• 식물조직배양학회지
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• 제28권3호
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• pp.129-134
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• 2001

Winter bud explants from 15 individual angelica tree (Aralia elata) were cultured in vitro to find out optimal conditions for somatic embryo induction as well as plant regeneration. Calli are induced and grown on MS medium supplemented with 1.0 mg/L 2,4-D for 4 weeks and subcultured on a half-strength MS medium without phytohormones to induce somatic embryos. Inter-simple sequence repeat (I-SSR) markers were analyzed with total DNAs extracted from the trees. Genotype effects on somatic embryo induction were examined by cluster analysis. Callus induction rate varied from 58.5 to 100% among the genotypes. Somatic embryo induction rate also greatly varied from 0 to 100% among the genotypes. There was a significant difference in somatic embryo induction rate even among the individual trees that showed close genetic relationships each other. This suggested that somatic embryo induction rate in Aralia elata be influenced by a few major specific genes rather than whole genomic similarity among individual trees. Four individuals of Ulneong-7, Cheju-1, Shingu and China, which are recalcitrant to somatic embryo induction, turned out to have a close genetic relationship, suggesting that both physiological and genetic factors affect somatic embryo induction. The results suggest that genotype selection be the most important factor to achieve an efficient propagation, although cultural optimization through medium and explant manipulation may also play crucial roles in somatic embryogensis as well as plant regeneration of these species.

• Journal of Plant Biotechnology
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• 제42권4호
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• pp.388-395
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• 2015

본 연구는 약용 및 식용으로 유망한 음나무의 대량생산 및 생식질 보존을 위한 기초 자료를 제공하기 위해 배발생 조직을 재료로 저온저장 기간별 SE 유도 및 식물체 재생을 시험하였다. 배발생 캘러스는 저온저장 6개월까지 정상적인 체세포배 형성이 가능하였으나 유도 빈도는 저장 기간에 따라 감소하는 경향을 보였다. 배발생능이 가장 좋은 ZE 10 세포주은 체세포배 유도 빈도에 있어 저장 기간에 따른 감소폭이 비교적 적은 것으로 나타났다. 체세포배의 발아 및 식물체 재생은 캘러스의 저장기간에 따라 다소 감소하는 경향을 보였으나 발아율은 93% 이상, 식물체 재생율은 91% 이상으로 저온저장에 따른 큰 문제는 없었다. 조직검경 결과 저장 기간에 따라 세포의 갈변화 및 활력 저하가 나타났으며, 아세토카민과 에반스블루의 염색을 통해서도 확인되었다. 이상의 결과로 볼 때 음나무 배발샐 캘러스의 저온저장은 계대배양 없이 $4^{\circ}C$의 냉장고에서 6개월 까지 가능한 것으로 나타났으며, 저장 후 체세포배 유도효율을 위해 배발생능이 양호한 세포주의 저장이 필요한 것으로 관찰되었다.

• Journal of Plant Biotechnology
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• 제7권3호
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• pp.187-193
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• 2005

A protocol was developed for somatic embryogenesis with 100% induction rate from immature zygotic embryo axes of peanut (Arachis hypogaea L. cv. DRG-12) cultured on MS medium containing $18.09\;{\mu}M$ 2,4-D. The frequency of somatic embryogenesis (31.7%) as well as the number of somatic embryos induced per explant (6.6) decreased when the concentration of 2,4-D was increased to $72.4\;{\mu}M$. Morphologically abnormal somatic embryos were observed at a frequency of 43.3% on MS medium containing $72.4\;{\mu}M$ 2,4-D. Somatic embryos isolated from 30-day-old cultures of immature zygotic embryo axes exhibited precocious germination with varied responses when placed on MS basal medium with 3% sucrose. Maximum shoot induction (80.0%) was observed from somatic embryos isolated from 60-day-old cultures of immature zygotic embryo axes when placed as a clump rather than individually on MS medium supplemented with $26.63\;{\mu}M$ BA and $0.54\;{\mu}M$ NAA. Shoots developed from somatic embryos rooted with higher frequency (93.3%) on Blaydes' medium containing $5.4\;{\mu}M$ NAA.

• 한국산림과학회지
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• 제97권5호
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• pp.547-551
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• 2008

The study aimed to develop a cryopreservation method for long-term storage using mature somatic embryo of Japanese larch. In this study, desiccation treatments significantly affected re-induction rates of embryogenic tissue (ET) from dried somatic embryos. In the effect of different dehydration temperature and duration on the re-initiation ET. the highest frequency was shown when somatic embryos were dehydrated at $25^{\circ}C$ for 2 (45.5%) or 1 day (43.3%), respectively. In addition, low temperatures [$4^{\circ}C$, 2 days (44.2%) or 3 days (43.5%)] were marked higher ET initiation. After that, the initiation value was declined with dehydration duration. For comparison of different relative humidity on re-induction frequency of ET, the best re-induction (43.5%) was obtained from somatic embryos pre-dried at $(NH_4)_2SO_4$ (RH 79%). Both $Na_2HPO_4$ (RH 97%) and $Na_2CO_3$ (RH 88%) treatments were showed the similar rate, 34.6, 34.2%, respectively. However the lowest rate (19.6%) was observed in distilled water (RH 100%). In comparison of the various storage temperatures and duration of the dried somatic embryos, the highest frequency (66.9%) of re-initiation was obtained when somatic embryos were cryopreserved for one day. However, the frequency was gradually decreased as the time length of storage increased regardless of types of storage. None of ET re-initiated when stored at $4^{\circ}C$ for 1, 2 and 84 days.

• Journal of Plant Biotechnology
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• 제6권1호
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• pp.45-50
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• 2004

Somatic embryos were induced from immature cotyledons and cultured on a MS medium containing 40mg/L 2,4-D. The maximum induction of embryos was obtained from immature cotyledons in a size of 3-4mm, and the highest frequency was obtained in the induction medium at pH 7.0. For embryo development, embryogenic tissues were transferred to a MSM6AC and MSM6 media. Developing embryos were placed at 27$^{\circ}C$with dim light (20$\mu$$molm^{-2}$$s^{-1}$) provided by cool fluorescent tubes (3-D wavelength light is better than standard light). Somatic embryos were clearly developed from globular stage to cotyledonary stages. The color of embryo may be a useful parameter for estimation of embryo quality. When the embryo becomes mature, embryo will be ready for desiccation in order to induce roots and shoots of embryos.

• Journal of Plant Biotechnology
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• 제3권1호
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• pp.19-25
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• 2001

Induction of somatic embryogenesis from Brazilian eggplant variety F-100 was studied in response to four auxin types. NAA, at the optimal concentration of 54 $\mu\textrm{m}$, was the only one that resulted in the induction of somatic embryos in either leaf and cotyledon explant and, at murk lower intensity and frequency, in hypocotyl and epicotyl explants. The optimal temperatures for embryo induction were 28 and 35$^{\circ}C$ for cotyledon and leaf explants. Incubation at 22$^{\circ}C$ caused a significant reduction both in the frequency and intensity of induction. This system was used to study the effects of position and orientation of the tissue on the culture medium as well as of antibiotics and explant co-cultivation with Agrobacterium on the efficiency of somatic embryo induction. The intensity of embryo induction was greater in the midsections of cotyledons relative to apical and basal regions, when the abaxial surface was in contact with the culture medium. The presence of antibiotics resulted in approximately 40-60% reduction of embryo induction relative to control explants, which originated 335$\pm$26.6 embryos. Co-cultivation with Agrobacterium before treatment with antibiotics caused a more drastic reduction (80-99%). Ampicilin treatment after cocultivalion with Agrobacterium caused the least inhibitory effect, allowing the production of 60 embryos/explant.

• 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.

• 한국산림과학회지
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• 제87권1호
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• pp.57-61
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• 1998

10년생 두릅나무의 동아(冬芽)를 절편으로 체세포배(體細胞胚)를 유도하고 식물체(植物體)를 재생(再生)시켜서 단지(團地)에 활착시키는 실험을 수행하였다. 배발생(胚發生) 캘러스는 MS 배지(培地)에 2, 4-D를 첨가하여 유도(誘導)되었다. 다량의 체세포배가 MS 배지에 2, 4-D와 BA를 동시에 첨가한 조건에서 발생(發生)되었다. 배발생 캘러스에서는 기형의 체세포배가 흔히 관찰되었으나 대부분은 전형적(全形的)인 배(胚) 형태(形態)를 보였다. 어뢰형에서 자엽(子葉) 단계(段階)에 이르는 성숙(成熟)한 체세포(體細胞) 배(胚)는 모두 발아(發芽)가 가능하였으나 완전한 식물체로의 재분화(再分化)는 MS 기본 배지에서 평균(平均) 14%이었다. 재분화된 식물체는 온실(溫室)에서 순화(馴化) 후(後) 단지(團地)에서 95% 이상 활착(活着)되었고, 이식(移植) 후(後) 3년까지 정상적인 생장(生長)을 보였다.

• Journal of Plant Biology
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• 제37권3호
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• pp.333-341
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• 1994

Effective plant regeneration from immature cotyledons of soybean [Glycine max (L.) Merr.] cultivars was achieved via somatic embryogenesis. Somatic embryogenesis was performed with the cotyledons of immature embryos 14-20 d after flowering. Immature cotyledons of cv. Whangkeum were placed abaxial or adaxial side down on modified MS medium containing 20mg/L 2,4-D. The greatest number of somatic embryos, 1.2 per cotyledon, was produced from those of 4.0-4.9 mm in length which had been placed abaxial side down. Among cvs. Pecking, Whangkeum and Baekwoon, Pecking had the highest embryo induction efficiency with 4.3 somatic embryos per cotyledon in 20mg/L 2,4-D treatment and with 1.0 embryo per cotyledon in 8mg/L NAA treatment. Germinable globular somatic embryos were induced with the highest efficiency, 27.6%, in 20mg/L 2,4-D and were proliferated efficiently on liquid medium containing 10mg/L 2,4-D. The globular somatic embryos developed into germinable mature somatic embryos on medium containing 10 $\mu$M CoCl2, 9% sucrose, and 0.5% activated charcoal. These mature somatic embryos germinated on hormone-free mediu. After transfer to the soil, regenerated plants with seeds were obtained.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 추계학술대회
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• pp.80-80
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• 2018

Hosta is a genus of the family Asparagaceae and distributed in East Asia. There are six Hosta species (Hosta capitata (Koidz.) Nakai, H. clausa Nakai, H. jonesii M.G.Chung, H. minor (Baker) Nakai, H. venusta F.Maek., and H. yingeri S.B.Jones) native to Korea and among them, four species (H. minor, H. jonesii, H. venusta and H. yingeri) are endemic to the Korea peninsula. Hosta is generally propagated by seed, crown division or tissue culture. However, tissue culture is a more efficient method to mass proliferation, a new cultivar development and disease-free plantlet production in a limit time. Hence, we conducted this study to evaluate the influence of various plant growth regulators (PGRs) treatments on the induction of callus and somatic embryo of the six Hosta species. Leaf, petiole and root were used to select optimum tissue culture explants. Petiole explants thus only were used for callus induction and somatic embryogenesis with TDZ (0.1, 0.5 or 1.0mg/L) and NAA (0.1 or 0.5 mg/L) combinations. After 12 weeks of culture, the highest rate of somatic embryogenesis was achieved on modificated MS medium containing 1.0 mg/L TDZ and 0.1 mg/L NAA in H. capitata and H. minor (15.5%, respectively), 0.1 or 0.5 mg/L TDZ and 0.1 mg/L NAA in H. jonesii (22.2%), 1.0 mg/L TDZ and 0.5 mg/L NAA in H. yingeri (26.7%), and 0.1 mg/L TDZ and 0.5 mg/L NAA in H. venusta (53.3%). H. clausa showed very low effect on somatic embryogenesis by PGRs; 2.2%. There was interspecies difference to PGRs respond for callus and somatic embryo induction. Regenerated multiple shoots and plantlet of H. minor, H. jonesii, H. venusta and H. yingeri were obtained via somatic embryogenesis.