• Journal of Microbiology and Biotechnology
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• v.19 no.1
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• pp.11-16
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• 2009

Magnaporthe oryzae, the causal agent of rice blast, forms a specialized infection structure, called an appressorium, which is crucial for penetration and infection of the host plant. Pharmacological data suggest that calcium/calmodulindependent signaling is involved in appressorium formation in this fungus. To understand the role of the calcium/calmodulin-activated protein phosphatase on appressorium formation at the molecular level, MCNA, a gene encoding the catalytic subunit of calcineurin, was functionally characterized in M. oryzae. Transformants expressing sense/antisense RNA of MCNA exhibited significant reductions in mycelial growth, conidiation, appressorium formation, and pathogenicity. cDNA of MCNA functionally complemented a calcineurin disruptant strain (cmp1::LEU2 cmp2::HIS3) of Saccharomyces cerevisiae. These data suggest that calcineurin A plays important roles in signal transduction pathways involved in the infection-related morphogenesis and pathogenicity of M. oryzae.

• Research in Plant Disease
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• v.25 no.4
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• pp.205-212
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• 2019

Analysis of differentially expressed genes has assisted discovery of gene sets involved in particular biological processes. The purpose of this study was to identify genes involved in appressorium formation in the rice blast fungus Magnaporthe oryzae via analysis of cDNA-amplified fragment length polymorphisms. Amplification of appressorial and vegetative mycelial cDNAs using 28 primer combinations generated over 200 differentially expressed transcript-derived fragments (TDFs). TDFs were excised from gels, re-amplified by PCR, cloned, and sequenced. Forty-four of 52 clones analyzed corresponded to 42 genes. Quantitative real-time PCR showed that expression of 23 genes was up-regulated during appressorium formation, one of which was the MCK1 gene that had been shown to be involved in appressorium formation. This study will be providing valuable resources for identifying the genes such as pathogenicity-related genes in M. oryzae.

• The Plant Pathology Journal
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• v.26 no.1
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• pp.17-24
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• 2010

A logistic model for describing combined effects of both temperature and wetness period on appressorium formation was developed using laboratory data on percent appressorium formation of Colletotrichum acutatum. In addition, the possible use of the logistic model for forecasting infection risks was also evaluated as compared with a first-order linear model. A simplified equilibrium model for enzymatic reactions was applied to obtain a temperature function for asymptote parameter (A) of logistic model. For the position (B) and the rate (k) parameters, a reciprocal model was used to calculate the respective temperature functions. The nonlinear logistic model described successfully the response of appressorium formation to the combined effects of temperature and wetness period. Especially the temperature function for asymptote parameter A reflected the response of upper limit of appressorium formation to temperature, which showed the typical temperature response of enzymatic reactions in the cells. By having both temperature and wetness period as independent variables, the nonlinear logistic model can be used to determine the length of wetness periods required for certain levels of appressorium formation under different temperature conditions. The infection model derived from the nonlinear logistic model can be used to calculate infection risks using hourly temperature and wetness period data monitored by automated weather stations in the fields. Compared with the nonlinear infection model, the linear infection model always predicted a shorter wetness period for appressorium formation, and resulted in significantly under- and over-estimation of response at low and high temperatures, respectively.

• The Plant Pathology Journal
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• v.37 no.2
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• pp.87-98
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• 2021

To establish an infection, fungal pathogens must recognize diverse signals from host surfaces. The rice blast fungus, Magnaporthe oryzae, is one of the best models studying host-pathogen interactions. This fungus recognizes physical or chemical signals from the host surfaces and initiates the development of an infection structure called appressorium. Here, we found that protein MoAfo1(appressorium formation, MGG_10422) was involved in sensing signal molecules such as cutin monomers and long chain primary alcohols required for appressorium formation. The knockout mutant (ΔMoafo1) formed a few abnormal appressoria on the onion and rice sheath surfaces. However, it produced normal appressoria on the surface of rice leaves. MoAfo1 localized to the membranes of the cytoplasm and vacuole-like organelles in conidia and appressoria. Additionally, the ΔMoafo1 mutant showed defects in appressorium morphology, appressorium penetration, invasive growth, and pathogenicity. These multiple defects might be partially due to failure to respond properly to oxidative stress. These findings broaden our understanding of the fungal mechanisms at play in the recognition of the host surface during rice blast infection.

• Journal of Microbiology and Biotechnology
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• v.16 no.7
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• pp.1158-1162
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• 2006

The glyoxylate cycle can conserve carbons and adequately supply tricarboxylic acid (TCA) cycle intermediates for biosynthesis when microorganisms grow on $C_{2}$ carbon sources. It has been reported that isocitrate lyase (ICL1), a key enzyme of the glyoxylate cycle, is highly induced when Magnaporthe grisea, the causal agent of rice blast, infects its host. Therefore, the glyoxylate cycle is considered as a new target for antifungal agents. A 1.6-kb DNA fragment encoding the ICL1 from M. grisea KJ201 was amplified by PCR, cloned into a vector providing His-tag at the N-terminus, expressed in Escherichia coli, and purified using Ni-NTA affinity chromatography. The molecular mass of the purified ICL1 was approximately 60 kDa, as determined by SDS-PAGE. The ICL1 inhibitory effects of TCA cycle intermediates and their analogs were investigated. Among them, 3-nitropropionate was found to be the strongest inhibitor with an $IC_{50}$ value of $11.0{\mu}g/ml$. 3-Nitropropionate inhibited the appressorium development in M. grisea at the ${\mu}M$ level, whereas conidia germination remained unaffected. This compound also inhibited the mycelial growth of the fungus on minimal medium containing acetate as a $C_{2}$ carbon source. These results suggest that ICL1 plays a crucial role in appressorium formation of M. grisea and is a new target for the control of phytopathogenic fungal infection.

• 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.

• Journal of Microbiology and Biotechnology
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• v.22 no.7
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• pp.889-893
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• 2012

Appressorium is a specialized infection structure of filamentous pathogenic fungi and plays an important role in establishing a pathogenic relationship with the host. The Egh16/Egh16H family members are involved in appressorium formation and pathogenesis in pathogenic filamentous fungi. In this study, a homolog of Egh16H, Magas1, was identified from an entomopathogenic fungus, Metarhizium acridum. The Magas1 protein shared a number of conserved motifs with other Egh16/Egh16H family members and specifically expressed during the appressorium development period. Magas1-EGFP fusion expression showed that Magas1 protein was not localized inside the cell. Deletion of the Magas1 gene had no impact on vegetative growth, conidiation and appressorium formation, but resulted in a decreased mortality of host insect when topically inoculated. However, the mortality was not significant between the Magas1 deletion mutant and wild-type treatment when the cuticle was bypassed by injecting conidia directly into the hemocoel. Our results suggested that Magas1 may influence virulence by affecting the penetration of the insects' cuticle.

• The Korean Journal of Mycology
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• v.21 no.3
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• pp.224-229
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• 1993

The optimum temperature for germination of conidia and germ tube elongation were between $20\;and\;30^{\circ}C$ in C. dematium and C. gloeosporioides. Appressoria were fairly formed well at $20^{\circ}C$ despite the delay of conidial germination. At $30^{\circ}C$, both the germination and germ tube elongation are favored, but appressoria were poorly detected to be formed. In C. acutatum, the optimum temperature for germination of conidia was from $20\;to\;30^{\circ}C$, but at $25^{\circ}C$, germ tube elongation are accelerated. The conidia become septate and one or both doughter cells become conidiogenous instead of producing germ tubes and a secondary conidia produced, resulting in an arborescent type of connected conidia. Appressoria are infrequently formed by germinating conida. At $20\;to\;25^{\circ}C$ was the optimum for appressorium formation. But conidia that germinated at $30^{\circ}C$ seemed to lose the ability to form appressoria. The relation of temperature to germination of conidia and appressorium formation in Colletotrichum acutatum, C. dematium and C. gloeosporioides are discussed.

• 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.

• Korean Journal Plant Pathology
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• v.3 no.2
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• pp.100-107
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• 1987

Pre-pentration activity of Pyricularia oryzae Cav. from the stage of conidia germination to appressorium formation was examined on rice leaf epidermis under light and scanning electron microscopes to determine the causes· for differences in blast susceptibility between plants grown under three different soil moisture conditions in the greenhouse. No significant differences were found in the external shape of leaf epidermal cells including bulliform cells between plants grown under different soil moisture conditions. Growth and orientation of germ tube and morphology and size of appressorium of P. oryzae did not vary with soil moisture treatment. Site of appressorium formation was consistent over soil moisture treatment with the highest frequency of bulliform cell $(35\~48\%)$, followed by short cell $(19\~27\%)$, and long and guard cells $(13\~20\%)$. No appressorium was formed on trichome. This result suggests that the observed differences in blast susceptibility between plants grown under different soil moisture conditions were not due to the differences in the pre-pentration activity of P. oryzae on those plants.