• 한국균학회지
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• 제50권4호
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• pp.361-365
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• 2022

Here, Podospora leporina was isolated and cultured from Cypripedium guttatum Sw. (spotted lady's slipper), which was designated as an endangered wildlife class 1 by the Ministry of Environment. Further, Podospora sp., which is found in Calanthe discolor Lindl. (Japanese hardy orchid) root is recorded as an endophytic fungus in Jeju Island, Korea. However, there is no existing record of Podospora spp. Thus, in this study, we analyzed the morphological characteristics and two molecular markers, internal transcribed spacer (ITS) and large ribosomal subunit (LSU), of P. leporina and compared these data with other relative species. Consequently, this endophytic fungal species was confirmed as an unrecorded species in Korea, so we presented cultural characteristics and microstructures, such as the conidia of P. leporina.

• 한국환경복원기술학회지
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• 제23권4호
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• pp.1-11
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• 2020

This study aimed to develop the HSI (Habitat Suitability Index) model of Cypripedium guttatum. and to verify this model by applying to the candidate sites for replacement habitat. The development of HSI and SI (Suitability Index) model was conducted based on the existing literature, field surveys, and expert opinions for information on ecological habitat characteristics. Seven variables were selected as habitat variables including mean maximum temperature in Jul.-Aug., lighting, slope, altitude, effective soil depth, soil texture, and artificial overexploitation (i.e. protected areas). HSI model was developed for C. guttaum based on these variables. This HSI model showed high applicability to selection and evaluation of replacement habitats for C. guttaum. Our findings could provide the basic information on habitat assessment to prevent the extinction of endangered C. guttatum. However, since there is a limitation that the survey data were insufficient, further field surveys should be conducted on several habitat types to improve the accuracy of the HSI model.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2020년도 춘계학술대회
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• pp.11-11
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• 2020

Background : The genus Cypripediums are typically adapted to the low temperature and cold climates of the high land in Korea. The species which were reported in Korean peninsula are C.japonicum, C.macranthos, C.calceolus and C.guttatum. We had already reported the successful germination of the C.macranthos and C.guttatum. The small spotted lady's slipper orchid(C.guttatum) is most endangered plants to extinction of which population was reported less than 100 in Korea. The objective of the present research is restoration of the C.guttatum in ex situ and adaptation in high land and nursery. Methods and Results : The 350 seedlings of C.guttatum were transplanted in soil at alpine area(altitude 750m) and the nursery of Korea National Arboretum(KNA) in YangPyeong city on May 2019 and 2020 in order to verify the possibility of ex situ restoration. The sprouted ratio of the seedlings were less than 10% in the alpine area and 83% in the nursery of KNA. The reasons of the diverse survival ratio are presumed as the low moisture content in the soil and heavy shade of the alpine area. The 15 cypripedium species and 29 hybrids were also effectively adapted in the KNA rare plant garden. Conclusion : These results demonstrated the difficulties of the restoration for small spotted lady's slipper orchid in the natural habitation. From the above results, we may conclude that the critical factors of the genus Cypripedium are intensive care for the fragile seedlings during the 1^st year acclimatization.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2019년도 춘계학술대회
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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.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2019년도 추계학술대회
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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.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2010년도 정기총회 및 춘계학술발표회
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• pp.13-13
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• 2010

복주머니난(Cypripedium)속 식물은 우리나라에 광릉요강꽃을 비롯하여 털복주머니난, 흰털복주머니란, 복주머니란, 노랑복주머니란등 5종이 분포하는 것으로 알려져 있다. 광릉요강꽃과 털복주머니란 두 종은 환경부에서 지정한 멸종위기 식물 1급과 2급에 지정되어 보호를 받고 있고 나머지 3종은 제도적으로도 보호를 받지 못하는 실정이다. 본 연구에서는 충분히 성숙한 털복주머니난과 복주머니난의 종자에 NaOCl처리를 하여 발아율을 향상시킬수 있었는데 이러한 전처리가 발아에 미치는 원인에 대한 연구를 실시하였다. 털복주머니란의 무균적인 종자발아는 1.0% NaOCl 처리구에서 70% 이상의 종자발아율을 보였으며, POM배지가 MS배지보다 신초분화가 양호했다. GA3와 활성탄(Activated charcoal)의 혼합첨가는 신초증식에 효과적이었다. 신초분화 후 저온처리는 신초의 증식 율을 증가시켰다. 한편 NaOCl 농도(0, 1, 3, 5, 10%)와 NaOCl 처리시간(0, 5, 15, 30, 45, 60분)에 따라서 복주머니란의 종자발아를 확인한 결과, NaOCl 1%를 30분간 처리하였을 때 가장 높은 발아율을 나타냈다. NaOCl을 처리하여 종자의 종피상태를 SEM과 TEM으로 관찰한 결과 NaOCl의 처리는 종피 세포벽의 부분적 해리 및 작은 구멍을 만들게 하였는데 이러한 종피의 물리화학적 변화가 물과 양분의 이동을 원활히 하여 종자의 발아를 촉진하는 것으로 사료된다. 복주머니난의 신초분화에 미치는 casein과 활성탄의 효과를 알아본 결과 casein 200 mg/L와 활성탄 200 mg/L를 혼합 첨가한 실험구에서 가장 높은 신초분화율을 보였다. 토양순화 후 생존률은 극히 저조했으며 30 개체중에 5 개체만이 다음해 어린동아를 싹틔우는 것을 확인 하였다. 본 결과들을 종합하여 보면 멸종위기식물, 특히 털복주머니란과 복주머니란의 조직배양을 통해서 서식지외 보존방안(기내증식)에 관해 가능성을 제시하였다고 생각되어 진다.