• Mycobiology
• /
• v.51 no.5
• /
• pp.273-280
• /
• 2023

The nucleolus is the largest, membrane-less organelle within the nucleus of eukaryotic cell that plays a critical role in rRNA transcription and assembly of ribosomes. Recently, the nucleolus has been shown to be implicated in an array of processes including the formation of signal recognition particles and response to cellular stress. Such diverse functions of nucleolus are mediated by nucleolar proteins. In this study, we characterized a gene coding a putative protein containing a nucleolar localization sequence (NoLS) in the rice blast fungus, Magnaporthe oryzae. Phylogenetic and domain analysis suggested that the protein is orthologous to Rrp8 in Saccharomyces cerevisiae. MoRRP8-GFP (translational fusion of MoRRP8 with green fluorescence protein) co-localizes with a nucleolar marker protein, MoNOP1 fused to red fluorescence protein (RFP), indicating that MoRRP8 is a nucleolar protein. Deletion of the MoRRP8 gene caused a reduction in vegetative growth and impinged largely on asexual sporulation. Although the asexual spores of DMorrp8 were morphologically indistinguishable from those of wild-type, they showed delay in germination and reduction in appressorium formation. Our pathogenicity assay revealed that the MoRRP8 is required for full virulence and growth within host plants. Taken together, these results suggest that nucleolar processes mediated by MoRRP8 is pivotal for fungal development and pathogenesis.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.5 no.2
• /
• pp.183-188
• /
• 2002

Anti-fungal potential of 5 plant extracts viz., Eucalyptus citriodora, Allium sativum, Cassia sophera, Chromolaena odorata and Datura metel on the growth of mulberry brown leaf spot pathogen Myrothecium roridum were examined. Except fur the aqueous extract of Allium bulb, ethanolic leaf extract of all other plants more efficiently reduced the colony growth of the fungus on potato-dextrose-agar, Of which, Allium and Eucalyptus extracts were more effective. Initiation of radial growth of M. roridum on solid media was deferred maximum 6 days by ethanolic Eucalyptus extract and 4 days by aqueous Allium extract at $0.4 mg.ml^{-1}$. In the liquid media amended with Eucalyptus extract ($0.4 mg.ml^{-1}$) complete inhibition of sporulation was noticed upto 8 days, and initial inhibition of mycelial bio-mass generation was considerably diminished with time and reduction was 1.3 fold 14 days after application. While, complete inhibition of mycelial growth for 6-14 days was recorded with $\geq$0.1 mg.ml$^{-1}$ commercial eucalyptus oil. However, rejuvenation of growth appeared when fungus was re-inoculated in fresh media. Post-inoculate application of different doses Of Eucalyptus and Allium extracts significantly (p < 0.05) reduced the disease severity in pot-ted mulberry. However, persistence of the effect up to 28 days was apparent at $\geq$ 1.0 mg.ml$^{-1}$ and effectively was on par with carbendazim (1 mg.ml$^{-1}$ ). Almost equal control ability of 1.0 mg.ml$^{-1}$ Eucalyptus extracts can be achieved by ca. 10 times lowered dose of commercial eucalyptus oil. It seems, the toxic principle of E. citrodora to M. roridum is fungistatic in nature and may have essential oil based origin.

• Journal of Microbiology and Biotechnology
• /
• v.11 no.4
• /
• pp.552-557
• /
• 2001

By UV-irradiation of Eremothecium ashbyii DTl, a higWy flavinogenic mutant (UV-18-57) and a nonflavinogenic mutant (UV -85) were obtained. The physio-morphological characteristics of these three strains were studied on glucose medium in submerged fermentation. Glucose utilization and mycelial growth occurred in 0 - 2 days of fermentation. By the third day, the biomass had declined. Extracellular riboflavin excretion was distinct from the second day, reaching a maximum rate by the fourth day. The hyphae of the highly flavinogenic mutant UV-18-57 were broader than DTl, while the nonflavinogenic UV-85 hyphae were very thin. Riboflavin accumulation was high in UV-18-57 (extracellular riboflavin,$825\mu\textrm{g}/ml$ , and intracellular, $490\mu\textrm{g}/ml$) and caused the mycelia to swell into bulbous forms. Riboflavin accumulation was less in DTl ($108\mu\textrm{g}/ml$ extracellular and $24\mu\textrm{g}/ml$ intracellular) and correspondingly its hyphae were thinner than those of UV-18-57 and swollen bulbous mycelia were not prominent. UV-85 was nonflavinogenic and, accordingly, its mOlphological characteristics included long thin filaments with no intracellular riboflavin accumulation. A large number of greenish fluorescence spores were seen in UV-18-57, whereas DTI had less spores and UV-85 was nonsporulating. Sporulation is correlated with riboflavin production. UV-18-57 had better mycelial integrity and lysis started only by the seventh day, whereas DTI and UV -85 started to lyze earlier by 4 -5 days. By the late stage of fermentation (eighth day), DTl had a few long, thin filaments indicating some secondary growth, whereas UV -85 showed a compact pellet form of mycelia. Most mycelia of UV-18-57 still appeared intact.

• Proceedings of the Korean Society of Plant Pathology Conference
• /
• 2003.10a
• /
• pp.117.3-118
• /
• 2003

We tested for the presence of double-stranded RNA (dsRNA) mycovirus in 827 Fusarium graminearum isolated from diseased barley and maize. dsRNA mycoviruses with various sizes were isolated. Of them, it was previously reported that dsRNA from DK2l isolate had pronounced morphological changes, including reduction in mycelial growth, increased to red pigmentation, reduced virulence and sporulation. (Chu et al., Appl. Environ. Microbiol. 2002). For better understanding of this hypovirulence associated with DK2l dsRNA virus, we determined the complete nucleotide sequence of dsRNA genome and named Fusarium hypovirus DK2l strain (Fhv-DK2l ). Genomic RNA of Fhv-DK2l was determined to be 6625 nucleotides in length excluding the poly (A) tail and contained three putative open reading frame. RNA-dependent RNA polymerase (RdRp) and helicase domain were expected in ORF A, 54 to 4709 nucleotide position. ORE B, 4752 to 5216 nucleotide position, and ORF C, 5475 to 6578 nucleotide position, were predicted to encode 16.7kDa and 41.3kDa protein respectively each. We could not detect any conserved domains from these two proteins. Phylogenetic analysis showed Fhv-DK2l was related to Cryphonectria hypovirus 3. Ten additional isolates were found that were infected with dsRNA mycoviruses. These mycoviruses contain 2 to 4 different segments of dsRNAs with the size range of approximately 1.7 to 10-kbp in length. The presence of dsRNAs isolates did not affect colony morphology and were transmissible through conidia and ascospore with incidence of 30-100％. These results indicate that there is genomic diversity of dsRNA mycoviruses that infect F. graminearum isolates and that impact of virus infection on host's morphology and virulence is determined by the interaction between dsRNAs and the fungal host, not by the mere presence of the dsRNAs

• Korean journal of applied entomology
• /
• v.16 no.4 s.33
• /
• pp.211-215
• /
• 1977

The Sporulation of Didymella bryomiae were observed under diurnal cycles of light/darkness of near ultraviolet light (NUV) and artificial daylight (ADL) and continous darkness in eight isolates growing on PDA and V-8 juice agar. Light stimulated pycindial and perithecial formation of this fungus on potato dextrose agar and V-8 juice agar. Sprulation was poor in darkness, but some isolates were able to produce pycnidia and perithecia in the absence of light. Perithecial formation was much better under artificial daylight (ADL) on V-8 juice agar than those grown under near ultraviolet light (NUV). In general, cultures grown on V-8 juice agar sporulated better than cultures grown on PDA under three setsof light condition. Most of the pycnidiospores obtained from each isolates of this fungus grown on PDA were non-septate and microtype, but macrotype of non-septate and uniseptate pycnidiospores were produced on V-8 juice agar. Pycnidiospore produced on V-8 juice agar were similar to those produced on the radicle of naturally infected seeds. The appearance of perithecia were quite distinctive from pycnidia. The mature perithecia were darker than pycnidia and whitish spore masses formed on the ostiole of perithecia.

• CELLMED
• /
• v.4 no.1
• /
• pp.6.1-6.12
• /
• 2014

Anthrax is the deadly disease for human being caused by Bacillus anthracis. Instantaneous research work on the mode of infection of the organism revealed that different proteases are involved in different steps of pathogenesis. Present study reports the in silico characterization and the detection of pathogenic proteases involved in anthrax infection through protein-protein interaction. A total of 13 acid, 9 neutral, and 1 alkaline protease of Bacillus anthracis were selected for analysing the physicochemical parameter, the protein superfamily and family search, multiple sequence alignment, phylogenetic tree construction, protein-protein interactions and motif finding. Among the 13 acid proteases, 10 were found as extracellular enzymes that interact with immune inhibitor A (InhA) and help the organism to cross the blood brain barrier during the process of infection. Multiple sequence alignment of above acid proteases revealed the position 368, 489, and 498-contained 100% conserved amino acids which could be used to deactivate the protease. Among the groups analyzed, only acid protease were found to interact with InhA, which indicated that metalloproteases of acid protease group have the capability to develop pathogenesis during B. anthracis infection. Deactivation of conserved amino acid position of germination protease can stop the sporulation and germination of B anthracis cell. The detailed interaction study of neutral and alkaline proteases could also be helpful to design the interaction network for the better understanding of anthrax disease.

• The Korean Journal of Mycology
• /
• v.25 no.1 s.80
• /
• pp.30-34
• /
• 1997

Glomerella cingulata (conidial state: Colletotrichum gloeosporioides) was identified as the causal organism of anthracnose of citrus on the basis of morphological characteristics of the conidial state of the fungus isolated from infected leaves of Satsuma mandarin and its ascigerous state isolated from diseased twigs. The pathogen infected the leaves of Satsuma mandarin, citron and Natsu daidai only by wound inoculation. The optimum temperature range for mycelial growth and sporulation of conidia of the strain was $25{\sim}30^{\circ}C$, respectively. The characteristics of anthracnose strain of Satsuma mandarin such as growth rate and color of colony, shape and size of conidia, and appressoria were similar to those of FGG strain. However, the strain isolated from infected leaves and twigs of Satsuma mandarin was different from FGG strain to cause postharvest anthracnose of citrus, because some of morphological and pathological characteristics of the strain isolated did not correspond to those of FGG strain.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.16 no.1
• /
• pp.37-43
• /
• 1983

The effects of pH, sodium chloride and potassium sorbate on the germination of Bacillus cereus spores in the medium of cooked rice homogenate were studied. At the range of pH $4.5{\sim}10.0$, the germination of spores were observed. Germinated spores were reached to the number of $10^7/ml$ within 5 hours at $32^{\circ}C$ under the condition of pH 7.0, which was found as optimum pH of germination. In the range of sodium chloride $2{\sim}10\%$, the maximum growth were exhibited under $2\%$ concentration, while it proportionally decreased under the salinity condition higher than $5\%$. The growth of Bacillus cereus were inversely related to the concentration of potassium sorbate within the range of $0{\sim}0.2\%$. Maximum sporulation ratio was observed under the culturing condition: $10\%$ NaCl and $0.2\%$ potassium sorbate in the medium of cooked rice homogenate.

• The Plant Pathology Journal
• /
• v.24 no.2
• /
• pp.191-201
• /
• 2008

Bacillus sp. SB-007 was isolated from pea leaves harvested from the southwestern parts of South Korea through screening on a minimal medium containing 0.2% purified cutin for its ability to induce the cutinase production. However, no cutinase was produced when it was grown in a minimal medium containing 0.2% glucose. A proteomic approach was applied to separate and characterize these differentially secreted proteins. The expression level of 83 extracellular proteins of the cutinase-producing Bacillus sp. strain SB-007 incubated in a cutinase-induced medium increased significantly as compared with that cultured in a non cutinase-induced medium containing glucose. The extracellular proteome of Bacillus sp. SB-007 includes proteins from different functional classes, such as enzymes for the degradation of various macromolecules, proteins involved in energy metabolism, sporulation, transport/binding proteins and lipoproteins, stress inducible proteins, several cellular molecule biosynthetic pathways and catabolism, and some proteins with an as yet unknown function. In addition, the two protein spots showed little similarities with the known lipolytic enzymes in the database. These secreted proteome analysis results are expected to be useful in improving the Bacillus strains for the production of industrial cutinases.

• Microbiology and Biotechnology Letters
• /
• v.25 no.6
• /
• pp.566-570
• /
• 1997

During sporulation, Bacillus thuringiensis strains produce crystals consist of toxin proteins highly specific against insect pests. Their host specificities are desirable from a standpoint of environmental safety, but also limit market potential. Thus, development of improved Bacillus thuringiensis strains having broad host spectrum will contribute to increase its use. For the construction of Bacillus thuringiensis strain having broad host spectrum, we cloned cry1C gene encoding a toxin protein highly toxic against Spodoptera exigua from a B. thuringiensis isolate and constructed two recombinant plasmids, pUBClC and plC60. The plasmid PUBC1C has a replication origin of the natural plasmid pBC16 from B. cereus which is closely related species to B. thuringiensis, and the pBC16 was known to be replicated by rolling-circle mechanism. The plasmid pIC60 has a replication origin of a resident 60 MDa plasmid from B. thuringiensis subsp. kurstaki HD263, and it is believed that the pIC60 is replicated in a theta mode. The two plasmids were introduced into B. thuringiensis subsp. kurstaki cryB strain, and the transformed strains produced well-shaped bipyramidal crystals. We confirmed the expression of the cry1C gene by SDS-PAGE, and Western blotting. By investigating the segregational stability, it was found that the plasmid pIC60 is more stable than the pUBC1C.