• Title/Summary/Keyword: spore analysis

Search Result 186, Processing Time 0.027 seconds

"A La Recherche" of Functions for the Spore Protein SASP-E from Bacillus subtilis

  • Ruzal, Sandra M.;Bustos, Patricia L.;Sanchez-Rivas, Carmen
    • Journal of Microbiology and Biotechnology
    • /
    • v.23 no.1
    • /
    • pp.15-21
    • /
    • 2013
  • We previously observed that Bacillus subtilis spores from sspE mutants presented a lower germination capacity in media containing high salt concentrations (0.9M NaCl). This deficiency was attributed to the absence of SASP-E (gamma-type small-acid-soluble protein), rich in osmocompatible amino acids released by degradation. Herein we observed that, in addition, this mutant spore presented a reduced capacity to use L-alanine as germinant (L-ala pathway), required longer times to germinate in calcium dipicolinate ($Ca^{2+}$-DPA), but germinated well in asparagine, glucose, fructose, and potassium chloride (AGFK pathway). Moreover, mild sonic treatment of mutant spores partially recovered their germination capacity in L-ala. Spore qualities were also altered, since sporulating colonies from the sspE mutant showed a pale brownish color, a higher adherence to agar plates, and lower autofluorescence, properties related to their spore coat content. Furthermore, biochemical analysis showed a reduced partition in hexadecane and a higher content of $Ca^{2+}$-DPA when compared with its isogenic wild-type control. Coat protein preparations showed a different electrophoretic pattern, in particular when detected with antibodies against CotG and CotE. The complementation with a wild-type sspE gene in a plasmid allowed for recovering the wild-type coat phenotype. This is the first report of a direct involvement of SASP-E in the spore coat assembly during the differentiation program of sporulation.

Isolation and Analysis of Temperature Sensitive Mutants in Aspergillus nidulans (Aspergillus nidulans 온도감수성 돌연변이주의 분리 및 분석)

  • 박찬규;강현삼
    • Korean Journal of Microbiology
    • /
    • v.16 no.3
    • /
    • pp.111-121
    • /
    • 1978
  • About 40 temperature-sensitive mutants have been isolated as a preliminary step to study the spore germination, the cell cycle, and the control of macromolecular synthesis in Aspergillus nidulans. To obtain temperature-sensitive mutants rapidly and effectively, the selective enrichment method using antifungal antibiotic nystatin was developed. Based on the data which had applied to the concentration of auxotrophic mutants by the earlier investigators, the optimal concentration and the time of treatment at the nonpermissive temeprature were determined as 50 to 100 units per ml and 4.5 hr., respectively. Out of 41 ts mutants assigned to the strain symbol PK, thirteen that seemed to be arrested at the earlystage of spore germination were subjected to the further cytological and genetic analysis. Elght of these mutants are able to form germ tube and five not. Staining with acid fuchsin for the 5PK strains shows that one irreversible mutant, PK6 strain able to form germ tube, accumulate mitotic spindle, being arrested in mitosis. Another PK15 and PK23 strain have more than one intact nucleolus without germ tube formation at the restrictive temperature. the temperature-senstive mutation in PK12 strain, the onlystrain which is able occurred in certain gene specific for the germination of spore. All of the ts markers are recessive and complement each other in heterokaryon between two different ts markers at the restrictive temperature.

  • PDF

Spore-Pollen Specters of Deed Turnn Lake Terrace, Darkhad Depression, Mongolia

  • Punsalpaamuu, G.
    • The Korean Journal of Quaternary Research
    • /
    • v.17 no.2
    • /
    • pp.39-41
    • /
    • 2003
  • Pollen and spores as well as their morphological characteristics are biological stability materials in which contains historical information about vegetation and climate changes. As noted researcher Neishtadt in 1971, the lake and swamp deposits were developed only in Holocene period. The geological history of Darkhad depression which is situated in the Northern end of Mongolia from ancient time become under scientific interest of many researchers. Our investigation was focused on pollen analysis of lake sediment of Darkhad depression which caused bottom of paleolake. In Mongolia the palynology science is beginning to develop since 1990 and in 2000 the laboratory of palynology was established at the department of Biology, State Pedagogical University of Mongolia. Researchers from this laboratory working on pollen morphology of present flora in order to classify plants taxonomy. Another part of our investigation is mellitopalynology, which is studying of pollen grains in a honey. Although, this research has been extending by pollen analysis of sediment and lake deposits in past few years on the basis of high sensitive microscope and modem technology.

  • PDF

Spore Display Using Bacillus thuringiensis Exosporium Protein InhA

  • Park, Tae-Jung;Choi, Soo-Keun;Jung, Heung-Chae;Lee, Sang-Yup;Pan, Jae-Gu
    • Journal of Microbiology and Biotechnology
    • /
    • v.19 no.5
    • /
    • pp.495-501
    • /
    • 2009
  • A new spore display method is presented that enables recombinant proteins to be displayed on the surface of Bacillus spores via fusion with InhA, an exosporium component of Bacillus thuringiensis. The green fluorescent protein and $\beta$-galactosidase as model proteins were fused to the C-terminal region of InhA, respectively. The surface expression of the proteins on the spores was confirmed by flow cytometry, confocal laser scanning microscopy, measurement of the enzyme activity, and an immunogold electron microscopy analysis. InhA-mediated anchoring of foreign proteins in the exosporium of Bacillus spores can provide a new method of microbial display, thereby broadening the potential for novel applications of microbial display.

The Environmental Change at Kimpo Alluvial Plain during the Upper Holocene (김포충적평야의 홀로세 후기 환경변화)

  • 윤순옥;김혜령
    • The Korean Journal of Quaternary Research
    • /
    • v.15 no.2
    • /
    • pp.83-91
    • /
    • 2001
  • The environmental change during the upper Holocene was restored at Kimpo alluvial plain on the left bank of Han-river near the Yellow Sea according to the results of pollen analysis and carbon dating, based on the sea-level change from Ilsan area. Pollenzone I (5.8~7.0 m.a.s.l.) was the period of AP-dominance, including mostly Alnus. Study area was under the lagoon or swamp environment influenced by Flandrian transgression during 5,000~3,200 BP. Pollenzone II (7.0~7.4 m.a.s.l.) was the period of Spore & NAP-dominance. It represents that the vegetational environment was changed to drier condition by falling underwater surface caused by sea-level regression, and influenced by human activity during 3,200~2,300 BP. Subzone I a represented the characteristics of the climax of transgression. During subzone I b, herbs with the dominant Alnus were increased gradually, and it was correlated to the stagnation of high sea-level. Subzone II a was the dominant period of Spore by the gradual falling of sea-level. After that, the study area in swamp was emerged to the lowland by the fallen sea-level and herbs, especially Gramineae increased suddenly during Subzone II b. The sudden increase of NAP such as Artemisia, Chenopodiaceae and Umbelliferae with Gramineae during this period indicates the beginning of agriculture at this study area.

  • PDF

Halotolerant Spore-Forming Gram-Positive Bacterial Diversity Associated with Blutaparon portulacoides (St. Hill.) Mears, a Pioneer Species in Brazilian Coastal Dunes

  • Barbosa Deyvison Clacino;Irene Von Der Weid;Vaisman Natalie;Seldin Lucy
    • Journal of Microbiology and Biotechnology
    • /
    • v.16 no.2
    • /
    • pp.193-199
    • /
    • 2006
  • Halotolerant spore-forming Gram-positive bacteria were isolated from the root, rhizosphere, and non-rhizosphere soil of Blutaparon portulacoides. The different isolates were characterized genetically using an amplified ribosomal DNA restriction analysis (ARDRA), and phenotypically based on their colonial morphology, physiology, and nutritional requirements. Three different 16S rRNA gene-based genotypes were observed at a 100% similarity using the enzymes HinfI, MspI, and RsaI, and the phenotypic results also followed the ARDRA groupings. Selected strains, representing the different ARDRA groups, were analyzed by 16S rDNA sequencing, and members of the genera Halobaeillus, Virgibacillus, and Oceanobacillus were found. Two isolates showed low 16S rDNA sequence similarities with the closest related species of Halobacillus, indicating the presence of new species among the isolates. The majority of the strains isolated in this study seemed to belong to the species O. iheyensis and were compared using an AP-PCR to determine whether they had a clonal origin or not. Different patterns allowed the grouping of the strains according to Pearson's coefficient, and the resulting dendrogram revealed the formation of two main clusters, denoted as A and B. All the strains isolated from the soil were grouped into cluster A, whereas cluster B was exclusively composed of the strains associated with the B. portulacoides roots. This is the first report on the isolation and characterization of halotolerant spore-forming Gram-positive bacteria that coexist with B. portulacoides. As such, these new strains may be a potential source for the discovery of bioactive compounds with industrial value.

Role of Sugars in Early Stage of Spore Germination in Filamentous Fungi, Aspergillus nidulans (사상균인 Aspergillus nidulans의 무성포자 발아와 당의 역할)

  • Chung, Kwang-Hee;Kim, Jae Won
    • The Korean Journal of Mycology
    • /
    • v.46 no.4
    • /
    • pp.511-518
    • /
    • 2018
  • Initiation of spore germination in filamentous fungi such as Aspergillus nidulans and Botrytis cinerea requires the presence of nutrients. In this study, involvement of sugar sensing machinery was suggested in the germination of A. nidulans spores. Germination did not occur when the spores of A. nidulans were incubated in distilled water, whereas they were successfully germinated in the presence of 5% glucose with a germination rate of over 98% after 6hr incubation. Similar results were obtained when the spores were incubated in the presence of various sugars such as fructose, sucrose, and starch. Interestingly, spore germination was not observed in the presence of D-arabinose, whereas L-arabinose could induce germination as determined by the formation of germ tubes, indicating the presence of sugar sensing machinery that distinguish between the enantiomers of sugars. This inference was further supported by a decrease in germination rate (less than 25%) upon treatment of spores with trypsin. Subsequent MALDI-TOF mass spectrometry analysis of the surface proteins of spores identified ten proteins among which eight were involved in sugar metabolism. Taken together, our results suggest that spore germination in A. nidulans is initiated by the interaction of sugars with sugar binding proteins on the surface of spores.

Characteristics of Transformants in Pleurotus florida (사철느타리버섯 형질전환주(形質轉換珠)의 특성(特性))

  • Byun, Myung-Ok;Kim, Kyung-Soo;Cha, Dong-Yeol
    • The Korean Journal of Mycology
    • /
    • v.20 no.3
    • /
    • pp.222-228
    • /
    • 1992
  • Pleurotus florida was transformed by complementation of auxotrophic mutant using chimeric plasmid containing Flammulina velutipes leu 2 gene and pBR 322 replicon. Mycelial morphology of transformants was grown and compared on mushroom complete and minimal medium. Transformants were mated with monokaryon and their genetic recombination was investigated for the morphology of fruitbody and spore analysis. $Leu^+$ transformant showed same mating type of $A_1B_1$ as to the untransformed mutant. The transformant and the untransformed mutant were mated with monokaryon of which mating type is $A_2B_1$, respectively. Although fruitbody of the untransformed mutant was not produced, $leu^+$ transformant produced fruitbody. Spore analysis showed that leucine requiring spores from fruitbody of $leu^+$ transformant were diminished when compared with those of untransformed mutant.

  • PDF

Anti-aging & Skin Hydration Effects of Spore oil Extracted from Ganoderma lucidum (영지버섯에서 추출한 포자오일의 항노화 및 보습 효능)

  • Song, Hwan;Kim, Myun Soo
    • Journal of Convergence for Information Technology
    • /
    • v.10 no.5
    • /
    • pp.232-238
    • /
    • 2020
  • This study evaluated the anti-aging activity with antioxidant, anti-inflammatory and moisture activity of Ganoderma lucidum spore oil(GLS). GLS increased DPPH radical scavenging activity in a dose-dependent manners. Anti-inflammatory assay measured the inhibitory effect of GLS on NO, TNF-α and IL-6 production in LPS-stimulated RAW264.7 cells. As a result GLS inhibited NO and pro-inflammatory cytokine, TNF-α, IL-6 production. Also using human fibroblast cell to the procollagen production analysis and COL1A1 mRNA expression level analysis for defining, and for AQP-3 mRNA expression level analysis, used human keratinocyte cell. GLS increased procollagen production and COL1A1, AQP-3 mRNA expression. Our results suggest that the GLS have potential anti-inflammatory and wrinkle improves, skin moisture effect.

Inhibitory Activity of Oak Pyroligneous Liquor against Coleosporium Plectranthi, an Obligate Parasite Responsible for the Rust Disease on Perilla Leaf

  • Kumar, Varun;Chauhan, Anil Kumar;Baek, Kwang-Hyun;Kang, Sun-Chul
    • Korean Journal of Environmental Agriculture
    • /
    • v.30 no.4
    • /
    • pp.453-458
    • /
    • 2011
  • BACKGROUND: Coleosporium plectranthi, an obligate parasite, which is responsible for the rust disease of Perilla frutescens, a plant in Korea, commonly known as Perilla. All rusts are obligate parasites, meaning that they require a living host to complete their life cycle. They generally do not kill the host plant but can severely reduce growth and yield. Food and feed spoilage fungi cause great economic losses worldwide. It is estimated that between 5 and 10% of the world food production is wasted due to fungal deterioration. Rust disease of Perilla is highly frequent and is widely spread in Korea. The present study was designed to investigate a novel media for the urediniospore germination in vitro and anti-rust activity as well as GC-MS analysis of oak pyroligneous liquor. METHOD AND RESULTS: Urediniospores were collected from the infected leaf of Perilla. Spore suspension was made and the suspension was inoculated in the 2% water agar media with proper humidity, then they were incubated at $26^{\circ}C$ for 56 hrs. The GC-MS analysis of the oak pyroligneous liquor was also done to check the chemical composition. GC-MS analysis of the wood vinegar was found 15 compounds, among them o-mthoxyphenol (25.93%), 2,6-dimethoxyphenol (16.06%), 4-methylenecyclohexanone (10.69%), 2,3-dihydroxytoluene (7.84%), levoglucosane (6.14%) and propanoic acid (5.32%) were the major components. Different concentration of the oak pyroligneous liquor was used, and spore inhibition was recorded on the basis of spore counting. The best results were noted at the concentration of 50% solution where 31.8% spores were inhibited. CONCLUSION: On the basis of the chemical composition of the oak pyroligneous liquor and the activity recorded we can use it as an anti-rust agent.