• Journal of Microbiology and Biotechnology
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• v.20 no.1
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• pp.208-216
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• 2010

Fusarium verticillioides is an important pathogen of maize, being responsible for ear rots, stalk rots, and seedling blight worldwide. During the past decade, F. verticillioides has caused several severe epidemics of maize seedling blight in many areas of China, which lead to significant losses. In order to understand the molecular mechanisms regulating fungal development and pathogenicity in this pathogen, we isolated and characterized the gene fpk1 (GenBank Accession No. EF405959) encoding a homolog of the cAMP-dependent protein kinase catalytic subunit, which included a 1,854-bp DNA sequence from ATG to TAA, with a 1,680-bp coding region, and three introns (lengths: 66 bp, 54 bp, and 54 bp), and the predicated protein precursor had 559 aa. The mutant ${\Delta}fpk1$, which was disrupted of the fpkl gene, showed reduced vegetative growth, fewer and shorter aerial mycelia, strongly impaired conidiation, and reduced spore germination rate. After germinating, the fresh hypha was stubby and lacking of branch. When inoculated in susceptible maize varieties, the infection of the mutant ${\Delta}fpk1$ was delayed and the infection efficiency was reduced compared with that of the wild-type strain. AU this indicated that gene fpk1 participated in hyphal growth, conidiophore production, spore germination, and virulence in F. verticillioides.

• The Plant Pathology Journal
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• v.38 no.3
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• pp.254-260
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• 2022

Fusarium incarnatum-equiseti species complex (FIESC) contain over 40 members. The primer pair Smibo1FM/Semi1RM on the RPB2 partial gene has been reported to be able to identify Fusarium semitectum. The F. fujikuroi species complex (FFSC) contains more than 50 members. The F. verticillioides as a member of this complex can be identified by using VER1/VER2 primer pair on the CaM partial gene. In this research, the Smibo1FM/Semi1RM can amplify F. sulawesiense, F. hainanense, F. bubalinum, and F. tanahbumbuense, members of FIESC at 424 bp. The VER1/VER2 can amplify F. verticillioides, F. andiyazi, and F. pseudocircinatum, members of FFSC at 578 bp. Polymerase chain reaction-restriction fragment length polymorphism by using the combination of three restriction enzymes EcoRV, MspI, and HpyAV can differentiate each species of FIESC used. The two restriction enzymes HpaII and NspI can distinguish each species of FFSC used. The proper identification process is required for pathogen control in the field in order to reduce crop yield loss.

• Journal of Microbiology and Biotechnology
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• v.18 no.4
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• pp.648-657
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• 2008

In this study, we used functional genomic strategies, proteomics and quantitative real-time (qRT)-PCR, to advance our understanding of genes associated with fumonisin production in the fungus Fusarium verticillioides. Earlier studies have demonstrated that deletion of the FCC1 gene, which encodes a C-type cyclin, leads to a drastic reduction in fumonisin production and conidiation in the mutant strain (FT536). The premise of our research was that comparative analysis of F. verticillioides wild-type and FT536 proteomes will reveal putative proteins, and ultimately corresponding genes, that are important for fumonisin biosynthesis. We isolated proteins that were significantly upregulated in either the wild type or FT536 via two-dimensional polyacrylamide gel electrophoresis, and subsequently obtained sequences by mass spectrometry. Homologs of identified proteins, e.g., carboxypeptidase, laccase, and nitrogen metabolite repression protein, are known to have functions involved in fungal secondary metabolism and development. We also identified gene sequences corresponding to the selected proteins and investigated their transcriptional profiles via quantitative real-time (qRT)-PCR in order to identify genes that show concomitant expression patterns during fumonisin biosynthesis. These genes can be selected as targets for functional analysis to further verify their roles in $FB_1$ biosynthesis.

• Research in Plant Disease
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• v.21 no.4
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• pp.326-329
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• 2015

Fusaric acid is a mycotoxin produced by species of the fungus Fusarium and can act synergistically with other Fusarium toxins. In order to develop a specific detection method for fusaric acid-producing fungus, PCR primers were designed to amplify FUB10, a transcription factor gene in fusaric acid biosynthetic gene cluster. When PCR with Fub10-f and Fub10-r was performed, a single band (~550 bp) was amplified from F. oxysporum, F. proliferatum, F. verticillioides, F. anthophilum, F. bulbicola, F. circinatum, F. fujikuroi, F. redolens, F. sacchari, F. subglutinans, and F. thapsinum, all of which were known for fusaric acid production. Whereas the FUB10 specific band was not amplified from Fusarium species known to be trichothecene producer. Because production of fusaric acid can co-occur in species that also produce fumonisin mycotoxins, we developed a multiplex PCR assay using the FUB10 primers as well as primers for the fumonisin biosynthetic gene FUM1. The assay yielded amplicons from fumonisin producers such as F. proliferatum and F. verticillioides, allowing for the simultaneous detection of species with the genetic potential to produce both types of mycotoxins.

• The Korean Journal of Mycology
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• v.39 no.3
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• pp.217-222
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• 2011

Job's tears (Coix lacryma-jobi) is native to East Asia, and grains of the plant are used as health food and medicinal material. Head blight symptoms of the plant were frequently observed during disease surveys in Korea from 2006 to 2008. The symptoms were characterized as discoloration of husks, and subsequently inside of mature grains were shriveled or emptied. One hundred fifty nine isolates of Fusarium species were obtained from the disease symptoms of the plant collected from several locations in the country. Out of the isolates, the most frequently isolated Fusarium species were F. graminearum (34%), F. proliferatum (14.5%), F. verticillioides (10.1%), F. equiseti (6.9%), and F. fujikuroi (6.3%). Other Fusarium species isolated were F. subglutinans, F. semitectum, F. poae, and F. sporodochioides. Elongation factor 1 alpha gene sequences of the isolates were used for phylogenetic analysis. Analyses of the sequences revealed that the isolates were confirmed to be identical with each reference species of NCBI GenBank. Pathogenicity tests showed that F. graminearum, F. proliferatum and F. verticillioides were strongly virulent to grains of Job's tears. The present study is the first report of head blight of Job's tears caused by Fusarium species in Korea.

• 한국균학회소식:학술대회논문집
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• 2015.05a
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• pp.14-14
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• 2015

Fungi are of particular interest due to their capacity to produce an extensive array of secondary metabolites. While many secondary metabolites have no known functions to the producing fungal organisms, these metabolites have tremendous importance to humans with beneficial (e.g., antibiotics) or detrimental (e.g., mycotoxins) properties. In this study, two important filamentous fungi, Fusarium verticillioides and Mycosphaerella graminicola were selected as target species and the genes regulatory functions on the biosynthesis of secondary metabolisms were studied. Functional genomics including forward and reverse genetics, and proteomics were utilized to better understand the complex secondary metabolism regulations in both F. verticillioides and M. graminicola. Identified genes in either F. verticillioides or M. graminicola background were CPP1 (a putative protein phosphatase gene), GAC1 (encoding a GTPase activating protein), MCC1(encoding c-type cyclin), and the velvet gene, MVE1. Our data suggest that there are diverse regulatory genes on fungal secondary metabolites with distinct or overlapping functional roles.

• 한국균학회소식:학술대회논문집
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• 2014.10a
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• pp.25-25
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• 2014

Three species of Fusarium, F. fujikuroi, F. verticillioides and F. proliferatum, are known to be associated with bakanae disease of rice [1, 2]. F. fujikuroi infects rice flowers and survive in endosperm and embryo of the seeds. Infected seed is an important source of primary inoculum of pathogens [3]. Seeds of rice (Oryza sativa cv. Boramchan) collected from bakanae-infected field were found to be 96% infected with Fusarium sp., 52% with F. fujikuroi, 42% with F. verticillioides, and 12% with F. proliferatum as determined by incubation method and species-specific PCR assays. F. fujikuroi was detected at lemma/palea, endosperm and embryo whereas F. verticillioides and F. proliferatum were recovered only from lemma/palea by means of component plating test. Seed disinfection methods have been developed to control bakanae disease and prochloraz has been most widely used for rice seeds. Two chemicals formulated with prochloraz (PC 1) and prochloraz + hexaconazole (PC 2) that inhibit biosynthesis of ergosterol strongly reduced the incidence of Fusarium spp. on selective media to 4.7% and 2.0%, respectively. Disease symptoms of rice seedlings in nursery soil were alleviated by chemical treatment; seedlings with elongated leaves or wide angle between leaf and stem were strikingly reduced from 15.6 to 3.2% (PC 1) and 0 (PC 2), stem rots were reduced from 56.9 to 26.2% (PC 1) and 32.1% (PC 2), and normal seedling increased from 0.4 to 13.3% (PC 2). Prochloraz has some disadvantages and risks such as the occurrence of tolerant pathogens [4] and effects on the sterol synthesis in animals and humans [5]. For these reasons, it is necessary to develop new disinfection method that do not induce fungal tolerance and are safe to humans and animals. Chlorine dioxide ($ClO_2$), that is less toxic, produces no harmful byproducts, and has high oxidizing power, has been reported to be effective at disinfection of several phytopathogenic fungi including Colletotrichum spp. and Alternaria spp. [6]. Gaseous $ClO_2$ applied to rice seeds at a concentration of 20 ppm strongly suppressed mycelial growth of Fusarium fujikuroi, F. verticillioides and F. proliferatum. The incidence of Fusarium spp. in dry seed with 8.7% seed moisture content (SMC) tended to decrease as the concentration of $ClO_2$ increased from 20 to 40 ppm. Applying 40 ppm $ClO_2$ at 90% relative humidity, incidence was reduced to 5.3% and resulted in significant reduction of disease symptoms on MS media. In nursery soil, stem rot was reduced from 56.9 to 15.4% and the number of normal seedlings increased from 0.4 to 25.5%. With water-soaked seeds (33.1% SMC) holding moisture in the endosperm and embryo, the effectiveness of disinfection using $ClO_2$ increased, even when treated with only 20 ppm for four hours. This suggests that moisture was a key element for action of $ClO_2$. Removal of the palea and lemma from seeds significantly decreased the incidence of Fusarium spp. to 3.0%. Seed germination appeared to decrease slightly by water-soaking at $30^{\circ}C$ because of increased SMC and by physical damage of embryos from hulling. These results indicate that the use of gaseous $ClO_2$ was effective as a means to disinfect rice seeds infected with Fusarium spp. and that moisture around the pathogens in the seed was an important factor for the action of $ClO_2$. Further investigations should be conducted to ascertain the best conditions for complete disinfection of Fusarium spp. that infect deep site of rice seeds.

• Research in Plant Disease
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• v.18 no.4
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• pp.310-316
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• 2012

Gibberellea fujikuroi species (Gf) complex comprises at least 15 species, most of which not only causes serious plant diseases, but also produces mycotoxins including fumonisins. Here, we focused on the abilities of the field isolates belonging to the Gf complex associated with rice and corn, respectively in Korea to produce fumonisin, all of which were confirmed to carry FUM1, the polyketide synthase gene essential for fumonisin biosynthesis. A total of 88 Gf complex isolates (55 F. fujikuroi, 10 F. verticillioides, 20 F. proliferatum, 2 F. subglutinans, and 1 F. concentricum), and 4 isolates of F. commune, which is a non-member of Gf complex, were grown on rice substrate and determined for their production levels of fumonisins by a HPLC method. Most isolates of F. verticillioides and F. proliferatum, regardless of host origins, produced fumonisin $B_1$ and $B_2$ at diverse ranges of levels ($0.5-2,686.4{\mu}g/g$, and $0.7-1,497.6{\mu}g/g$, respectively). In contrast, all the isolates of F. fujikuroi and other Fusarium species examined produced no fumonisins or only trace amounts ($<10{\mu}g/g$) of fumonisins. Interestingly, the frequencies of relatively high fumonisin-producers among the F. proliferatum and F. fujikuroi isolates derived from corn were higher than those among the fungal isolates from rice. In addition, it is a first report demonstrating the ability of the FUM1-carrying F. commune isolates from rice to produce fumonisins.

• Mycobiology
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• v.41 no.4
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• pp.225-233
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• 2013

Gibberella fujikuroi species complex (GFSC) was isolated from rice (Oryza sativa L.) seed samples from ten Asian countries and investigated for incidence of GFSC, molecular characteristics, and pathogenicity. Regardless of geographic origin, GFSC was detected with incidences ranging from 3% to 80%. Four species, Fusarium fujikuroi, F. concentricum, F. proliferatum, and F. verticillioides, were found to show an association with rice seeds, with F. fujikuroi being the predominant species. In phylogenetic analyses of DNA sequences, no relationship was found between species, isolates, and geographic sources of samples. Unidentified fragments of the ${\beta}$-tubulin gene were observed in ten isolates of F. fujikuroi and F. verticillioides. With the exception of three isolates of F. fujikuroi, F. fujikuroi, F. proliferatum, and F. verticillioides were found to have FUM1 (the fumonisin biosynthetic gene); however, FUM1 was not found in isolates of F. concentricum. Results of pathogenicity testing showed that all isolates caused reduced germination of rice seed. In addition, F. fujikuroi and F. concentricum caused typical symptoms of bakanae, leaf elongation and chlorosis, whereas F. proliferatum and F. verticillioides only caused stunting of seedlings. These findings provide insight into the characteristics of GFSC associated with rice seeds and might be helpful in development of strategies for management of bakanae.

• Research in Plant Disease
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• v.23 no.3
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• pp.256-261
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• 2017

The effects of insecticide and fungicide treatment were investigated to reduce mycotoxin contamination of corn (Zea mays L.) seeds. Deoxynivalenol and zearalenone contents were reduced in the treated seeds, but aflatoxin, ochratoxin A, fumonisin, and T-2 toxin were not effective by chemical treatments. The chemical treatment did not affect the growth of saprophyte, but inhibited the pathogenic fungi such as Fusarium verticillioides, F. graminearum and F. equiseti. Myotoxin contents at different harvesting time were compared. As the harvest time was delayed, both levels of deoxynivalenol and zearalenone and frequency of Fusarium spp. increased. However, the major nutrient contents of corn seeds were not affected by harvesting period. These results show that chemical treatments are necessary to reduce the fungal contamination of corn and harvest without delay is important as well.