• The Plant Pathology Journal
• /
• v.28 no.3
• /
• pp.259-269
• /
• 2012

Protein phosphatase 2A (PP2A), a family of serine/threonine protein phosphatases, plays an important role in balancing the phosphorylation status of cellular proteins for regulating diverse biological functions in eukaryotic organisms. Despite intensive studies in mammals, limited information on its role is available in filamentous fungi. Here, we investigated the functional roles of genes for a putative B' delta regulatory subunit (FgPP2AR) and a catalytic subunit (FgPP2AC) of PP2A in a filamentous ascomycete, Fusarium graminearum. Molecular characterization of an insertional mutant of this plant pathogenic fungus allowed us to identify the roles of FgPP2AR. Targeted gene replacement and complementation analyses demonstrated that the deletion of FgPP2AR, which was constitutively expressed in all growth stages, caused drastic changes in hyphal growth, conidia morphology/germination, gene expression for mycotoxin production, sexual development and pathogenicity. In particular, overproduction of aberrant cylindrical-shaped conidia is suggestive of arthroconidial induction in the ${\Delta}FgPP2AR$ strain, which has never been described in F. graminearum. In contrast, the ${\Delta}FgPP2AC$ strain was not significantly different from its wild-type progenitor in conidiation, trichothecene gene expression, and pathogenicity; however, it showed reduced hyphal growth and no perithecial formation. The double-deletion ${\Delta}FgPP2AR;{\Delta}FgPP2AC$ strain had more severe defects than single-deletion strains in all examined phenotypes. Taken together, our results indicate that both the putative regulatory and catalytic subunits of PP2A are involved in various cellular processes for fungal development in F. graminearum.

• Journal of Microbiology and Biotechnology
• /
• v.26 no.5
• /
• pp.918-927
• /
• 2016

Cryptococcus neoformans is a life-threatening pathogenic yeast that causes devastating meningoencephalitis. The mechanism of cryptococcal brain invasion is largely unknown, and recent studies suggest that its extracellular microvesicles may be involved in the invasion process. The 14-3-3 protein is abundant in the extracellular microvesicles of C. neoformans, and the 14-3-3-GFP fusion has been used as the microvesicle's marker. However, the physiological role of 14-3-3 has not been explored. In this report, we have found that C. neoformans contains a single 14-3-3 gene that apparently is an essential gene. To explore the functions of 14-3-3, we substituted the promoter region of the 14-3-3 with the copper-controllable promoter CTR4. The CTR4 regulatory strain showed an enlarged cell size, drastic changes in morphology, and a decrease in the thickness of the capsule under copper-enriched conditions. Furthermore, the mutant cells produced a lower amount of total proteins in their extracellular microvesicles and reduced adhesion to human brain microvascular endothelial cells in vitro. Proteomic analyses of the protein components under 14-3-3-overexpressed and -suppressed conditions revealed that the 14-3-3 function(s) might be associated with the microvesicle biogenesis. Our results support that 14-3-3 has diverse pertinent roles in both physiology and pathogenesis in C. neoformans. Its gene functions are closely relevant to the pathogenesis of this fungus.

• Journal of Microbiology and Biotechnology
• /
• v.12 no.2
• /
• pp.249-257
• /
• 2002

Pseudomonas fluorescens GL20 is a plant growth-promoting rhizobacterium that produces a large amount of hydroxamate siderophore under iron-limited conditions. The strain GL20 considerably inhibited the spore germination and hyphal growth of a plant pathogenic fungus, Fusarium solani, when iron was limited, significantly suppressed the root-rot disease on beans caused by F. solani, and enhanced the plant growth. The mechanism for the beneficial effect of strain GL20 on the disease suppression was due to the siderophore production, evidenced by mutant strains derived from the strain. Analysis of the outer membrane protein profile revealed that the growth of strain GL20 induced the synthesis of specific iron-regulated outer membrane proteins with molecular masses of 85- and 90 kDa as the high-affinity receptors for the ferric siderophore. In addition, a cross-feeding assay revealed the presence of multiple inducible receptors for heterologous siderophores in the strain. In order to induce increased efficacy and potential in biological control of plant disease, a siderophore-overproducing mutant, GL20-S207, was prepared by NTG mutagenesis. The mutant GL20-S207 produced nearly 2.3 times more siderophore than the parent strain. In pot trials of beans with F. solani, the mutant increased plant growth up to 1.5 times compared with that of the parent strain. These results suggest that the plant growth-promoting P. fluorescens GL20 and the genetically bred P. fluorescens GL20-S207 can play an important role in the biological control of soil-borne plant diseases in the rhizosphere.

• Journal of Life Science
• /
• v.20 no.2
• /
• pp.176-182
• /
• 2010

Magnaporthe oryzae is destructive plant-pathogenic fungus and causes rice blast. The pathogen uses several mechanisms to circumvent the inhibitory actions of fungicides. ATP-binding cassette (ABC) transporters are known to provide protection against toxic compounds in the environment. PC facilitated bioinformatic analysis, particularly with respect to accessing and extracting database information and domain identification. We predicted ABC transporter genes from the M. oryzae genome sequence with computation and bioinformatics tools. A total of thirty three genes were predicted to encode ABC transporters. Three of thirty three putative genes corresponded to three known ABC transporter genes (ABC1, ABC2 and ABC3). Copy numbers of the ABC transporter genes were proven by Southern blot analysis, which revealed that twenty genes tested exist as a single copy. We amplified the DNA complementary to RNA corresponding to eleven of these by reverse transcriptase polymerase chain reaction.

• Korean Journal of Agricultural Science
• /
• v.9 no.1
• /
• pp.275-283
• /
• 1982

One hundred and sixteen seed samples of 16 different crops were tested by routine seed health testing methods. Seeds of rice, soybean and barley were found to be associated with 18, 15 and 7 species of fungi, respectively. Discoloration and spotting of various kinds were observed in rice seeds. Epicoccum purpurascens, Pyricularia oryzae and Trichoconiella pedwickii were higher in these discoloured than in normal seeds of rice. Macrophomina phaseolina, the charcoal-rot fungus is a new record for soybean in Korea. Seeds of sesame were infected by 13 fungi Alternaria sesami, A. sesarmicola and Corynespora cassiicola were detected predominantly. Twenty one species of fungi were detected in seed samples of 12 commercial vegetable crops. Alternaria, Colletotrichum and Fusarium were found predominantly. Some seed samples of commercial vegetable crops were heavily infected with pathogenic fungi and showed heavy seed and seedling rot.

• Journal of Microbiology
• /
• v.43 no.3
• /
• pp.308-312
• /
• 2005

The global RNA transcription profiles of Bacillus lentimorbus WJ5 under an in vitro co-culture with Colletotrichum gloeosporioides were analyzed in order to study the antagonistic bacteria-fungi interactions. Using a filter membrane system, B. lentimorhus WJ5 was exposed to the spores of C. gloeosporioides at the late exponential stage. The transcription profiles of the B. lentimorhus WJ5, both with and without a challenge from C. gloeosporioides, were analyzed using custom DNA chips containing 2,000 genome fragments. A total of 337 genes were expressed, with 87 and 47 up- and down-regulated, respectively. Of these, 12 genes, which were involved in central carbon metabolisms, and 7 from minor catabolism were relatively highly up-regulated (> 10 fold) and down-regulated (< 0.2 fold), respectively. Nine genes, which were thought to be related to the antifungal activity, were also up-regulated, but their levels were not so high (2.0 - 9.7 folds). From the results, during the early stage of the co-culture of B. lentimorbus WJ5 and C. gloeosporioides, nutrient competition seemed to occur; therefore, the genes from central carbon metabolisms could be up-regulated, while those from minor catabolism could be down-regulated.

• Journal of Microbiology and Biotechnology
• /
• v.22 no.1
• /
• pp.141-146
• /
• 2012

Malassezia globosa is a common pathogenic fungus that causes skin diseases including dandruff and seborrheic dermatitis in humans. Analysis of its genome identified a gene (MGL_1677) coding for a putative NADPH-P450 reductase (NPR) to support the fungal cytochrome P450 enzymes. The heterologously expressed recombinant M. globosa NPR protein was purified, and its functional features were characterized. The purified protein generated a single band on SDS-PAGE at 80.74 kDa and had an absorption maximum at 452 nm, indicating its possible function as an oxidized flavin cofactor. It evidenced NADPH-dependent reducing activity for cytochrome c or nitroblue tetrazolium. Human P450 1A2 and 2A6 were able to successfully catalyze the O-deethylation of 7-ethoxyresorufin and the 7-hydroxylation of coumarin, respectively, with the support of the purified NPR. These results demonstrate that purified NPR is an orthologous reductase protein that supports cytochrome P450 enzymes in M. globosa.

• Korean Journal Plant Pathology
• /
• v.11 no.1
• /
• pp.9-16
• /
• 1995

Undiluted neoasozin solution (6.5% a.i.), an arsenic fungicide, was sprayed on 169 cankers of apple trees from early March to September in 1987 twice at intervals of one week without scraping off the affected barks. Among the treated cankers, 79.9% ceased to grow within 1∼7 weeks, 13.0% showed partial development, and 7.1% grew continuously to girdle the branches. The partially developed cankers, however, could also be cured by an additional spray after slightly piercing at the edge of cankers to facilitate the penetration of the chemical. When the canker growth was blocked, cankers were encircled by cracks developed at the marginal area of the cankers. If the cracks developed once, very few cankers grew beyond them. The above results suggest that the crack development may be the consequence of the host defense activity to wall off the pathogen. In addition to the curative efficacy, the neoasozin solution inhibited sporulation of the pathogenic fungus almost completely. However, the pathogen survived for more than three months in some cankers that externally appeared to be cured, suggesting that an indirect mode of action of the chemical against apple Valsa canker seems to be still more persuasive than the direct fungicidal effect. In the final examination conducted in the mid April of the next year, 72.7% of the cankers were completely cured by the two successive neoasozin treatments. Moreover the cure rate became 83.1% if that of partially developed cankers which were also completely cured by an additional treatment was also taken into account. Since 1989 when this method was widely applied in apple orchards in Korea, apple Valsa canker has been effectively controlled to reach a tolerable level.

• Research in Plant Disease
• /
• v.7 no.2
• /
• pp.93-101
• /
• 2001

Occurrence of Fusarium wilt was surveyed in fields of cruciferous vegetable crops in Korea from 1996 to 1998. The disease severely occurred up to 40% in fields of Chinese cabbage and radish but slightly in Fields of cabbage. A total of 123 isolates was obtained from roots of the diseased plants and identified as Fusarium oxysporum based on the morphological and cultural characteristics. Pathogenicity of nine isolates selected from the isolates was tested by artificial inoculation to the hosts. All the isolates had similar virulence on Chinese cabbage and cabbage, although there were some differences in virulence on cultivars tested among the isolates. The isolates from radish were more virulent to radish than those from Chinese cabbage and cabbage. All isolates from the crucifers were not virulent to eight species of vegetable crops except the crucifers. The results of pathogenicity tests showed that the pathotype of Chinese cabbage-infecting isolates was identical to that of cabbage-infecting isolates, but somewhat different from that of radish-infecting isolates.

• Journal of Microbiology and Biotechnology
• /
• v.4 no.4
• /
• pp.296-304
• /
• 1994

Antibiotic substances were produced by Bacillus subtilis YB-70, a potential biocontrol agent found to suppress root-rot of eggplant (Solanum melonggena L) caused by Fusarium solani, in a dextrose glutamate medium and isolated by isoelectric precipitation. Partial purification was performed by column chromatography on silica gel with two solvent systems: chloroform-methanol and methanol-chloroform-water as eluting solvents, This active fraction YBS-1 s contained antifungal activity were soluble in ethanol, methanol, and water, but were not soluble in other solvents including acetone, butanol, ethyl ether, dimethylformamide, propanol, and etc. High performance liquid chromatography and thin layer chromatographic separation of YBS-1s showed that they have been composed of three biological active bands that were named YBS-1A, -1B, and -1C. The substances were stable to heat and resistant to protease. YBS-1s were active against a wide range of plant pathogenic fungi but did not inhibit the growth of bacteria and yeasts. They were not only fungicidal but also fungistatic against chlamydospores of F. solani. The $ED_{50}$ values for the chlamydospore germination and the germ-tube growth of F. solani were $O.725\mu\textrm{m}/ml\;and\;O.562\mu\textrm{m}/ml$, respectively. Microscopic observations proved the substances restricted the growth of phytopathogenic fungus F. solani by spore burst followed by dissolving of its germ-tube, and caused abnormal hyphal swelling after application to chlamydospores or growing hyphae. Cultural filtrate of B; subtilis YB-70 also suppressed the development of root-rot of eggplant in pot tests.