• Journal of Plant Biotechnology
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• v.6 no.3
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• pp.193-197
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• 2004

In vitro multiplication of Curculigo orchioides was achieved by direct somatic embryogenesis in young leaf segments. Immature leaf segments of about 0.5 cm in length were cultured on MS medium supplemented with different concentrations of BAP (2-10 $\mu{M}$) or Kin (2-10 $\mu{M}$). Optimum response in terms of per cent cultures responding (89%) and the number of embryos per explant (16) were observed on MS medium supplemented with 8 $\mu$M BAP. The emergence of several somatic embryos on the adaxial side of the leaf segments was observed one month after the culture. Germinated somatic embryos were grown up to about 1.5 cm length before transferring to maturation medium. For maturation, the individual embryos were isolated and transferred to MS medium supplemented with BAP (5 $\mu{M}$) and NAA (0.5 $\mu{M}$). The plantlets emerged from the embryos were transferred to soil containing 1 peat: 1 sand with 90% success. The embryos were formed directly on the leaf segments without any callus phase. Direct regeneration of somatic embryos is important for the conservation of this endangered species, as rare somaclonal variants are likely to arise than from indirect regeneration.

• Plant Biotechnology Reports
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• v.5 no.4
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• pp.367-373
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• 2011

Linum album accumulates anti-tumor podophyllotoxin (PTOX) and its related lignans, which were originally isolated from an endangered species Podophyllum. In the present study, we examined the effects of five fungal extracts on the production of lignans in L. album cell cultures. Fusarium graminearum extract induced the highest increase of PTOX [$143{\mu}g\;g^{-1}$ dry weight (DW) of the L. album cell culture], while Rhizopus stolonifer extract enhanced the accumulation of lariciresinol up to $364{\mu}g\;g^{-1}$ DW, instead of PTOX. Typical elicitors, such as chitin, chitosan, or methyl jasmonate (MeJA), were shown to be less effective in lignan production in L. album cell cultures. These results verified the advantages of fungal extracts to increase lignan production in L. album cell culture, and suggested potential on-demand metabolic engineering of lignan biosynthesis using differential fungal extracts.

• Korean Journal of Medicinal Crop Science
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• v.12 no.5
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• pp.372-377
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• 2004

Scopolia parviflora Nakai, a rare and endangered species, is the sole plant producing tropane alkaloids (TA) among the Korean native species. In order to enhance TA productivity the SP72 root line was selected by screening 100 of root line, and the optimal culture media for root growth and TA production were investigated with the SP72 roots. Based on the several media, SH and 2B5 medium were determined as growth medium and White and NN medium as production medium. Among the four combinations of two-stage culture, 2BN (2B5 as growth medium plus NN as production medium) showed more enhanced root growth and TA production as compared with production media of White and NN medium and growth media of SH and 2B5 medium, respectively. However, bubble column bioreactor (BCB) cultures applying two-stage culture did not reveal the effective results despite of the each successful operation of two-stage culture in conical flasks and BCB cultures.

• Journal of Plant Biotechnology
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• v.43 no.1
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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.

• Journal of Plant Biotechnology
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• v.31 no.1
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• pp.31-35
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• 2004

In order to develop an efficient micropropagation technique for an endangered species, Stellera rosea N., stem node cultures were conducted on MS medium supplemented with cytokinins. Generally, BA was better than zeatin on shoot proliferation from stem nodes, whereas zeatin showed more effective on shoot elongation. In vitro rooting of shoots was achieved by application of an auxin pre-culturing method. Overall rooting rate was relatively low and differed depending on the culture period. Pre-culturing of shoots for 15 days at 1.0mg/L IBA revealed a slightly better rooting efficiency reaching 30％ rooting rate than NAA. Root induction rate by NAA also varied with concentration of NAA and culture periods. Total 51％ of the rooted plantlets survived on artificial soil mixture and grew normally without any distinct morphological variation. The results suggest that the endangered Stetllera plants are propagated via in vitro culture system, but still need to more study for the improvement of rooting and acclimatization of the plantlets in soil.

• Plant Biotechnology Reports
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• v.2 no.3
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• pp.213-218
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• 2008

Swertia chirata is an endangered gentian species that prefers to grow at higher altitudes. This ethnomedicinal herb is known primarily for its bitter taste caused by the presence of important phytochemicals that are directly associated with human health benefits. Due to a continuous loss of habitat and inherent problems of seed viability and seed germination, alternative strategies for propagation and conservation are urgently required to prevent the possible extinction of this species. We have formulated a reproducible protocol for the rapid propagation and conservation of this plant using leaves taken from in vitro shoot cultures. Direct induction of more than seven shoot buds per explant was achieved for the first time when the explants were placed on MS medium supplemented with $2.22{\mu}M$ N-6-benzyladenine, $11.6{\mu}M$ kinetin, and $0.5{\mu}M$ ${\alpha}-naphthalene$ acetic acid. Direct organogenesis was noted exclusively from the adaxial surface of the basal segments of leaves. Leaves closer to the apical meristem were more responsive than those farther away from the meristem. Plants raised through direct organogenesis were evaluated for their clonal fidelity by chromosomal analysis and DNA fingerprinting. Complete plants were successfully transferred to the field condition and produced viable seeds. Given the enormous potential of this age-old medicinal plant in terms of potential health-benefitting drugs, this protocol can be used for commercial propagation purposes and to initiate future genetic improvement studies.

• Korean Journal of Plant Tissue Culture
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• v.26 no.2
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• pp.133-136
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• 1999

Different kinds of cytokinins and auxins were tested for both shoot induction and rooting in a rare species, Abeliophyllum distichum Nakai. BA in WPM medium was the most effective in shoot induction, whereas zeatin seemed to be the most suitable for shoot elongation. Kinetin, at the concentration of 2.0~5.0 mg/L showed an effect in shoot induction, but the effect was inferior to BA and zeatin. Rapid shoot elongation could be achieved when the cultures were maintained on the diffuse light condition (below 500 lux) regardless of cytokinin treatments. For in vitro rooting, IBA was investigated as the best type of auxin tested when half strength GD medium was incorporated. The frequency of rooting using the plant growth regulator and medium just mentioned above was revealed as approximately 90%. In addition, the survival rate of rooted plantlets was almost 100% in an artificial soil mixture.

• SUVANNABHUMI
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• v.16 no.2
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• pp.215-261
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• 2024

In the 1960s, the Aoheng, a small tribal group with immense territories on the upper Mahakam River, began out-migrating to downstream settlements in search of better living conditions. A trickle of young men, then their families, and more sizable groups, they settled in various towns along the river. In Samarinda, the provincial capital, they came to form a community of several hundred. When the powerful forest product boom (c. 1990) for the P. R. China market opened up the hinterland to extractive ventures, many Aoheng returned home to protect their rich natural resources from forceful outsiders. After 1998, decentralization policies established scores of new provinces, regencies, and districts across the country. Soon, West-Kutai was created as the interior "Dayak" regency, upstream and autonomous from the Moslem-Malay coastal regions. Coal mining and oilpalm plantations massively intensified, while Sendawar, its capital, offered hundreds of civil-service jobs and business opportunities. In 2012, West-Kutai was split to create yet another regency, Upper-Mahakam, prompting robust Aoheng reflux/return moves toward its upstream capital, Ujoh-Bilang. Already open to wild-frontier-style inroads by outsiders, it will soon be flooded by industrial ventures. The Aoheng, bound to become a minority in their own district, are struggling to defer their inevitable final dissolution.

• Journal of Plant Biotechnology
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• v.37 no.4
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• pp.343-356
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• 2010

Plant micropropagation techniques include bud cultures using apical or axillary buds, organogenesis through callus culture or adventitious bud induction, and somatic embryogenesis. In Korea Forest Research Institute (KFRI), the first tissue culture trial in woody plant was initiated from the bud culture of hybrid poplars (Populus alba x P. glandulosa) in 1978. Since then several mass propagation techniques have developed from conifer and hardwood species, resulting in allowing practical application to Poplars, Birches and some oak species. In addition, useful micropropagation and genetic resources conservation techniques were established in some rare and endangered tree species including Abeliophyllum distichum. Among various in vitro propagation techniques, somatic embryogenesis is known to be the most efficient plant regeneration system. Since the first somatic embryo induction was reported in Tilia amurensis by KFRI in 1986, various protocols for direct or indirect somatic embryogenesis systems have developed in conifer and hardwood species including Larix leptolepis, Pinus rigida x P. taeda F1, Kalopanax septemlobus and Liliodendron tulipifera, etc. However, most of these technologies have been developed using juvenile tissues, i.e. immature zygotic embryos or mature embryos. Therefore it has been difficult to directly application to tree breeding program due to their unproven genetic background. Recently remarkable progresses and new approaches have been achieved in mature tree somatic embryogenesis. In this article we reviewed several micropropagation techniques, which have been mainly developed by KFRI and recent international progresses.

• 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.