• Title/Summary/Keyword: effective micropropagation

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Effect of Medium Components and Culture Methods on Prothallus Propagation of Pteridium aquilinum var. latiusculum (Desv.) Underw. ex Hell. (고사리 전엽체의 증식에 미치는 배지구성물질과 배양방법의 영향)

  • Shin, So-Lim;Lee, Moo-Yeul;Choi, Jae-Sun;Lee, Cheol-Hee
    • Korean Journal of Plant Resources
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    • v.22 no.4
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    • pp.337-342
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    • 2009
  • Present studies were conducted to evaluate the effects of medium strength(MS and Hyponex), carbon sources and their concentrations, agar concentrations, and inoculation amounts on prothallus propagation of Pterdium aquilinum var. latiusculum(Desv.) Underw. ex Hell in vitro. The optimum MS medium strength for prothallus propagation was 2MS concentration. Phosphate source was most effective for prothallus growth of P. aquilinum var. latisculum. The addition of 1% sucrose or glucose to MS medium promoted prothallus multiplication. Growth of prothallus was not affected by agar concentration. Propagation of homogenized prothallus was vigorous even in liquid medium. Chopped gametophytes(100 and 200 mg) were inoculated on 250 ml ${\Delta}$flask with 100 mL of 2MS concentration medium and suspension culture was done at 100 rpm for 22 days. After 20 days, prothallus multiplication slowed down, so 100 mg of chopped prothalli is recommended for initial inoculation, since initial amount of inoculum did not affect subsequent prothallus multiplication. Consequently after 20 days of suspension culture, prothallus should be subcultured or transplanted outside of growing vessels.

Plant Regeneration Via Direct Adventitious Roots from Free Root Segments of Ulmus davidiana Planch (당느릅나무(Ulmus davidiana Planch) 부정근 절편으로 부터 부정아 유도를 통한 식물체 생산)

  • Kim, Ji-Ah;You, Xiang-Ling;Ahn, Chang-Ho;Lee, Jae-Seon;Choi, Yong-Eui
    • Journal of Korean Society of Forest Science
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    • v.96 no.1
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    • pp.83-88
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    • 2007
  • Micropropagation of Ulmus davidiana Planch was established via adventitious shoot formation from the segments of adventitious roots. Adventitious roots were produced directly from root segments of seedlings on a 1/2 SH medium plus various concentrations of IBA. The maximum growth of adventitious roots was observed in the presence of 2.0 mg/L IBA. After the segments of adventitious roots were cultured on various cytokinins (zeatin, 2-iP, BA, kinetin) and cytokinins plus auxin (IBA), formation of adventitious shoot was investigated. Among cytokinin treated, kinetin was the most effective on both adventitious shoot induction and number of shoots. Especially, 2.0 mg/L kinetin was the best to increase adventitious shoot induction (95.8%) and a number of shoots (8.4). Adventitious shoots were rooted on 1/2 WPM medium and the plantlets were acclimated 100% on composed soil (peatmoss : vermiculite 1 : 1).

Effect of Mother Trees and Dark Culture Condition Affecting on Somatic Embryogenesis of Liriodendron tulipifera L. (백합나무(Liriodendron tulipifera L.) 체세포배(體細胞胚) 발생(發生)에 미치는 모수(母樹) 및 암배양(暗培養) 효과(效果))

  • Son, Seog-Gu;Moon, Heung-Kyu;Kim, Yong-Wook;Kim, Ji-Ah
    • Journal of Korean Society of Forest Science
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    • v.94 no.1 s.158
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    • pp.39-44
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    • 2005
  • An effective micropropagation system for Liriodendron tulipifera L via somatic embryogenesis was established using immature seeds. Immature seeds from five individual trees were bisected longitudinally and cultured on two basal media (MS and B5) containing different combinations of 2,4-D and TDZ to induce callus and embryogenic tissue under light ($40{\mu}mol\;m^{-2}s^{-1}$, 16 hr/day) or complete darkness at $25{\pm}2^{\circ}C$. There was no distinctive difference on callus and embryogenic tissue induction between the two basal media with PGRs. Optimum culture medium appeared to be MS medium supplemented with 1.0mg/L 2,4-D and 0.01mg/L TDZ plus 3% sucrose. Nonembryogenic callus induction rate was not significantly different among the genotypes. However, However, the embryogenic callus induction frequency differed greatly by the genotypes ranging from 55% to 72% when cultured in the dark. Generally, the cultures maintained in the dark tended to show normal somatic embryo development as well as embryogenic tissue formation and this was confirmed by histological examination. Above results suggest that a proper selection of mother tree and dark culture condition are necessary to optimize somatic embryogenesis system of Liriodendron tulipifera.

Mass Propagation by In Vitro Culture of Bupleurum latissimum Nakai (섬시호(Bupleurum latissimum Nakai)의 조직배양을 통한 대량생산)

  • Cho, Han-Jik;Kim, Ee-Youp;Kim, Mu-Yeol;Park, Hark-Bong;Kim, Hyo-Jin
    • Korean Journal of Plant Resources
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    • v.20 no.4
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    • pp.367-374
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    • 2007
  • This study was carried out to establish the micropropagation system of Bupleurum latissimum Nakai that is a Korean native endangered species. Callus were induced from the leaf, petiole and floral bud and the percentage of callus formation was highest in the floral bud on the MS medium containing 2.0 mg ${\cdot}$ $L^{-1}$ 2,4-D. Especially, callus induced from floral bud was formed 77.8% and the percentage of shoot formation was 42.6% on the MS medium containing 2.0 mg ${\cdot}$ $L^{-1}$ 2,4-D plus 1.0 mg ${\cdot}$ $L^{-1}$ TDZ. For simultaneously callus formation and shoot regeneration, 1/2 MS medium was more effective than MS medium. The percentage callus formation, shoot regeneration and rooting were 46.3%, 13.0%, 13.0% in 1/2 MS medium, respectively. Soot regeneration from callus was good in 1/2 MS medium supplemented with 2.0 mg ${\cdot}$ $L^{-1}$ 2,4-D plus 1.0 mg ${\cdot}$ $L^{-1}$ BA where percentage of shoot regeneration was 74.1 %, and the number of shoot per explant was 2.4. The percentage of rooting was lowest (57.8%) in control while it was highest (97.8%) in 1.5 mg ${\cdot}$ $L^{-1}$ NAA. In acclimatization of regenerated plantlets, the percentage of survived plantlets was highest (86.1%), and plant height, root length and fresh weight were good in the soil for horticulture.

Factors Effecting Agrobacterium Mediated Transformation and Regeneration of Populus nigra × P. maximowiczii (Agrobacterium tumefaciens에 의한 양황철나무의 형질전환(形質轉換) 요인(要因))

  • Park, Young Goo;Shin, Dong Won;Kim, Joung Hee
    • Journal of Korean Society of Forest Science
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    • v.79 no.3
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    • pp.278-284
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    • 1990
  • We have demonstrated expression of bacterial genes transferred into cells of Populus nigra ${\times}$ P. maximowiczii by A. tumefaciens strain 6044 (pGA 472). We determined the optimum concentration of kanamycin sulfate for effective selection of punctured leaf transformed using Agrobacterium binary vector pGA 472 containing a neomycine phosphotransferase gene (NPT-II) which confers kanamycin resistance. The combination of cefotaxime (200mg/l) and carbenicillin (300mg/l) showed good performance of discarding Agrobacterium from inoculated punctured leaf. A relatively low concentration (10mg/l) of kanamycin sulfate inhibited callus and shoots induction from punctured leaf. Number of shoots regenerated from co-cultured punctured leaf was 3.0 on MS basal medium supplemented with 10 mg/l kanamycin sulfate, while that of not co-cultured punctured leaf was none. The regeneration rate was 10% from the punctured leaf co-cultured on MS medium with 10 mg/l kanamycin. Regenerated shoots are developing from micropropagation for Southern blot analysis and inheritance of the kanamycin resistance trait (NPT-II).

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Effects of cytokinins, GA, and IBA on in vitro propagation of Vitex negundo var. insica (좀목형 (Vitex negundo var. insica) 신초의 기내증식에 미치는 cytokinin, GA 및 IBA의 영향)

  • Han, Mu-Seok;Moon, Heung-Kyu;Park, So-Young;Kim, Yong-Wook;Son, Suk-Gu
    • Journal of agriculture & life science
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    • v.45 no.3
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    • pp.53-58
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    • 2011
  • To develop an efficient micropropagation technique for Vitex negundo var. insica, which is known as aromatic and medicinal tree, the effects of various plant growth regulators (PGRs) on in vitro shoot proliferation and rooting were evaluated using the newly-developed shoots of a 3-year-old tree. Multiple shoot induction was achieved effectively on WPM (woody plant medium) supplemented with 0.5-2.0 mg/L BA, and the highest shoot number (7.9/explant) was obtained at the concentration of 1.0 mg/L BA. Typically 1 or 2 superior shoots (about 3.4 cm) were induced on hormone-free WPM. Combined treatment of BA 2.0 + GA 0.5 mg/L appeared to effective on shoot proliferation and rooting. Plant growth regulators added in shoot proliferation medium had strong impact on subsequent rooting as well. Overall, shoots induced by BA treatment resulted in high rooting rates while the effect was reduced gradually by ascending BA levels. TDZ of low concentration also revealed a similar tendency as BA, but the rooting ability was strongly inhibited at the concentration of 0.5 mg/L, and rooting was never observed at the concentrations higher than 0.5 mg/L. Combined treatment of BA and IBA had positive influence in both shoot proliferation and rooting. These results suggest that Vitex negundo var. insica could be effectively micropropagated via axillary bud cultures.

The Photoautotrophic Culture System Promotes Photosynthesis and Growth of Somatic Embryo-derived Plantlets of Kalopanax septemlobus (독립영양방식 액체대량배양 시스템하에서 배양한 체세포배 유래 음나무 기내묘의 생장과 광합성)

  • Park, So-Young;Moon, Heung-Kyu;Kim, Yong-Wook
    • Journal of Korean Society of Forest Science
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    • v.100 no.2
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    • pp.212-217
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    • 2011
  • A photoautotrophic micropropagation methodology in liquid culture medium has a number of advantages for large-scale propagation of plants. This paper describes an improved system for the mass propagation via somatic embryogenesis of the medicinal plant Kalopanax septemlobus Nakai. Somatic embryo-derived young plantlets of K. septemlobus were cultured either under heterotrophic conditions with sucrose on half-strength MS medium with $30gL^{-1}$ sucrose, under heterotrophic conditions without sucrose, or under photoautotrophic conditions (MS liquid medium without sucrose under forced aeration) for four weeks before transferring the plantlets for acclimatization. Plantlets grown under photoautotrophic conditions had more leaves, higher chlorophyll content, a higher net photosynthetic rate (NPR), and a higher survival rate. The results indicate that the photoautotrophic conditions with a forced ventilation system are effective in enhancing the growth of plantlets and the rate of net photosynthesis. The plantlets grown under photoautotrophic conditions had a high survival rate (92%) upon ex vitro transplantation. Our study shows that autotrophically produced plantlets acclimatize better and sooner upon ex vitro transplantation than conventionally cultured plants.

Effect of Medium Composition on in vitro Plant Root Regeneration from Axillary Buds of Cassava (Manihot esculenta Crantz) (카사바 액아배양 시 배지조성이 기내 식물체 발근에 미치는 영향)

  • Young Hee Kwon;Won IL Choi;Hee Kyu Kim;Kyung Ok Kim;Ju Hyoung Kim;Yong Sup Song
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2021.04a
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    • pp.24-24
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    • 2021
  • The Cassava (Manihot esculenta Crantz) is one of the major food crops in the tropical or subtropical regions. Recently, clean planting materials of improved cassava cultivars are in high demand. Problems in the propagation of cassava are virus vulnerable and low rates of seed germination. Thus, the study was undertaken to develop an efficient in vitro mass propagation protocol of Manihot esculenta Crantz. So we tried to optimize protocols for mass production from axillary buds of Cassava. Young and actively growing stem segments were excised from adult plants of cassava. Samples were cut into a 3~4 cm nodal segments with axillary buds, and cultivated in the different medium supplemented with various plant growth regulators for 4 weeks. For shoot multiplication, axillary buds approximately 1 cm in length were taken from in vitro derived shoots and subcultured. After 4~6 weeks, the shoot generation rate showed 55.6%. The shoot number and its length was 1.0/explant and 2.3 cm in the most favorable medium composition. The auxin β-indolebutyric acid(IBA) 0~2.0 mg/L was proved to be effective on root development. Plantlets with fibrous roots easily generated tuberous roots in vitro. The tuberous roots were induced only when both kinetin and IBA were used in combination. after 8 weeks, the root generation rate showed 100%. The root number and its length was 17.2/explant and 2.2 cm in the most promising medium composition. Our experiments confirmed that in vitro growth and multiplication of plantlets could depend on its reaction to the different medium composition, and this micropropagation techniques could be a useful system for healthy and vigorous plant production.

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The Effect of Nutrient Solution Concentration on Growth of Potato Plantlet in Microponic System (Microponic system에서 배양액의 농도변화가 감자 소식물체 생육에 미치는 영향)

  • Ko, Sun A;Choi, Ki Young;Lee, Yong-Beom
    • Journal of Bio-Environment Control
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    • v.23 no.2
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    • pp.144-147
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    • 2014
  • It was intended to closely examine an effect that a change in the concentration of culture medium had on the potato(Solanum tuberosum L.) plantlet growth in the microponic system so as to mass-produce the virus-free plant of new variety 'Saebong' for potato processing. The adjusted concentration of potato culture medium was 0.2, 0.6, 1.0, 1.4, 1.8, and $14.0dS{\cdot}m^{-1}$. And potato seedling was cut into pieces of 1.5 cm in length, which included 2 growth points and leaves. And each was explanted in glass vial of 50 mL. And experiments were carried out twice for 18 days or 21days. Culture medium of 2ml was put in the container respectively. And 1 mL was added after 10 days. And in terms of cultivation environment, the experiment was carried out at the day length of 16 hours at the temperature of $23{\pm}1^{\circ}C$ under the white LED light of $40{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$. The concentration of culture medium in the experiment I was EC 0.2, 1.0, $14dS{\cdot}m^{-1}$ and was adjusted to 0.6, 1.0, 1.4, $1.8dS{\cdot}m^{-1}$ in the experiment II. The results showed that the survival rate of plantlet was 90% at $0.2dS^2m^{-1}$, 100% at $0.6dS^2m^{-1}$, 100% at $1.0dS^2m^{-1}$. 0% at $1.4dS{\cdot}m^{-1}$, 0% at $1.8dS{\cdot}m^{-1}$. and 0% at $14.0dS{\cdot}m^{-1}$ after 7 days. With regard to the explanted potato seedling, in case of the treatment where the electrical conductivity of culture medium was adjusted to $1.0dS{\cdot}m^{-1}$, root developed 2 days after transplantation. And the plantlet vigorously grew into strong plant that had 7 leaves, length of 5cm, and fresh weight of 0.5 g after 18 days. In case of the treatment where the concentration of culture medium was adjusted to $0.6dS{\cdot}m^{-1}$, the root plantlets developed 4 days after transplantation. And those grew into plant that had 7 leaves and fresh weight of 0.2 g after 21 days. Therefore, we found that it is effective to control potato culture medium by adjusting its electrical conductivity to $0.6{\sim}1.0dS{\cdot}m^{-1}$ for the mass production of virus-free potato seedling in the microponic system.

Effects of Different Natural Extracts and Plant Growth Regulators on Plant Regeneration and Callus Induction from Pseudobulbs Explants through in vitro Seed Germination of Endangered Orchid Bulbophyllum auricomum Lindl. (멸종 위기에 처한 Bulbophyllum auricomum Lindl. orchid의 시험관 내 종자 발아를 통한 구근 절편체의 식물 재생 및 캘러스 유도에 대한 천연 추출물 및 식물 성장 조절제(PGR)의 효과)

  • Aung, Win Theingi;Bang, Keuk Soo;Yoon, Seo A;Ko, Baul;Bae, Jong Hyang
    • Journal of Bio-Environment Control
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    • v.31 no.2
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    • pp.133-141
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    • 2022
  • Bulbophyllum auricomum Lindl. is a rare orchid and has flowers with an attractive fragrance. The present study investigated the tissue culture method for micropropagation. Capsules derived from artificial self-pollination were obtained for the best seed germination in MS basal medium. Plant growth regulators (1.0 mg·L-1 of BAP and 2.0 mg·L-1 of NAA) were affected by callus induction from subcultured pseudobulb explants. For the callus subculture, different natural plant extracts were tested in 11 treatment media. Among them, MS medium with 150 mL·L-1 of coconut water was generally effective in fresh weight (1.75 ± 0.08) and (3.01 ± 0.20) of callus proliferation and PLBs induction at 1 and 2 months, respectively, followed by an MS combination of 30 g·L-1 of banana and 20 g·L-1 of potato extract. The results of a comparative study of different MS mediums containing plant growth regulators with a natural extract combination and MS medium supplemented with natural extract only showed that MS medium supplemented with a combination of natural extracts (150 mL·L-1 of coconut water) and plant growth regulators (2.0 mg·L-1 of BAP and 1.0 mg·L-1 of NAA) obtained the highest shoot regeneration (3.37 ± 0.17) and (6.41 ± 0.68) after 1 month and 2 months of culturing, respectively.