• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2003.04a
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• pp.93-93
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• 2003

• Korean Journal of Plant Resources
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• v.21 no.4
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• pp.341-345
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• 2008

Two orchid species of Calanthe discolor Lindl. and C. discolor for. sieboldii (Decne.) Ohwi, which have different form flower color and size. They were crossed in mid April by artificial pollination, and the F1 hybrid seeds were collected mid October. Germination of seeds was investigated on pre-treatment of seeds and under the various environmental conditions. Germination was promoted by moisture absorption and ultrasonic treatment of seeds. Dark culture of F1 hybrid seeds enhanced germination and protocorm formation, and development into seedlings compared with light culture. Although, plant growth regulators such as NAA and BA had a slightly promotive effect on seed germination and protocorm growth, regenerated seeding were showed abnormal growth patterns. Regenerated F1 hybrid plantlets were successfully transferred to pot.

• Horticultural Science & Technology
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• v.16 no.2
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• pp.236-238
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• 1998

This study was conducted to investigate the effective media and optimal days after pollination for germination of seeds which were crossed by selfing or reciprocal pollination in Cymbidium spp. The elongation of ovary stopped about 110 days after pollination regardless of cross combinations. The increase of ovary width stopped about 140 days after pollination showing some differences among cross combinations. The cross combinations expressing high capsule number and viability showed good germination when they were sown in vitro. The capsules collected 150 to 180 days after pollination showed good germination performance and short days of germination. Hyponex media was the most effective for protocorm development and germination.

• Journal of Plant Biotechnology
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• v.46 no.1
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• pp.9-16
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• 2019

Dendrobium candidum Wallich ex Lindley is a traditional Chinese medicine plant and has been widely used for medicinal and ornamental purposes. In this study, several different factors affecting micro propagation of protocorm-like bodies (PLBs) such as basal media, plant growth regulators, and carbon sources. The proliferation PLB derived from seeds was the best in $H_3P_4$ basal medium containing $0.1mg{\cdot}L^{-1}$ NAA and $0.1mg{\cdot}L^{-1}$ Kinetin. PLB growth was the best when $10g{\cdot}L^{-1}$ sucrose was added to the carbon atoms in the medium. The rate of shoot formation from the propagated PLB was the highest in 1/4 MS or $H_1P_2$ medium containing $10g{\cdot}L^{-1}$ sucrose, and the shoot length was longer than the others.

• Journal of Plant Biology
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• v.33 no.2
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• pp.105-110
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• 1990

Physiological gradient of IAA-oxidizing enzyme activities was investigated in order to elucidate the mechanism of shoot differentiation in Cymbidium sp. (‘Jungfrau’) protocorms by using phenolic compounds (2, 4-dichlorophenol, catechol), auxin-inhibitors (PCIB, TIBA), and hormones (GA3, ABA, BA). The activity of IAA oxidase was decreased in protocorms treated with catechol decreased the catalytic activity of IAA oxidase or TIBA but this enzyme activity was increased after a temporary decrease at initial stages in the presence of 2, 4-dichlorophenol or PCIB. The activity of IAA oxidase in BA-treated protocorms (white and crown gall-like) was the highest of all. However, the catalytic activity of peroxidase increased after a temporary decrease at initial period. These results suggest that shoot differentiation and growth may be influenced by effective IAA levels in the protocorms causing IAA-oxidizing enzyme and phenolic compounds.

• Plant Resources
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• v.5 no.3
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• pp.176-180
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• 2002

This study was conducted to develop the clonal propagation technique through in vitro culture using by leaf sheath explants of Phalaenopsis grown in vitro. The highest frequency of protocorm-like body (PLB) formation was obtained when explants of leaf sheath were cultured on VW medium containing 30g/L sucrose, 500 mg/L activated charcoal, 150 mVL coconut water, 1 mg/L NAA, 1 mg/L 2ip and 2.5 g/L gelrite. The PLB formation rate of VW medium was highest followed by modified Hyponex medium, and lowest in MS medium. Plantlets induced from PLBs transferred to plastic pots including spagnum moss were well developed.

• Plant Resources
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• v.6 no.2
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• pp.89-93
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• 2003

This study was conducted to develop the clonal propagation technique through in vitro culture using basal stem explants in Phalaenopsis hybrid grown in vitro. The highest frequency of protocorm-like body (PLB) formation was obtained when basal stem explants were cultured on VW medium containing 30g/L sucrose, 500 mg/L activated charcoal, 150 ml/L coconut water, 1 mg/L NAA, 5 mg/L 2iP and 2.5g/L gel rite. PLBs transferred to Hyponex medium were regenerated to plantlets. Plantlets transferred to plastic pots containing spagnum moss were developed and successfully acclimatized under greenhouse. The flower was bloomingly opened in plants regenerated from basal stem explants. The flower was not different from both mother plant and plant induced through clonal propagation of Phalaenopsis hybrid.

• Journal of Plant Biology
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• v.33 no.1
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• pp.41-48
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• 1990

Changes in polamine titers during shoot differentiation in Cymbidium sp. (Jungfrau) protocorms were studied in order to investigate the mechanism of shoot differentiation by using auxin-inhibitors(PCIB, TIBA), hormones(GA3, ABA, BA), and phenolic compounds (2,4-dichlorophenol, catechol). The shoot differentiation and propagation of protocorms were promoted by PCIB or 2,4-dichlorophenol, and the growth of differentiated shoot were promoted by TIBA or catechol. In BA-treated protocorms, white or brown protocorms were observed. Putrescine was the most abundant polyamine during the propagation and differentiation processes. As compared with putrescine, spermidine did not show significant changes and spermine was not detected at all. Putrescine titers decreased after a temporary increase, and then again increased in the presence of GA3, ABA, 2,4-dichlorophenol, and then again increased in the presence of GA3, ABA, 2,4-dichlorophenol, catechol, or PCIB. But, in BA-treated protocorms, putrescine level was much lower than spermidine.

• Journal of Plant Biotechnology
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• v.44 no.1
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• pp.56-60
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• 2017

This study investigated the in vitro symbiotic seed germination of the achlorophyllous orchid, Gastrodia elata, using a new Mycena species. A leaf-disc ($2{\times}2cm$) of Quercus accutissima was inoculated with either of the two fungal species, NIFOS101 (NCBI accession number KY449288) or KFRI1212 (HQ662845), following which it was placed on water agar medium, prior to sowing seeds of G. elata. After 20 days of symbiotic culture, NIFOS101 and KFRI1212 germinated 94% and 70% of G. elata seeds, respectively, although the initiation of protocorm development was induced much earlier by KFRI 1212 than by NIFOS101. Furthermore, the NIFOS101 mycelia grew much faster than KFRI 1212 at all temperatures tested. A phylogenetic analysis using the internal transcribed spacer (ITS) sequences showed that NIFOS101 belonged to a clade with M. purpureofusca, which completely differed from the clade with KFRI1212. This study not only identified a new fungal species, NIFOS101, which improved the rate of symbiotic seed germination up to 94% as compared to KFRI1212 (70%), but also revealed that G. elata required a broad taxonomic range of fungi for its symbiotic germination.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.04a
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• pp.53-53
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• 2019

Cypripediums, popularly called lady's slippers or moccasin flowers, are the showiest and most sought after hardy terrestrial orchids, collected and grown by orchid and alpine plant enthusiasts alike. In Korea, 4 species of cypripedium are reported as Cypripedium japonicum, C. macranthos, C. guttatum, and C. calceolus. We had already reported the feasibilities of C. macranthos and C. guttatum with in vitro germination methods from immature seeds. The seeds of white lady's slipper orchid (Cypripedium macranthos Sw. alba) were collected 65 days after pollination in 2018. The green pods were sterilized with flame and sowed immediately on the POM(Phytomax orchid maintenance media(R), Sigma) supplemented with BAP 0, 0.5, 1.0 mg/L and NAA 0, 1, 2mg/L. The germination of seed was observed 90 days after sowing, and the plantlets were subcultured to the same media according to the size of the protocorm with 1~2, 2~3, 3~4, 5~6, 7~8mm. The time of the subculture to the new media seems to be critical factors of forming rhizoids which is the hairy root of the cypripediums. As a results, the protorms of the white lady's slipper orchid was successfully germinated in the POM media supplemented BAP 0.5 and NAA 1.0 mg/L. The roots and rhizoids were formed in 5~6mm protocorms subculture over 95% survival ratio. We also tried to subculture to liquid medium without activated charcoal, however the browning or malformation of the roots was observed in the root. The formation of shoots from the protocorm was effectively enhanced in the POM media with non-additives of plant growth regulators. These results indicate the possibility of high and stable production and practical industrialization of endangered white lady's slipper orchids.