• 한국균학회소식:학술대회논문집
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• 한국균학회 2018년도 춘계학술대회 및 임시총회
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• pp.51-51
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• 2018

The application of recombinant DNA technology has been remarkable and nearly replaced commonly used traditional methods. Traditional industrial microbiology long depended on the discovery of valuable strains and mutagenesis of such strains to improve its secretion capacity of enzymes and secondary metabolites on the industrial scale. Commodities included industrial enzymes and biopharmaceuticals. The purpose of genome manipulation by the crossing of different strains or genetic recombination of naked DNA to the genome is of increased production of valuable metabolites. We optimized a transformation method to either for removal of innate genes, introduction of heterologous genes, or combination of both. We have been used selected whole or partial genes to manipulate target fungi toward the development of strains overproducing invaluable proteins. We have also used the whole genome sequence information of fungal genomes in public databases and functional genomics approach to select genes to manipulate and eventually contributing greatly to the development of overproducing industrial strains overproducing proteins or secondary metabolites. I will briefly review 1) filamentous fungi as a host for production of recombinant proteins and secondary metabolites, 2) markets of industrial metabolites, 3) a new approach to manipulate up to five genes at the same time in the system that ProxEnrem uses.

• Mycobiology
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• 제49권2호
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• pp.122-132
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• 2021

The reexamination of the fungal genus Botryosphaeria on 12 plant species of 10 families was carried out based on molecular phylogenetic analyses using the regions of translation elongation factor 1-α, β-tubulin, DNA-directed RNA polymerase II subunit, and internal transcribed spacer region of rDNA and morphological characteristics. Japanese isolates were divided into five clades and include Botryosphaeria dothidea, B. qingyuanensis, B. sinensis, and Botryosphaeria spp. Two species, B. qingyuanensis and B. sinensis have been newly added to the Japanese mycoflora, but their host plants are not specified. Botryosphaeria tenuispora isolated from Leucothoe fontanesiana and insect galls on fruits of Aucuba japonica has been proposed as a new species.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.344-344
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• 2017

The glycoprotein known as glomalin-related soil protein (GRSP) is abundantly produced on the hyphae and spores of arbuscular mycorrhizal fungi (AMF) in soil and roots. GRSP play a decisive role in the soil aggregation, but GRSP was also sensitive to agricultural managements. Thus, our objectives were to assess the effect of different cover crop incorporation on the GRSP content in soil and growth of subsequent soybean and maize. Pot experiments with the incorporation of four cover crops were set up. The same amount (666g) of aboveground plant parts of wheat (AMF host), hairy vetch (AMF host), mustard (non-host) and rapeseed (non-host) was separately incorporated into soils. The aboveground plant parts and roots of soybean and maize were grown in each incorporated pots and sampled at 6 and 9 weeks after sowing. Our results showed that the different cover crops incorporation affected soil biological and chemical properties such as EC, $NO_3-N$ content, ${\beta}-glucosidase$ activity, alkaline phosphatase (ALP) activity and GRSP content. The soil EC and $NO_3-N$ content in the hairy vetch, mustard and rapeseed was higher compared to the wheat. The ${\beta}-glucosidase$ activity in the wheat and hairy vetch was significantly higher than that in the mustard and rapeseed, and the ALP activity in the wheat was significantly higher than that in the hairy vetch, mustard, and rapeseed. The GRSP content in the mustard and rapeseed was significantly lower than that of the hairy vetch and wheat. Moreover, The top dry weight and leaf area of soybean and maize in the hairy vetch at 6 weeks were significantly higher compared to the other treatments. Our results indicated that the incorporation of mustard and rapeseed may cause indirectly the decrease of GRSP content and soil enzyme activity in soil. One possible explanation for the decrease of GRSP in non-AMF host crop treatments may be the decrease of AMF density in the soil. AMF are not able to form a symbiotic relationship with Brassicaceae roots due to the release of anti-fungal compounds. This means the AMF may not be able to produce GRSP in the soil. However, the differences in the benefit of cover crop incorporation were shown only by a pot experiment. Comparative investigations of crop residue managements would be applied to both pot experiment and field study to clarify a better selection of cover crops in rotation to encourage GRSP production.

• The Plant Pathology Journal
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• 제22권3호
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• pp.255-259
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• 2006

The pathogenicity to pear trees and other experimental hosts of the Apple stem grooving virus Korean isolate (ASGV-K) carried by a fungal vector, Talaromyces flavus was examined. ASGV-harboring T. flavus induced mild symptoms on virus-free pears. Symptom severity was intermediate between pears showing typical PBNLS and virus-free pears. Ten cultivars of Phaseolus vulgaris showed 35%-90% infectivity by direct infiltration into leaves and roots by ASGV-harboring T. flavus. Application of fungal cultures to soils showed 0%-70% infectivity depending on the P. vulgaris cultivar. Sap extracted from ASGV-infected Chenopodium quinoa induced similar symptoms on P. vulgaris at 25 days after inoculation. Similar symptoms were also detected on P. vulgaris which were inoculated with ASGV-harboring T.flavus. When healthy P. vulgaris leaves were challenged with sap extracted from P. vulgaris leaves infected with ASGV-harboring T. flavus, typical symptoms were observed. These data suggest that T. flavus mediates the transfer of ASGV to host plants.

• The Plant Pathology Journal
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• 제25권4호
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• pp.333-343
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• 2009

Here, we show that an endophytic bacterial strain, Enterobacter asburiae B1 exhibits the ability to elicit ISR in cucumber, tobacco and Arabidopsis thaliana. This indicates that strain B1 has a widespread ability to elicit ISR on various host plants. In this study, E. asburiae strain B1 did not show antifungal activity against tested major fungal pathogens, Colletotrichum orbiculare, Botrytis cinerea, Phytophthora capsici, Rhizoctonia solani, and Fusarium oxysporum. Moreover, the siderophore production by E. asburiae strain B1 was observed under in vitro condition. In greenhouse experiments, the root treatment of strain B1 significantly reduced disease severity of cucumber anthracnose caused by fungal pathogen C. orbiculare compared to nontreated control plants. By root treatment of strain B1 more than 50% disease control against anthracnose on cucumber was observed in all greenhouse experiments. Simultaneously, under the greenhouse condition, the soil drench of strain B1 and a chemical inducer benzothiadiazole (BTH) to tobacco plants induced GUS activity which is linked with activation of PR promoter gene. Furthermore, in Arabidopsis thaliana plants the soil drench of strain B1 induced the defense gene expression of PR1 and PDF1.2 related to salicylic acid and jasmonic acid/ethylene signaling pathways, respectively. In this study, for the main focus on root colonization by strain B1 associated with defense responses, bacterial cells of strain B1 was tagged with the gfp gene encoding the green fluorescent protein in order to determine the colonization pattern of strain B1 in cucumber. The gfp-tagged B1 cells were found on root surface and internal colonization in root, stem, and leaf. In addition to this, the scanning electron microscopy observation showed that E. asburiae strain B1 was able to colonized cucumber root surface.

• Journal of Microbiology and Biotechnology
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• 제27권3호
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• pp.439-449
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• 2017

Beauveria bassiana infects a number of pest species and is known to produce insecticidal substances, such as the nonribosomal peptides (NRPs) beauvericin and bassianolide. However, most NRPs and their biological roles in B. bassiana remain undiscovered. To identify NRPs that potentially contribute to pathogenesis, the 21 predicted NRP synthetases (NRPSs) or NRPS-like proteins of B. bassiana ARSEF 2860 were primarily ranked into three functional groups: basic metabolism (7 NRPSs), pathogenicity (12 NRPSs), and unknown function (2 NRPSs). Based on the transcript levels during in vivo growth on diamondback moth (Plutella xylostella (Linnaeus)), half of the Group II NRPSs were likely to be involved in infection. Given that the metabolites biosynthesized by these NRPSs remain to be determined, our result underlines the importance of the NRPSome in fungal pathogenesis, and will serve as a guide for future genomic mining projects to discover functionally essential and structurally diverse NRPs in fungal genomes.

• International Journal of Industrial Entomology and Biomaterials
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• 제35권1호
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• pp.58-62
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• 2017

The two-spotted spider mite (Tetranychus urticae) has sustained damage on more than 200 host plants worldwide. Many farmers have relied on chemical acaricides to control mite, but the abuse of acaricides has caused serious resistance to mite. To overcome this problem, microbial control using entomopathogenic fungi have been studied. Entomopathogenic fungi have been an important role against the control of pest, and most of their culture products have been demonstrated to have virulence against pest population. In this study, we evaluated and compared the virulence of culture filtrates, aerial conidia and blastospores of selected Metarhizium anisopliae 4-2 and Beauveria bassiana 2R-3-3-1, respectively, among two-spotted spider mite-pathogenic fungi. As a result, the virulence was confirmed in all treatments, and the accumulated mortality rates were between 77 and 100% within 7 days. Especially, treatment with the fungal culture filtrate alone exhibited quite high virulence, and combined treatment with aerial conidia or blastospores enhanced activity. However, the median lethal time of treatments was not significantly different. When two isolates were compared, M. anisopliae 4-2 showed higher virulence than B. bassiana 2R-3-3-1. These results suggest that the selected two fungal isolates and their culture products could be used effectively for the control of two-spotted spider mite.

• The Plant Pathology Journal
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• 제28권1호
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• pp.55-59
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• 2012

Northern stem canker caused by $Diaporthe$ $phaseolorum$ var. $caulivora$ ($Dpc$) has become a serious disease in soybean. The objectives of this study were to survey the existence of $Dpc$ on soybean in Korea, and to examine the potential pathogenicity of $Dpc$ in seed decay. One such isolate, SSLP-4, isolated from a field-grown plant of the Korean soybean cultivar Danbaekkong, was identified as $Dpc$, based on its morphological and molecular characteristics by sequences of internal transcribed spacer (ITS), translation elongation factor (TEF) 1-${\alpha}$ and ${\beta}$-tubulin regions, as well as pathogenic analyses. Moreover, morphological and molecular analyses revealed that isolate SSLP-4 was nearly identical to $Dpc$ strains from the United States. Pathogenicity tests on hypocotyls of soybean seedlings and detached leaves resulted in typical symptoms of soybean northern stem canker and inoculation on plants at R5-R7 stage caused seed decay. All results suggest that the $Dpc$ strain SSLP-4 can cause both stem canker and seed decay on soybean. Thus, the SSLP-4 isolate has the potential to contribute greatly to understanding of host plant resistance mechanisms, both at vegetative and reproductive growth stages in soybean.

• The Plant Pathology Journal
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• 제23권3호
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• pp.203-209
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• 2007

Infection structures were observed using a fluorescence microscope at the penetration sites on the leaves of potato plants pre-inoculated with the bacterial strains Pseudomonas putida TRL2-3, Micrococcus luteus TRK2-2, and Flexibacteraceae bacterium MRL412, which mediated an induced systemic resistance on potato plants against late blight disease caused by Phytophthora infestans. In order to compare the infection structures on the leaves expressing systemic acquired resistance, the leaves of potato plants pre-treated with DL-3-amino butyric acid (BABA) were also observed after challenge inoculation with the same pathogen. The infection structures were investigated. The total number of germination and appressorium formation of P. infestans were counted. Furthermore, the frequencies of fluorescent epidermal cells at the penetration sites, which indicate a defense response of plant cell, were estimated. There were no differences on the germination rates of the fungal cysts among the untreated control, BABA pre-treated, and bacterial strains pre-inoculated plants. However, appressorium formation was slightly decreased on the leaves of BABA pre-treated plants compared to those of untreated as well as bacterial strains pre-inoculated plants. Furthermore, the frequencies of fluorescent cells of BABA pre-treated and bacterial strains pre-inoculated were higher than that of untreated plants, indicating an active defense reaction of the host cells against the fungal attack. On the other hand, the pre-treatment with BABA caused a stronger fluorescent of epidermal cells at the penetration sites compared to the pre-inoculation with the bacterial strains. Interestingly, the frequency of fluorescent cells by BABA, however, was lower than that by the bacterial strains. Based on the results it is suggested that the infection structures showing resistance reaction on the leaves of potato plants were different between by pre-inoculation with bacterial strains and by pre-treatment with BABA against the late blight pathogen.

• Journal of Microbiology and Biotechnology
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• 제15권1호
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• pp.147-154
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• 2005

A survey for vesicular-arbuscular mycorrhizae (VAM) status was undertaken in three different industrially polluted sites at Uyyakondan channel of Senthanneerpuram area in Trichy, India. The soils and the effluents were acidic, and contained higher Zn (621 to 711 ppm) than the other heavy metals, such as Cu, Pb, and Ni. Eighteen plant species were collected from the rhizosphere soils, and 13 species were positive for VAM colonization. Fifteen VAM fungal species were isolated from the plant species. The number of VAM fungal spores from the soils ranged from 45 to 640 per 100 g of soil. There was a significant correlation observed between the number of spores and percentage root colonization, as exemplified by Acalypha indica (45 and 20%, respectively) and Paspalum vaginatum (640 and 98%, respectively). Hostspecific and site-specific associations were observed in site 2; particular VAM species, Gigaspora gigantea and Glomus fasciculatum, were specific to particular host plants, Phyllanthus maderaspatensis and A. indica, respectively, even though Eclipta prostrata and Physalis minima were maximally associated with 8 VAM species. G. fasciculatum was found in 11 plant species and predominant VAM species. These results led us to conclude that VAM fungi are associated with a majority of the plants in the industrial polluted sites and support the plants to survive in the acidic soils, polluted with heavy metals of the industrial effluents.