• The Korean Journal of Mycology
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• v.43 no.1
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• pp.20-25
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• 2015

This study was conducted to investigate diversity of orchid endophytic fungi from roots of two terrestrial orchids, Cypripedium japonicum and Cypripedium macranthum in Korea. The endophytic fungi were identified using morphological and DNA sequences isolated from the roots. Totally, 11 taxa of the endophytic fungi were identified from the roots of C. japonicum and 15 taxa from C. macranthum. Three species of the fungi were common in both host species; Leptodontidium orchidicola, Humicola fuscoatra var. fuscoatra, Umbelopsis dimorpha. Six species of the fungal isolates were the first report in Korea; Oidiodendron echinulatum, Pseudogymnoascus roseus, Geomyces vinaceus, Cryptosporiopsis ericae, U. dimorpha, Chaetomium cupreum.

• The Korean Journal of Mycology
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• v.38 no.1
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• pp.1-4
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• 2010

Orchid mycorrhizal fungi(OMF) were examined in roots of the two threatened orchid species, Cypripedium japonicum and C. macranthum. The morphological characteristics of mycorrhizal colonization in the roots of two orchid species were observed. OMF colonized in the roots of two species were identified using molecular analysis. DNA from the root was extracted and amplified internal transcribed spacer(ITS) region using basidiomycete ITS primers, ITS1-OF and ITS4-OF. Four species belonging to Tulasnellaceae in roots of C. japonicum and two species of Tulasnellaceae and one basidiomycetous species was found in roots of C. macranthum.

• Mycobiology
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• v.48 no.5
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• pp.423-426
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• 2020

Cypripedium japonicum is known to be the indigenous plant to Korea, Japan, and China. However, C. japonicum represents the most critically endangered plant species in South Korea. The plant is esthetically pleasing due to its flower, which is larger than any other orchidaceous species. Disease symptoms relating to gray mold were observed on C. japonicum in May 2019. The suspected pathogen was successfully isolated from the symptomatic leaf tissue and conducted a pure culture of the fungi. The conidia formed consisted of a colorless or light brown single cell, which was either egg or oval-shaped with a size of 7.1 to 13.4 × 5.2 to 8.6 ㎛. Molecular phylogenetic relationship analysis was also confirmed that the pathogen concerned belonging to the family of Botrytis cinerea. Therefore, the findings confirmed that the pathogen isolated from C. japonicum was consistent with the unique properties of B. cinerea.

• Mycobiology
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• v.45 no.2
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• pp.97-100
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• 2017

Endangered native plant habitats and populations are rapidly disappearing because of climate and environmental changes. As a representative, the abundance of the first-class endangered wild plant, Cypripedium japonicum, has been rapidly decreasing in Korea. The purpose of this study was to evaluate the distribution of rhizosphere and endophytic fungi on C. japonicum in its native habitat. A total of 440 rhizosphere and 79 endosphere fungi isolates were isolated and identified on the basis of their molecular characteristics. Sixty-five genera and 119 fungi species were identified in this study. The genus Trichoderma showed the highest abundance among both rhizosphere and endosphere fungi. Mortierella, Hypocrea, and Penicillium spp. were also relatively dominant species on C. japonicum. The community structures of rhizosphere and endosphere fungi were similar, but endosphere fungi showed greater diversity.

• Mycobiology
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• v.50 no.2
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• pp.142-149
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• 2022

Orchids live with mycorrhizal fungi in mutualism. This symbiotic relationship plays an essential role in the overall life cycle of orchids from germination, growth, settlement, and reproduction. Among the 1000 species of the orchid, the Korean lady's slipper, Cypripedium japonicum, is known as an endangered species. Currently, only five natural habitats of the Korean lady's slipper remain in South Korea, and the population of Korean lady's slipper in their natural habitat is not increasing. To prevent extinction, this study was designed to understand the fungal community interacting in the rhizosphere of the Korean lady's slipper living in the native and artificial habitats. In-depth analyses were performed to discover the vital mycorrhizal fungi contributing to habitat expansion and cultivation of the endangered orchid species. Our results suggested that Lycoperdon nigrescens contributed most to the increase in natural habitats and Russula violeipes as a characteristic of successful cultivation. And the fungi that helped L. nigrescens and R. violeipes to fit into the rhizosphere community in Korean lady's slipper native place were Paraboeremia selaginellae and Metarhizium anisopliae, respectively. The findings will contribute to restoring and maintaining the endangered orchid population in natural habitats.

• Korean Journal of Ecology and Environment
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• v.48 no.4
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• pp.253-262
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• 2015

The sustainability of Cypripedium japonicum, a rare plant designated by the Korea Forest Service, is threatened due to artificial factors such as habitat loss and climate change etc. and internal factors such as changes in biological properties of the habitat etc. but conservation research has not been performed in South Korea. The objective of this study is to establish the species conservation strategies by analyzing the characteristics of their habitats, including: 1) Population characteristics, and 2) habitat analysis of the vegetation and abiotic environments. From April to September, 2014, population characteristics [density (stems $m^{-2}$), flowering rate (%), and leaf area ($cm^2$)] in Cypripedium japonicum habitats such as Chuncheon (CC), Hwacheon (HC), Muju (MJ), and Gwangyang (GY) and vegetation characteristics (plant sociological research and ordination analysis), and abiotic environments [temperature ($^{\circ}C$), relative humidity (%), transmitted light ($mol{\cdot}m^{-2}{\cdot}d^{-1}$) and canopy openness (%)] were measured. Cypripedium japonicum was mainly distributed at elevation 450 to 990 m and 5 to $30^{\circ}$ slope. Slope direction was shown as 0 to $110^{\circ}$. Habitats temperature (mean $18.94^{\circ}C$) was well matched to seasonal changes. Differences among sites showed greater level according to latitude difference. It showed the highest in habitat, GY located in the South. On the other hand, relative humidity (77.38%) didn't show much difference among sites. The average degree of canopy openness was 18.17%. It showed the highest at HC (22.1%) and the lowest at MJ (16.1%). The average degree of transmitted light was $9.1mol{\cdot}m^{-2}{\cdot}d^{-1}$. It showed the highest at CC ($10.6mol{\cdot}m^{-2}{\cdot}d^{-1}$) and the lowest at GY ($6.87mol{\cdot}m^{-2}{\cdot}d^{-1}$). Chlorophyll content showed average 26.12 SPAD. It showed the highest at MJ (30.64 SPAD value) and the lowest at HC (23.69 SPAD value). Leaf area was average $253.35cm^2$. It showed the highest at CC ($281.51cm^2$) and the lowest at HC ($238.23cm^2$).

• Korean Journal of Environment and Ecology
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• v.35 no.5
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• pp.548-557
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• 2021

Cypripedium japonicum Thunb. is an endemic plant in East Asia, distributed only in Korea, China, and Japan. At the global level, the IUCN Red List evaluates it as "Endangered Species (EN)," and at the national level in Korea, it is evaluated as "Critically Endangered Species (CR)." In this study, we investigated the characteristics of the age structure and the sustainability of the population based on the data obtained by demographic monitoring conducted for seven years in the natural habitat. C. japonicum habitats were observed in 7 regions of Korea (Pochoen, Gapyeong, Hwacheon, Chuncheon, Yeongdong, Muju, Gwangyang), and 4,356 individuals in 15 subpopulations were identified. The population size and structure differed from region to region, and artificial management had a very important effect on the size and structural change of the population. Population viability analysis (PVA) based on changes in the number of individuals of C. japonicum showed a very diverse tendency by region. And the probability of population extinction in the next 100 years was 0.00% for Pocheon, 10.90% for Gwangyang, 24.05% for Chuncheon, and 79.50% for Hwacheon. Since the above monitored study sites were located within the conservation shelters, which restricted access by humans, unauthorized collection of C. japonicum, the biggest threat to the species, was not reflected in the individual viability. So, the risk of extinction in Korea is expected to be significantly higher than that estimated in this study. Therefore, it is necessary to reflect population information in several regions that may represent various threats to determine the extinction risk of the C. japonicum population objectively. In the future, we should expand the demographic monitoring of the C. japonicum population known in Korea.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.10a
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• pp.23-23
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• 2019

Cypripedium is an orchid genus of over 50 species widely distributed in the northern temperate zone, with a few taxa extending central America and the Himalayas and the southwesternmost tip of China in Asia. Of the known species approximately two thirds are found in China. In Korea, 4 species were reported to find with rare popularity ; C. macranthos, C. guttatum, C. japonicum, and C. calceolus. The seed has a firm brown fusiform testa with an opening at one end where the micropyle of the ovule was situated. Cypripedium has small, fusiform, wind-dispersed seeds that weigh about $1{\sim}2{\mu}g$ and range from 0.1 to 2 mm long and 0.07 to 0.4 mm across. The germination of Cypripedium was previously thought to be hard to cultivate in one's garden and impossible to propagate artificially, nowadays lab-propagated seedlings are readily available in EU or United States. We had already reported the successful micropropagation of lady's slipper orchids with green pod sowing methods. The collected Cypripedium species and hybrids were planted in Korea National Arboretum (KNA) on Oct. 2018. The 11 species including C. candidum and the 31 hybrids such as C. Ingrid were introduced from Germany with CITES certification. The lady's slipper orchids bloomed on May to June, and the flowers were pollinated in the same species or hybrids to get seeds for in vitro germination on 2~3 days after fully flowering. The green pods were collected after 49~70 days after pollination. The 6 species of Cypripedium seeds were easily germinated on the POM within 70 days after sowing, and the 12 hybrids PLBs were observed with small roots. We had established the scheme of lady's slipper orchids germination in 3 months with green pods, and get the possibility of seedling in 10 months of sowing in vitro.

• Korean Journal of Plant Taxonomy
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• v.36 no.4
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• pp.335-360
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• 2006

The distribution on 88 taxa of wild orchids in the Korean Peninsula was investigated by the examinations of specimens and the distribution maps were presented. The species numbers distributed at each eight floral regions in the Korean Peninsula are as the followings; Gapsan Province 27 taxa, Gwanbuk 21, Gwanseo 13, Middle 37, South 39, Southern-coast 33, Jeju 64 and Ulleung 19. Most species (72.7%) of Korean wild orchids are found in the Jeju Island. Eighteen taxa of them are restricted to the island in the Korean Peninsula. Among Korean species, 30 taxa grow at evergreen broad-leaved forests, and 16 are northern elements distributed at high mountains or northern part. In terms of distribution, the Korean wild orchid species are classified into IUCN Red List Categories by a modified criterion for Korean plants. No orchid species included EX or EW categories is found in the Korean Peninsula. Ten species are designated to be in CR category; Cymbidium kanran, C. lancifolium, C. ensifolium, Cypripedium japonicum, Cyrtosia septentrionalis, Dendrobium moniliforme, Habenaria chejuensis, H. radiata, Neofinetia falcata and Sedirea japonica, of which C. ensifolium and H. chejuensis are regarded as CR species for the first time. On the other hand, 22 taxa are classified into EN category, and the following nine taxa are newly proposed to be EN species; Gastrochilus japonicum, G. fuscopunctatus, Gastrodia verrucosa, Habenaria flagellifera, Herminium lanceum var. longicrure, Chamaegastrodia sikokiana, Lecanorchis kiusiana, Neottia hypocastanoptica and Tipularia japonica.

• 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.