• KOREAN JOURNAL OF CROP SCIENCE
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• v.51 no.spc1
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• pp.242-246
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• 2006

This study was showed into the pollen development with in vivo by bud size and genotype. Microspores of buds from 2.0 mm to 2.5 mm of all genotypes were composed of mainly tetrad cells and early uninucleate stage cells. Microspores derived from buds of 2.5-3.0 mm were exposed cells of early uninucleate, middle uninucleate, and late uninucleate. Microspores from buds of 3.0-3.5 mm contained mostly late uninucleate stage cells and showed some early binucleate stage cells. Microspores of buds with 3.5-4.0 mm in length were composed of mainly binucleate stage cells and decreased late uninucleate stage cells. Microspore with more than 4.0 mm were entered into binucleate stage cells of divided generative nucleus and vegetative nucleus. In 'Tamlayuchae', microspores derived from buds of 3.5-4.0 mm were observed cells of late uninucleate stage and early binucleate stage because of late microspore development. In MS-maintainer, the spring type, microspore derived from buds of 2.5-3.0 mm were observed tetrad stage cells.

• FLOWER RESEARCH JOURNAL
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• v.16 no.1
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• pp.12-16
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• 2008

This study was carried out to produce haploid plants to verify a systematic breeding program and genetic analysis. Effect of developmental stage of pollen grains and pre-treatment temperature on production of haploid plants was investigated. Microscopic investigation of the explants (Lilium Asiatic hybrid 'reamland' revealed that the length of flower bud at 23.0-24.9, 25.0-26.9, and 27.0-28.9 mm long coincided with tetrad, uninucleate, and binucleate, respectively. When the efficiency of the anther culture from microgametogenetic stages was tested, late uninucleate to early binucleate stage, having the length of 23.0 to 28.0 mm long flower bud, was the best. The frequencies of the callus induction and plant regeneration from the stage mentioned above were 17.8 and 6.7%, respectively. When calli were cultured on the MS medium containing picloram and zeatin at $25^{\circ}C$, shoots were obtained. Roots of regenerated plantlets were confirmed as haploid through an microscopic observation.

• ALGAE
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• v.19 no.2
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• pp.79-90
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• 2004

Nuclear DNA contents of spermatia from eight ceramiacean and four dasyacean algae (Ceramiales, Rhodophyta) and microspores from two land plants were estimated by fluorescence microscopic image processing and their nuclear SSU rDNA sequence data were analyzed. In frequency distribution patterns, the DAPI-stained nuclear volume (NV) of spermatia showed two peaks corresponding to 1C and 2C. Nuclear 2C DNA contents estimated from NV were 0.45-2.31 pg in ceramiacean and 0.40-0.57 pg in dasyacean algae and 8.42-9.51 pg in two land plants, Capsicum annuum and Nicotiana tabacum. By nuclear patterning of vegetative cells derived from an apical cell, 2C DNA contents of spermatia were 2.31 pg in an alga having uninucleate and non-polyploid nucleus (Aglaothamnion callophyllidicola), 0.45-1.94 pg in algae having uninucleate and polyploid nucleus (Antithamnion spp. and Pterothamnion yezoense), and 0.40-0.62 pg in algae having multinucleate and non-polyploid nuclei (Griffithsia japonica and dasyacean algae). Each mature spermatium and microspore (pollen grain) seemed to have a 2C nucleus, which may provide a genetic buffering system to protect the genetic content of a spermatium and microspore from potentially lethal mutations. Nuclear DNA content and SSU rDNA sequence of Antithamnion sparsum from Korea were reasonably different from those of Antithamnion densum from France. The data did not support the previous taxonomic studies that these two taxa could be conspecific.

• The Plant Pathology Journal
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• v.17 no.2
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• pp.94-100
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• 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 Plant Tissue Culture
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• v.21 no.5
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• pp.293-297
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• 1994

In order to induce immature pollen derived plants, anthers of Ranunculus japonicus Thunb. were cultured on Murashige and Skoog's medium supplemented with various combinations of auxins and cytokinins. The combinations of NAA and BA were more effective than those of 2,4-D and kinetin in the formation of calli and embryos. Up to 5t5% of the anthers cultured on medium containing 0.5 mg/L NAA and 1.0 mg/L BA gave rise to plantlets. The most suitable stage for anther culture in the induction of calli and/or embryos from immature pollens was at the uninucleate and early binucleate stage (3 days before anthesis). Immature pollens developed into embryos by repeated division of the vegetative nucleate after 60days of culture.

• Horticultural Science & Technology
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• v.32 no.3
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• pp.382-389
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• 2014

Raphanus sativus L. cv. Taebaek, a efficiently microspore-derived embryo (MDE)-forming cultivar, and 'Chungwoon', a non-MDE-forming cultivar were selected as donor plants for isolated microspore culture. Radish flower bud of 2.0 (small, S), 4.0 (medium, M), and 6.0 (large, L) ${\pm}$ 0.5 mm in length were isolated to determine the temporal relationship between flower bud size and MED yield. Anatomical observations revealed no difference in the structure of the flower buds between the two cultivars. In both cultivars, the stigmas were much longer than the floral leaf in M-sized flower buds. The MDE yields for 'Taebaek' per petri dish were 6.6 and 1.3 for M- and L-sized of flower buds, respectively, but MDE formation was not induced in the S flower buds. On the other hand, 'Chungwoon' failed to form MDEs in all flower buds. The microspore density of 'Taebaek' was 1.3 times more than that of 'Chungwoon' for M sized flower buds. Of the M-sized buds from 'Taebaek' and 'Chungwoon', 92.1 and 81.6%, respectively, were in the late uninucleate microspore stage, which is characterized by the highest frequency of MDE formation. Anatomical observations of MDE formation revealed that the microspores were able to divide to form a primordium from which cell division took place continuously in the 'Teabeak' cultivar. However, the microspores of 'Chungwoon' failed to progress beyond the primodium stage, resulting in lack of MDE formation. By contrast, after the formation of the primordium, various developmental stages of embyos from microspore were observed in the 'Taebaek' cultivar. These results can be used to determine MDE forming potentials of radish cultivars.

• Proceedings of the Botanical Society of Korea Conference
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• 1996.07a
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• pp.48-60
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• 1996

We previously identified and characterized a predominantly pollen-expressed gene of Petunia inflata that encodes a receptor-like kinase named PRK1. The extracellular domain of PRK1 contains leucine-rich repeats which have been implicated in protein-protein interactions, and the cytoplasmic domain was found to autophosphorylate on serine and tyrosine. To investigate the function PRK1 in pollen development, we transformed P. inflata plants with a construct containing the promoter of a predominantly pollen-expressed gene of tomato, LAT52, fused to an antisense PRK1 cDNA corresponding to part of the extracellular domain of PRK1, There transgenic plants were found to each produce approximately equal amounts of normal and aborted pollen. Analysis of the inheritance of the transgene inserts in two of the transgenic plants, ASRK-13 and ASRK-20, to their progeny revealed that certain transgene inserts cosegregated with the pollen abortion phenotype. Microscopic examination of the aborted pollen grains showed that their outer wall, the exine, was essentially normal, but that their cytoplasm contained only starch-like granules. Staining of the nuclei of the microspores at different stages of uninucleate stage. However, at subsequent stages half of the microspores completed mitosis and developed into normal binucleate pollen, but the other half initially remained uninucleate, then lost their nucleio. Analysis of the amounts of PRK1 mRNA and the antisense PRK1 transcript suggested that the pollen abortion phenotype most likely resulted from down-regulation of the PRK1 gene by the antisense PRK1 transgene. These results suggest that PRK1 plays an essential role in a signal transduction pathway that mediates post-meiotic development of microspores.

• Journal of Plant Biology
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• v.36 no.3
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• pp.241-249
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• 1993

The pattern of RNA synthesis during male gametogenesis of Brassica napus was studied using 3H-uridine autoradiography. No incorporation of isotope occurred in the newly released microspore and the nonvacuolate, furrowed microspore. Peak incorporation of label during male gametogenesis occurred in the uninucleate, furrowed microspores showing various degrees of vacuolation. In this microspore stage, silver grains were localized in the nucleus and cytoplasm. Moderate incorporation of the isotope occurred in the nulceus of the vacuolated microspore. After the microspore mitosis, isotope incorporation occurred predominantly in the nucleus of the vegetative cell with little or no incorporation in the generative cell. In tricellular pollen, no incorporation of isotope was observed in both the vegetative nucleus and the sperms. Silver grains almost completely disappeared from tricellular mature pollen grains ready to germinate.

• Journal of Plant Biology
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• v.36 no.2
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• pp.183-192
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• 1993

Efficient plant regeneration system was established through the culture of rice (Oryza sativa L.) microspores. Microspores released by anther shedding culture developed into proembryos and calluses in B5 medium within two weeks of culture. The optimal hormone and carbon sources were dependent on the genotypes used. Microspore's viability decreased rapidly in culture time, therefore less than 3% of the total microspores were viable at the 9th day when the first microspore division was observed. Of two types of microspores (pollen dimorphism) observed in culture, only the P-grain, larger microspores than normal one was able to divide. Using 4',6-diamidino-2-phenylindole (DAPI) fluorescence staining, it was confirmed that the symmetrical division of uninucleate microspore was the first step leading to continuous microspore development. Microspore-derived proembryos and calluses were regenerated to plants in N6 medium containing 1 mg/L NAA and 5 mg/L kinetin, and 78.3% of the regenerated plants were haploids.

• Applied Microscopy
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• v.12 no.1
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• pp.49-56
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• 1982

A case of systemic cryptococcosis developed in 4 year old boy was described and illustrated by light and electron microscope. Light microscopically, the upper dermis of the skin showed chronic nonspecific inflammation with numerous spherical spores surrounded by a clear halo created by the wide gelatinous capsule. Ultrastructurally, the C. neoformans showed the wide capsule containing microfibrils that appeared to radiate from the cell wall and to coil and interwine in various directions. The cell was uninucleate with a single nucleolus. Along the inner nuclear envelope, numerous small vesicles were present. In addition, C. neoformans presented membranous organelles derived from the plasma membrane and comparable to bacterial mesosomes.