• Journal of Life Science
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• v.25 no.2
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• pp.158-163
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• 2015

Rice embryo is more abundant than endosperms in nutrients such as proteins, lipids, and vitamin B1. In this study, we constructed p53 plasmid that could be expressed in a plant system, and investigated optimal germination conditions in a variety of media. For construction of p53 plasmid, we performed p53 amplification from pCDNA-p53, subcloned to TA cloning vector, and then reconstructed into pGEM-CaMV plant expression vector. On the other hand, we prepared a variety of imbibition buffers and complete media for efficient germination of the rice embryo. Imbibition buffers prepared with different concentrations of salt or detergent showed no significant effect on germination efficiency. We prepared further culture media, such as solid agar, liquid media, and paper towel to establish the optimal conditions. Rice embryo showed germination rates of more than 70% in the solid medium, more than 60% in the paper towel medium, but less than 25% in liquid media, although germination rate did not differ with varying concentrations of salt and sucrose in culture media. Under the optimal germination conditions, we introduced the p53 plasmid using imbibition method, and finally detected human p53 gene expression in the germinated rice embryo. This method might present a novel, practical approach for evaluating efficient gene expression utilizing imbibition method in rice embryo.

• Journal of Plant Biotechnology
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• v.6 no.4
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• pp.213-219
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• 2004

Zygotic embryos were excised from immature and mature seeds of the Himalayan yew, Taxus wallichiana. The embryos germinated precociously when kept in darkness for 5 weeks and developed into full seedlings within 10-12 weeks. The highest rate of embryo germination ($81\%$) was obtained in modified Lloyd & McCown' s woody plant medium containing macro and micronutrients at half strength supplemented with $1\%$ activated charcoal, which supported both the best embryonic growth ($43\%$) and seedling development ($32\%$). However, the supplementation of basal media with kinetin, thidiazuron, 6-benzyl aminopurine or $GA_3$ had no effect on the germination of the embryos. The embryos derived from immature seeds germinated but the frequency of embryonic growth was better in mature seeds. Stratification of seeds effected precocious germination of embryos. Seeds kept at $4^{\circ}C$ for 1 week germinated earlier and at a higher frequency irrespective of the stage of seed maturity, while the germination rate declined with prolonged cold treatment for 1 month at that same temperature. Analysis of taxanes in germinating seedlings revealed that root tissues contained high levels of taxol, 10-deacetyl-baccatin ill and baccatin ill as compared to shoots. Thus embryo culture technique appears to overcome the lengthy dormancy requirement of T. wallichiana seeds.

• Journal of Plant Biotechnology
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• v.2 no.1
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• pp.29-33
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• 2000

To develop an efficient propagation method for yew tree, zygotic embryos were cultured under various conditions. When dissected embryos were cultured on GA$_3$ containing media, the highest germination frequency was observed on WPM medium contaning 1.0 mg/L GA$_3$. For germination of the embryos, two different conditions were compared; culturing embryos with endosperm (Method I), and 2) culturing embryos only (Method II). Maximum germination was achieved in 0.5 mg/L GA$_3$ when embryos with endosperm were cultured on the media. Of the media tested, White and WPM medium were the most suitable on germination of embryos. The abnormality of yew embryos found was observed when it cultured on GA$_3$ or culture media. About 40% of the precociously germinated embryos could be developed into full seedlings. Seedlings contained taxol in high quantity (535 $\mu\textrm{g}$/g dry weight). In vitro techniques will be sewed as a useful tool for the development of transformed root cultures and biosynthesis studies.

• Journal of Ginseng Research
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• v.25 no.4
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• pp.167-170
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• 2001

Experiments were conducted to study the optimum chilling period for breaking physiological dormancy of dehisced ginseng(Panax ginseng C. A. Meyer) seed. Embryo of ginseng seed is too small to be noticed with naked eyed on harvesting time. Embryo grew to half size of endosperm after seeds were stratified for 3 months. It takes 6 months for this embryo to reach the size enough for germination. And it grew faster indoors than outdoors. Dehisced ginseng seed with full-size embryo did not germinate at room temperature and required chilling treatment for 75days in outdoors and 90 days in cold chamber. While seed receiving sufficient chilling treatment were left to be in room temperature, the chilling effect decreased remarkably.

• Korean Journal of Plant Tissue Culture
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• v.27 no.4
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• pp.299-307
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• 2000

A large amount of information is currently available for somatic embryogenesis of plants. However, one common problem related to somatic embryos is that the conversion rate can be low for some species. Apical meristems are responsible for post-embryonic growth of the embryo. The low percentage observed is most likely a result of poor apical meristem development or defects in the meristem organization during somatic embryogenesis. In flowering plants, apical meristems are well developed by the late heart stage of zygotic embryo development. In conifers, such as white spruce, apical meristems are formed at the pre-cotyledon stage. Thus, apical meristem development occurs very early, prior to the maturation stage of embryo development. Once formed, meristems are stably determined. In the somatic embryo, as exemplified by white spruce, since embryo development is not synchronous, tissue differentiation including apical meristem formation occurs throughout the“maturation”stage. Different apical meristem organizations can be found among different individuals within a population. In contrast to their zygotic counterparts, the apical meristems appear not to be stably determined as their organization, as the shoot apical meristem especially, can be easily modified or disrupted. Precocious germination seldom results in functional plantlets. All these observations suggest that the conditions for somatic embryo maturation have not been optimized or are not suitable for meristem formation and development. The following strategies could improve meristem development and hence conversion: 1. Simulate in ouuio conditions to promote meristem development prior to the“maturation”treatment.2. Prevent deterioration of apical meristem organization during somatic embryo maturation.3. Promote further meristem development during embryo germination.

• Journal of Ginseng Research
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• v.9 no.2
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• pp.146-153
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• 1985

This study was conducted to know the effect of vinyl mulching from the sowing to germination on the seedbed for germination and production of seedling. Embryo growth was restrained with decreasing the water content in seed and stopped below 10% water content. Germination was possible over 55% water content but radicle growth was stopped at 55% water content. Ratioes of embryo / emdosperm length were about 50% at seeding time, and about 80% at just before freeing season, and the ratio was increased from the thawing season again. Vinyl mulching increased the soil water content and soil temperature. Germination rate and number of available seedling in vinyl mulching were increased 10% and 12%, respectively.

• Korean Journal of Agricultural Science
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• v.15 no.1
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• pp.47-68
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• 1988

This study was conducted to define the optimal conditions for embryo growth during seed stratification and for breaking dormancy as well as seed germination of stratified ginseng seeds. The experiments were also carried out to detect some materials which were expected to induce seed dormancy in the ginseng seeds. The results summarized as follows; 1. The growth of embryo during seed stratification was significantly inhibited by the existence of endocarp. The fastest embryo growth was resulted at $15^{\circ}C$ and an estimated optimal temperature for embryo growth was about $18^{\circ}C$. 2. There was no significant difference between the embryo growth and germination ratio of ginseng seeds which were sown in seed bed at Aug-5 without seed stratification and that of artificial seed stratification. 3. Embryo growth and germination ratio was significantly inhibited by high temperature treatment at $30^{\circ}C$ for 24 hours or respiration stress by immersing seeds in water for 10 days or more. 4. When the seed stratification was started at $10^{\circ}C$, growth of embryo in the ginseng seeds were almost stopped. But, when the seeds were stratified first at $20^{\circ}C$ for 50 days and next at $10^{\circ}C$ for 50 days, the embryo growth was significantly promoted compared with the embryo growth in the seeds which were stratified at $20^{\circ}C$ for 100 days. 5. The successive embryo growth after seed stratification was significantly accelerated at $10^{\circ}C$ but the seeds chilled at $5^{\circ}C$ for 100 days were resulted in the highest germination ratio as well as the shortest days for germination. 6. The successive embryo growth during chilling treatment and seed germination were significantly inhibited by immersing seeds in water just before chilling treatment or during chilling treatment and by interruption of chilling treatment with raising temperature to $20^{\circ}C$ for 20 days during chilling treatment. 7. The germination ratio of ginseng seeds which finished chilling treatment was highest at $10^{\circ}C$ and 62.5% was the estimated soil moisture for the best germination of ginseng seeds. The ginseng seeds were found to require high amount of oxygen for germination. 8. Only water soluble material in homogenized ginseng seeds showed a significant inhibiting effect on the seed germination of sesame, millet and soybean. Water soluble material dissolved from undehisced ginseng seeds showed stronger inhibiting effect on the seedling growth of sesame than material from dehisced ginseng seeds. Extraction temperature did not influence the inhibiting effect of the material dissolved from ginseng seeds on the seedling growth of sesame. 9. Water soluble materials dissolved from the berry pulps, leaves, fresh roots and dried roots also showed a significant inhibiting effect on the seedling growth of sesame. 10. Water soluble materials dissolved from the ginseng seeds, leaves and fresh roots showed a significant inhibiting effect on the germination of true fungi and the growth of spawn but the growth of phytopathogenic bacteria was not. 11. Among the water soluble materials dissolved from ginseng seeds, the materials of low molecular weight less than 3,000 were resulted a significant inhibiting effect on the seedling growth of sesame and the materials of high molecular weight also showed an inhibiting effect.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.22 no.1
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• pp.45-51
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• 1977

The studies were carried to know the effects of GA$_3$, Ethrel and $H_2O$$_2$ on dormancy and germination in ginseng seeds. GA$_3$ stimulated the embryo growth and increased dehiscent (Kaekapp) ratio of the seeds for more than Ethrel and $H_2O$$_2$ GA$_3$ not only increased germination ratio but also shortened the period of germination. Ethrel and $H_2O$$_2$ showed no effects on the germination and there were no significant differences among the treatment levels of GA$_3$. The slow germination of ginseng seeds seemed to be mainly due to the dormancy of endosperm or seed coat rather than of embryo.

• Korean Journal of Plant Resources
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• v.29 no.3
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• pp.339-346
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• 2016

This study was carried out to develop an effective seed propagation method for Thalictrum rochebrunianum var. grandisepalum (H. Lev.) Nakai by analyzing seed dormancy types and germination characteristics. Seeds were collected between September to October at Gangwon province, and well-selected seeds were used while being dry-stored at 4±1℃. The seed size ranged 4.52 × 1.58 ㎜ and the weight of thousand seeds were 1,603.5 ± 0.02 ㎎. The moisture content was 7.2%. Seeds were achene type, and morphology characters showed an elliptical shape and rough texture, and light brown in color. Moist-chilling treatment was conducted for dormancy breaking because the seeds had an undeveloped embryo of liner type. The embryo had developed during a moist-chilling period, constantly, and fully developed in 10 weeks. Consequently, it seemed to be non-deep complex or intermediate complex type of morphophysiological dormancy, and embryo dormancy was broken by wet-chilling for 10 weeks. After 10 weeks of wet-chilling treatment, seed germination began. Germination percentage was higher in dark condition raher than light condition and recorded the maximum at 25℃ in the dark (16.3%). A pre-soaking treatment with a combined plant growth hormones promoted germination and shortened T₅₀. Specifically, seed germination of 84.5% was achieved by pre-soaking of seeds with a combined solution of 500 ㎎/L GA₃ and 10 ㎎/L kinetin for 24 h after a wet-chilling treatment for 10 weeks. Thus the effect of plant growth hormones coupled with chilling temperature on seed breaking dormancy provide asubsequent growth of seedlings for successful plantation.

• Journal of Plant Biotechnology
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• v.45 no.4
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• pp.347-356
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• 2018

Chionanthus retusus is a small deciduous tree that is widely used in landscaping due to its beautiful white spring flowers and ornamental value. Conventional propagation through seeds requires one to two years of breaking dormancy. The objective of this study was to determine the conditions of in vitro germination in C. retusus. In vitro embryo culture was carried out to investigate the effects of six factors: basal media (McCown Woody Plant Medium (WPM) and Murashige and Skoog (MS)); plant growth regulators (different combinations and concentrations of naphthaleneacetic acid (NAA), 6-Benzylaminopurine (BA), and gibberellic acid ($GA_3$)); embryo age (collected weekly beginning 36 days after fruit setting); low temperature pretreatment (storing $4^{\circ}C$ for 1, 2, 3, and 4 weeks); coconut additives (100, 200, and $300ml{\cdot}L^{-1}$); and genotype (grouping plants depending on their flowering nature). The basal medium used in this study was WPM with $2mg{\cdot}L^{-1-1}\;GA_3$, $20g{\cdot}L^{-1}$ sucrose, and $6g{\cdot}L^{-1}$ Agar. WPM medium mixed with $GA_3$, resulted in higher germination rate as compared to when using a combination of auxin and cytokinin. $GA_3$ at $2mg{\cdot}L^{-1}$ was the most effective of all combinations and concentrations of PGRs. WPM medium with $2mg{\cdot}L^{-1}GA_3$ resulted in better and faster germination (75.93%). Embryos collected at 57 days after fruit setting had the highest percent of germinated seeds (87.04%) while low-temperature pretreatment of fruits at $4^{\circ}C$ for two weeks produced the highest germination (95.37%). These results of this study could be an open ground for development of an efficient protocol for commercial production of the ornamental tree.