• Title/Summary/Keyword: Conidia formation

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Effects of Isocitrate Lyase Inhibitors on Spore Germination and Appressorium Development in Magnaporthe grisea

  • Kim Seung-Young;Park Jin-Soo;Oh Ki-Bong
    • 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.

Antifungal Activity of Nor-securinine Against Some Phytopathogenic Fungi

  • Sahni, Sangita;Maurya, S.;Singh, U.P.;Singh, A.K.;Singh, V.P.;Pandey, V.B.
    • Mycobiology
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    • v.33 no.2
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    • pp.97-103
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    • 2005
  • Crude extracts and active principles from medicinal plants have shown potential role in controlling plant diseases in glasshouses as well as in fields as one of the safest and ecofriendly methods. The effect of nor-securinine (an alkaloid) isolated from Phyllanthus amarus has been seen against spore germination of some fungi (Alternaria brassicae, A. solani, Curvularia pennisetti, Curvularia sp., Erysiphe pisi, Helminthosporium frumentacei) as well as pea powdery mildew (Erysiphe pisi) under glasshouse conditions. The sensitivity of fungi to nor-securinine varied considerably. Nor-securinine was effective against most of the fungi. H. frumentacei was more sensitive even at the lowest concentration ($1,000\;{\mu}g/ml$). Likewise conidia of E. pisi were also inhibited in partially or completely appressorium formation. Pre-inoculation treatment showed greater efficacy than post-inoculation in inhibiting powdery mildew development on pea plants in a glasshouse. Maximum inhibition occurred at $2000\;{\mu}g/ml$.

Microscopic Examination of the Suppressive Action of Antifungal Substances from Pseudomonas aeruginosa on Asexual Sporulation of Fungi

  • Kwon S. Yoon;Bu Y. Min;Park, Hyoung T.;Lee, Jong K.;Kim, Kun W.
    • Journal of Microbiology
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    • v.37 no.1
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    • pp.27-34
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    • 1999
  • Two fractions with unusual antifungal activity that suppress asexual sporulation of several fungi were obtained from culture filtrate of Pseudomonas aeruginosa and were partially purified through the repeated silicagel flash column chromatographies. The sporulation-suppressive actions of these fractions in Aspergillus nidulans, Rhizopus stolonifer, and Coprinus cinereus, were analyzed by light and electron microscopes. The germination ability of the spores produced in the presence of these fractions were also checked to determine the persistent effects of these antifungal substances on the next generation. Light microscopic observation of developing sporangia of R. stolonifer grown in the presence of both fractions revealed that the significant number of sporangia failed to reach maturity, and frequently, uncontrolled growths of hyphae and rhizoids from the sporangiophores were found. In A. nidulans addition of these fractions appeared to cause different classes of morphological abnormality in conidia development, which included aborted formation of conidiogenous cells from the apex of conidiophores and enhanced hyphal growths either at the tip or middle of the conidiophores. Germination abilities of spores obtained from the cultures grown in the presence of antifungal fractions were 40∼60% in Aspergillus, 50∼80% in Coprinus (thallic spores), and 30∼40% in Rhizopus compared to those of normal spores.

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Effects of Temperature and Culture Media Composition on Sporulation, Mycelial Growth, and Antifungal Activity of Isaria javanica pf185

  • Lee, Jang Hoon;Lee, Yong Seong;Kim, Young Cheol
    • Research in Plant Disease
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    • v.27 no.3
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    • pp.99-106
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    • 2021
  • The fungal isolate Isaria javanica pf185 has potential as a mycopesticide because it demonstrates insecticidal activity against the green peach aphid and antifungal activity against Colletotrichum gloeosporioides. For commercialization of this isolate, determination of the optimal and least expensive culture conditions is required; however, these data are not currently available. This study describes the conditions for optimal development of conidia and production of metabolites for the biocontrol of the fungal pathogen. The optimal culture conditions were examined using cultures on solid agar and liquid media. High growth temperature enhanced spore formation but reduced antifungal activity in both solid and liquid media. The highest spore yield was obtained in a medium containing glucose as a carbon source and yeast extract as a nitrogen source. Soybean powder and wheat bran were effective nitrogen sources that promoted spore production and antifungal activity of the isolate. These results revealed the basic, cost-effective growth media for commercial production of a biopesticide with insecticidal and antifungal properties for use in integrated pest management.

Functions of PUF Family RNA-Binding Proteins in Aspergillus nidulans

  • Son, Sung-Hun;Jang, Seo-Yeong;Park, Hee-Soo
    • Journal of Microbiology and Biotechnology
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    • v.31 no.5
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    • pp.676-685
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    • 2021
  • RNA-binding proteins are involved in RNA metabolism and posttranscriptional regulation of various fundamental biological processes. The PUF family of RNA-binding proteins is highly conserved in eukaryotes, and its members regulate gene expression, mitochondrial biogenesis, and RNA processing. However, their biological functions in Aspergillus species remain mostly unknown in filamentous fungi. Here we have characterized the puf genes in the model organism Aspergillus nidulans. We generated deletion mutant strains for the five putative puf genes present in the A. nidulans genome and investigated their developmental phenotypes. Deletion of pufA or pufE affected fungal growth and asexual development. pufA mutants exhibited decreased production of asexual spores and reduced mRNA expression of genes regulating asexual development. The pufE deletion reduced colony growth, increased formation of asexual spores, and delayed production of sexual fruiting bodies. In addition, the absence of pufE reduced both sterigmatocystin production and the mRNA levels of genes in the sterigmatocystin cluster. Finally, pufE deletion mutants showed reduced trehalose production and lower resistance to thermal stress. Overall, these results demonstrate that PufA and PufE play roles in the development and sterigmatocystin metabolism in A. nidulans.

The Hybrid Formation between Aspergillus oryzae var. oryzae and Penicillium chrysogenum by Nuclear Transfer and the Production of Alkaline Protease. (핵전이에 의한 Aspergillus oryzae var. oryzae와 Penicillium chrysogenum의 잡종형성 및 Alkaline Protease생성)

  • 양영기;강희경;임채영;문명님
    • Microbiology and Biotechnology Letters
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    • v.26 no.4
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    • pp.290-296
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    • 1998
  • Interspecific hybrids between Aspergillus oryzae var. oryzae and Penicillium chrysogenum (Tyr$\^$-/), high alkaline protease producing fungi, were obtained by nuclear transfer technique. Nuclei isolated from the wild type Aspergillus oryzae var. oryzae strain were transferred into auxotrophic Penicillium chrysogenum mutants and selected the new strains showing an increased protein degrading capability. Maximum production of protoplasts were obtained by 1% Novozym 234 at $30^{\circ}C$ for 3 hours and the most effective osmotic stabilizers for the isolation of protoplasts were 0.6M KCl. Frequencies of hybrid formation by nuclear transfer were 1.3${\times}$10$\^$-3/∼2.8${\times}$10$\^$-3/. They could be suggested as an aneuploid by the observation of genetic stability, conidial size, DNA content, and nuclear strain. The hybrids showed 1.1~2.2 fold higher alkaline pretense activities than parental strains.

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Effect of Soil Moisture on the Pre-Penetration Activity of Pyricularia oryzae Cav. on Rice Leaf Epidermis (벼 잎 표피에서 도열병균의 침입전 행훈에 대한 토양수분의 효과)

  • Kim Choong Hoe
    • 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.

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Effect of light on conidial formation in several anthracnose fungi (몇가지 탄저병균의 분생포자형성에 미치는 광선의 영향)

  • Chung H. S.
    • Korean journal of applied entomology
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    • v.7
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    • pp.57-59
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    • 1969
  • Effect of light on conidial formation of Colletotrichum panacicola, C. gloeosporioides, C. runcatum and C. nicotianae was examined on potato dextrose agar on which each conidial suspension was seeded. The light source was Mitsubishi day light fluorescent lamps wi th an intensity of 100-120 ft-c at the colony surface. The treatments were (i)constant light, (ii) constant darkness, and (iii) alternating light (11hr.) and darkness {13hr.) at $26\{\circ}C\pm2$. With the exception of C. nicotianae, constant light gave the greatest sporulation over that obtained in the other two treatments; there were on significant differences in number of sporulation between under constant darkness and alternating light and darkness. C. nicotianae resulted in maximum. inter mediate and minimum sporulation under alternating light and darkness. constant darkness and constant light, respectively. These differences were statistically significant at $5\%$ level. C. truncatum and C. nicotianae produced abundant conidia in the entire test; C. gloeosporioides was intermediate, and C. panacicola was the least. Differences in number of sporulation due to light. organisms. and t he interaction of these two were all statistically significant.

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Structural and Functional Analysis of a Forkhead Gene, fkhF, in a Filamentous Fungus Aspergillus nidulans (사상성 진균 Aspergillus nidulans에서 forkhead 유전자인 fkhF의 구조와 기능 분석)

  • Park, Mi-Hye;Kim, Hyoun-Young;Kim, Jong-Hwa;Han, Kap-Hoon
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.312-317
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    • 2009
  • Genome analysis of a model filamentous fungus, Aspergillus nidulans, revealed that there are six putative forkhead genes. Among them, fkhF (AN8949.2) showed A. nidulans-specific. fkhF gene is located in chromosome VII and composed of 2,337 bp coding region for 778 amino acid. Since little is known about the involvement of the forkhead proteins in the developmental process of the filamentous fungi, including A. nidulans, we generated a deletion mutant of fkhF gene and analyzed. Deletion of fkhF resulted in less-dense conidiophore formation in a solid culture. However, the sexual developmental process or cleistothecia formation was normal. Furthermore, fkhF deletion mutant produced conidiophores and conidia under the submerged culture, suggesting that the fkhF gene is involved in repression of inappropriated induction and maturation of asexual developmental process but not in sexual development.

The Magas1 Gene is Involved in Pathogenesis by Affecting Penetration in Metarhizium acridum

  • Cao, Yueqing;Zhu, Xiangxian;Jiao, Run;Xia, Yuxian
    • 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.