• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.04a
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• pp.45-45
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• 2018

We had already reported the successful germination for green pods of purple lady's slipper orchid (Cypripedium macranthos Sw.). The green pod methods is to take immature seeds in green capsules, sterilize the capsule, and take out the sterile seeds. This method, however, needs very critical time of harvest. The critical time of seed harvest changes depending upon the species, condition of the specimen, and climatic influence, and the right time lies between 5 and 12 weeks after fertilization. In this study, the mature seeds were collected after 120-130 days with hand-polination of lady's slipper orchids. Mature seeds are usually dormant and it has to be overcome, either with hormone or storing the seeds near freezing for two or three months to break dormancy. The seeds were first surface sterilized with 70% ethanol and then transferred 1% NaOCl for 10-15 minutes, followed by rinses 3 times with sterilized distilled water. The cypripedium seeds consists of an embryo within a seed coat known as a testa. The testa is water repellent and the seed has a large air space between the embryo and testa so the seed tends to float on water. We had resolved the problems with vacuum pump to soak water into the testa before sterilization. The seeds were placed on liquid or agar solidified germination media. Cultures were incubated at $24{\pm}1^{\circ}C$ in dark. The seeds were germinated in 6-8 weeks in liquid suspension culture (germination percentage over 18%); however, the seeds on agar solidified media took more than 5 months to germinate and the germination percentage less than 5%. The most effective media for liquid culture was 1/4 strength Murashige and Skoog (MS) medium with 50 ml/l coconut water ($4brix^{\circ}$) at pH 5.8.

• FLOWER RESEARCH JOURNAL
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• v.17 no.4
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• pp.221-225
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• 2009

Interspecific crossing was conducted to obtain hybrid lilly with better quality of adapting to the unfavorable environment using oriental Lilium spp. as a female parent. Average interspecific crossing rate between oriental and asiatic lines by cut-style pollination was as low as 29%. Although the average germination rate of zygotic hybrid embryo between oriental 'Rodolfa' ${\times}$ asiatic 'Toronto' was increased from 76.6% to 78.3%. on 114 or 1/6 strength MS medium compared to the control, the rate of zygotic embryos formation between oriental 'Farolito' ${\times}$ asiatic 'Buff Pixie' was significantly enhanced up about 86~90% on 1/2 strength MS medium. Meanwhile, germination rate of interspecific hybrid embryo between oriental 'Snow Cristal' ${\times}$ asiatic 'Royal Trinity' showed up 100%. Germination rate of interspecific hybrids slightly increased by addition of $0.5-1.0mg{\cdot}L^{-1}$ GA. Germination rate of zygotic embryos between oriental 'Farolito' ${\times}$ asiatic 'Buff Pixie' and oriental 'Belcanto' ${\times}$ L. callosum was highest on the MS medium containing $0.01mg{\cdot}L^{-1}$ NAA and $0.1mg{\cdot}L^{-1}$ kinetin, as 66.6% and 45.2% respectively.

• Korean Journal of Breeding Science
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• v.42 no.5
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• pp.463-469
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• 2010

Rice is a staple food for more than 50% of the world's population. Embryo comprises only 2 to 3% of the weight distribution of the entire pericarp but has higher concentration of vitamins, proteins, and essential fatty acids than the other parts of grains. Moreover, ${\alpha}$-tocoperol, ${\gamma}$-oryzanol, phytic acid and ${\gamma}$-aminobutric acid that have nutraceutical value are abundant. Increasing the volume of embryo assures the fortification of nutritional value of rice grain. We developed new black waxy giant embryo rice, Milyang 263 by crossing Josaengheugchal, a black waxy rice variety, and $ge^t$, a giant embryo mutant generated by tissue culture. The nutrient contents and physical properties of Milyang 263 were compared with several giant embryo mutants and normal embryo rice varieties. Changes in the nutrient properties after germination were also observed. Results indicated that this new black waxy giant embryo rice, Milyang 263, offers a promising source for improving nutritional quality of rice especially anthocyanin, essential minerals, and GABA.

• Korean Journal of Plant Tissue Culture
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• v.21 no.4
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• pp.215-220
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• 1994

Pollen-derived embryos cultured on the hormone-free medium showed a low germination frequency (12.5%) and poor growth response after germination. The greatest frequency of germination (81.3%) was obtained from the embryos cultured on medium with 0.3mg/L GA$_3$.The greatest frequency of generation (81.3) was obtained from embryos cultured on medium with 0.3mg/L GA$_3$. The embryos precultured for 20 days on medium with 0.3mg/L GA$_3$were transferred to the medium with various combination of hormones such as IAA, kinetin, zeatin, 6-benzylaminopurin (BA) and Gh$_3$. The germination frequency of cotyledonary stage embryos showed above 72% on media with all of the hormonal combinations, but the embryos germinated on medium with 2mg/L BA or 0.1mg/L kinetin and 0.3mg/L GA$_3$ developed more vigorously into plantlets than those of other hormonal combinations. Torpedo-stage embryos cultured on medium with 0.3 mg/L Gh$_3$ were pretreated for 8 weeks at 2-week intervals at 4$^{\circ}C$, The germination frequency of the cold-preheated embryos increased with the increment of pretreatment period from 2 to 8 weeks. The greatest frequency of germination (73.3%) was obtained from the embryos pretreated for 8 weeks at 4$^{\circ}C$. The chromosomes of the root-tip cells of W plane grown for 40 days after germination were observed. Most of the regenerated plants were haploid (55.8%) or diploid (315%), but triploid (1.3%), tetraploid (5.2%), or aneuploid (6.5%) were also detected among them.

• Journal of Plant Biotechnology
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• v.6 no.2
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• pp.91-96
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• 2004

The use of liquid-medium-based procedure relative to the solid media led to a 4.5-fold increase in the number of cotyledon-stage embryos. The most efficient system for multiplication and regeneration of somatic embryos was CP6 procedure with the media MSD40/MSD20/MSM6AC/FNL0S3S3GM. However, the rate of regeneration was lower. About 71％ of the embryos with dicotyledon were continued to develop the roots after desiccation treatment and 92％ of the germinated embryos produced shoots in 10 days. Of the four morphologically different types of embryos, dicotyledonous ones showed a high frequency of conversion, while only a few with fused and horn type cotyledon developed shoots. Mature somatic embryos were desiccated in empty petri dishes for 12-72 h. Embryo survival rate was the highest after 12 h of desiccation, but maximal germination was observed at 24 h. After desiccation, they were placed on MS medium without growth regulators for germination. Germinating embryos were transferred to small pots with vermiculite for plant regeneration. The etiolating the plants during the growth was resolved to add 1％ activated charcoal on hormone-free MS medium.

• Korean Journal of Plant Tissue Culture
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• v.27 no.3
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• pp.227-232
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• 2000

This study was carried out to elucidate the effect of cadmium on somatic embryogenesis and plant regeneration from cultured cells of Daucus carota L. Embryogenic calli were induced from cotyledon explants of carrot seedlings cultured on MS solid medium supplemente with 1 mg/L 2,4-D Embryogenic cells proliferated on medium supplemented with 1 mg/L 2,4-D were also cultured in liquid MS medium containing various concentrations (50, 100, 200, 500, 1000 $\mu$M) of cadmium for one week and then transferred to MS basal medium. Somatic embryogenesis occurred in suspension culture treated with 50 $\mu$M and 100 $\mu$M cadmium or untreated with cadmium. When cadmium was treated in suspension culture, production of two and four cotyledonary somatic embryos was reduced, but that of three cotyledonary somatic embryo was increased. Two cotyledonary embryos showed higher regeneration frequency than abnormal somatic embryo with one, three and four cotyledon. Regardless of cotyledonary variation, germination frequency of somatic embryos treated with cadmium was decreased in compared with that of embryos in basal medium.

• Journal of Forest and Environmental Science
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• v.38 no.3
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• pp.152-158
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• 2022

A cryopreservation is an essential tool for preservation of germplasm. In this study, the possibility for cryopreservation of embryogenic cells of Siberian ginseng (Eleutherococcus senticosus) in liquid nitrogen (-196℃) was evaluated. The effects of glycerol and dimethyl sulfoxide (DMSO) at different concentrations (5%, 10% and 20%) as cryoprotectants on regrowth of cryopreserved E. senticosus embryogenic cells were tested. There was significant effect of cryoprotectants on regrowth of embryogenic cells (p=0.0019). The highest and lowest fresh mass gain were achieved when embryogenic cells were frozen with 10% DMSO and 5% glycerol (138.2±5.9 and 61.3±14.6, respectively). The effect of the cryoprotectants on the frequency embryo germination was tested. There was no significant difference between glycerol and DMSO (p=0.846). Three different concentrations of cryoprotectants did not significantly affect the frequency embryo germination (p=0.534). Finally, the genetic fidelity of the plantlets regenerated from non-cryopreserved and cryopreserved embryogenic cells was tested by random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analysis. RAPD and ISSR analysises showed that there was no genetic variation among regenerants.

• Journal of Plant Biotechnology
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• v.35 no.1
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• pp.47-53
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• 2008

Effects of growth regulators and culture conditions on seed germination, haustorium development, and callus formation of Korean mistletoe (Viscum album var. coloratum (Kom.) Ohwi) were described. Histological examination showed that seed of V. album contained one or two zygotic embryos with rod shape, and actively dividing cells were mainly distributed in radicle region rather than cotyledon of zygotic embryo. The most significant factor for seed germination and haustorium development of V. album was the requirement of the light. Various growth regulators examined in this study failed to substitute the effect of the light on seed germination. The frequency of callus formation was highest at 27.3% when flower buds were cultured onto B5 medium containing $0.1\;mgl^{-1}$ IAA. Explants from other organs were recalcitrant in forming calluses. Culture conditions described in this study could be applied for production of useful metabolites and multiplication of V. album in future.

• Korean Journal of Breeding Science
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• v.41 no.4
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• pp.403-411
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• 2009

We inspected seed sizes of 353 genetic accessions of watermelon to diversify functional utility related to seed size and classified them into six representative groups based on their seed sizes. Each group was named as giant seed (GS), big seed (BS), medium size (NS), small size (SS), micro seed (MS) and tomato seed (TS) from the biggest. As the seed size was getting smaller, decreased seed length and seed width, increased seed number per fruit, and decreased seed weight per fruit were observed, but seed shape did not change significantly. In order to study the effect of seed size on fruit weight and seed germination, we developed three near isogenic lines (NILs) with three different seed sizes, SS, MS and TS, from crossing between two accessions 'NT' and 'TDR', and one NIL with seed size of TS from crossing between two accessions 'S55' and 'TDR'. In the study on the fruit weight of NILs with various seed sizes, NS, SS, MS, and TS NILs produced an average of 6.4, 6.3, 5.9, and 4.2 kg fruits, respectively. The bigger seed types showed the better germination rate. NS type showed the highest germination percentage, while TS showed very low germination percentage. Fermentation treatment for 48 hrs increased the germination percentage on TS type seed, but still remained at a low level. In NS, SS, and MS material, the ratio of embryo/whole seed weight was over 50%, meanwhile that of TS was only 44.4% of which low embryo percentage would be one of the reason of low germination percentage. From this study, we concluded that watermelon has very wide genetic diversity on seed size which is somehow related to fruit sizes and germination rate.

• Journal of The Korean Society of Grassland and Forage Science
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• v.19 no.4
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• pp.297-302
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• 1999

Cotyledonary abnormalities were observed in secondary somatic embryos which were developed from calli cultured on MS medium with various concentrations of 2,4-D. In MS medium containing hormone-free or $0.1mg/{\ell}$ 2,4-D, the frequency of normal embryo formation with two cotyledons were above 57%. According to concentration of 2,4-D increment the frequency of normal embryos were decreased. In MS medium containing $4mg/{\ell}$ 2,4-D, the frequency of normal embryo formation was just 10%. The rate of germination was as follows; 37.7% of somatic embryos had one cotyledon, 85% two cotyledons, 38% three cotyledons, 35% four cotyledons, 25% five cotyledons and 29% trumpet-like cotyledons. About 80% of the embryos with two cotyledons were converted into normal plants, but one, three or four cotyledons were only 6.8~10%. The five or trumpet-like embryos were not developed into normal plants.