• Title/Summary/Keyword: plant regeneration.

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Plant Regeneration of Soybean Cultivars via Somatic Embryogenesis

  • Kwang-Woong Lee
    • Journal of Plant Biology
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    • v.37 no.3
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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.

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Plant Regeneration from Shoot Tip-Derived Embryogenic Callus of Dianthus superbus

  • Lee, Eun-Ae;Kim, Joon-Chul;Kim, Won-Bae;Kim, Byeong-Hyeon;Kim, Jeong-Kan
    • Journal of Plant Biology
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    • v.37 no.3
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    • pp.381-385
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    • 1994
  • The highest degree of callus formation was obtained from the shoot tips of Dianthus superbus when cultured on the MS medium supplemented with 2.0 mg/L NAA and 0.5 mg/L BAP. Embryogenic calluses were obtained from the seperated friable calluses on MS medium containing 2.0 mg/L 2,4-D after 7-8 wk of culture. For plant regeneration, embryogenic calluses were selected and cultured on te proliferation medium. After 3 wk, somatic embryos appeared on MSK medium (0.5 mg/L NAA, 2.0 mg/L kinetin) and N6 medium (2.0 mg/L kinetin, 0.1 mg/LNAA, 0.1 mg/L 2,4-D and 2.0 g/L casein hydrolysate). When these somatic embryos were kept under continuous illumination, shoots were successfully regenerated on the both media. The shoots were rooted on MS medium supplemented with 2.0 mg/L NAA.

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Plant Regeneration from Rice Microspore Cultures (벼의 소포자 배양을 통한 식물체 재분화)

  • 김영수
    • Journal of Plant Biology
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    • v.36 no.2
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    • pp.183-192
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    • 1993
  • Efficient plant regeneration system was established through the culture of rice (Oryza sativa L.) microspores. Microspores released by anther shedding culture developed into proembryos and calluses in B5 medium within two weeks of culture. The optimal hormone and carbon sources were dependent on the genotypes used. Microspore's viability decreased rapidly in culture time, therefore less than 3% of the total microspores were viable at the 9th day when the first microspore division was observed. Of two types of microspores (pollen dimorphism) observed in culture, only the P-grain, larger microspores than normal one was able to divide. Using 4',6-diamidino-2-phenylindole (DAPI) fluorescence staining, it was confirmed that the symmetrical division of uninucleate microspore was the first step leading to continuous microspore development. Microspore-derived proembryos and calluses were regenerated to plants in N6 medium containing 1 mg/L NAA and 5 mg/L kinetin, and 78.3% of the regenerated plants were haploids.

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Transformation of Birdsfoot trefoil by BcHSP17.6 Gene using Agrobacterium tumefaciens (BcHSP17.6 유전자 도입에 의한 버즈풋 트레포일의 형질전환)

  • 김기용;성병렬;임용우;최기준;임영철;장요순;정의수;김원호;김종근
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.21 no.3
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    • pp.145-150
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    • 2001
  • This study was conducted to obtain the transformed birdsfoot trefoil (Lotus corniculatus L.) plants with BcHSP17.6 gene using Agrobacterium turnefaciens LBA4404 and we confirmed transformed gene from the regenerated birdsfoot trefoil plants. The expression vector, pBKH4 vector, harboring BcHSP17.6 gene was used for production of transgenic birdsfoot trefoil plants. The callus of birdsfoot trefoil was cocultivated with Agrobacteriurn turnefaciens and transformed calli were selected on kanamycin-containing SH-kc medium to regenerate into plants. The transformed birdsfoot trefoil plants were produced 4 momths after cultivation on BOi2Y medium. The transgenic birdsfoot trefoil plants were analyzed by isolation of genomic DNA and genomic Southern hybridization using a -32P labelled BcHSPl7.6 fragments. (Key words : Birdsfoot trefoil, Transgenic plant. BcHSP17.6 gene, Callus induction, Plant regeneration)

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Optimization of Shoot Induction, Histological Study and Genetic Stability of in vitro Cultured Pisum sativum cv. 'Sparkle'

  • Kantayos, Vipada;Bae, Chang-Hyu
    • Korean Journal of Plant Resources
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    • v.32 no.1
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    • pp.19-28
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    • 2019
  • An efficient shoot regeneration condition for pea cv. 'Sparkle' was developed by using optimum explant, plant growth regulator concentrations, and pretreatment of BA onto explant. The average shoot number per explant showed the highest on two kinds of shoot induction media (MSB5 media containing 2 mg/L BA and a combination of 2 mg/L BA and 1 mg/L TDZ) when cotyledonary node explants were cultured. Moreover, the pretreatment of explant in 200 mg/L BA solution was found to be more effective in shoot induction than that of non-pretreatment. By histological study, cell division and proto-meristem were formed near the surface of the sub-epidermal and epidermal cell layers of cotyledonary node in earlier than 3 days after culture. The analysis of genetic stability of regenerants by using thirteen ISSR markers showed that in vitro regenerated plants showed polymorphism with 8.3% compared with their mother plants.

Callus Induction and Plant Regeneration from Mature Seeds of Timothy (티모시 성숙종자로부터 캘러스 유도 및 식물체 재분화)

  • Lee, Ki-Won;Kim, Ki-Yong;Choi, Gi-Jun;Lim, Young-Chul;Kim, Won-Ho;Jung, Min-Wong;Seo, Sung;Lee, Byung-Hyun;Lee, Sang-Hoon
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.28 no.3
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    • pp.165-170
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    • 2008
  • Timothy (Phleum pratense L.) is an important grass species as forage. In order to optimize tissue culture conditions of timothy, the effects of plant growth regulators on callus induction and plant regeneration was investigated with mature seeds of colt cultivar. The optimal concentration of 2,4-D for the induction of primary callus from mature seeds was 3 mg/L. The highest embryogenic callus frequenc (25%) was observed when the mature seed were cultured on MS medium supplemented with 3 mg/L 2,4-D and 0.1 mg/L BA. The highest plant regeneration frequency was observed when type B callus was transferred to N6 medium supplemented with 1 mg/L 2,4-D and 3 mg/L BA. Regenerated plants were grown normally when shoots were transplanted to the soil. A short tissue culture period and regeneration system would be beneficial for molecular breeding of timothy by the production of transgenic plant.

Effect of Embryo Maturity and Medium on Callus Formation and Plant Regeneration from Immature Embryo of Eleutherococcus senticosus (가시오갈피 미숙배로부터 Callus 형성 및 식물체 재분화에 미치는 배의 성숙정도 및 배지의 효과)

  • 유창연
    • Korean Journal of Plant Resources
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    • v.10 no.2
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    • pp.122-127
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    • 1997
  • This study was conducted to establish mass propagation system from the tissue culture using immature embroys in Eleutherococus senticosus. Immature embroys from seeds were removed under the microscope and placed on modified SH and WPM medium containing several plant growth regulators. The calli were well formed on media containing 1mg/l of 2,4-D on modified SH medium and 1mg/l of 2,4-D and 3mg/l of TDZ on WPM medium. Shoot regeneration was better on modified SH or WPM medium with combination of high concentration of TDZ and low concentration of 2,4-D. Treatment of 2,4-D alone was better than treatment of TDZ alone in callus induction on modified SH medium but plant regeneration reversed. Treatment of 2.4-D and TDZ combination was better than treatment of 2,4-D alone in callus induction on WPM medium. The results of callus formation and shoot regeneration on WPM media differed to those of SH media. The rate of callus formation was nearly 83% when 2,4-D was added to SH medium on concentration of 1mg/l. The rate of callus formation was nearly 38% when combination treatment of 2,4-D 1mg/l and TDZ 3mg/l was added to WPM medium. Also, plant regeneration differed depending on the mature degree of immature embryo.

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Effect of Thidiazuron on the Formation of Micro-tubers and Plantlet Regeneration of Pinellia ternata T. (Thidiazuron이 반하(半夏)의 자구(子球) 및 식물체 재생에 미치는 영향)

  • Kim, Jae-Kwang;Cho, Hye-Kyoung;Yu, Chang-Yeon;Ahn, Sang-Deuk;Lim, Hak-Tae
    • Korean Journal of Medicinal Crop Science
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    • v.5 no.1
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    • pp.21-27
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    • 1997
  • This experiments were conducted to determine the effect of thidiazuron on forming tuberlets and plant regeneration of Pinellia ternata T. by tissue culture. The addition of $5\;{\mu}M$ TDZ to the medium had better regeneration than that of any other treatments of NAA and TDZ. At the combination treatments of NAA and TDZ, as the level of thidiazuron increased, the rate of shoot regeneration was incresed while the increment of NAA concentration inhibited the rate of shoot regeneration. The supplement of $5\;{\mu}M$ thidiazuron produced the best number of micro-tubers per explant and the number of micro-tuber formed was 25 in MS medium and 29 in MG medium on 30 day culture, respectively. Microtuber formation was the best on MG medium with 1.0 mg/l NAA and $5\;{\mu}M$ thidiazuron. MG medium was superior to MS and B5 medium for the growth of tuberlets. Half strength of MS medium with NAA 2 mg/l was the most effective for root formation. Rooting ability on nursery soil of plantlets produced in in vjtro was good as a 80% after 3 weeks.

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