• Molecules and Cells
• /
• v.21 no.3
• /
• pp.411-417
• /
• 2006

We have developed a polymerase chain reactionrestriction fragment length polymorphism (PCR-RFLP) marker that can distinguish male-fertile (N) and male-sterile (S) cytoplasm in onions. The PCR-RFLP marker was located in a chloroplast psbA gene amplicon. Digesting the amplicons from different cytoplasm-containing varieties with the restriction enzyme MspI revealed that N-cytoplasm plants have a functional MspI site (CCGG), whereas the S-cytoplasm plants has a substitution in that site (CTGG), and thus no MspI target. The results obtained using this PCR-RFLP marker to distinguish between cytoplasmic male sterile factors in 35 onion varieties corresponded with those using a CMS-specific sequence-characterized amplified region (SCAR) marker. Moreover, the PCR-RFLP marker can identify N- ot S-cytoplasms in DNA sample mixtures in which they are in up to a 10-fold minority, indicating that use of the marker has high diagnostic precision. We also demonstrated the usefulness of the SNP detected in the psbA gene for high-throughput discrimination of CMS factors using Real-time PCR and a TaqMan probe assay.

• Food Science and Preservation
• /
• v.7 no.1
• /
• pp.124-131
• /
• 2000

Antibacterial activities of powdered spices(garlic , ginger, cinnamon and clove) against pathogenic Escherichia coli )157:H7 and Staphyloccus auresus were investigated. Spice powder was added in was exponetial phase of each bacterial culture . Growth inhibition was determined by the absorbance at 660nm and morphological changes of the cells were observed by transmission electron microscope (TEM). Ginger powder has the highest antibacterial activity, following cinnamon , clove and garlic has the least activity.Growth of Escherichia coli O157:H7 and Staphyloccus aureus were completely inhibited within 5 hours after addition of 1 % of garlic , 0.3% of ginger or cinnamon , 0.5% of clove powder on the exponential phase of the cells. Spice untreated cells of E. coli and S. aureus, the cytoplasm was entirely surrounded by rigid cell wall and cell walls formed a smooth layer well attached to the plasma membrane. In the cells of E. coli and S. aureus treated with spice powder, cell wall and plasma membrane were lysed and severely damaged. E.coli cells growth in the presence of spice powder showed plammolysis, the loss of electron dense material, the formation of extra cellular blebs and cytoplasm burst out from the cell. S .sureus cells grown in the presence of spice powder showed swell of cell wall, the loss of electron dense material , coagulation of cell cytoplasm and formation of extra cellular blebs. Severely damaged cells of S. aureus lost whole cytoplasm and left as ghost of the cell. Spice powder stimulated autolyssi and induced cell death.

• Mycobiology
• /
• v.39 no.2
• /
• pp.79-84
• /
• 2011

Nuclear distribution within the extra-radical fungal structures and during spore production in the arbuscular mycorrhizae fungus Glomus intraradices was examined using an in vitro monoxenic culture system. A di-compartmental monoxenic culture system was modified using a nitrocellulose membrane and a coverglass slip for detailed observations. Nuclear distribution was observed using the fluorescent DNA binding probes SYBR Green I and DAPI. Both septate and non-septate mycelial regions were observed, but cytoplasmic contents were only found within non-septate mycelia. Nuclear fluorescent staining revealed that the non-septate hyphal region contained nuclei only with cytoplasm, and that nuclear distribution was limited by septa. Swollen hyphal bodies were often associated with septate and empty-looking hyphae. Cytoplasmic contents filled the swollen hyphal body from the non-septate hyphal region following removal of the septa. As a consequence, the swollen body developed into a new spore. These observations provide understanding about the distribution of AM fungal nuclei within extra-radical mycelia and during spore formation. The results suggest a mechanism by which the development of a cytoplasm-containing mycelium is controlled by the formation or removal of septa to efficiently maintain and proliferate essential contents. This mechanism may provide a survival strategy to the fungus.

• The Plant Pathology Journal
• /
• v.17 no.2
• /
• pp.94-100
• /
• 2001

Roots of Chinese cabbage (Brassica campestris var. chinensis) seedlings infected with Plasmodiophora brassicae were examined by light and electron microscopy to reveal histopathological changes related to pathogenesis in the susceptible host. The pathogen colonized the cortex and partly the stele as well, invading up to the xylem. Gall tissues could be differentiated from the initially infected tissues, involving less compact organization and new vascular development. The infected cells were much hypertrophied, and contained one to several plasmodia. Except cellular hypertrophy, no pathological ultrastructural modification was noted in the infected calls. Infected cytoplasm became dense with ground cytoplasm, inconspicuous central vacuole, and increased cellular organelles such as mitochondria and dictyosomes. There were two types of nuclear states of plasmodium, uninucleate and multinucleate. Both plasmodia were structurally similar, filled with lipid droplets, bounded with envelope, and containing mitochondria, endo-plasmic reticulum, and sometimes small vacuoles. Plasmodial fragmentation, which may be regarded as a way to discharge plasmodial materials into host cytoplasm, commonly occurred, forming plasmodial fragments by outgrowth of plasmodial cytoplasm and regional compartmentalization. Plasmodial fragments were degenerated sometimes followed by forming chains of spherical vesicles especially in the uninucleate plasmodial state. These ultrastructural features indicate the biotrophic nature of the pathogen associated with its pathogenesis in the susceptible host.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.43 no.2
• /
• pp.59-63
• /
• 1998

Three different male-sterile cytoplasmic lines and their common maintainer 'Zhenshan 97B' and two elite restorer lines were used to study cytoplasmic effects on agronomic trait manifestation per se under different nitrogen supply levels. The result showed that cytoplasmic effects could be modified by nitrogen environments. The cytoplasmic effect on grain yield under 150 kg N/ha varied depending on crosses, while it was significantly negative in most crosses under both 60 and 330 kg N/ha. The correlation and path-coefficient analyses suggested that it was expected to improve cytoplasmic effects through reducing maximum tillers and increasing the percentage of productive tillers, leading to increased productive tillers and higher yield in hybrid rice by the aid of cultural practice and genetic transformation. This study also revealed that the same cytoplasm in different combinations had differential effect under the same nitrogen environment, indicating that cytoplasmic effect was produced by interaction of nuclear genes with cytoplasm rather than cytoplasm per se. These results indicated the usefulness of evaluating diverse cytoplasmic sources in various nuclear genotypes bred for hybrid rice breeding program. The finding also suggested that negative cytoplasmic effect could be effectively overcome by elite restorer lines through the interaction of nuclear genes with female cytoplasm.

• Applied Microscopy
• /
• v.30 no.3
• /
• pp.303-310
• /
• 2000

Ultrastructures on the integumentary epidermis of the marbled sole, Limanda yokahamae, were examined by means of the light and transmission electron microscope. Epidermal layer consists of supporting cells, unicellular glands and accessory cells. The supporting cells were classified into superficial cell, intermediated cell and basal cell. The cytoplasm of supporting cells is divided into cortex and medullar part. In the cortex and medullar part, microfilaments and cell organelles are well developed, respectively. Gland cells are present in the superficial and middle epidermis. The cytoplasm of mucous cell reacted to blue in AB-PAS (pH 2.5). Club cell has a roundish central vacuole and well-developed microfilaments in the cytoplasm. Granular cells are occurs in the middle and basal epidermis , and the cytoplasm is occupied with membrane-bounded granules of electron dense. Chloride cells are present in the superficial epidermis , and the cytoplasm is occupied with tubular mitochondria. Three types of pigment cells can be distinguished by electron density of cytoplasmic inclusions.

• Applied Microscopy
• /
• v.25 no.3
• /
• pp.63-74
• /
• 1995

The development of pale chub oocyte from the immature oogonium to mature oocyte was investigated by light and electron microscope. The cytoplasm of pale chub oogonia was acidic and many vesicles were located at inner side of nuclear membrane. In primary oocytes, yolk vesicles were distributed in cytoplasm. Also, fibrous materials and protuberances were distributed on the surface of zona radiata. The nucleus of secondary oocyte was enlarged and yolk vesicles in cytoplasm migrated to zona radiata. In early egg, yolk mass are formed and yolk vesicles were located at inner side of zona radiata. Three-layered zona radiata was about $3{\mu}m$ in thickness. The three layers were an outer fibrous material layer, a middle nurse cell layer in which microvilli of early egg cytoplasm contact with processes of nurse cells, and an inner layer with high electron density. In mature egg, euchromatin and a germinal vesicle were developed, mitochondria, free ribosomes, and yolk mass were distributed in cytoplasm. But, yolk vesicles were disappeared. Specially, zona radiata of matured eggs were better thin than the one of immature eggs In conclusion, it is summerized that the oogenesis of pale chub were the increase of cell size, the formation and accumulation of yolk, the decrease in nucleat electron density, changes of zona radiata, and the development of microvilli.

• Asian Pacific Journal of Cancer Prevention
• /
• v.14 no.10
• /
• pp.5789-5795
• /
• 2013

Background: The high mobility group box 1 (HMGB1) protein is a widespread nuclear protein present in most cell types. It typically locates in the nucleus and functions as a nuclear cofactor in transcription regulation. However, HMGB1 can also localize in the cytoplasm and be released into extracellular matrix, where it plays critical roles in carcinogenesis and inflammation. However, it remains elusive whether HMGB1 is relocated to cytoplasm in clear cell renal cell carcinoma (ccRCC). Methods: Nuclear and cytoplasmic proteins were extracted by different protocols from 20 ccRCC samples and corresponding adjacent renal tissues. Western blotting and immunohistochemistry were used to identify the expression of HMGB1 in ccRCC. To elucidate the potential mechanism of HMGB1 cytoplasmic translocation, HMGB1 proteins were enriched by immunoprecipitation and analyzed by mass spectrometry (MS). Results: The HMGB1 protein was overexpressed and partially localized in cytoplasm in ccRCC samples (12/20, 60%, p<0.05). Immunohistochemistry results indicated that ccRCC of high nuclear grade possess more HMGB1 relocation than those with low grade (p<0.05). Methylation of HMGB1 at lysine 112 in ccRCC was detected by MS. Bioinformatics analysis showed that post-translational modification might affect the binding ability to DNA and mediate its translocation. Conclusion: Relocation of HMGB1 to cytoplasm was confirmed in ccRCC. Methylation of HMGB1 at lysine 112 might the redistribution of this cofactor protein.

• Research in Plant Disease
• /
• v.12 no.1
• /
• pp.25-27
• /
• 2006

Rice dwarf phytoreovirus (RDV), a member of the family Reoviridae has a genome composed of 12 segmented dsRNAs designated as 51 to 512 with an increasing order of mobility in polyacrylamide gel electrophoresis (PAGE). RDV encode 12 structural and non-structural proteins, $P1{\sim}P12$ which are encoded by the $S1{\sim}S12$ segments of the dsRNA genome, respectively. In this experiment, we confirmed in situ localization of RDV particles and P12 in cytoplasm of infected rice plant. We observed specific reaction of the gold particles using virus particle and P12 protein specific antiserum with protein A-gold immunolabelling in electron microscope. It was observed that gold particles specifically react to virus particles in cytoplasm in case using the antiserum for virus particles. In the case of antiserum for P12 protein, gold particles sporadically existing on cytoplasm without existing in organelle of cytoplasm specifically. As this result, RDV P12 protein encoded by S12 located in cytoplasm.

• Applied Microscopy
• /
• v.25 no.1
• /
• pp.75-85
• /
• 1995

The goal of the present study is to investigate the precise variation of tubulin substances in the cytoplasm of oviductal ciliated cells and the morphological changes in cytoplasmic organelles of ciliated cells for ciliogenesis by estrous cycles. The animals used in this study were female rat (Sprague Dawley strain), weighing approximately 200 gm. The ampulla oviducts of these animals (at each of estrous cycle) were rapidly excised. At each stage of estrous cycle, the tissues were used for immunocytochemical study and other were used for electron microscopical study. All specimens were observed by the light and electron microscope. The results obtained are as follows: 1. In the ciliated cells at proestrus, Golgi complex showed $5{\sim}7$ stacked cisternae with numerous saccules and vacuoles. Large amount of fibrous granules were located near the Golgi complex. But at metestrus and diestrus, few fibrous granules were seen. 2. A moderate number of rough endoplasmic reticulum and polyribosomes were scattered in the cytoplasm of ciliated cells at proestrus, but were decrease in number at metestrus and diestrus. 3. At proestrus and estrus, there were a large amount of vesicles in the apical cytoplasm of ciliated cells. 4. Numerous mitochondria were located in the apical cytoplasm at proestrus and estrus, but only a few at metestrus and diestrus. 5. At proestrus and estrus, tubulin substances showed strong reactions in the cytoplasm but weak reactions at metestrus and diestrus. It is suggested consequently that the ciliated cells of the rat oviducts showed no morphological changes of cilia but the ultrastructural organelles of the cells were changed in its shape and location during the entire estrous cycle.