• 한국식물병리학회지
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• 제14권5호
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• pp.413-417
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• 1998

Magnaporthe grisea, the casual agent of rice blast, requires formation of an appressorium, a dome-shaped and well melanized infection structure, to penetrate its host. Environmental cues that induce appressorium formation include hydrophobicity and hardness of contact surface and chemicals from its host. Artificial surfaces are widely used to induce appressorium formation, but frequencies of appressorium induction are not always consistent. To understand variable induction of appressorium formation in M. grisea, several factors were tested on GelBond. High levels of appressorium formation were induced over a wide range of temperature (20~3$0^{\circ}C$) and pH (4~7). spore age up to 3-week-old did not significantly affect appressorium formation, but only a few apressoria on GelBond. However, adenosine specifically inhibited appressorium formation. Adenosine inhibition of appressorium formation was restored by exogenous addition of cAMP. Germ tube tips of M. grisea maintained the ability to differentiate appressoria by chemical inducers on GelBond at least up to 16 h after conidia germination. These results suggest that environmental factors have little effect on the variable induction of appressorium formation on the artificial surface in M. grisea.

• The Plant Pathology Journal
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• 제27권3호
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• pp.225-231
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• 2011

Previously, we identified the 20S proteasome ${\alpha}$-subunit of Magnaporthe oryzae (M. oryzae) induced during appressorium formation, and detected an increase in multiple protein ubiquitination during the early appressorium formation process (Kim et al., 2004). In this study, we further attempted to determine whether the proteasome is involved in the appressorium formation of M. oryzae both in vitro and in planta, using proteasome inhibitors. A significant increase in 20S proteasome during fungal germination and appressorium formation was observed using Western blot analysis with 20S proteasome antibody, demonstrating that proteasome-mediated protein degradation was involved in appressorium formation. Pharmacological analysis using proteasome inhibitors, MG-132, proteasome inhibitor I (PI) and proteasome inhibitor II (PII) revealed that germination and appressorium formation were delayed for 4 to 6 h on rice leaf wax-coated plates. Similarly, the treatment of proteasome inhibitors with fungal conidia on the rice leaf surface delayed appressorium formation and host infection processes as well. Additionally, fungal pathogenicity was strongly reduced at 4 days' postfungal infection. These data indicated that the fungal 20S proteasome might be involved in the pathogenicity of M. oryzae by the suppression of germination and appressorium formation.

• Journal of Microbiology and Biotechnology
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• 제17권7호
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• pp.1198-1203
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• 2007

Conidial adhesion and appressorium formation of Magnaporthe oryzae on the rice surface are important early events in the infection process. As an initiative step to understand the mechanisms underlying these cellular processes at a biochemical level, the effect of a human fibronectin antibody (HFA) and RGD peptides on conidial adhesion and appressorium formation was evaluated. HFA inhibited conidial adhesion and appressorium formation in a dosage-dependent manner. RGD peptides also inhibited these cellular events. Conidial adhesion and appressorium formation inhibited by RGD peptides were restored by chemicals involved in the cyclic AMP-dependent signaling pathway. These results suggest that extracellular matrix proteins might be involved in conidial adhesion and appressorium formation through integrin-like receptor mediation and modulation of cAMP-dependent signaling in the cells.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.84.1-84
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• 2003

Magnaporthe grisea, the casual agent of rice blast, forms an appressorium to penetrate its host. Much has been learned about environmental cues and signal transduction pathways, especially those involving CAMP and MAP kinases, on appressorium formation during the last decade. More recently, pharmacological data suggest that calcium/calmodulin-dependent signaling system is involved in its appressorium formation. To determine the role of phosphoinositide-specific phospholipase C (PI-PLC) on appressorium formation, a gene (WPLCl) encoding PI-PLC was cloned and characterized from M. grisea strain 70-15. Sequence analysis showed that MPLCl has alt five conserved domains present in other phospholipase C genes from several filamentous fungi and mammals. Null mutants (mplcl) generated by targeted gene disruption exhibited pleiotropic effects on conidial morphology, appressorium formation, fertility and pathogenicity. mplcl mutants developed nonfunctional appressoria and are also defective in infectious growth in host tissues. Defects in appressorium formation and pathogenicity in mplcl mutants were complemented by a mouse PLCdelta-1 cDNA under the control of the MPLCl promoter. These results suggest that cellular signaling mediated by MPLCl plays crucial and diverse roles in development and pathogenicity of M. grisea, and functional conservation between fungal and mammalian Pl-PLCs.

• 한국균학회지
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• 제15권3호
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• pp.142-148
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• 1987

대두(大豆) 탄저병(炭疽病)(C. truncatum)을 공시(供試)하여 In vitro에서 광(光), pH 및 영양원(營養源)과 In vivo에서 시비수준(施肥水準)(P, K 및 Ca)이 부착기형성(附着器形成)에 미치는 영향(影響)을 조사(調査)한 바는 다음과 같다. 1. 3시간(時間) 및 6시간(時間)이 지난후의 조사(調査)에 따르면 광처리(光處理)를 한 것이 암처리(暗處理)를 한 것보다 부착기형성률(附着器形成率)이 양호(良好)하였다. 그러나 시간이 진행(進行)됨에 따라서는 각처리(各處理)에서 부착기형성률(附着器形成率)이 높아졌다. 2. 부착기형성(附着器形成)에 적정 pH는 $pH\;6{\sim}8$이었다. 3. 탄소원(炭素源) 및 질소원(窒素源) 공(供)히 무처리구(無處理區)에서 부착기형성률(附着器形成率)이 양호(良好)하였고 농도(濃度)가 높을수록 억제되는 경향이었으며 질소원(窒素源)에 비(比)하여 탄소원(炭素源)에서의 부착기형성률(附着器形成率)이 양호(良好)하였다. 그리고 시간(時間)이 경과함에 따라서 부착기형성률(附着器形成率)이 상승했다. 4. 3시간(時間) 경과후의 조사(調査)에서는 인산 및 칼륨처리(處理)는 증시(增施)했을 경우 다소 부착기형성(附着器形成)이 억제되었으나, 칼슘처리(處理)에 있어서는 무비구(無肥區)<표준비구(標準肥區)<2배비구(培肥區)의 순(順)으로 부착기형성(附着器形成)이 상당히 억제되는 경향이었다. 그러나 시간(時間)이 경과함에 따라서는 부착기형성률(附着器形成率)이 시비수준(施肥水準)과 관계없이 상당히 증가(增加)됨을 보였다.

• The Plant Pathology Journal
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• 제21권4호
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• pp.322-327
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• 2005

Regression models for determining infection periods of apple white rot were developed based on conidial germination and appressorium formation of Botryosphaeria dothidea. A total of 120 apple fruits were inoculated with the fungal conidial suspension and subjected to 6 temperatures and 10 wetness periods. Conidia germinated and produced appressoria, exhibiting swollen tips of germ tubes on the fruit surface. Conidial germination (G) increased with temperature (T) and wetness period (W), and was described as $G=-89.273+7.649T+7.056W-0.109T^{2}-0.085W^{2}-0.066TW(R^{2}=0.75)$. Less than 2 hr of wetness period were enough for conidia to germinate at 25 to $30^{\circ}C$. Effects of temperature and wetness period on appressorium formation (A) could be explained as $A=-1.540-2.375W+0.045W^{2}+0.213TW(R^{2}=0.77)$. The relationship between conidial germination and appressorium formation ($A_g$) was described as$A_g=0.381-0.227G+0.005G^{2}(R^{2}=0.67)$, suggesting that conidial germination may have to reach approximately $43.7\%$ to initiate appressorium formation. Using the regression equation for conidial germination and the criterion of $43.7\%$ conidial germination, an infection model was developed to determine infection periods based on temperature and wetness period. The infection model with the criterion of $43.7\%$ conidial germination was apparently more conservative than the appressorium formation model in determining possibility of apple infection. The infection model seemed sensitive to variable weather conditions, suggesting possible use of the model for timing fungicide sprays to control white rot of apples in practice.

• 한국식물병리학회지
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• 제10권4호
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• pp.254-260
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• 1994

Magnaporthe grisea, a causal agent of blast, forms a specialized infection structure, an appressorium, to infect host. Hydrophobicity of contact surface and cAMP have been suggested as a primary environmental signal and a second messenger to trigger and mediate appressorium formation in this fungus, respectively. To generalize these factors in field isolates of M. girsea, twenty isolates originated from rice and other gramineous hosts were tested. Seventeen including rice and non-rice isolates formed appressoria on hydrophobic surface, but none of isolates formed appressoria on hydrophilic surface. Eighteen isolates formed appressoria on hydrophilic surface in the presence of IBMX, an inhibitor of phosphodiesterase, except two rice isolates. These results strongly support the hypothesis that appressorium formation by M. grisea is induced by hydrophobic hard surface and regulated by the endogenous level of cAMP in the cells. Understanding fungal development is not only of biological interest but provides new targets for novel disease control strategies.

• 식물병연구
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• 제25권4호
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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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• 제19권1호
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• pp.34-42
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• 2003

The ascomycete Magnaporthe grisea is a pathogen of rice blast and is known to form specialized infection structures called appressoria for successful infection into host cells. To understand the molecular mechanism underlying infection process, appressorium-related genes were identified through global approaches including EST sequencing, differential hybridization, and sup-pression subtractive hybridization. EST database was generated on >2,000 cDNA clones randomly selected from appressorium stage cDNA library. Large number of ESTs showed homology to known proteins possibly involved in infection-related cellular development (attachment, germination, appressorium formation, and colonization) of rice blast fungus. The 1051 ESTs showing significant homology to known genes were assigned to 11 functional categories. Differential hybridization and suppression subtractive hybridization were applied to identify genes showing an appressorium stage specific expression pattern. A number of genes were selected as up-regulated during appressorium formation compared with the vegetative growing stage. Clones from various cDNA libraries constructed in different developmental stages were arrayed on slide glass for further expression profiling study. functional characterization of genes identified from these global approaches may lead to a better understand-ing of the infection process of this devastating plant disease, and the development of novel ways to protect host plant.

• The Plant Pathology Journal
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• 제26권1호
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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.