• Journal of Plant Biology
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• v.14 no.2
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• pp.7-10
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• 1971

Haploid cell obta-ined from microspores of Solanum nigrum were cultured on two kinds of medium, "Callus-inducing medium" and "Differentiation medium", in order to conduct histological studies of callus and examine differentiation of plantlets. On the callus-inducing medium the calli grew rapidly. The bulk of callus mass was light brown colored "Wet callus" covered on the surface with thin layers of rough and gleaming "White callus". The wet callus was consisted of parenchyma and meristematic tissues, while the white callus had no meristematic tissues. Large parenchyma cells, by successive divisions, became multicellular or poly nucleate cells which developed later to be meristematic tissues. The calli embedded on the differentiation medium quickly turned to dark brown color. Plantlets, however, came out later from these blackened callus mass. In the callus sectioned about ten weeks after imbedding on the differentiation medium, radially elongated tissue, concentric tissue, epidermis, tracheid-like structure, and plant jprimordia were observed.ure, and plant jprimordia were observed.

• Applied Microscopy
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• v.8 no.1
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• pp.67-76
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• 1978

A comparative investigation of fine structure of callus cells derived from tissue culture of Panax ginseng was made by electron microscope. Callus was consisted of large superficial cells and small inner zone cells derived from shoot apex tissue cultured for 16 weeks. Large superficial cells were contained the clusters of starch grains surrounded by a double plastid membrane. Especially, electron dense particles were deposited just inside and outside of plastid membrane and also deposited on mitochondria-like and endoplasmic reticulum-like structures. Crystalline body was also found in superficial cells. Small inner zone cells were characterized by presence of proplastids sheathed by short endoplasmic reticulum profiles. presence of spiral configuration of ribosomes and absence of crystalline body. Organ primordia was consisted of a dense cytoplasm and notable nucleate cells derived from nodal tissue cultured for 67 weeks. Proplastids containing starch grains and crystalline bodies were frequently observed; starch grains are of small quantity and does not form the clusters as in inner zone cells; hexagonal crystalline body itself does not have always limiting membrane. Remarkably. in a few cells of primordia, particles resembling the presumptive virus were observed mainly in condensed nuclear chromatin and also in cytoplasm, in mitochondrion-like organelle.

• Korean Journal of Plant Tissue Culture
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• v.24 no.3
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• pp.183-188
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• 1997

Callus from scale segments of Lilium longiflorum 'Gelia' was effectively induced and maintained from unorganized tissue on the semi-solid medium by 0.42% Bacto agar with MS basal salts and vitamins of SH medium supplemented with 0.5 mg/L 2, 4-D, 1.0 mg/L NAA, 0.3 mg/L BA, and 3% sucrose. More than 5% of high sucrose level had inhibiting effect on regeneration capacity of formed callus and decreased callus growth. Various combinations of nitrogen did not effective to proliferate the ELC (Embryogenic-like callus), but friability of callus was increased in the medium containing only nitrate as nitrogen source. 5 mL conditioned medium into 30 mL fresh medium was good for cell growth. However friable cell aggregates during suspension culture had to form hard callus which hindered to establish suspention culture system. Addition of 2 g/L casein hydrolysate increased callus growth and friability of the hard callus. As a result of anatomical observation of callus, organogenesis such as shoots, roots and bulblets was independently induced from callus tissue. Somatic embryogenesis from callus tissue could be observed with low frequency.

• Journal of Plant Biology
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• v.13 no.3
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• pp.17-19
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• 1970

Histological observation of micropore-originated haploid rice callus was reported previously. Present study was attempted to clarify the growth or development of the calli when they were transferred to differentiation media prepared exclusively for differentiation of plantlets. When the callus was transferred to differentiation medium, the cells and tissues became radially elongated. Meristematic tissues were present but few in number, and their structures were quite different from those grown in the propagaton medium. Differentiation of tracheid, chloroplast, and epidermis-like cell layer, and formation of gap in the callus tissue were more conspicuous in differentiation media. Approximately ten days after transfer of callus to differentiation medium, plantlet was formed.

• Korean Journal of Plant Tissue Culture
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• v.27 no.3
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• pp.233-238
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• 2000

To obtain a basic information of the development of Genus Viola, morphological and histological observation of in vitro calli and cells in Viola culture cells were investigated. There were two callus types obtained by long term subculture of wild viola (Viola partrinii DC. ) petiole callus. One was friable callus - soft and pale green in color and small cells in size, and the other was compact callus - compact and deep bluish green in color, large cells in size. In scanning electron microscopic observation, friable callus was composed of voculated cell around small. cell clump, while compact callus was composed of cells filled with protoplasm Somatic embryogenesis was observed from suspension culture of the compact callus.

• Journal of Plant Biology
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• v.19 no.4
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• pp.100-106
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• 1976

The fine structure of the callus induced from epidermis of Panax ginseng leaves cultured on Murashige & Skoog medium plus kinetin 0.1mg/l, NAA 0.2mg/l and 2.4-D 0.5mg/l was observed. The cells composing callus tissue are mononucleus. Three types of cells were identified; cells with abundant cytoplasm, cells with relatively differentiated vacuoles and with numerous starch grains in the plastids and ones with highly differentiated vacuoles and with unsaturated lipid granules. Prolamellar body, plastid lamellae, plastid globules, stromacenter, fine tubules, crystal-containing body and DNA-like structures were observed in the stroma of the plastids. The chromoplasts were identified in some cells believed as the mother cells of secretory cells in secretory ducts. Curved or straight micro-fibrils of 100~150A in diameter were observed in the cytoplasm. And the characteristics of cell organelles and cell inclusions and the vacuole formation in callus tissues were discussed.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.38 no.4
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• pp.366-370
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• 1993

This study was conducted to determine the optimum culture conditions for inducing callus and regenerating plantlets from cultured leaf tissues of Aralia elata. Young leaf tissues(1cm) of A. elata plant were cultured on MS medium supplemented with 2, 4-D and Thidiazuron. Embryogenic callus was induced along the leaf veins, more efficiently on the medium containing 1.0mg /1 Thidiazuron in 4 weeks after culture initiation. Calli were subcultured to proliferate on MS media containing 2, 4-D, Dicamba, Picloram, and Thidiazuron. Callus was better proliferated on the medium containing Dicamba than on the others.. However, callus subcultured on the medium containing Thidiazuron was more embryogenic and light green-colored, of which some showed embryoid-like structure on the surface. Hormone-free medium was more efficient to regenerate plantlets than media supplemented with Kinetin, BA, and Thidiazuron.

• The Plant Pathology Journal
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• v.15 no.1
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• pp.38-43
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• 1999

Mutants of the monopartite geminivirus beet curly top virus (BCTV) have been screened for infectivity, systemic movement, replication and symptom development in Arabidopsis thaliana. As known by coding for coat protein, R1 mutant was not infectious and did not move systemically. R2, R3 and L2/L3 mutants produced milder symptoms compared to wild type BCTV but the infectivity was reduced by 40% to 60%. R2 ORF is thought to be involved in the regulation of ssDNA and dsDNA accumulation because only dsDNA was accumulated on R2-infected organs. Disruption of ORF L4 resulted in reduced infections, but the viral DNA was accumulated in infected organs from roots to shoot tips as much as wild type BCTV on Sei-O. In addition, 4 mutants did not produce callus-like tissues on infected organs, suggesting that L4 ORF may play a role in the induction of host cell divisions by virus infection. This result was supported by the patterns of mRNA expression and promoter analysis of the cell cycle marker gene, cycl, on Arabidopsis. cycl mRNA was accumulated on symptomatic organs by wild type BCTV infections but not by L4 mutant. We conclude that the BCTV L4 ORF is essential for symptom developments, specially callus-like formation on infected organs.

• Journal of the Korean Society of Tobacco Science
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• v.17 no.1
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• pp.33-40
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• 1995

Structures of the adventitious root meristem induced from callus culture of tobaco (Nicotiana tabacum cv. NC 82) leaves were investigated by light and transmission electron microscopy. Structural differences between in vitro root and callus cells were also examined by the microscopy. The submicroscopic features of the in vitro root cells were as follows. Intercellular spaces were not developed and nuclei with two nucleoli were observed occasionally. Plasmodesmate were found in groups or sing1y on transverse and longitudinal walls. Amyloplast solely filled with starch grains, with one to five electron - dense bands, was surrounded by single membrane. in the callus cells, vacuolization of central part in the cytoplasm having mitochondria with swollen cristae and starch grains like those of in vitro root cells was a distinct feature. Vesicles which were found between cell wall and plasma membrane may be arisen by a process of protoplasmic invagination. By comparing of ultrastructures between the cells of callus and in vitro roots we found that the distinct differences lied on thickened cell walls and hypertrophed vacuoles in the former, and less thickened cell walls and several small vacuoles in the later.

• Journal of Plant Biotechnology
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• v.35 no.2
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• pp.133-140
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• 2008

The modification of DNA and histone plays an important role for gene expression in plant development. The objective of this research is to observe the effects of methylation on the gene expression during dedifferentiation from rice mature seeds to callus and differentiation from callus to shoots. The embryogenic callus with ability to shoot regeneration was not induced on the N6A medium supplemented with 5-azacytidine and abnormal callus with brown color was formed. When the normal rice callus was placed on the regeneration MSRA medium supplemented with 5-azacytidine, the shoot regeneration was inhibited. The results showed that 5-azacytidine, DNA demethylating agent, had negative effects on normal embryogenic callus formation and shoot regeneration. This suggested that DNA methylation of some genes was required for normal cell dedifferentiation and differentiation in tissue culture. The microarray and $GeneFishig^{TM}$ DEG screening were used to observe the gene transcript profile in callus induction and regeneration on N6A (N6 medium + 5-azaC) and MSRA (MS regeneration medium + 5-azaC). Subsets of genes were up-regulated or down-regulated in response to 5-azaC treatments. The genes related with epigenetic regulation, electron transport, nucleic acid metabolism and response to stress were up and down regulated. The different expression of some genes (germin like protein etc.) during callus induction and shoot regeneration was confirmed using RT-PCR and northern blot analysis.