• Journal of the Korean Society of International Agriculture
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• v.23 no.5
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• pp.465-474
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• 2011

Agricultural environment of Lam Dong province, which is located in central highland area, is totally different from that of other lower areas in Vietnam. In Lam Dong province, abundant plant resources were naturally grown such as pine trees, taxus, and wild orchids, which can grow in high mountainous area. In Lam Dong, the field proportion of perennial crops was higher than that of annual crops. However, the field proportion and yields of vegetables were highest among the all cultivated crops, estimating 38% (36,552ha) and 72% (993,082MT), respectively. Especially in Da Lat, vegetables, flowers, orchids, and industrial crops were mainly produced because this area is geographically close to Ho Chi Minh city. And also in Da Lat, 64% (8,447ha) and 36% (4,777ha) of farm fields were used for producing annual and perennial crops, respectively, and the yields of fresh vegetables in this area was estimated to 213,478MT which was 21.5% of the whole yields in Lam Dong province. Thus Korea, Taiwan, Japan, France, and Holland have invested to agriculture in Da Lat for producing and exporting flowers, vegetables, and tea. In 2009, flower cultivation area of Da Lat was over 55% in Lam Dong province and average amount of values were 9,781 million USD, which was higher than that of al other crops. Thus following strategies could be suggested for the development of agriculture in Lam Dong province. The first, agricultural cooperation with Da Lat, Lam Dong, should be characterized to horticulture and floriculture, followed by supporting both appropriate R&D techniques and equipments. And then agricultural system should be made in relationship with the local companies. Finally, agricultural cooperation program should be conducted toward the direction for both donor and recipient countries.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.10a
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• pp.37-37
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• 2018

We use two different methods for laboratory propagation from seed of lady's slipper orchid(Cypripedium macranthos Sw.); immature seed which also called green capsule or fully mature seed about 120~130 days from pollination. In green capsule culture, the seed pods should be collected within precisely right time. The right time of seed collection could be diverse under the wether conditions or nutritional factors of the plants. In fully matured seed culture, the more complicated procedures are needed to break the dormancy of the seed; thermal or chemical treatment. The seedlings in this study were easily germinated from immature seeds in Harvais medium; 53 days after pollination(DAP) in Cypripedium pubescens, DAP 65 in C. parviflorum and C. macranthos. The germinated seedlings were transplanted to hormone free media immediately to avoid abnormal growth of seedlings. When the seedlings have roots with a minimum length of around 2-3cm and have visible dormant buds, the seedlings were removed from the flask and stored in refrigerator for vernalization. To examine the correlation of seedlings and maternal plants, the 125 seedlings of C. macranthos were transplanted in the soil bed at a distance of 20-100 cm from mother plants on April 20. The survival rate of seedlings were 92% in 20 cm distance from the ripe plants, and 56 % in 100 cm distance. The seedlings which were transplanted near mother plants showed vigorous growth in plant height, leaf width, and especially dormant buds. Considering the existence of mycorrhiza which is a symbiotic association between a fungus and the roots of a orchid vascular, the various fungus from mother plants could affect the growth of the seedlings. These results indicate the possibility of high and stable production and practical industrialization of endangered lady's slipper orchids.

• Journal of Plant Biotechnology
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• v.33 no.4
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• pp.297-302
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• 2006

In general, the proliferation of orchids via somatic embryos has been used for mass production of somatic clones because of high propagation efficiency. In spite of high propagation rate, this method often brings somaclonal variation, especially polyploid frequency. Therefor we here concentrated to investigate the relationship between endopolyploidization patterns of explants and the occurrence of tetraploid variant in clonally proliferated Doritaenopsis via somatic embryo regeneration system. In the fully developed somatic embryo, upper part contained 2C to 16C while middle and lower parts showed 2C to 32C DNA content. Two-week-old embryo contained 2C to 16C, whereas those regenerated after 4 to 10-week-old contained 2C to 64C nuclei. Results showed that endoreduplication was variable depending upon tissue types, ages, and parts in one species. lower part of somatic embryo having high endoreduplication degree increased the regeneration of tetraploid variants by about 3-fold comparing to upper part of somatic embryo culture. polyploid frequency occurrence might be closely related to the high levels of endoreduplication of somatic embryos used as explant. It suggested that the upper part of somatic embryo having comparatively low endoreduplication degree is suitable for the stable in vitro propagation system.

• Journal of Plant Biotechnology
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• v.39 no.3
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• pp.163-168
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• 2012

An optimization in vitro seed germination was established by using triphenol tetrazolium chloride (TTC) test, which has been known as two rare orchids (Gastrodia verrucosa Blume and Hetaeria sikokiana Tuyama) in Jeju island. We have established proper NaOCl treatment for in vitro germination of G. verrucosa and H. sikokiana through TTC test. In the case of H. sikokiana, seed viability through TTC test was high with 95% in control. However, NaOCl 1% treatment for 30 minutes showed the highest embryo swelling rate to seed viability. Likewise, swelling formation of embryos, diameter of embryos, protocorm formation and diameter of protocorms of G. verrucosa was 87%, $59{\mu}m$, 91% and $138{\mu}m$ through NaOCl 1% treatment for 30 minutes. This result will be applied on the basic information for improving in vitro seed germination rate of G. verrucosa and H. sikokiana.

• Korean Journal of Plant Taxonomy
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• v.45 no.4
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• pp.353-361
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• 2015

Spatio-temporal variation in fruit set in orchids would affect long-term population viability and will influence genetic diversity over many generations. The aim of this study was to examine the breeding system of the nectariferous terrestrial orchid Epipactis thunbergii, to specifically determine levels of fruit set in terms of time and space under natural conditions. We examined pollination under natural conditions and conducted hand pollination experiments during a 2-year survey in four populations located along 1.5 km of coastal line in Jinguiri (rual village) [Jeollanam-do (province), southern Korea]. We found that, over a 2-year period, levels of percentage of fruit set were similar within patches of the four populations. By contrast, we detected significant differences in the percentage of fruit set among patches. We also found that plants with larger inflorescence size produced significantly more fruits than plants with fewer flowers. Over a 2-year period, the percentage of fruit set for E. thunbergii was similar but low (14.1%) compared to that averaged for eighty-four rewarding species (37.1%). However, an increase in fruit set was achieved by hand-pollinations: artificial self-pollination (90.5-95.2%), artificial geitonogamy (94.7-95.0%), and cross-pollination (artificial xenogamy, 91.3-91.4%). No emasculated flowers produced fruits and no automatic pollination was found in E. thunbergii. Our findings suggest that E. thunbergii is a self-compatible terrestrial orchid that depends on pollinators (insects) to achieve fruit set in natural habitats, and that local environmental conditions were similar over a period of 2 years in the study area. Our results also highlight the cryptic variation of fruit production in time, but more pronounced variability in space.

• Korean Journal of Plant Taxonomy
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• v.47 no.4
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• pp.289-296
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• 2017

The three self-compatible, terrestrial orchids Cephalanthera erecta, C. falcata, and C. longibracteata flower synchronously in sympatric populations. Cephalanthera erecta and C. longibracteata, which have white flowers that do not fully open, are predominantly autogamous, whereas the food-deceptive C. falcata, whose bright yellow flowers open completely, is predominantly outcrossing. The formation of hybrids rarely occurs between species owing to strong prepollination barriers (floral isolation). If these three species have evolved toward the prevention of interspecific hybridization, we can expect that the spatial distribution patterns of the three species would be characterized as spatial segregation (i.e., 'spatial repulsion') from each other. To test this prediction, we studied the three Cephalanthera species in sympatric populations showing coincident flowering within Yeonwhasan Provincial Park (YPP, Gyeongsangnam Province, South Korea). We found strong spatial aggregation in each population and spatial independence in the interspecific spatial distribution, differing from previous studies. We further hypothesize that Cephalanthera species in sympatry within YPP are distributed somewhat randomly in space, perhaps due to the sharing of similar mycorrhizal fungi.

• 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.

• Korean journal of applied entomology
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• v.54 no.4
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• pp.345-349
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• 2015

The pest species occurring on the aboveground parts of Phalaenopsis orchids were studied for a period of two years (Aug. 2013 to June 2015) at 9 Phalaenopsis farms in Korea. In total, 10 species of insect pest were identified : Tenuipalpus pacificus Baker, Tyrophagus putrescentiae Schrank, Frankliniella occidentalis Pergande, Frankliniella intonsa Trybom, Thrips tabaci Lindeman, Scirtothrips dorsalis Hood, Pseudococcus dendrobiorum Williams, Pseudococcus longispinus Targioni Tozzetti, Coccus hesperidum L., Limax marginatus $M{\ddot{u}}ller$. T. pacificus occurred in most of the farms surveyed in this study (6). The observation frequency of T. pacificus and F. intonsa were highest, they were observed in 10 out of 96 surveys. Although P. longispinus was only observed to occur on two farms, their observation frequency (9 observations, 9.4%) and density (2-289 per 20 plants) were high. Thus, it seems that T. pacificus, F. intonsa, and P. longispinus are the major pests affecting Phalaenopsis orchid farms in Korea.

• Journal of Plant Biotechnology
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• v.41 no.4
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• pp.216-222
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• 2014

This study was carried out to develop an efficient transformation protocol via particle bombardment with PLBs (protocorm-like bodies) in Phalaenopsis. To achieve this aim, osmoticum treatment and an increasing shooting chances in particle bombardment process were applied for this study. In addition, pCAMBIA3301: ORE7 vector which contains a herbicide-resistance bar gene as a selectable marker and ORE7 gene as a gene of interests were employed. With regard to the increasing chances of shooting in particle bombardment, double shooting was the best results with 1.5 ~ 2.5 times higher than those of a single or triple shooting treatment in the productioon of PPT (D-L-phosphinothricin)-resistant PLBs. However, regeneration rate of shoots in double shooting was not high as a single shooting. Further, double shooting showed 35 ~ 40% higher than that of a single shooting in the frequency of browning. Regarding effects of different osmotic treatments, combination of 0.2 M sorbitol with 0.2 M mannitol showed the best results in transformation efficiency, regeneration of transformants and reduction of browning. Putative transgenic Phalaenopsis plants were analyzed by PCR analysis and confirmed the presence of bar and ORE 7 gene. Also, real-time PCR was conducted by using 21 transgenic plants and showed only 4 plants had one copy of transgene; whereas, the other 17 plants had more than 2 copies of transgene. Transgenic phalaenopsis plants produced in this study were transferred to pots and flowered normally without morphological variations in flower and leaf.

• Journal of Plant Biotechnology
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• v.41 no.4
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• pp.223-228
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• 2014

To establish an efficient proliferation and regeneration of PLBs (protocorm-like bodies) of Phalaenopsis plants, a variety of propagation medium types, various concentraions of sucrose as well as liquid and solid type were tested in this study. Further, activated charcoal, citric acid and ascorbic acid were compared whether these agents are suppose to reduce the browning in culture process using PLBs of Phalanopsis plants. With regard to the proper propagation medium, VW medium showed 1.3 ~ 2 times highr than those of other medium in an index of increasing for fresh weight and 50% higher than those of other medium in the frequency of shoot regeneration. However, regarding liquid and solid types of culture, there were no significant differences in the proliferation of PLBs and regeneration of shoots from PLBs. In the experiment for a variety of sucrose concentrations (0 ~ 50 g/l), 10 g of sucrose showed 30 ~ 50% higher than other concentrations in increasing index and 10 ~ 50% higher in the regeneration of shoots from PLBs. Regarding the reduction of browning in tissue culture via PLBs of Phalaenopsis plants, 1 g of activated charcoal showed only 1.5% browning of PLBs cultured. Whereas, other treatments including citric acid and ascorbic acid showed 6 ~ 16% of browning of PLBs. Therefore, activated charcoal was selected as an efficient anti-browning agents for the culture of PLBs in Phalaenopsis plants. Using above-described results can be contibuted to the establishment of mass propagation system using PLBs of Phalaenopsis plants in the future.