• Journal of Plant Biotechnology
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• 제3권2호
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• pp.101-106
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• 2001

In order to establish efficient plant regeneration from embryogenic suspension cultures of soybean, Glycine max L, we examined the effects of auxin type and concentration, cytokinin type and concentration, and amino acid type and concentration on the growth of embryogenic clumps from induced callus, and the effect of desiccation of mature somatic embryos obtained from these clumps on the frequency of somatic embryo germination. Embryogenic callus was induced from the edge of the cotyledons cultured on MS medium containing 6% sucrose, 40 mg/L 2,4-D, 0.2% gelrite and pH 5.7. The growth of embryogenic clumps was best in early staged, embryogenic callus that was placed in suspension culture of MS medium containing 5 mg/L 2,4-D and 0.5 mg/L asparagine. Single somatic embryos were isolated from the clumps and plated on the same medium for maturation. When the mature single somatic embryos were desiccated for 96 h, somatic embryo germination came up to approximately 90%. The plantlets germinated after embryos desiccation for 2 weeks were transfered to MS medium containing 3% sucrose,0.2% gelrite and pH 5.7.

• Journal of Plant Biotechnology
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• 제6권1호
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• pp.55-61
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• 2004

Callus induction and somatic embryogenesis are fundamental to cotton tissue culture biotechnology. An efficient protocol for callus induction, somatic embryogenesis and their maturation have been developed to regenerate plantlets from cotton (Gossypium hirsutum L.) variety coker 312. Embryogenic callus was initiated from hypo-cotyl region that was used as an explant at seedling stage when it was about 7-8 days old. Callus induction was achieved through culturing hypocotyls (5-7mm) on $MS_{1a} medium supplemented with 2,4-D (0.1 mg/L) and KT (0.5 mg/L) for six weeks. A friable, colorless, bulky and well proliferating callus becomes greenish with the addition of NAA (2.0 mg/L), ZT (0.1 mg/L) and removal of 2,4-D (M $S_{1b}$) cultured for two weeks then again transferred to $MS_{1a}. 2,4-dichlorophenoxyacetic acid (2,4-D) promoted the proliferation of embryogenic callus, but had a negative effect on the differentiation and germination of somatic embryos. ZT (0.1mg/L) and activated charcoal (2g/L), both hormones play an important role in differentiation and germination of somatic embryos in hypocotyls derived embryogenic callus but in case of cotton, such a capability have been observed on MS medium with 1.92 g/L $KNO_3$, but it is considered to attain somewhat more improvement. High embryogenesis frequency was achieved through nutrient deficient stress treatment. The frequency of globular embryogenesis (two-three folds) was achieved when well proliferating callus was (from $MS_{1a}$ media) cultured on MS (1/5 strength) medium for four weeks. Here the development of anthocyanins is the best indicator for somatic embryogenesis. However, when embryoid callus was cultured on MS (full strength) medium, the globular embryos were developed into normal plantlets immediately. In this procedure 27.49% cotyledenary embryos were developed. Of that 70% cotyledenary embryos were developed not only into normal plantlets but rooted simultaneously, when cultured on MS (with 0.05 mgg/L giberrelic acid) medium. So complete plants could be regenerated through somatic embryogenesis from hypocotyl explants within 6 months.s.

• 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.

• 한국약용작물학회지
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• 제1권2호
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• pp.184-190
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• 1993

1. 지황의 잎 조직에서 캘러스 유도는 옥신류와 사이토키닌류의 단독처리보다는 BA와 NAA를 혼합처리하는 경우에 캘러스 유도가 잘 되었다. 2. 옥신류의 embryogenic 캘러스 유도는 5% 정도로 극히 낮았으나 사이토키닌류를 처리한 경우는 $40{\sim}55%$로 높아졌다. 3. BA 4mg/1 NAA 0.5mg/1 처리에서 embryogenic 캘러스 유도가 가장 양호하였다. 4. 배지 종류별로는 Linsmaier-Skoog배지가 embryogenic 캘러스 유도와 유도량이 가장 많았다. 5. Embryogenic 캘러스 유지도 BA 4mg/1과 NAA 0.5mg /1 처리 에서 가장 효과적 이었다. 6. 계대배양 회수가 증가할수록 캘러스 생장량은 증가하였지만 embryogenic 캘러스 빈도는 감소되는 경향을 보였다. 7. 현탁배양시 체세포배 생산은 BA 1mg/1처리 에서 가장 많았다 8. 체세포배에서 식물체 분화는 식물 생장조절제가 처리되지 않은 1/2 LS기본배지에서 가장 잘 되었으며 토양활착율은 75% 정도이었다.

• 식물조직배양학회지
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• 제21권1호
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• pp.1-6
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• 1994

멜론의 배발생 캘러스의 유도와 식물체 재분화 시스템을 확립하기 위해서 자엽과 배축의 절편을 여러 농도의 2,4-D 와 BA가 첨가된 MS 기본배지에 치상하여 관찰한 결과, 초기배양에서는 4 mg/L 2,4-D와 1 mg/L BA 그리고 2 mg/L 2,4-D+O.1 mg/L BA가 첨가된 배지에서 연한 노란색의 배발생 캘러스가 형성되었고, 위의 조건에서 형성된 캘러스로부터 배발생 캘러스는 0.5와 1 mg/L 2,4-D가 첨가된 동일배지에서 약 75%가 형성되었다. 그리고 생장조절제를 첨가하지 않은 기본배지에서 대부분의 체세포배는 식물체로 재분화되었으며, 두개의 자엽을 갖춘 정상적인 체세포배보다 비정상적인 나팔형의 자엽을 가진 것이 3배 높은 빈도로 나타났다. 이러한 체세포배들은 MS 기본배지에서 식물체로 발아되었으며 생육상에서 생장하여 견실한 열매를 맺었다.

• 식물조직배양학회지
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• 제26권1호
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• pp.15-19
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• 1999

Corydalis remota for. peatinata의 꽃자루에서 캘러스를 유도한 후 MS기본배지에 생장조절물질을 첨가하여 체세포 배발생 및 재분화를 시도하였다. 생장조절 물질에 따른 캘러스 유도율을 비교하면 2,4-D 2.0 mg/L 에 zeatin 2.0 mg/L를 혼용 처리한 배지에서 아주 양호한 캘러스 유도 및 생장율을 나타냈다. 식물생장조절물질 또는 호르몬 첨가에 의한 체세포 배의 발생율은 광조건에서 cytokinin류의 농도가 0.5mg/L 단독 첨가한 배지에서 가장 높았다. 또한 발생된 부정배중 MS기본배지에 zeatin 1.0mg/L을 단독처리한 경우가 가장 쌍자엽형성율이 높은 것으로 나타났다. 또한 2,4-D를 미량 첨가하고 cytokinin류를 혼용첨가한 배지나 BAP를 단독 처리한 배지에서는 소식물체의 배지 접지면인 뿌리부분에 활발한 2차 배발생이 나타났다.

• 한국약용작물학회지
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• 제10권3호
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• pp.217-221
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• 2002

가시오갈피의 꽃봉오리 엽병, 줄기, 줄기마디, 뿌리, 꽃대, 잎 등 조직으로 부터 캘러스가 유기 되었으며 캘러스 유기재료로는 꽃봉오리, 엽병, 줄기, 줄기 마디, 뿌리 등이 적합하였다. 캘러스 유기율은 한국산 가시오갈피가 일본산과 러시아산보나 높게 나타났다. 캘러스유기에는 2.4-D와 TDZ조합처리가 효과적이었으며 한국가시오갈피의 엽병, 줄기, 액아, 뿌리절편으로부터 형성된 캘러스에서 배발생 캘러스가 형성되었으며 줄기에서 형성된 캘러스로부터 한국, 일본, 러시아산 가시오갈피 모두에서 배발생캘러스가 형성되었다. 엽병에서 형성된 캘러스로부터 배발생캘러스의 유기율이 가장 높았으며 2.4-D 1mg/l 처리가 가장 효과적이었다. 액체배지에서의 체세포배의 생장에는 생장조절물질을 첨가하지 않은 MS기본배지가 적합하였다. 기내에서 잎과 뿌리 분화가 이루어지고 건실하게 자란 식물체를 온실에 옮겨 순화하였을 경우 99.1%의 생존율을 보였다.

• 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.

• Plant Biotechnology Reports
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• 제3권3호
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• pp.243-250
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• 2009

In the present study, a simple one medium formulation protocol for callus culture, somatic embryogenesis and in vitro production of ${\beta}-carboline$ alkaloids and diosgenin in Tribulus terrestris L. was developed. Extensive callus induction and proliferation was obtained in leaf explant on Murashige and Skoog (MS) medium supplemented with $5.0{\mu}M$ 6 benzyl adenine (BA) and $2.5{\mu}M$ ${\alpha}-naphthaleneacetic$ acid (NAA). The embryogenic callus was maintained on subculture to fresh parental medium at 4-week intervals over a period of 28 months. The frequency of embryo formation was at a maximum ($18.1{\pm}0.9$ per g of callus) on MS medium containing $5.0{\mu}M$ BA and $2.5{\mu}M$ NAA together with $75mg\;1^{-1}$ casein hydrolysate. Globular embryo developed into torpedo stage embryo under the influence of starvation. The accumulation of ${\beta}-carboline$ alkaloids (harmaline and harmine) and steroidal saponin (diosgenin) in non-embryogenic and embryogenic callus culture derived from leaf explant was compared with root, leaf, stem, and fruit of the mother plant. The embryogenic callus accumulated equivalent amounts of harmaline ($66.4{\pm}0.5{\mu}g/g$ dry weight), harmine ($82.7{\pm}0.6{\mu}g/g$ dry weight), and diosgenin ($170.7{\pm}1.0{\mu}g/g$ dry weight) to that of the fruit of T. terrestris. The embryogenic callus culture of this species might offer a potential source for production of important pharmaceuticals.

• Plant Biotechnology Reports
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• 제4권4호
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• pp.261-267
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• 2010

We describe culture conditions for a high-efficiency in vitro regeneration system of Papaver nudicaule through somatic embryogenesis and secondary somatic embryogenesis. The embryogenic callus induction rate was highest when petiole explants were cultured on Murashige and Skoog (MS) medium containing 1.0 mg $1^{-1}$ ${\alpha}$-naphthaleneacetic acid (NAA) and 0.1 mg $1^{-1}$ 6-benzyladenine (BA) (36.7%). When transferred to plant growth regulator (PGR)-free medium, 430 somatic embryos formed asynchronously from 90 mg of embryogenic callus in each 100-ml flask. Early-stage somatic embryos were transferred to MS medium containing 1.0 mg $1^{-1}$ BA and 1.0 mg $1^{-1}$ NAA to germinate at high frequency (97.6%). One-third-strength MS medium with 1.0% sucrose and 1.0 mg $1^{-1}$ $GA_3$ had the highest frequency of plantlet conversion from somatic embryos (91.2%). Over 90% of regenerated plantlets were successfully acclimated in the greenhouse. Secondary somatic embryos were frequently induced directly when the excised hypocotyls of the primary somatic embryos were cultured on MS medium without PGRs. Sucrose concentration significantly affected the induction of secondary embryos. The highest induction rate (89.5) and number of secondary somatic embryos per explant (9.3) were obtained by 1% sucrose. Most secondary embryos (87.2-94.3%) developed into the cotyledonary stage on induction medium. All cotyledonary secondary embryos were converted into plantlets both in liquid and on semisolid 1/3-strength MS medium with 1.0% sucrose.