• Journal of Plant Biotechnology
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• 제41권2호
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• pp.89-93
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• 2014

장미 형질전환체를 개발하기 위한 유전자 도입 재료로써 5 ~ 6년 이상 장기간 유지 증식된 장미 배발생캘러스의 이용 가능성이 확인되었다. 2007년 및 2009년에 각각 처음으로 기내 뿌리유래 캘러스로부터 유도된 후 유지 증식배지에서 계대배양 해 온 장미 '스위트 옐로우' 품종 및 KR056002와 KR056006 2계통 유래 배발생캘러스로부터 식물체의 재분화 능력을 확인하였다. 장기간 계대배양 된 배발생캘러스로부터 신초의 모습을 갖춘 식물체로 재분화 되기까지 소요기간 및 재분화율은 '스위트옐로우' 품종은 3 ~ 4개월, KR056002 및 KR056006은 4 ~ 5개월로, 이들 배발생캘러스가 기내뿌리로부터 처음으로 배발생 된 후 최초 신초 재분화 때와 동일한 양상이었다. 또한 체세포배발생캘러스의 유지 증식을 위한 계대배양과정에서 발생되는 비정형체 위에 새로운 배 및 배발생캘러스가 유도되었다. 이 비정형체는 새로운 배발생캘러스를 유도할 수 있는 재료로서 이용될 수 있을 것이다.

• Journal of Plant Biotechnology
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• 제44권1호
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• pp.49-55
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• 2017

옥수수의 최적 조직 배양 조건을 확립하기 위하여 옥수수 국내 5 계통과 국외 11 계통 총 16 계통을 포트와 포장 재배하여 미성숙 배를 분리하여 배발생 캘러스 유도 및 식물체 재분화율을 조사하였다. MS 배지에 auxin으로 1.5 mg/L Dicamba와 0.5 mg/L 2,4-D 가 첨가된 배지에서 캘러스 형성은 본 실험에 사용된 옥수수 계통 모두에서 높은 빈도로 유도되었으며, 캘러스로부터 식물체 재분화는 5mg/L zeatin이 첨가된 재분화 배지에서 높은 재분화율을 보였다. 또한 포장에서 재배된 옥수수로부터 미성숙 배를 분리하여 사용하였을 때 캘러스 유기 및 식물체 재분화 효율이 높았던 것으로 보아 미성숙 배를 분리하기 위한 옥수수 상태 및 genotype이 중요한 영향을 준다는 것을 알 수 있었다. 본 실험을 통하여 배 발생 캘러스 형성 및 식물체 재분화 효율이 조사된 옥수수 계통들은 생명공학 기술을 활용한 신품종 개발을 위한 형질전환 시스템 개발에 유전자원으로 활용될 수 있는 정보를 제공할 것으로 사료된다.

• Journal of Plant Biotechnology
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• 제37권4호
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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.

• Journal of Forest and Environmental Science
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• 제23권1호
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• pp.5-9
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• 2007

Zygotic embryos just after harvest of seeds were immature globular to heart stage. Maturation of zygotic embryos rapidly proceed when zygotic embryos together with small excised parts of endosperm were cultured on 1/3-strength MS solid medium with 2% sucrose, and the zygotic embryos were germinated within two months. Embryogenic callus was formed from the excised segments of germinating zygotic embryos of Eleutherococcus pedunclus on Murashige and Skoog (MS) medium with $4.5{\mu}M$ 2,4-D. The embryogenic callus formation occurred at a low frequency (less than 7%) from hypocotyl segments. The embryogenic calli were maintained on the same medium as primary medium. High frequency somatic embryogenesis was obtained after the cells were transferred to medium lacking 2,4-D. Cotyledonary embryos were germinated and converted into plantlets on medium with $20{\mu}M$ $GA_3$. Embryogenic callus and somatic embryos were produced spontaneously on the surfaces of roots and/or hypocotyls of plantlets. The frequency of embryogenic callus formation was 85% in roots and 34% in hypocotyls. Therefore maintain of cell lines performed very easily. Plantlets with developed epicotyls at more than 3 cm acclimatized at high frequency (89%). While plantlets with small epicotyls (less than 1 cm) were acclimatized at low rate (32%). The soil survived plantlets produced new sprouts after over wintering in the field.

• Journal of Ginseng Research
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• 제43권1호
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• pp.38-48
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• 2019

Background: Interspecific ginseng hybrid, Panax ginseng ${\times}$ Panax quenquifolius (Pgq) has vigorous growth and produces larger roots than its parents. However, F1 progenies are complete male sterile. Plant tissue culture technology can circumvent the issue and propagate the hybrid. Methods: Murashige and Skoog (MS) medium with different concentrations (0, 2, 4, and 6 mg/L) of 2,4-dichlorophenoxyacetic acid (2,4-D) was used for callus induction and somatic embryogenesis (SE). The embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 Schenk and Hildebrandt (SH) medium. The developed taproots with dormant buds were treated with $GA_3$ to break the bud dormancy, and transferred to soil. Hybrid Pgq plants were verified by random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analyses and by LC-IT-TOF-MS. Results: We conducted a comparative study of somatic embryogenesis (SE) in Pgq and its parents, and attempted to establish the soil transfer of in vitro propagated Pgq tap roots. The Pgq explants showed higher rate of embryogenesis (~56% at 2 mg/L 2,4-D concentration) as well as higher number of embryos per explants (~7 at the same 2,4-D concentration) compared to its either parents. The germinated embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 SH medium to support the continued growth and kept until nutrient depletion induced senescence (NuDIS) of leaf defoliation occurred (4 months). By that time, thickened tap roots with well-developed lateral roots and dormant buds were obtained. All Pgq tap roots pretreated with 20 mg/L $GA_3$ for at least a week produced new shoots after soil transfer. We selected the discriminatory RAPD and ISSR markers to find the interspecific ginseng hybrid among its parents. The $F_1$ hybrid (Pgq) contained species specific 2 ginsenosides (ginsenoside Rf in P. ginseng and pseudoginsenosides $F_{11}$ in P. quinquefolius), and higher amount of other ginsenosides than its parents. Conclusion: Micropropagation of interspecific hybrid ginseng can give an opportunity for continuous production of plants.

• Journal of Ginseng Research
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• 제38권3호
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• pp.220-225
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• 2014

An efficient in vitro protocol has been established for somatic embryogenesis and plantlet conversion of Korean wild ginseng (Panax ginseng Meyer). Wild-type and mutant adventitious roots derived from the ginseng produced calluses on Murashige and Skoog (MS) medium supplemented with 0.5 mg/L 2,4-dichlorophenoxyacetic acid and 0.3 mg/L kinetin; 53.3% of the explants formed callus. Embryogenic callus proliferation and somatic embryo induction occurred on MS medium containing 0.5 mg/L 2,4-dichlorophenoxyacetic acid. The induced somatic embryos further developed to maturity on MS medium with 5 mg/L gibberellic acid, and 85% of them germinated. The germinated embryos were developed to shoots and elongated on MS medium with 5 mg/L gibberellic acid. The shoots developed into plants with well-developed taproots on one-third strength Schenk and Hildebrandt basal medium supplemented with 0.25 mg/L 1-naphthaleneacetic acid. When the plants were transferred to soil, about 30% of the regenerated plants developed into normal plants.

• Journal of Plant Biotechnology
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• 제35권1호
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• pp.81-86
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• 2008

감귤에 있어서 원형질체 융합에 의한 체세포잡종체 생산을 위해서는 우선 캘러스 원형질체로부터의 식물체 재분화가 가능하여야 하기 때문에 온주밀감의 캘러스 원형질체로 부터 배형성과정을 통한 식물체 재분화에 관한 실험을 수행하였다. 흥진조생의 어린 미성숙 배주의 주심조직으로부터 유기된 배형성 캘러스를 사용하여 건강한 원형질체를 분리하고 0.6 M $BH_3$ 배지를 배양배지로 사용하여 배양방법에 따른 plating efficiency를 비교해 본 결과 liquid over solid 배양시 원형질체로부터의 미소괴 형성율이 더 높음을 알 수 있었으며, 적정 배형성 배지 선발에서는 1500 mg/L malt extract 첨가 배지에서 배형성율이 높았다. 자엽형 배로부터의 신초유기를 위한 배지비교에서는 1.0 mg/L GAB 첨가배지에서 발근과 함께 정상적으로 재분화된 신초를 얻을 수 있었다. 원형질체 배양으로부터 재분화된 식물체들은 순화과정을 거쳐 온실 육묘중에 있으며, 생장상이나 형태적인 특성에 있어서 모본 식물체와 차이가 없음을 확인할 수 있었다. 본 실험으로 얻어진 원형질체 배양에 의한 식물체 재분화 체계를 바탕으로 이종속간 감귤의 원형질체 융합 기술을 이용하여 교배육종이 불가능한 품종들로부터 우수한 형질을 지닌 감귤 품종 및 대목용 품종 생산에 활용하고자 한다.