• 제목/요약/키워드: Direct shoot regeneration

검색결과 40건 처리시간 0.019초

Optimized Shoot Induction and Histological Study of in vitro Cultured Korean Soybean Cultivars

  • Kantayos, Vipada;Bae, Chang-Hyu
    • 한국자원식물학회지
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    • 제32권3호
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    • pp.237-243
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    • 2019
  • Soybean is the one of recalcitrant legume species for shoot induction. Shoot regeneration via direct organogenesis was investigated in five soybean cultivars, 'Dawon', 'Pungsan', 'Daewon', 'Taekwang' and 'Chongdoo 1' by using cotyledonary node explants. Out of 5 soybean cultivars, an efficient shoot regeneration condition was developed in the two soybean cultivars, 'Dawon' and 'Pungsan'. When various kinds of plant growth regulators with different concentration were estimated, the optimum medium condition for shoot induction in both soybean cultivars was MS + B5 vitamin supplemented with BA at concentration 2 mg/L. In addition, shoot formation efficiency was increased with 97.09% and 93.88% by the pretreatment of BA onto the explants before in vitro culture in both cultivars. Shoot induction in 'Dawon' cultivar was originated from epidermal tissue and sub-epidermal layers when histological changes were investigated under shoot regeneration after culturing cotyledonary node segments on shoot induction medium for 0 to 21 days. Especially, cell dedifferentiation was observed from parenchyma cells to meristematic cell in 3-day cultured segments.

Comparison of chrysanthemum cultivars based on direct shoot regeneration rates in tissue culture

  • Han, Bong-Hee;Lee, Su-Young;Park, Byoung-Mo
    • Journal of Plant Biotechnology
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    • 제36권3호
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    • pp.275-280
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    • 2009
  • Direct shoot regeneration from leaf or internode or petiole segments was conducted in 33 cultivars of chrysanthemum. Shoot regeneration rates varied according to cultivars, culture media, and explant types. The high shoot regeneration rate of more than 70% in 15 cultivars (‘Pink Pangpang’, ‘Orange Memory’, ‘Relance’, ‘Zinba’, ‘Beakma’, ‘Innocence’, ‘Sunny Pangpang’, ‘Euro Yellow’, ‘Dublin’, ‘Boramae’, ‘Peak’, ‘Euro White’, ‘Vesuvio White’, ‘Linneker Salmon’ and ‘Pink Pride’) and 2 ones (‘Forward’ and ‘Agason’) was obtained from the segments of leaves and internodes, respectively, cultured on MS medium containing 1.0 mg-$L^{-1}$ BAP, 0.5 mg-$L^{-1}$ IAA and 30 g-$L^{-1}$ sucrose. That in 6 cultivars (‘Shuhonochikara’, ‘Hakunosen’, ‘Whitney Pangpang’, ‘Plaisir D’Amour’, ‘Grace’ and ‘Kumsu’) was observed from the segments of leaves or internodes cultured on 1/2 MS medium 1.0 mg-$L^{-1}$ BAP, 0.5 mg-$L^{-1}$ IAA and 15 g-$L^{-1}$ sucrose In case of 3 cultivars (‘Ilweol’, ‘Puma White’ and ‘Sharon’), when internode explants excised from mother plants, which were pre-cultured on MS medium containing 2 g-$L^{-1}$ activated charcoal and 30 g-$L^{-1}$ sucrose for two months in the dark, and cultured on MS medium containing 1.0 mg-$L^{-1}$ BAP, 0.5 mg-$L^{-1}$ IAA and 30 g-$L^{-1}$ sucrose, that was shown. Seven cultivars including ‘Puma Yellow’, ‘Argus’, ‘Yes Morning’, ‘Whiparam’, ‘Hakunohikari’, ‘Charming Eye’ and ‘Moon light’ requires more improved culture conditions. Tissues with the highest shoot regeneration rate were in descending order, leaf, petiole, and internode segments.

High Frequency Plant Regeneration from Leaf, Petiole and Internode Explants of Codonopsis lanceolata Benth.

  • Ghimire, Bimal Kumar;Shin, Chul-Min;Li, Cheng Hao;Kim, Na-Young;Chung, III-Min;Lim, Jung-Dae;Kim, Jae-Kwang;Kim, Myong-Jo;Cho, Dong-Ha;Yu, Chang-Yeon
    • 한국약용작물학회지
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    • 제15권2호
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    • pp.73-81
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    • 2007
  • An efficient regeneration system was developed using leaf, petiole, and internode explants. Highly embryogenic callus was obtained following cultivation on MS basal nutrient supplemented with 2 $mg/{\ell}$ 2,4-D. Globular, heart, torpedo and cotyledon shaped somatic embryo were produced from the surface of embryogenic callus. Direct shoot regeneration without intermediate callus formation has been achieved on MS medium supplemented NAA and BAP. The percentage of response varies with different concentration of auxin and cytokinin treated individually or in combination. The best shoot regeneration response (54.28%) and number of shoot per explant (12.67) were achieved on the medium supplemented with 0.1 $mg/{\ell}$ NAA and 1 $mg/{\ell}$ BAP. The regenerated shoot transformed into young plant when cultured into elongation and root induction medium. More than 90% of in vitro propagated plants could survive when transferred to the greenhouse for acclimation. This optimized regeneration system can be used for rapid shoot proliferation and genetic transformation.

Plant regeneration via direct and indirect adventitious shoot formation and chromosome-doubled somaclonal variation in Titanotrichum oldhamii (Hemsl.) Solereder

  • Takagi, Hiroki;Sugawara, Shintaro;Saito, Tomoka;Tasaki, Haruka;Yuanxue, Lu;Kaiyun, Guan;Han, Dong-Sheng;Godo, Toshinari;Nakano, Masaru
    • Plant Biotechnology Reports
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    • 제5권2호
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    • pp.187-195
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    • 2011
  • The gesneriaceous perennial plant Titanotrichum oldhamii has beautiful foliage and attractive bright yellow flowers. However, breeding of T. oldhamii by conventional sexual hybridization may be difficult because sexual reproduction of this species is very rare. In the present study, plant regeneration systems via both direct and indirect formation of adventitious shoots from leaf explants were established as the first step toward breeding T. oldhamii by using biotechnological techniques. Adventitious shoots were formed efficiently on medium containing $0.1mg\;l^{-1}$ benzyladenine. Histological observation showed that shoot formation on this medium occurred directly from leaf epidermal cells without callus formation. On the other hand, leaf explants formed calluses on medium containing $0.1mg\;l^{-1}$ 2,4-dichlorophenoxyacetic acid. The calluses could be maintained by monthly subculturing to fresh medium of the same composition. When the calluses were transferred to plant growth regulator-free medium, they formed adventitious shoots. Directly and indirectly formed shoots rooted well on medium containing $0.1mg\;l^{-1}$ indole-3-butyric acid. Plantlets thus obtained were successfully acclimatized and grew vigorously in the greenhouse. Flow cytometry analysis indicated that no variation in the ploidy level was observed in plants regenerated via direct shoot formation, whereas chromosome doubling occurred in several plants regenerated via indirect shoot formation. Regenerated plants with the same ploidy level as the mother plants showed almost the same phenotype as the mother plants, whereas chromosome-doubled plants showed apparent morphological alterations: they had small and crispate flowers, and round and deep green leaves.

High frequency direct plant regeneration from leaf, internode, and root segments of Eastern Cottonwood (Populus deltoides)

  • Yadav, Rakesh;Arora, Pooja;Kumar, Dharmendar;Katyal, Dinesh;Dilbaghi, Neeraj;Chaudhury, Ashok
    • Plant Biotechnology Reports
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    • 제3권3호
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    • pp.175-182
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    • 2009
  • Simple, reproducible, high frequency, improved plant regeneration protocol in Eastern Cottonwood (Populus deltoides) clones, WIMCO199 and L34, has been reported. Initially, aseptic cultures established from axillary buds of nodal segments from mature plus trees on MS liquid medium supplemented with $0.25mg\;1^{-1}$ KIN and $0.25mg\;1^{-1}$ IAA. Nodal and internodal segments were found to be extra-prolific over shoot apices during course of aseptic culture establishment, while $0.25mg\;1^{-1}$ KIN concentration played a stimulatory role in high frequency plant regeneration. Diverse explants, such as various leaf segments, internodes, and roots from in vitro raised cultures, were employed. Direct plant regeneration was at high frequency of 92% in internodes, 88% in leaf segments, and 43% in root segments. This led to the formation of multiple shoot clusters on established culture media with rapid proliferation rates. Many-fold enhanced shoot elongation and growth of the clusters could be achieved on liquid MS medium supplemented with borosilicate glass beads, which offer physical support for proliferating shoots leading to faster growth in comparison to semi-solid agar or direct liquid medium. SEM examination of initial cultures confirmed direct plant regeneration events without intervening calli. In vitro regenerated plants induced roots on half-strength MS medium with $0.15mg\;1^{-1}$ IAA. Rooted 5- to 6-week-old in vitro regenerated plants were transferred into a transgenic greenhouse in pots containing 1:1 mixture of vermicompost and soil at $27{\pm}2^{\circ}C$ for hardening and acclimatization. 14- to 15-week-old well-established hardened plants were transplanted to the field and grown to maturity. The mature in vitro raised poplar trees exhibited a high survival rate of 85%; 4-year-old healthy trees attained an average height of 8 m and an average trunk diameter of 25 cm and have performed well under field conditions. The regeneration protocol presented here will be very useful for undertaking genetic manipulation, providing a value addition to Eastern Cottonwood propagation in future.

High Frequency Shoot Regeneration from leaf Explants of Cucumber

  • Seo, Seung-Hee;Bai, Dong-Gyu;Park, Hyeon-Yong
    • Journal of Plant Biotechnology
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    • 제2권1호
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    • pp.51-54
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    • 2000
  • Leaf explants of the cucumber (Cucumis sativus L.) were cultured on Murashige and Skoog's (MS) medium supplemented with various concentrations of $\alpha$-naphthalene acetic acid (NAA) and 6-benzylaminopurine (BAP). Direct shoot orgnogenesis as well as callus formation with somatic embryos and multiple shoots was observed from leaf explants of cvs. Shinhukjinju and Chungjang. The highest frequency of shoot formation 80% was observed on MS medium supplemented with NAA/BAP (5.0 ${\mu}{\textrm}{m}$/2.5 ${\mu}{\textrm}{m}$), with explants forming 3-7 shoots. Shoots formation occured within 3 to 4 weeks. Only one subculture of calli was required for plant regeneration on normal growth regulator-free medium. Plantlets transferred to soil developed into plants of normal appearance, which flowered and set fruits.

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High-frequency plant regeneration from leaf-disc cultures of Jatropha curcas L.: an important biodiesel plant

  • Deore, Ajay C.;Johnson, T. Sudhakar
    • Plant Biotechnology Reports
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    • 제2권1호
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    • pp.7-11
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    • 2008
  • A simple, high-frequency and reproducible protocol for induction of adventitious shoot buds and plant regeneration from leaf-disc cultures of Jatropha curcas L. has been developed. Adventitious shoot buds were induced from very young leaf explants of in vitro germinated seedlings as well as mature field-grown plants cultured on Murashige and Skoog's (MS) medium supplemented with thidiazuron (TDZ) ($2.27{\mu}M$), 6-benzylaminopurine (BA) ($2.22{\mu}M$) and indole-3-butyric acid (IBA) ($0.49{\mu}M$). The presence of TDZ in the induction medium has greater influence on the induction of adventitious shoot buds, whereas BA in the absence of TDZ promoted callus induction rather than shoot buds. Induced shoot buds were multiplied and elongated into shoots following transfer to the MS medium supplemented with BA ($4.44{\mu}M$), kinetin (Kn) ($2.33{\mu}M$), indole-3-acetic acid (IAA) ($1.43{\mu}M$), and gibberellic acid ($GA_3$) ($0.72{\mu}M$). Well-developed shoots were rooted on MS medium supplemented with IBA ($0.5{\mu}M$) after 30 days. Regenerated plants after 2 months of acclimatization were successfully transferred to the field without visible morphological variation. This protocol might find use in mass production of true-to-type plants and in production of transgenic plants through Agrobacterium/biolistic-mediated transformation.

제초제(除草劑) BIALAPHOS에 대(對)한 연초(煙草)의 내성(耐性) 증대(增大) (Development of Herbicide(BIALAPHOS) Tolerant Tobacco through Tissue Culture)

  • 배윤주;김길웅
    • 한국잡초학회지
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    • 제8권2호
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    • pp.182-186
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    • 1988
  • 제초제(除草劑) bialaphos 처리(處理)에서 살아남은 개체(個體)로부터 다시 bialaphos가 처리(處理)된 배지(培地)에서 callus의 유도(誘導), 생장(生長) 및 direct shooting, ammonia 함량(含量) 등(等)으로 내성(耐性)을 재검정(再檢定)하여 얻어진 약간의 결과(結果)를 요약(要約)하면 다음과 같다. 1. Bialaphos가 0.5~0.75 ppm의 저농도(低濃度)로 처리(處理)된 배지(培地)에서 생장(生長)한 callus로부터 분화(分化)된 식물체(植物體) 수(數)는 품종(品種)에 관계없이 분화(分化)가 잘 되었으나 1.0 ppm 처리구(處理區)에서 계대배양(繼代培養)된 것 가운데서는 KA 101만이 분화(分化)되었다. 2. 분화식물체(分化植物體) 가운데 bialaphos에 내성(耐性)인 개체(個體)를 재선발(再選拔)하기 위해 bialaphos 100.0 ppm 처리구(處理區)에 치상시(置床時) 모두 고사(枯死)하였으며 bialaphos 10.0 ppm 처리구(處理區)에서는 NC82 품종(品種) 2.43%. KA 101 2.76%. BY4 0.78%가 살아남아 품종간(品種間)에 다소 차이(差異)가 있었다. 3. Bialaphos 10.0 및 100.0 ppm의 처리구(處理區)에서 고사(枯死)한 shoot에 함유(含有)된 ammonia 함량(含量)은 무처리구(無處理區)보다 약 15배(倍) 정도(程度) 높아 shoot 의 고사원인(枯死原因)이 ammonia 축적(蓄積)에 의한 것임을 입증(立證)할 수 있었다. 4. NC 82 품종(品種)의 TP 1 (내성식물체(耐性植物體) 개체(個體)가 비선발개체(非選拔個體)보다 bialaphos가 2.5 ppm 첨가(添加)된 callus 증식배지(增殖培地)에서 callus 증식률(增殖率)이 2.14배(倍)나 높았고, bialaphos가 10.0 ppm 첨가(添加)된 shoot 분화배지(分化培地)에서는 샤레당(當) 11 개체(個體)의 shoot를 분화(分化)시켜 다른 개체(個體)보다 내성(耐性)이 컸다.

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삽주(Atractylodes ovata (Thunb.) DC.)의 효율적인 기내 줄기 재분화 (An efficient in vitro shoot regeneration system for Atractylodes ovata (Thunb.) DC. )

  • 정희영 ;김지아
    • Journal of Plant Biotechnology
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    • 제49권4호
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    • pp.325-330
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    • 2022
  • 본 연구는 국내에서 소비되는 약용작물 중 경제적 가치는 높으나 국내 생산량이 적고 수입 의존도가 높은 Atractylodes의 식물 조직 배양 시스템을 구축하기 위해 수행되었다. 삽주는 A. ovata를 사용하였고, 4가지 cytokinins류, 6-benzylaminopurine (BA), zeatin, kinetin, thidiazuron (TDZ)을 2가지 농도(0.5, 1.0 mg/L)로 처리하였다. 4가지 유형의 cytokinin 중 BA처리는 A. ovata의 신초유도와 뿌리 생육에 효과적이었다. 0.5 mg/L 및 1.0 mg/L BA 모두 BA 처리에서 유사한 결과를 나타내었지만 1.0 mg/L BA가 신초와 뿌리 생육을 촉진하는데 더 효과적이었다. 처리 중 신초의 개수와 뿌리의 생중량(FW)을 제외하고는 TDZ 처리가 신초와 뿌리생육에 효과적이지 않아 본 수종에서는 적합하지 않았다. 본 연구에서는 A. ovata의 아배양을 이용한 기내증식 시스템을 구축하였다. 위 결과는 기내 재분화를 이용한 산림약용자원 A. ovata의 안정적인 생산 및 증식을 위한 기반 기술로 사용될 것으로 사료된다.

Plant Regeneration from Callus and Adventitious Root Segments of Pulsatilla Koreana Nakai

  • Jung, Su-Jin;Jeong, Jae-Hun;Yoon, Eui-Soo;Choi, Yong-Eui
    • Journal of Plant Biotechnology
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    • 제34권2호
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    • pp.153-159
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    • 2007
  • Plant regeneration of Pulsatilla koreana was achieved via adventitious shoot formation indirectly from callus and directly from adventitious root segments. For the callus induction from leaf or petiole explants, combination of 2,4- dichlorophenoxyaceticacid (2,4-D) with $2.22\;{\mu}M$ 6-benzyladenine (BA) was effective. Adventitious shoot induction from callus was enhanced by the combined treatment with $0.1\;{\mu}M$ polyvinylpyrrolidone (PVP) compared to cytokinin treatment alone. Adventitious roots were induced from the petiole segments on 1/2 MS medium with $4.93\;{\mu}M$ IBA. High frequency direct adventitious shoot formation from the segments of adventitious roots was achieved on medium with $4.92\;{\mu}M$ 2-isopentenyladenine (2-ip). Elongated shoots were rooted on half-strength MS medium containing $5.71\;{\mu}M$ indole acetic acid (IAA). Regenerated plantlets with well-developed shoots and roots were successfully transferred to soil. This in vitro propagation protocol might be useful for mass propagation as well as conservation of this plant.