• Title/Summary/Keyword: in vitro rooting

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In vitro Mass Propagation of Ardisia pusilla DC. (산호수 (Ardisia pusilla DC.)의 기내 대량번식)

  • Kang Gwan-Ho;Oh Owel-Sun;Goo Dae-Hoe;Eun Jong-Seon;Kim Hyung-Moo
    • Journal of Plant Biotechnology
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    • v.32 no.4
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    • pp.281-285
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    • 2005
  • To establish the mass proliferation system of Ardisia pusilla DC, the shoot tips of Ardisia pusilla DC were cultured on the MS and half-strength MS medium supplemented with $0{\sim}5.0$ mg/L BA or $0{\sim}0.5$ mg/L thidiazuron(TDZ), respectively. A few multiple shoot formation observed when the shoots were cultured on MS medium containing TDZ. However, the frequency of multiple shoot formation was reached up to 82.4%, when the shoots were cultured on half-strength MS medium supplemented with 0.5 mg/L BA. Also the number of shoot per explant was 7.1. To promote rooting from multiple shoot, newly formed shoots were transferred to half-strength MS medium containing 0.5 mg/L IBA or 0.5 mg/L NAA, respectively. Regenerated plantlets were grown to normal mature plants in soil.

Callus induction and in vitro plant regeneration of Polygonatum stenophyllum Maxim. (층층둥굴레(Polygonatum stenophyllum Maxim.)의 캘러스 유도 및 기내 식물체 분화)

  • Park, Min Wan;Ryu, Shi Hyun;Lee, Yoon Young;Song, Jae-Mo;Kim, Jin Ho;Ahn, Young-Hee;Bae, Kee Hwa
    • Journal of Plant Biotechnology
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    • v.45 no.3
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    • pp.266-272
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    • 2018
  • A Polygonatum stenophyllum Maxim. is an important endangered plant belonging to the family Liliaceae. A method was developed for the rapid micropropagation of P. stenophyllum through plant regeneration from rhizome (1-year, 3-years, and 5-years) explant-derived calli. The rhizome segments were cultured in Murashige and Skoog (MS) medium supplemented with varying concentrations of 2,4-D (0, 0.5, 1.0, $1.5mg{\cdot}L^{-1}$) for callus induction. In media supplemented with $0.5mg{\cdot}L^{-1}$ of 2,4-D, 87% of 3-years rhizome produced callus. Subsequently, the callus was transferred to 1/2MS medium supplemented with various concentrations of IAA, IBA, NAA, and 2,4-D (0, 0.1, 0.5 and $1.0mg{\cdot}L^{-1}$) for adventitious shoot formation. The highest percentage of adventitious shoot induction (57%) was observed in 1/2MS medium containing $0.5mg{\cdot}L^{-1}$ of NAA. Elongation of the adventitious shoot was achieved in 1/2MS medium supplemented with $0.1mg{\cdot}L^{-1}$ of BA. Rooting was achieved in 1/2MS medium without any hormones. It is hypothesized that the stated in vitro propagation protocol will be useful for conservation and mass propagation of the endangered Polygonatum stenophyllum Maxim. for bioresources.

Hormonal Study to Induce Direct Organ Differentiation of Kalanchoe pinnata by Tissue Culture (조직배양으로 Kalanchoe pinnata의 직접기관분화를 유도하기 위한 호르몬 연구)

  • Kim, Donggiun
    • The Journal of the Convergence on Culture Technology
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    • v.7 no.4
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    • pp.721-726
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    • 2021
  • Plant cells have a totipotencial capacity, the ability of each cell to produce a new complete individual through development. By applying this, several technologies are being developed for widespread application of somatic embryogenesis by processing hormones in vitro as a method of propagation of plants. In order to use this technology, in Kalanchoe pinnata, a plant capable of asexual reproduction with more regular cell division, kinetin belonging to cytokinin and picloram among hormones belonging to auxin were added in combination and treated for 8 weeks, and then the typical performance was evaluated. As a result of our experiment, the rooting effect in leaf slices showed a 70% incidence rate at a picloram concentration of 0.1 mg/L. It has been proven that a concentration difference of 1:5-1:10 in the ratio of kinetin and picloram is effective. It is the experimental result that the effect of auxin is essential for the development of Kalanchoe roots. As for the effect of shooting, the incidence rate was 60% at the picloram concentration of 0.5 mg/L. The kinetin concentration from 0.5 and 1.0 mg/L and has a significant effect on development. It has been proven that the ratio of kinetin to picloram is effective with a concentration difference of 1:1-1:2. These results show that the combination of cytokinin and auxin is crucially important for shooting. It is thought that it can be the basis of a technology for inducing mass proliferation in vitro by inducing direct organogenesis with a combination of hormones.

In Vitro Plantlet Regeneration from Axillary Buds of Tilia amurensis Mature Trees and Clonal Variation in Tissue Culturability (피나무 성숙목(成熟木)의 액아배양(腋芽培養)에 의한 유식물체(幼植物體) 재생(再生)과 조직배양능력(組織培養能力)에 있어서의 클론간(間) 변이(變異))

  • Youn, Yang;Ohba, Kihachiro
    • Journal of Korean Society of Forest Science
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    • v.79 no.2
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    • pp.109-114
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    • 1990
  • The axillary buds of 15-year-old Tilia amurensis were cultured on Saito and Ide (IS), Murashige and Skoog (MS) media and woody plant medium (WPM) to establish an effective micropropagation method. Five levels of 6-benzylaminopurine (BAP) were tested. On IS medium and WPM addition of 1.0/l BAP enhanced shoot development and shoot elongation, whereas addition of 0.5/l BAP was effective on MS medium. A better results were obtained from WPM with 1.0/l BAP and MS with 0.1/l BAP. Developed shoots were subcultured on each basal media but with 0.2/l BAP, Multiple shoots were almost doubled in a month. Root formation could be enhanced at higher concentration of indole-3-butyric acid (IBA). Better rooting rate (83.3%) was achieved on a half-strength MS medium with 3.0 /l IBA. Regenerated plantlets were successfully transferred to soil. To investigate the clonal variation in shoot development and shoot elongation by axillary bud culturing, seven plus tree clones were tested, Clonal variation in tissue culturability among plus trees was recognized by the Duncan's multiple range test at the 5% level. Kang Won No. 12 showed the best response on WPM with 1.0/l BAP.

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Culture and Regeneration of Populus alba × glandulosa Leaf Protoplasts Isolated from in vitro Cultured Explant (현사시나무 기내배양(器內培養) 엽육조직(葉肉組織)에서 분리(分離)된 원형질체(原形質体) 배양(培養) 및 식물체(植物体) 재분화(再分化))

  • Park, Young Goo;Son, Sung Ho
    • Journal of Korean Society of Forest Science
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    • v.77 no.2
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    • pp.208-215
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    • 1988
  • The leaf mesophyll protoplasts of Populus alba ${\times}$ glandulosa were isolated from leaf of plantlet in vitro and cultured for plant regeneration. The MS medium (minus $NH_4NO_3$) with 0.5 mg/l BAP and 2.0 mg/l 2, 4-D showed the moderate frequency of dividing protoplasts cultured by the liquid plating method during the first week of culture. The percentage of colony formation was revealed the highest frequency by the gauze contained semi-solid agar plating method after 5 weeks cultured. Ridding out the gauze, the micro-callus was formed on the same semi-solid medium in 8 weeks after protoplasts culture. For proliferation of callus, mini-callus was transferred on the MS solid medium with 0.5 mg/l 2, 4-D and 0.1 mg/l BAP 12 weeks after culture. Shoot regeneration occurred when the calli derived from protoplasts were cultured on MS medium with 1.0 mg/l zeatin and such shoots could be readily rooted on the one half strengthen MS medium with non-phytohormone. Rooting shoots were planted in green-house 22 weeks after protoplast culture.

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Effect of BA Concentrations and Culture Methods on in Vitro Plant Multiplication from Shoot-Tip Culture of Wasabia japonica (고추냉이 정단배양에 있어서 BA 농도 및 배양방법에 따른 기내증식 효과)

  • Park, Yun-Young;Cho, Moon-Soo;Lee, Young-Deuk;Chung, Jong-Bae;Park, Shin;Jeong, Byeong-Ryong;Park, Sang-Gyu
    • Journal of Plant Biotechnology
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    • v.34 no.1
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    • pp.1-6
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    • 2007
  • Effect of BA concentrations and culture methods on in vitro plant multiplication from shoot-tip cultures of Wasabia japonica was studied. Shoot-tips with leaf primordia and apical meristem were cultured on MS basal medium for all the experiments. Liquid medium for 2 weeks followed by semi-solid medium for 4 weeks containing 1.0 mg/L BA was the best to number of shoots (22.8) and shoot length (3.5 cm). Shoots proliferated could be divided into ca. 5 to 11 of cultures for the multiplication of plantlets. Divided plantlets showed root formation (90%) well onto MS basal medium without growth regulators like IBA and NAA. After rooting, all the plantlets transferred into the pots containing composed soil (bio-media Co., peatmoss $8{\sim}10%$, coir dust $66{\sim}70%$, zeolite $13{\sim}17%$, vermiculite $3{\sim}7%$, perlite $2{\sim}4%$) and grown well into whole plants with multiple shoots.

Establishment of tissue culture and acclimatization method for in vitro mass propagation of Echeveria laui and Echeveria elegans (에케베리아 라우이(Echeveria laui)와 엘레강스(Echeveria elegans)의 대량증식을 위한 조직배양 및 순화 조건 확립)

  • Kim, Youn Hee;Lee, Gee Young;Kim, Hye Hyeong;Lee, Jae Hong;Jung, Jae Hong;Lee, Sang Deok
    • Journal of Plant Biotechnology
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    • v.46 no.1
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    • pp.22-31
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    • 2019
  • The objective of this study was to investigate the suitable parts for callus induction and optimal concentrations of growth regulators contained in the medium affecting shooting and rooting Echeveria laui and Echeveria elegans for in vitro mass production. To determine the suitable plant parts for callus induction, the leaves were divided into upper, medium and bottom parts and cultured on MS medium at different concentrations with $0{\sim}2mgL^{-1}\;NAA$ and $0{\sim}4 mgL^{-1}BA$. The upper and middle parts of leaves both showed 100% callus formation rate with $NAA\;1\;mgL^{-1}$ and $BA\;1\;mgL^{-1}$ treatment in E. laui. The middle parts of leaves showed 83.3% callus formation rate at $NAA\;2\;mgL^{-1}$ and BA 4 mgL-1 treatment in E. elegans. The shoot induction rate from callus was highest at $NAA\;0.1\;mgL^{-1}$ and $BA\;3\;mgL^{-1}$ treatment in E. laui and $NAA\;0.3\;mgL^{-1}$ in E. elegans. In addition, the number of shoots formation was 10.4 shoots high in $NAA\;1\;mgL^{-1}$ and $BA\;1\;mgL^{-1}$ treatment in E. laui and 12.0 shoots in most effective $NAA\;1\;mgL^{-1}$ and $BA\;0.1\;mgL^{-1}$ treatment in E. elegans. In the case of acclimatization of regenerated plant, growth characteristics did not show any significant difference (35 ~ 55%) shading with respect to the different ratio of substrate mixture, and it was determined that would be appropriate considered plant height and appearance preference of E. laui and E. elegans. It was established that the optimization of culture condition was responsible for the mass propagation in vitro cultures of E. laui and E. elegans.

In vitro mass propagation and acclimatization of Haworthia truncata (하월시아 옥선(Haworthia truncata)의 기내 대량 증식 및 순화 조건 구명)

  • Kim, Youn Hee;Lee, Gee Young;Kim, Hye Hyeong;Lee, Jae Hong;Jung, Jae Hong;Lee, Sang Deok
    • Journal of Plant Biotechnology
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    • v.46 no.2
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    • pp.127-135
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    • 2019
  • The purpose of this study was to investigate suitable parts for callus induction and optimal concentrations of growth regulators, contained in the medium affecting shoot and rooting for in vitro mass production of Haworthia truncata. Leaves and flower bud showed 100% callus formation rate at NAA $1{\sim}2mgL^{-1}$ treatment, and NAA $1mgL^{-1}$ + TDZ $2mgL^{-1}$ treatment. The flower stalk showed 75% callus formation rate, at NAA $2mgL^{-1}$ + TDZ $2mgL^{-1}$ treatment in H. truncata. While the rate of callus formation was high in leaves and flower bud, leaves were the most efficient in obtaining most culture parts. Shoot induction rate from callus was highest, at NAA $0.1mgL^{-1}$ treatment in H. truncata. Additionally, the number of shoots formation was 66.3 shoots high, in NAA $1mgL^{-1}$ + BA $0.1mgL^{-1}$ treatment in H. truncata. In the case of acclimatization of regenerated plant, growth characteristics did not show significant difference (95%) shading with respect to the different ratio of substrate mixture, and it was determined that would be appropriate, considering plant height and appearance preference of H. truncata. It was established that optimization of culture condition, was responsible for mass propagation in vitro cultures of H. truncata.

Plant regeneration and soil acclimatization through photoautotrophic culture from leaf explant of a rare species in Sedum tosaense Makino (희귀수종인 주걱비름(Sedum tosaense Makino)의 잎절편으로부터 기내 식물체 재분화 및 광독립배양을 통한 토양순화)

  • Ko, Myoung-Suk;Bae, Kee Hwa;Song, Gwanpil;So, In Sup
    • Journal of Plant Biotechnology
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    • v.40 no.2
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    • pp.79-87
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    • 2013
  • The aim of this study was to establish plant regeneration from leaf explants of Sedum tosaense Makino, which is globally rare and endangered species. The leaf explants of S. tosaense were cultured on the MS medium supplemented with different concentration of BA and NAA for callus induction. Callus induction was showed the highest (100%) on MS medium containing $2.0mg{\cdot}L^{-1}$ BA and $1.0mg{\cdot}L^{-1}$ NAA. The highest number of shoots were regenerated when callus were cultured on MS medium containing $2.0mg{\cdot}L^{-1}$ BA and $1.0mg{\cdot}L^{-1}$ NAA for 5 weeks. The axillary bud were cultured on the MS media supplemented with combination of BA and NAA for in vitro propagation. The highest number of adventitious shoot (7.9 per explants) formed at $1.0mg{\cdot}L^{-1}$ NAA and $2.0mg{\cdot}L^{-1}$ BA. For rooting, MS medium supplemented with or without $2.0g{\cdot}L^{-1}$ activated charcoal was tested. The optimal results were observed using MS medium supplemented with $2.0g{\cdot}L^{-1}$ activated charcoal, on which 85.7 (No. of root), 4.6 cm (length of root). 1,200 ppm $CO_2$ and 350 ppm $CO_2$ were supplied for make certain the effects of $CO_2$ on pre-acclimatization by photoautotrophic culture. 1,200 ppm $CO_2$ treatment was established higher than 350 ppm $CO_2$ treatment. Soil acclimatization of in vitro plantlets was the best in mixture soil consisted of peat moss and perlite with 100% survival rate and they showed the maximum growth.

Regeneration of adventitious root from Calystegia soldanella L. in Jeju island and mass proliferation method using bioreactor system (제주지역 갯메꽃(Calystegia soldanella L.) 유래 부정근 재분화 및 생물반응기 시스템 이용 대량증식법)

  • Jong-Du Lee;Eunbi Jang;Weon-Jong Yoon;Yong-Hwan Jung
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2021.04a
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    • pp.37-37
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    • 2021
  • Calystegia soldanella L. is a perennial herbaceous halophyte belonging to the convolvulaceae family, which mainly grows in coastal sand dunes in Korea. Shoots and rhizomes are edible, and roots called 'Hyoseon Chogeun' are known to have medicinal effects such as antipyretic, sterilization, and diuretic. In addition, physiological activities of antioxidant, anti-inflammatory, antiviral, antifungal and PTP-1B (protein tyrosine phosphate-1B) inhibition have been reported. In this study, in vitro induction cell lines of C. soldanella L. collected from the coastal sand dunes in Jeju island was redifferentiated into adventitious roots that can be used as medicinal resources. Also the biomass of mass-proliferated adventitious roots using a bioreactor were evaluated. Plants of C. soldanella L. were collected from the crevice of the seashore in the coastal area of Taeheung 2-ri, Namwon-eup, Seogwipo-si. Then, it was separated into leaves, stems, rhizomes, and roots, and surface sterilized with 70% ethyl alcohol and 2% NaOCl (sodium hypochlorite). After washing with sterilized water, each organ section was cultured in Hormone-free MS medium (Murashige & Skoog Medium). As a result, the induction response rates were evaluated at 85% and 55%, respectively, in terms of callus formation and shoot generation in the rhizome segment. In the case of the adventitious roots morphological characteristics induced by single-use treatment of auxin-based plant growth regulators IBA and NAA from redifferentiated shoots were compared. Most efficient adventitious root culture method as a rooting rate, number, length, and biomass proliferation in the bioreactor system was confirmed when treated by culturing in MS salts, Sucrose 30 g·L-1, and IBA 1mg·L-1 for 4 weeks. In this study, the medium composition and culture period were confirmed using a bioreactor system to mass-proliferate adventitious roots derived from C. soldanella L. in Jeju island. Also this adventitious root line developed a new medicinal material could increase value of the bio-industry ingredient through quantitative and qualitative screening of phyto-bioactive compounds.

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