• 한국약용작물학회지
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• 제17권4호
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• pp.238-242
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• 2009

Somatic embryos on growth regulator-free medium can be produced directly from cotyledon explants of Panax ginseng C. A. Meyer. When the embryo developmental stage was torpedo and cotyledon, the germination rate of embryos was quite high on MS medium supplemented with gibberellic acid ($GA_3$). However, the percentage of plantlet formation at the cotyledon stage was higher than that at the torpedo stage. This result demonstrates that the embryo at the cotyledon stage was the most appropriate for increasing germination by $GA_3$. Embryos cultured on medium including four levels of $GA_3$ concentrations (3, 5, 10, or 20 mg/$\ell$) showed all quite high germination rates (87-91%). When the well-developed embryos were continuously cultured on media including high concentrations of $GA_3$ from 10 to 20 mg/$\ell$, the percentage of formation of normal plantlets was lower than that seen under low concentrations from 3 to 5 mg/$\ell$. This treatment of high concentrations resulted in shoots with abnormal shape. The optimal $GA_3$ treatment provides a basis for the efficient method obtaining healthy plantlets derived from ginseng somatic embryos.

• 식물조직배양학회지
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• 제24권6호
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• pp.345-349
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• 1997

한라산에 자생하는 왕벚나무(Prunus yedoensis)의 미성숙 접합자배로부터 배발생 캘러스를 거쳐 체세포배를 유도할 수 있었으며, 이들 체세포배로부터 식물체를 재분화시킬 수 있었다. 배발생 캘러스는 1.0 mg/L 2,4-D와 0.1 mg/L BAP가 혼합 처리된 MS 배지에서 가장 효과적으로 유도되었으며, 그 중에서 90%가 배발생 캘러스였다. 또한 배발생 캘러스는 만개 후 45일된 종자의 접합자배에서 전체 60%가 발생되어 가장 양호하였다. 유도된 배발생 캘러스를 1.0 mg/L 2,4-D와 0.1 mg/L BAP가 혼합처리된 MS 배지에서 4주 간격으로 계대배양하여 구형 또는 심장형으로 발달한 체세포배를 얻을 수 있었고, 이들 체세포배는 식물생장조절제가 첨가되지 않은 MS 배지에서 배양하여 정상적인 자엽을 갖는 체세포배를 얻을 수 있었다. 이들 체세포배들은 배지의 종류에 따라 발아율은 0~49%로 맡은 차이를 보였으나, 식물생장조절제가 첨가되지 않은 1/2 MS 배지에서 49%의 발아율을 보여 가장 양호하게 식물체로 재분화되었다.

• Journal of Plant Biotechnology
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• 제7권4호
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• pp.259-265
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• 2005

Indian citrus ringspot virus (ICRSV)-free plants of Kinnow mandarin (Citrus nobilis Lour x C. deliciosa Tenora) were raised from virus-infected plants using unfertilised ovules as explants. Plants were tested by indirect ELISA and RT-PCR before using their explant. An amplified product of 539 bp was obtained by RT- PCR in ICRSV infected plants. Unfertilized ovules were excised from unopened flower buds of plants tested postive for virus and were cultured on Murashige and Skoog's (MS) basal medium supplemented with various concentrations of kinetin (KN) or malt extract (ME). Maximum induction (31.94%) of embryogenic callus was observed on MS medium supplemented with KN ($9.29\;{\mu}M$). Transfer of embryogenic calli to similar media composition resulted in somatic embryogenesis in all cultures, with an average number of 60.36 globular, 17.39 heart and 7.71 cotyledonary-shaped somatic embryos per culture. All cotyledonary shaped embryos developed into complete plantlets within 60 days on transfer to similar medium. Embryogenic callus induction, somatic embryo formation, maturation, germination and plantlet formation were achieved on MS medium supplemented with KN ($9.29\;{\mu}M$) alone. The plantlets derived from somatic embryos were transferred to sterilized soil, sand and vermiculite (3:1:1) mixture. After acclimatization, the plantlets were transferred to screen house and were indexed for ICRSV employing indirect ELISA and RT-PCR and found free of virus. A distinct feature of this study is the induction of somatic embryogenesis from unfertilised ovules to produce virus-free plants.

• Journal of Plant Biotechnology
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• 제29권4호
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• pp.253-257
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• 2002

To establish the optimum condition for plant regeneration from somatic embryos of Acanthopanax sessiliflorus Rupr. et Maxim, a medicinal plant, somatic embryos were induced from zygotic embryo-derived embryogenic callus in hormoen-free MS medium. To induce plantlet conversion, cotyledonary somatic embryos were cultured on MS solid medium with GA$_3$at various concentrations (0~10 mg/L) for three weeks. Plantlets were transferred to 1/3 MS solid medium with 0.5% charcoal for 7 weeks. Stem length was increased proportionally to the concentration and treatment period of GA$_3$. Also, the highest leaf width (8.9 mm) and leaf number (2.84) of plantlet were obtained when plantlets were converted on 5,10 mg/L GA$_3$pretreatments, respectively. The highest plant conversion frequency (66.7%) was obtained when the somatic embryos were cultured on medium containing 5 mg/L GA$_3$ for 3 weeks and then were transferred to 1/3 MS medium with 0.5% charcoal. The highest survival rate of soil transfer was 90% when plantlets were regenerated on medium with 5 mg/L GA$_3$ for 3 weeks and then transferred to plastic pots containing vermiculite and sand mixture for 4 weeks.

• Journal of Plant Biotechnology
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• 제5권3호
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• pp.169-172
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• 2003

Somatic embryogenesis was initiated from in vitro grown leaf explants of rose following an induction period of four weeks on MS basal medium supplemented with auxin and several subcultures on MS medium with cytokinin. '4th of July' showed the highest regeneration frequencies on 1 mg/L NAA followed by culture on medium with 4 mg/L zeatin. The embryogenic callus was propagated on MS medium with NAA, zeatin and $GA_3$. Germination of somatic embryos was achieved on MS medium with 1 mg/L BA. Somatic embryo derived plantlets were hardened and successfully transferred to the greenhouse.

• Journal of Ginseng Research
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• 제23권3호
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• pp.148-163
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• 1999

The efficacy of three auxins, viz. 2,4-0, NAA and dicamba, were compared for the induction of somatic embryogenesis in American ginseng (Panax quinquefolium L.). Somatic embryos (SEs) formed on ginseng cotyledonary, zygotic embryo and shoot explants after 8 weeks of induction by the auxin stimuli. Significantly more somatic embryos were induced by culture of any of the ginseng explants on media supplemented with $5{\mu}M$ 2,4-0 than any other auxin treatment. Shoots derived from somatic embryos had the greatest regenerative potential and zygotic embryos the least. Explants generated from green (unstratified) seeds gave similar or higher frequency of embryogenesis as the explants derived from stratified seeds. Histological and SEM studies confirmed that the regenerimts were somatic embryos. Somatic embryos germinated and developed into normal plants in $3\~6$ months. About $10\%$ of plantlets from second generation SEs formed flowers within 10 weeks, particularly on media supplemented with $GA_3$ The development of a regeneration system for ginseng through somatic embryogenesis is a necessary first step for mass propagation and genetic improvement of American ginseng.

• 식물조직배양학회지
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• 제22권4호
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• pp.189-194
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• 1995

유자의 배배양에 의한 캘러스의 유도는 미숙열매의 배로부터 가장 양호하게 유기되었다. 이들 캘러스는 3% sucrose 와 44.4 $\mu$M BA가 첨가된 1/2 MS배지에 치상하였을 때 가장 잘 유도되었으며 배양조건은 26$^{\circ}C$ 에서 2000 lx로 16 h의 광주기와 8 h의 암주기로 배양하였다. 이들 캘러스는 담황색으로 표면에 크고 작은 많은 돌기들을 가지고 있는 캘러스와 암갈색의 캘러스가 관찰되었다. 또한 캘러스 발달 단계와 기관분화의 정도에 따라서 세포배열 양상에 많은 차이가 있었는데, 배양 6주된 캘러스는 작은 세포들이 조밀하게 나타났으며 8주경부터 신장된 세포들이 분열하는 양상을 보여 주었으며 주된 캘러스 표면의 돌기들은 기관 발생의 초기에 구형의 세포돌기가 융기된 반면 주변의 세포들 은 액포화된 것으로 나타났다. 배양 16주 후에, 재분화된 shoot는 MS 배지에 이식하였을 때 발근이 되어 정상적으로 생장하였다. Shoo떼서는 유관속 형성층과 정유를 함유하고 있는 유낭의 초기 발생과정이 광학현미경으로 관찰되었다. 형성된 shoot를 화분에 이식하였을때 정상적으로 생장이 계속되는 것을 관찰하였다.

• 화훼연구
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• 제16권2호
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• pp.118-123
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• 2008

진한 붉은색을 띤 'Tiny Ghost'를 교배모본으로 Oriental group, Longiflorum group 및 Martagon group을 부본으로 하여 효율적인 종간잡종 생산과 상품성 있는 신품종을 획득하고자 실험을 수행하였다. 개화당일 채취한 화분의 발아율은 Oriental hybrid인 'Sorbonne'이 64%로 가장 높았다. 주두수분법과 화주 절단수분법을 이용하여 수분한 결과, 'Aktiva'를 부본으로 사용한 경우를 제외하고 모두 주두수분법을 사용한 교배에서 자방이 생성되었다. 또한 Longiflorum group인 'Norina'와 'Gelria' 중 화분발아율이 높은 'Norina' 가 모두 수정에 실패하여 같은 group 내에서도 종간의 유전적 친화성에 따라 수정률이 달랐다. 'Aktiva'를 부본으로 한 교배에서 1개의 embryo sac을 적출하여 1개의 구를 생성하였으며, 'Sorbonne'을 부본으로 한 교배에서는 4개의 ovule을 치상하여 1개의 구를 얻었다. 'Gelria'를 부본으로 한 교배에서는 1개의 자방에서 embryo 5개, embryo sac 15개, ovule 7개를 얻었으며 그 중 18개체가 발아하여 66.7%의 발아율을 나타내었다. Martagon group인 L. hansonii를 부본으로 한 교배에서는 5개의 ovule을 얻어 그 중 40%가 발아하였다.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1987년도 식물생명공학 심포지움 논문집 Proceedings of Symposia on Plant Biotechnology
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• pp.213-237
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• 1987

In vitro flowering system may minimize the confounded influence of non-floral meristem parts of plants in studying the relationship of a given treatment and flowering responses. We have induced flower buds from plantlets regenerated from zygotic embryo-derived somatic embryos of ginseng, which circumvented the normal 2-year juvenile period before flowering. The result suggests that the adulthood of ginseng root explants in the experiment previously conducted by Chang and Hsing (1980; Nature 284: 341-342) is not prerequired to flowering of plantlets regenerated through somatic embryogenesis. We have also induced flower buds from elongated axillary brandches from cotyledonary nodes by culturing ginseng zygotic embryos, seedlings, and excised cotyledonary nodes. It was found that 6-benzyladenine (BA) supplemented to the medium was essential for flowering, whereas abscisic acid (ABA) was inhibitory. Gibberellic acid(GA3) was also required for flowering when ABA was present with BA in the medium. The results suggest that cytokinins, gibberellins, and inhibitors play primary, permissive, and preventive roles, respective-ly, in the induction of flowering of ginseng. Tran Thanh Van (1980; Int. Rev. Cytol., Suppl. IIA: 175-194) has developed the "thin cell layer system" in which the induction of shoots, roots, or flower buds from epidermal layer explants were controlled by culture conditions and exogenous growth regulators in the medium, Utilizing the thin cell layer system, Meeks-Wagner et al. (1989; The Plant Cell 1: 25-35) have cloned genes specifically expressed during floral evocation. However, the system is too tedious for obtaining a sufficient amount of plant materials for biochmical and molecular biological studies of flowering. We have developed a garlic callus culture system and one obvious advantaging over the thin cell layer system is that an abundant cells committed to develope into flower buds proliferate. When the above cells were compared by two-dimensional gel electrophoresis with those which have just lost the competence for developing into flower buds, a few putative proteins specific to floral evocation were detected. The garlic callus culture system can be further explored for elucidation of the molecular biological mechanism of floral evocation and morphogenesis.hogenesis.

• 식물조직배양학회지
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• 제21권5호
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• pp.281-286
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• 1994

고구마 경단배양에 의한 우량종묘 대량증식 과정에서 분화 및 생육에 미치는 생장조절제의 효과를 구명하기 위하여 실험을 수행하였다. 고구마 경단조직은 1 mEm/L 2,4-D 첨가된 MS배지에서 배양 40일경 배발생 캘러스가 형성되었다. 또한 형성된 배발생 캘러스는 0.1 mg/L GA$_4$ 첨가된 배지에 배양한 경우, 효과적으로 배발생 캘러스를 유지 및 증식이 가능하였으며, 배발생캘러스로부터 shoot 재분화는 0.1mg/L BA가 유효하였다. 체세포배로부터 shoot 재분화는 0.01 ngh BA와 0.01 mg/L GA$_4$ 혹은 0.1 mg/L BA와 0.01mg/L GA$_4$ 혼합배지에서 양호하였으며, 0.1-1 mg/L BA를 첨가된 배지에서는 배발생 캘러스 및 체세포배로부터 녹색의 단단한 캘러스가 형성되었다. 체세포배를 배양하여 얻어진 5 mm 크기의 shoot 정단을 재배양 시, 0.1 mg/L jasmonic acid 또는 0.01 mg/L brassnlolide가 첨가된 배지에서 소식물체의 생육이 가장 양호하였다.