• Korean Journal of Microbiology
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• v.33 no.1
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• pp.31-37
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• 1997

Intergeneric hybrids between Aspergillus niger and Perricillium ch~y.sop~um(Tyr ), hyperlipolytic enzyne-producing fungi, were obtained by nuclear transfer technique:. Optimal conditions for formation of intergeneric hybrids were investigated. Maximum production of protoplasts were obtainrd by 1% Novozym 234 at $30^{\circ}C$ for 3 hrs and the most effective osmotic stabilizers for the isolation of protoplasts were 0.6 M KC]. Frequencies of hybrid formation by nuclear transfer were $1.3{\times}$10^{-3}$$ $-3.8{\times}$10^{-3}$$. From the chervation of genetic stability, conidial size, DNA content, ;md nuclear stain, it was suggested that their karyotypes are aneuploid. The hybrids showed 1.4-2.2 fold higher lipase activities than parental strains. It was strongly supported by results of this study that nuclear transfer technique is much more efficient in the formation of intergeneric hybrids than protoplast fusion and is very useful for the improvement of strains.

• Microbiology and Biotechnology Letters
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• v.30 no.2
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• pp.111-115
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• 2002

To produce yellow pigment selectively, mutants were induced from Monascus anka Nakazawa et Sato IFO 4478 (KCCM 11832 strain), and their characteristics were evaluated. Five kinds of auxotrophic mutants which required amino acids for growth and pigmentation, were isolated through a series of mutagenic treatments. Especially, asparagine auxotroph Y7 produced high ratio of yellow pigment. This mutant showed all the morphological characteristics of Monascuceae but the shape of colony and the diameter of conidia. Mutant Y7 was propagated by sexual reproduction more often than asexual reproduction, which could be effective in production of pigments. Yellow pigment produced extracellularly by the mutant Y7 was more soluble in polar solvents such as ethanol and water than in nonpolar solvents. Its productivity of yellow pigment was 2.2 times higher in the mutant Y7 than in parents. In addition, its yellow pigment showed characteristics of maximum absorption at 373 nm. Moreover, the hue of pigment produced by the mutant Y7 was bright yellow, and it was stable through the subculture over 10 generations.

• The Plant Pathology Journal
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• v.35 no.6
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• pp.564-574
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• 2019

Like many fungal pathogens, the conidium and appressorium play key roles during polycyclic dissemination and infection of Magnaporthe oryzae. Ran-binding protein microtubule-organizing center (RanBPM) is a highly conserved nucleocytoplasmic protein. In animalia, RanBPM has been implicated in apoptosis, cell morphology, and transcription. However, the functional roles of RanBPM, encoded by MGG_00753 (named MoRBP9) in M. oryzae, have not been elucidated. Here, the deletion mutant ΔMorbp9 for MoRBP9 was generated via homologous recombination to investigate the functions of this gene. The ΔMorbp9 exhibited normal conidial germination and vegetative growth but dramatically reduced conidiation compared with the wild type, suggesting that MoRBP9 is involved in conidial production. ΔMorbp9 conidia failed to produce appressoria on hydrophobic surfaces, whereas ΔMorbp9 still developed aberrantly shaped appressorium-like structures at hyphal tips on the same surface, suggesting that MoRBP9 is involved in the morphology of appressorium-like structures from hyphal tips and is critical for development of appressorium from germ tubes. Taken together, our results indicated that MoRBP9 played a pleiotropic role in polycyclic dissemination and infection-related morphogenesis of M. oryzae.

• Mycobiology
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• v.47 no.4
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• pp.473-482
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• 2019

Rice blast disease, caused by the ascomycete fungus Magnaporthe oryzae, is one of the most important diseases in rice production. PAS (period circadian protein, aryl hydrocarbon receptor nuclear translocator protein, single-minded protein) domains are known to be involved in signal transduction pathways, but their functional roles have not been well studied in fungi. In this study, targeted gene deletion was carried out to investigate the functional roles of the PAS-containing gene MoPAS1 (MGG_02665) in M. oryzae. The deletion mutant ΔMopas1 exhibited easily wettable mycelia, reduced conidiation, and defects in appressorium formation and disease development compared to the wild type and complemented transformant. Exogenous cAMP restored appressorium formation in ΔMopas1, but the shape of the restored appressorium was irregular, indicating that MoPAS1 is involved in sensing the hydrophobic surface. To examine the expression and localization of MoPAS1 in M. oryzae during appressorium development and plant infection, we constructed a MoPAS1:GFP fusion construct. MoPAS1:GFP was observed in conidia and germ tubes at 0 and 2 h post-infection (hpi) on hydrophobic cover slips. By 8 hpi, most of the GFP signal was observed in the appressoria. During invasive growth in host cells, MoPAS1:GFP was found to be fully expressed in not only the appressoria but also invasive hyphae, suggesting that MoPAS may contribute to disease development in host cells. These results expand our knowledge of the roles of PAS-containing regulatory genes in the plant-pathogenic fungus M. oryzae.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.469-472
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• 2002

Solid state fermentation was performed for the production of entomopathogenic fungus Metarhizium anisopliae HY-2 using wheat bran media containing rice bran. Fungal growth in a solid state fermentation system was estimated by viable cell count, spore count, and mycelial biomass. It was used chemical method measuring N-acetyl-glucosamine (chitin) content for estimating of mycelial biomass. In static flask culture, viable cell reached 2.40 ${\times}$ $10^8$ cfu/g at 23 days of culture at $27^{\circ}C$ and then mycelial biomass was 41.59 mg/g. Specific growth rate(${\mu}$ max) was 0.0418 $h^{-1}$ between 3 and 9 days when estimated by viable cell count and was 0.00976 $h^{-1}$ between 9 and 17 days when N-acetylglucosamine content was measured. Viable cells reached 1.12 ${\times}$ $10^8$ cfu/g in polypropylene-bag at 28 days of culture at $27^{\circ}C$. Formulated microbial pesticide containing M. anisopliae HY-2 were tested their bio-activity against Chestnut Brown Chafer (Adoretus tenuimaculatus). The protection rate of the liquid culture showed 13 ${\sim}$ 26 % with 1st to 3rd instar, and spore suspension of M. anisopliae HY-2 showed 56 ${\sim}$ 64%. Conidia produced by large scale solid-state fermentation showed 20 ${\sim}$ 27 % activity 60 ${\sim}$ 64 % with M. anisopliae HY-2.

• Mycobiology
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• v.41 no.4
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• pp.221-224
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• 2013

Aphids are one of the most destructive pests in crop production such as pepper, cucumber, and eggplants. The importance of entomopathogenic fungi as alternative pest control agents is increasing. Conidia of entomopathogenic fungi are influenced by environmental conditions, such as temperature and relative humidity, and cause slow and fluctuating mortality. These factors have prevented wider application and use of biocontrol agents. For investigation of means of mitigation of such problems, we conducted bioassays with 47 fungal culture filtrates in order to evaluate the potential of secondary metabolites produced by entomopathogenic fungi for use in aphid control. Among 47 culture filtrates cultured potato dextrose broth, filtrate of Beauveria bassiana Bb08 showed the highest mortality (78%) against green peach aphid three days after treatments. Filtrate of Bb08 cultured in Adamek's medium showed higher toxicity as 100% to third instar nymphs of the aphid compared with seven other filtrates cultured in different broths amended with colloidal chitin or oil. The culture filtrates and fungal cultures from media amended with colloidal chitin or oil had lower control efficacies than filtrates without these additives in three different media. These results indicate that the fungal culture fluid or culture filtrate of B. bassiana Bb08 cultured in Adamek's medium has potential for development as a mycopesticide for aphid control.

• The Plant Pathology Journal
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• v.34 no.6
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• pp.470-479
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• 2018

The rice blast pathogen Magnaporthe oryzae is a global threat to rice production. Here we characterized RHO2 gene (MGG_02457) that belongs to the Rho GTPase family, using a deletion mutant. This mutant ${\Delta}Morho2$ exhibited no defects in conidiation and germination but developed only 6% of appressoria in response to a hydrophobic surface when compared to the wild-type progenitor. This result indicates that MoRHO2 plays a role in appressorium development. Furthermore, exogenous cAMP treatment on the mutant led to appressoria that exhibited abnormal morphology on both hydrophobic and hydrophilic surfaces. These outcomes suggested the involvement of MoRHO2 in cAMP-mediated appressorium development. ${\Delta}Morho2$ mutation also delayed the development of appressorium-like structures (ALS) at hyphal tips on hydrophobic surface, which were also abnormally shaped. These results suggested that MoRHO2 is involved in morphological development of appressoria and ALS from conidia and hyphae, respectively. As expected, ${\Delta}Morho2$ mutant was defective in plant penetration, but was still able to cause lesions, albeit at a reduced rate on wounded plants. These results implied that MoRHO2 plays a role in M. oryzae virulence as well.

• The Plant Pathology Journal
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• v.38 no.3
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• pp.229-238
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• 2022

Fusarium wilt caused by Fusarium oxysporum f. sp. niveum (Fon) is the most serious soil-borne disease in the world and has become the main limiting factor of watermelon production. Reliable and quick detection and quantification of Fon are essential in the early stages of infection for control of watermelon Fusarium wilt. Traditional detection and identification tests are laborious and cannot efficiently quantify Fon isolates. In this work, a real-time polymerase chain reaction (PCR) assay has been described to accurately identify and quantify Fon in watermelon plants and soil. The FONRT-18 specific primer set which was designed based on identified specific sequence amplified a specific 172 bp band from Fon and no amplification from the other formae speciales of Fusarium oxysporum tested. The detection limits with primers were 1.26 pg/µl genomic DNA of Fon, 0.2 pg/ng total plant DNA in inoculated plant, and 50 conidia/g soil. The PCR assay could also evaluate the relationships between the disease index and Fon DNA quantity in watermelon plants and soil. The assay was further used to estimate the Fon content in soil after disinfection with CaCN₂. The real-time PCR method is rapid, accurate and reliable for monitoring and quantification analysis of Fon in watermelon plants and soil. It can be applied to the study of disease diagnosis, plant-pathogen interactions, and effective management.

• Korean journal of applied entomology
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• v.56 no.3
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• pp.301-308
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• 2017

Locusts, Locusta migratoria (Orthoptera: Acrididae) are periodical unpredictable agricultural pests worldwide and cause serious damage to crop production; however, little consideration has been given to the management of this pest. Herein, we constructed a locust-pathogenic fungal library and confirmed that some fungi could be used as resources for locust management. First, the entomopathogenic fungi were collected from sampled soils using a Tenebrio molitor-based baiting system. For the locust assay, a locust colony was obtained from the National Institute of Agricultural Science and Technology. A total of 34 entomopathogenic fungal granules, which were produced by solid cultures, were placed in the plastic insect-rearing boxes (2 g/box) and nymphs of locust were contained in the box. In 3-7 days, mycosis was observed on the membranous cuticles of the head, abdomen, and legs of locusts. In particular, Metarhizium anisopliae, M. lepidiotae, and Clonostachys rogersoniana exhibited high virulence against the locust. Given that the 34 isolates could be used in field applications, their conidial production and stability (thermotolerance) were further characterized. In the thermotolerance assay, Paecilomyces and Purpureocillium isolates had higher thermotolerance than the other isolates. Most of the fungal isolates produced ca. >$1{\times}10^8conidia/g$ on millet grain medium. In a greenhouse trial, the granular application of M. anisopliae isolate on the soil surface resulted in 85.7% control efficacy. This work suggests that entomopathogenic fungi in a granular form can be effectively used to control the migratory locust.

• Journal of Food Hygiene and Safety
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• v.37 no.1
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• pp.9-14
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• 2022

The purpose of this study was to analyze the hazard of fungi in Garaetteok (Korean rice cake) by isolating and identifying of fungi contaminated with Garaetteok and investigating the possibility of mycotoxin production. Garaetteok used in this study were the ones that were returned back to the manufacturers in Jeollanam-do due to the presence of foreign matters presumed to be fungi. The fungi foreign matter was collected and inoculated on Potato dextrose agar, Malt extract agar, and Czapek yeast extract agar, and then cultured at 25℃ for 7 days. The micro-structure was observed under an optical microscope for the colonies in which pure isolation was confirmed. The gene sequencing of the product of amplified PCR was analyzed using the ITS primer. Colony-1 and 2 maintained the same properties in each tray, confirming that they were purely isolated. Budding cells were observed from the Colony-1, thus, it was determined to be yeast. Colony-2 was determined to be a fungus that belongs to Fusarium spp. as fusiform conidia were observed. As a result of gene sequencing, a total of 76 cases of fungi of Fusarium spp. were found, among which Fusarium solani was the most observed cases (53 cases). From the morphological and genetic identification, Colony-2 was identified as Fusarium spp., specifically, Fusarium solani. The fungi found in Fusarium spp. produce mycotoxins such as nivalenol, zearalenone, and fumonisin, which may cause vomiting, diarrhea, and cancer. Conclusively, the results confirm the possibility of mycotoxin production by Fusarium spp. isolated from Garaetteok. Consequently, when an unknown fungus was found, it is necessary to isolate and identify the fungus, determine whether it is a mycotoxin producing species, and strengthen relative administrative measures, accordingly.