• Mycobiology
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• 제45권4호
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• pp.312-317
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• 2017

Orchidaceous plants have symbiotic relationships with endophytic fungi, including mycorrhizal fungi, which play important roles in the seed germination and growth of the host plants. In this study, endophytic fungal communities isolated from the roots of Cephalanthera longibracteata collected from three different sites in Korea were analyzed, and it was determined whether fungal communities were preferentially correlated with the sites. The fungal isolates were identified by sequence analysis of the internal transcribed spacer regions of rDNA. In total, 30 species of endophytic fungi, including two species of mycorrhizal fungi belonging to the genus Tulasnella, were identified. Leptodontidium orchidicola showed the highest frequency and was isolated from all root samples. Species diversity and richness were not significantly different among sites. However, the community structure of the endophytic fungi significantly differed among sites, suggesting that the site characteristics affected the community composition of the endophytic fungi colonizing the roots of C. longibracteata. Our findings will aid in developing methods involving the use of symbiotic fungi for orchid conservation and restoration in native habitats.

• Journal of Plant Biotechnology
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• 제8권1호
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• pp.37-41
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• 2006

In vitro mass multiplication of Vanilla planifolia was investigated using node as explant. Multiple shoots were developed in MS medium supplemented with $2.0mgl^{-1}$ 6-benzylaminopurine and $1.0mgl^{-1}$ $\alpha$-naphthalene acetic acid. Multiple shoots were maintained for 6-T weeks with regular subculturing at the end of $3^{rd}$ week onto fresh medium. The maximum number of shoots at the rate of 12.8 per node segment was achieved over a period of four weeks. The elongated shoots were separated from the shoot clusters and were transferred onto half strength MS medium supplemented with indole-3-acetic acid ($1.0mgl^{-1}$) over a period of 28 days for induction of roots. The development of roots was observed on $7^{th}$ day of incubation. The in vitro raised plantlets were transferred to poly-cups, covered with polyethylene sheets and maintained under shade net for 25 days for hardening. Finally these plants were transferred to field and recorded that 85 % of tissue cultured plants were survived. From the present study, a simple and efficient micropropagation protocol was developed for Vanilla planifolia using single node segments as explants.

• The Plant Pathology Journal
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• 제18권3호
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• pp.156-160
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• 2002

Cymbidium orchids with blight and rot symptoms were collected, and a total of 63 isolates of Fusarium app. was obtained from pseudobulbs, roots, and leaves of the diseased plants. The isolates were identified based on their morphological characteristics. Out of the 63 isolates of Fusatium sup., 51 isolates were identified as F. oxysporum, 10 isolates as F. solani, and the rest as F. proliferatum. F. oxysporum was isolated from all the Cymbidium spp., while F. solani and F. proliferatum were isolated only from Cymbidium ensifolium and C. ginatum, respectively. Isolates of the three Fusarium spp. were tested for pathogenicity to their hosts by artificial inoculation. The strongly pathogenic isolates of Fusarium spp. induced severe dry rot of pseudobulbs and roots of the host plants. The symptoms progressed up to the basal part of the leaves, which later caused blight of the entire plant. The dry root symptoms induced on the plants by artificial inoculation with the isolates of Fusarium app. were similar to those observed in the growers'greenhouses. This is the first report of dry rot of Cymbidium spp. caused by F. oxysporum, F. solani, and F. proliferatum in Korea.

• 한국균학회지
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• 제46권4호
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• pp.427-435
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• 2018

본 연구에서는 충남, 제주, 경남, 충북 등에서 새우난초(Calanthe discolor)와 은대난초(Cephalanthera longibracteata)의 뿌리를 채취하여 표면살균한 후 내생균을 순수분리 하였다. 분리된 내생균의 형태학 특징을 관찰하고 internaltranscribed spacer (ITS) rDNA영역의 염기서열을 ITS1F와 ITS4프라이머를 이용하여 분석하였다. 서로 다른 지역에서 채집한 새우난초와 은대난초이지만, 공통적으로 Leptodontidium orchidcola가 높은 상대수도와 빈도를 나타냈으며, 내생균류의 군집차이를 분석할 결과, 지역적 차이보다는 숙주에 의한 차이가 더 큰 영향을 미치는 것으로 나타났다.

• Journal of Plant Biotechnology
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• 제43권1호
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• pp.138-145
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• 2016

To find the optimal propagation condition for endangered Cypripedium macranthos Sw., also known as lady's slipper orchid, the effect of various organic additives on in vitro germination, protocorm formation and seedling growth was investigated during asymbiotic seed culture. When $100ml{\cdot}L^{-1}$ coconut water was added to the basal medium, the highest germination rate and protocorm formation rate were achieved, with 70.8% and 74.2% respectively. Supplementation of phloem sap from birch tree or maple tree also showed a facilitating effect to improve the germination and protocorm development. With $100ml{\cdot}L^{-1}$ birch sap or maple sap, both the germination and protocorm formation rates were roughly more than 65% and 68%. The roots and buds of the seedlings grew vigorously in the medium containing $100ml{\cdot}L^{-1}$ coconut water or phloem sap, in particular, their bud formation rates increased by more than 70%. Addition of banana powder and peptone could not create a more significantly favorable culture condition, and non-addition had the worst results. Our results demonstrated that proper organic amendments such as coconut water and phloem sap might be preferred to in vitro germination and the growth of seedlings developed from the protocorm of C. macranthos Sw. during asymbiotic seed culture.

• Journal of Plant Biotechnology
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• 제43권1호
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• pp.132-137
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• 2016

The genus Cypripedium, known as lady's slipper orchid, has been considered to have a considerable marketability as a potted and garden plant with beautiful flowers; however, this species is becoming endangered and, in some places extinct, due to habitat destruction and illegal collections. As such, an optimized artificial propagation system is necessary for its conservation and horticultural cultivation. For the establishment of the in vitro proliferation of the endangered Cypripedium macranthos Sw., native to Korea, the effects of medium strength and sucrose concentrations on germination, protocorm formation and seedling growth were investigated through asymbiotic seed culture. The highest germination rate and protocorm formation rate were obtained with a 1/4 MS medium; higher MS medium strengths did not generate the favorable conditions required for germination and protocorm growth. The overall growth characteristics of roots and buds developed from protocorms were best in cultures of 1/4 MS medium. On this medium containing $10g{\cdot}L^{-1}$ sucrose, both the germination rate and protocorm formation rate increased significantly. The general growth properties of seedlings developed on the medium with $10g{\cdot}L^{-1}$ sucrose were best, showing the highest bud formation rate and root number. Our results demonstrate that the 1/4 MS medium, supplemented with $10g{\cdot}L^{-1}$ sucrose, could improve in vitro germination, and facilitate the growth of seedlings developed from the protocorms of C. macranthos Sw.

• 식물병연구
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• 제13권1호
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• pp.6-14
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• 2007

2005년과 2006년에 걸쳐 경기도 남양주의 호접란(Phalaenopsis spp.), 풍란(Neofinetia falcata), 나도풍란(Aerides japonicum)을 재배하고 있는 온실재배농가에서 뿌리 및 줄기기부 썩음 증상이 발생되었다. 이들 난은 초기에는 시들음 증상을 나타내고 병든 개체의 잎들은 황화되거나 붉게 변하며 낙엽되고 결국 고사하였다. 병든 개체의 뿌리 및 하엽으로부터 59개 Furaium spp.를 분리하여 동정 한 결과 F. oxysporum, F. proliferatum, F. solani 3종으로 동정되었다. 이 중 F. oxysporum, F. proliferatum은 호접란과 풍란류에서 모두 분리되었고, F. solani는 호접란에서만 분리되었다. 병원성실험 결과 이들 3종 모두 병원성이 인정되었다. 비록 F. oxysporum, F. proliferatum, F. solani은 호접란에 대하여 뿌리썩음병원균으로 보고되어 있으나 풍란과 나도풍란에 대해서는 아직 보고된 바 없다. 따라서 본 연구를 통해 F. oxysporum, F. proliferatum에 의한 풍란과 나도풍란의 뿌리 및 줄기썩음병을 처음 보고하는 바이다.

• 한국균학회지
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• 제44권4호
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• pp.365-370
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• 2016

본 연구에서는 잣나무, 눈측백의 침엽과 자란의 뿌리에서 식물에 공생하는 내생균을 분리하여 동정하였다. 이를 위해 specific primer인 ITS1F와 ITS4를 이용하여 internal transcribed spacer (ITS) rDNA영역과 primer LR0R과 LR 16을 이용하여 large subunit rDNA 영역을, primer Bt2a과 Bt2b를 이용하여 ${\beta}$-tubulin 영역을, 그리고 EF-1 primer를 이용하여 translation elongation factor 지역의 염기서열을 동시에 분석하였다. 연구 결과 Colletotrichum simmondsii, Fusarium sterilihyphosum, Diatrypella pulvinata, Ochroconis globalis, Sphaeria chrysosperma 등 국내 미기록 균주 5종을 분리하여 형태적, 분자생물학적 동정 결과를 서술하였다.

• 원예과학기술지
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• 제17권5호
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• pp.578-580
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• 1999

춘란 뿌리에서 분리한 Rhizoctonia속 난 균근균 3종이 난의 생장에 미치는 영향을 알아보기 위하여 난 균근균을 기내에서 자라는 한란(제주한란${\times}$일본한란 '남국(南國)' 교잡종) 및 호접란 유묘와 2개월간 공동배양한 다음 온실에서 18개월간 재배하면서 식물체와 균근균과의 공생관계를 조사하였다. 기내에서 2개월간 공동배양한 결과 제주한란${\times}$일본한란 '남국(南國)' 교잡종과 호접란 모두 엽장, 엽수, 엽폭 등 지상부와 근수, 근장 등 지하부 및 생체중에 있어서 무처리구와 균근 균 처리구 간에 차이가 없었다. 4개월간 순화 재배한 결과는 제주한란${\times}$일본한란 '남국(南國)' 교잡종의 경우 P1 균(Rhizoctonia repens) 처리구에서 지상부와 지하부의 생육이 가장 양호하였고, 호접란은 무처리구와 균근균 처리간에 차이가 없었다. 온실에서 1년 6개월 동안 재배한 결과 제주한란${\times}$일본한란 '남국(南國)' 교잡종은 무처리구에 비해 P1 균 처리구에서 초장, 신초수, 엽수, 뿌리수 및 생체중이 증가하였으나, P2 균(R. endophytica) 처리구에서는 오히려 감소하였으며, P3 균(R. repens : P1 균과는 다른 균주임) 처리구에서는 차이가 없었다. 호접란의 경우 무처리구에 비해 P1 균 처리구에서 초장과 생체중이 증가하였다. 이상의 결과를 종합해 볼 때, P1 균이 제주한란${\times}$일본한란 '남국(南國)' 교잡종의 생육을 크게 촉진한 것으로 나타났다.

• 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.