• Korean Journal of Plant Tissue Culture
• /
• v.22 no.6
• /
• pp.323-327
• /
• 1995

This experiment was carried out to observe the origin and developmental pattern of somatic embryos of Oenanthe javanica ($B^{L}.$) DC. The experiment included observation of embryogenic cells and their development stages by light microscope, transmission electron microscope and scanning electron microscope. The embryogenic cells, which were smaller than non-embryogenic cells in size with expanded nucleus and dense cytoplasm. When stained with hematoxylin, the embryogenic cells were readily distinguished from the non-embryogenic cells of which cell walls were stained with safranin. It was observed at somatic embryos developed from single cells on the epidermis of developing embryos or in the surface or inside of embryogenic clumps by segmentation pattern. Observation with a transmission electron microscope revealed that the embryogenic cells had dense cytoplasm expanded nucleus, small vacuoles, large amyloplasts containing starch grains, and abundant organelles including lipid bodies. Under a scanning electron microscope, embryogenic callus was shown to consist of very smaller cells than non-embryogenic cells in an orderly arrangement and covered with a net-like structure, while the non-embryogenic callus consisted of large cells, irregular in size and arrangement, and covered with a gelatin-like material.

• Korean Journal of Plant Tissue Culture
• /
• v.26 no.3
• /
• pp.143-148
• /
• 1999

This study was conducted to elucidate the effects of ascorbic acid and dehydroascorbic acid on somatic embryogenesis from the cultured cells of carrot. Ascorbic acid in culture medium merely stimulated the proliferation of non-embryogenic cells but dehydroascorbic acid in medium induced embryogenic cells from non-embryogenic cells accompanying the inhibition of cell proliferation. Ascorbic acid in medium inhibited somatic embryogenesis from embryogenic cells while dehydroascorbic acid in medium enhanced somatic embryogenesis from the cells as well as non-embryogenic cells. This enhancement was limited to globular embryos and the maturation to cotyledonary embryos was inhibited by dehydroascorbic acid treatment. From the above results it is suggested that carrot callus cultures on medium containing dehydroascorbic acid could quickly induce embryogenic cells. In addition after brief culture of embryogenic cells on development medium containing dehydroascorbic there by acid the subculture of the cells to MS basal medium resulted in the high frequency production of somatic embryos.

• Applied Microscopy
• /
• v.29 no.1
• /
• pp.105-124
• /
• 1999

Histological and ultrastructural differentiations of the neuroepithelial cells in the mouse embryo during neurulation were observed. The neural plates and grooves consisted of pseudostratified columnar epithelium in the embryonic day (ED) 8 embryo were developed. In the ED 9 embryo, the neural tube was developed in all body length of embryo except both the cephalic and caudal ends. Secondary neurulation was shown at the tail bud of the ED 10 embryo. In the ED 8 embryo, the primitive streak was shown in the posterior end of the embryonic disc. The neuroepithelium, notochord and mesenchyme were well differentiated in the cephalic and cervical portions. In the ED 9 and 10 embryos, the roof plates of neural tubes were constituted of the closing of the surface ectodermal cells in the hindbrain and the neuroepithelial cells in the spinal cord. The floor plate of neural tube were consisted of the low pseudostratified columnar epithelium. The spinal motor nerve fibers were initially differentiated in the ED 10 embryo. According to the electron density of the cell and the differentiation of tell organelles, the neuroepithelial cells in the ED 9 and 10 embryos were classified into three types: dark, intermediate and light types. All types in the ED 9 embryo were observed but the dark cell in the ED 10 embryo was not done. The free ribosomes and polysomes in all neuroepithelial cells were developed. The RER and lipid droplets in the dark cell and the Golgi complex in the intermediate and light cells were observed. Many microfilaments in the cytoplasmic processes of intermediate cell and the microfilaments and microtubules in the light cell processes were observed to be well differentiated.

• Journal of Plant Biology
• /
• v.38 no.1
• /
• pp.55-62
• /
• 1995

Embryogenic cell (EC) suspension cultures derived from mature seed-embryo of rice (Oryza sativa L cv. Kye Hwa) were used for the expression patterns of isozyme and enzyme activity. EC suspension cultures were composed of cells that were densely cytoplasmic, potentially embryogenic. However, nonembryogenic cell (NEC) cultures were composed of large, elongated and vacuolated cells. These cells were analyzed for the isozyme pattern and enzyme activity of EC and NEC. Isozyme patterns of peroxidase, esterase, acid phosphatase and malate dehydrogenase exhibited striking difference in the total number of bands, specificity and intensity of band. Also, these isozymes showed very high activity in the EC. Specific band, band activity and higher enzyme activity of isozyme in EC was absent or low in NEC, which may indicate an association of these specific isozymes with morphological characterization and totipotency of embryogenic cells. These results indicate that specific pattern and activity of enzyme in EC could probably be used as a biochemical marker of EC in rice.n rice.

• Journal of Plant Biotechnology
• /
• v.36 no.1
• /
• pp.7-12
• /
• 2009

This study was conducted to establish the optimal suspension culture system for both the propagation of embryogenic cells (ECs) and the induction of somatic embryos (SEs) of Kalopanax septemlobus. The proliferation rate of ECs was reduced as the inoculum density was increased; the highest rate was obtained when 0.1 g/100 ml of cells was initially inoculated. According to the analysis of cell growth pattern and cell growth cycle (G1, Sand G2/M), the cell growth started in 5 days culture initiation, grew rapidly until 15 days and then decreased gradually. Distinctive changes of the cell growth cycle by the culture periods was also observed; the growth cycle was doubled from initial 5.6% to 11.7% of S stage in 5 days culture and then reached in stable stages again. Therefore, the results indicated that a 15-day-cycle was the optimal culture period for the propagation of the ECs through the suspension culture. Furthermore, the cell inoculum density was also important for the induction of SE; more than 65% of SEs at the torpedo stage was induced by using the low level of cell inoculum (0.5 g/L), while the higher inoculum densities were rapidly reduced the proportion of SEs at that stage. Although the higher inoculum density delayed the development of SE, it did not affect the proportion of SEs at the globular and heart stage. In conclusion, this study showed that the suspension culture of the Kalopanax septemlobus ECs through the control of inoculum density was an efficient way for both the propagation of ECs and the induction of SEs, suggesting that the development of this system might help to reduce the culture period for the somatic embryo production.

• Journal of Plant Biotechnology
• /
• v.31 no.3
• /
• pp.219-223
• /
• 2004

Embryogenic calli were induced from petioles of Aralia elata on MS solid medium supplemented with 1.0 mg/L 2,4-D. When embryogenic calli were transferred to MS liquid medium supplemented with 1.0 mg/L 2,4-D, embryogenic cells and embryogenic cell clusters were developed after 2 weeks of culture. Embryogenic cells were filtered through a 250 ${\mu}{\textrm}{m}$ sieve and the passed cells were proliferated and maintained in MS liquid medium supplemented with 1.0 mg/L 2,4-D. Embryogenic cell clusters entrapped on the sieve were transferred to 1/2 MS liquid medium without plant growth regulators, globular-shaped embryos were developed from embryogenic cell clusters after 2 weeks of culture. Numerous early stage somatic embryos could be developed to heart-shaped, torpedo-shaped, cotyledonary embryos and plantlets in 5 L bioreactor. Above results suggest that effective somatic embryo proliferation can be achieved via bioreactor culture systems in Aralia elata.

• Korean Journal of Plant Tissue Culture
• /
• v.24 no.6
• /
• pp.369-374
• /
• 1997

Immature flower buds of 'Desio' carnation were cultured on MS agar medium supplemented with 1 ㎎/L 2,L-D. Embryogenic calli were formed from 5-10% of the buds less than 20 ㎜ in length, but only non-embryogenic calli were produced from explants of shoot apex leaf, internode, and flowere buds larger than 20 ㎜. The same method was applied to 16 cultivars of cut Sower carnation and embryogenic calli were obtained in 7 cultivars. Several embryogenic callus lines were selected and maintained through subcultures over 120 weeks without loss of embryogenic competence. The embryogenic cultures were also proliferated rapidly in liquid agitation cultures using MS medium supplemented with 1mg/L 2,4-D. Numerous embryos were formed on the periphery of the cell aggregates upon transfer to auxin-free MS agar medium. Plantlets were transplanted in potting soil and grown to bloom in six months.

• Korean Journal of Plant Tissue Culture
• /
• v.24 no.2
• /
• pp.107-112
• /
• 1997

This study was carried out to establish a mass production of normal somatic embryos of Oenanthe iavanica (BL.) DC. including examination of nitrogen and sucrose sources, and ABA concentration. Embryogenic cell clumps and embryos were formed on the MS medium devoid of growth regulators. Proliferation of embryogenic cells and clumps was enhanced by 2, 4-D. Meanwhile embryo growth and development occurred on the media containing NAA and IBA. Growth of embryos was generally good in the media containing both 20 mM $KNO_3$ and 20 mM $NH_4NO_3$. The rate of shoot forming embryos was higher on the media containing on1y 20mM $NH_4NO_3$ than on the former. Addition of sucrose at 3-6% enhanced the embryo development, and normal embryos with short hypocotyl was observed on the medium containing $10\mu\textrm{M}$ ABA. Embryogenic cell clumps or globular embryos, when transferred to MS solid media devoid of growth regulators, developed into mature embryos and then into plantlets which had entire primary leaves like zygotic seedlings.

• Korean Journal of Plant Tissue Culture
• /
• v.25 no.1
• /
• pp.13-19
• /
• 1998

We have reported previously that intact embryogenic cells can be used instead of protoplasts for electroporation-mediated transformation of zoysiagrass and rice. In this study, conditions of the tissue electroporation were examined to optimize the procedures. Embryogenic cell suspensions were established in liquid MS medium containing 2 mg/L of 2,4-D with embryogenic calluses induced from mature embryos of Z. japonica. The suspension-cultured cell clumps were electroporated with 35S-gusA expression vector DNA, and degrees of DNA introduction into the cells were determined by histological expression rates of the gusA marker gene. DNA transfer into the cell clumps occurred in wide range of voltage (100-400 V) and capacitance (10-1980 $\mu\textrm{F}$), but more in the ranges of 200-300 V and 330-800 $\mu\textrm{F}$ DNA concentrations higher than 6 $\mu\textrm{g}$/mL were adequate for GUS expression of the electroporated cells. DNA transfers were confirmed in all three embryogenic cell lines but only in one out of eleven non-embryogenic lines. Positive GUS expressions occurred with DNAs added even 20-40 h after pulse treatments. As a promoter of gusA, Act1 and Ubi1 were effective 7 and 5 times than 35S respectively in number of GUS expression units on electroporated cell clumps. Embryogenic cell clumps survived and regenerated into plantlets after pulse treatments of wide range of conditions.

• Korean Journal of Medicinal Crop Science
• /
• v.14 no.4
• /
• pp.255-260
• /
• 2006

Culture conditions for high frequency plant regeneration via somatic embryogenesis from embryogenic cell suspension cultures of Hovenia dulcis are described. Germinated somatic embryos were selected for induction of secondary embryogenesis. Friable embryogenic cells were induced directly from somatic embryos when transfer to 1/3 MS solid or liquid medium lacking plant growth regulators. The temperature strongly effected on induction of secondary embryognesis than other conditions in culture. All somatic embryos produced friable embryogenic cell clumps within 10 days when germinated somatic embryos cultured in 1/3 MS medium at $30^{\circ}C$ in suspension culture. No somatic embryos formed from embryogenic cell suspension cultures at $18^{\circ}C$. Numerous somatic embryos were induced and subsequently developed uniformly into germination stage from suspended cell clumps after 4 weeks of culture on $18^{\circ}C$. Plantlets conversion were observed on $18^{\circ}C$ when germinated somatic embryos were transferred to 1/3 MS solid medium without plant growth regulators or supplemented with 0.1-0.5 mg/l benzyladenine.