivate the plants at $25^{\circ}C$ for 3 weeks.

• Research in Plant Disease
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• v.22 no.2
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• pp.72-80
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• 2016

Gummy stem blight, caused by the fungus Didymella bryoniae, is major disease of watermelons worldwide. The objective of the present study was to establish an efficient screening system to identify watermelon resistant to D. bryoniae. An GSB3 isolate was prepared from a watermelon plant showing typical symptoms of gummy stem blight in Haman-gun and identified as D. bryoniae based on molecular analysis of internal transcribed spacer sequence. A simple mass-production technique of inoculum was developed based on spore production of D. bryoniae GSB3 under several incubation conditions and their virulence on watermelon plants. Resistance degrees of 22 commercial watermelon cultivars to the GSB3 isolate were evaluated. Among them, four watermelon cultivars showing different degree of resistance response were selected for further study. Development of disease on the cultivars according to various conditions including inoculum concentrations, incubation periods in dew chamber, and incubation temperatures was investigated. From the results, we suggest an efficient screening method for resistant watermelon cultivars to gummy stem blight. Seeds of watermelon cultivar are sown and grown in a greenhouse until plant stage of 2-fully expanded leaves. Seedlings are inoculated with D. bryoniae by spraying spore suspension of the fungus at a concentration of $5.0{\times}10^5spores/ml$. The infected plants are incubated in humidity chamber at $25^{\circ}C$ for 48 hours and then transferred to a growth chamber at $25^{\circ}C$ and 80% relative humidity with 12-hour light a day. Three to four days after inoculation, disease severity of the plant are measured using percentage of infected leaf area.