• Title/Summary/Keyword: shoot tip

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Cryopreservation of in Vitro Grown Shoot Tips of Sweet Potato (Ipomoea batatas L.) by the Encapsulation-Vitrification Method

  • Yi, JungYoon;Lee, GiAn;Lee, YoungYi;Gwag, JaeGyun;Son, EunHo;Park, HongJae
    • Korean Journal of Plant Resources
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    • v.29 no.6
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    • pp.635-641
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    • 2016
  • Sweet potato (Ipomoea batatas L.) shoot tips grown in vitro were successfully cryopreserved by encapsulation-vitrification. Encapsulated explants are very easily manipulated, due to the relatively large size of the alginate beads, and a large number of samples can be treated simultaneously. In this study, the effects of sucrose preculture, cryoprotectant preculture, and post-warm recovery media on regrowth, following liquid nitrogen (LN) exposure, were investigated to establish an efficient encapsulation-vitrification protocol for sweet potato. Shoot tips of plants grown in vitro were precultured in 0.3 M sucrose for 2 d before encapsulation. Encapsulated shoot tips were pre-incubated in liquid MS (Murashige and Skoog) medium containing 0.5 M sucrose for 16 h, before preculturing in sucrose-enriched medium (0.7 M sucrose) for 8 h. Shoot tips were osmoprotected with 35% plant vitrification solution 3 (PVS3) for 3 h, before being dehydrated with PVS3 for 2 h at $25^{\circ}C$. The encapsulated and dehydrated shoot tips were transferred to 2 mL cryotubes, suspended in 0.5 mL PVS3, and plunged directly into liquid N. High levels of shoot formation were obtained for the cv. Yeulmi (65.7%) and Yeonwhangmi (80.3%). The regrowth rates of cryopreserved samples in Yeulmi (78.9%) and Yeonwhangmi (91.3%), following culture on ammonium-free MS medium for 5 d, were much higher than those cultured on standard MS medium (65.7% and 80.3%, respectively). This encapsulation-vitrification is a promising method for the long-term preservation of sweet potato.

Plant Regeneration via Organogenesis from Leaf and Stipule Segments of Strawberry (Fragaria ananassa Duch.) (딸기의 잎과 탁엽 절편체로부터 기관형성을 통한 식물체 재생)

  • 최준영;김현정;형남인
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.5
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    • pp.347-351
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    • 1998
  • Plant regeneration via organogenesis from leaf and stipule explants of micropropagated shoots of strawberry (Fragaria $\times$ ananassa cv. Suhong) was achieved. Leaf and stipule explants were detached from shoot-tip cultured shoots and cultured on MS medium with various combinations of BA and NAA under light or dark condition. Shoot regeneration from leaf explant was observed after 3 weeks in culture and was good at the high ratio of BA and NAA among various combination treatments. The highest shoot regeneration frequency from leaf explants was obtained with 1.0 mg/L BA and 0.1 mg/L NAA, in which 31.1% shoot regeneration frequency(1.7 shoots per leaf explant) was yielded. In case of stipule explants, shoot regeneration was largely affected by plant growth regulators during incubation under dark condition for initial 4 weeks but not under continuous light condition. The combination treatment with 2.0 mg/L BA and 0.1 mg/L NAA showed the most excellent shoot regeneration from stipule explants, where 44.4% regeneration frequency(4.0 shoots per explants) obtained. Regenerated shoots were rooted on MS medium with 0.1 mg/L NAA after shoot elongation, and the plantlets regenerated were transferred to soil mixtures with vermiculite and perlite for acclimation.

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Growth Responses of Petunia as Affected by Several Pollutants in Vitro (Petunia 조직배양시 몇가지 공해물질이 생장반응에 미치는 영향)

  • 백기엽;최성렬;이재욱;최주견
    • Journal of Plant Biology
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    • v.27 no.3
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    • pp.139-147
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    • 1984
  • This in vitro study was employed to clarify the effects of several pollutants i.e. $SO_2$, fluoride, cadmium(Cd), aluminum(Al) and NaCl, on the organogenesis and growth responses of shoot-tip, stem and multiple-buds segments derived from hypocotyl or cotyledon culture of petunia seedlings. ${Na_2}{SO_3}$levels of more than 200$\mu{g}$/ml had significantly reduced organogenesis, growth, and chlorophyll content. The injuries caused by ${Na_2}{SO_3}$, concentration of more than 400$\mu{g}$/ml were alleviated by increasing hydrogenion concentration of medium, indicating some relationship between two factors. Organogenesis was not affected by the fluoride concentration up to 100ppm in the media, but the growth and chlorophyll content were greatly reduced by the fluoride. The effect of Cd depended on the explant sources used for the culture; 1.0ppm was effective for fresh weight increase in shoot tip culture, and 3.0ppm in stem segments culture. Organogenesis and growth were greatly reduced by more than 10.0 Cd treatment. Growth and formation of shoots were better with Na conc. of 0.3% compared to control, but those of roots were inhibited. Na concentration goes over 1.0%, organogenesis and subsequent growth were inhibited, and chlorophyll synthesis was drastically reduced. Chlorophyll content was increased on the medium supplemented with Al 50$\mu{g}$/ml compared to control. However the formation and growth of shoots were greatly inhibited with more than 400$\mu{g}$/ml and roots were not produced at all.

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In vitro Propagation using Shoot Tip Culture in Gold Tree[Dendropanax morbifera $L_{EV}]$. (황칠나무의 경정배양에 의한 기내번식)

  • 최성규;윤경원
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.6
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    • pp.464-467
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    • 2001
  • In order to establish a in vitro propagation system for gold tree[Dendropanax morbifera $L_{EV}$], the effects of auxins and cytokinins on shoot multiplication and rooting were investigated. Germination rate was the best in MS medium. The fresh weight and number of shoot were the best on the medium containing 0.1 or 1.0 mg/l BAP and 0.5 or 1.0 mg/l NAA. Shoots were successfully rooted in MS medium with 1.0 mg/l NAA. Roots were easily formed by the addition of auxins, especially 0.1 or 1.0 mg/l BAP.P.

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Effect of Thidiazuron on Callus and Multiple Shoot Formation in Shoot-tip Culture of Hibiscus syriacus L. 'Honghwarang' (Thidiazuron이 무궁화 '홍화랑' 품종의 정단배양으로부터 Callus형성과 Multiple Shoot형성에 미치는 효과)

  • Kim, Eun Kyoung;Yoo, Yong Kweon;Kim, Ki Sun
    • Horticultural Science & Technology
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    • v.16 no.4
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    • pp.520-524
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    • 1998
  • This study was carried out to investigate the effect of thidiazuron(TDZ) on callus and shoot primordia formation, to determine the most optimum multiple shoot induction medium, and to obtain the plantlets on solid medium via shoot organogenesis. TDZ 0.01 mg/L in MS medium was most effective on callus formation, and BA 0.1 mg/L was most effective on shoot growth, while TDZ 0.01 mg/L was most effective on callus formation. TDZ 0.001 mg/L was most effective in shoot primordia formation. Shoot tips were cultured with TDZ 0.01 mg/L for 8 weeks and induced callus was transferred to regeneration medium containing TDZ 0.001 mg/L. After 4 weeks induced shoot primordia were resubcultured at growth regulator-free medium for 4 weeks. The induced multiple shoots rooted more efficiently at NAA 1.0, 5.0 mg/L, or IBA 5.0 mg/L.

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Shoot Primordium Culture for Multiplication of Carrot (당근의 다량증식을 위한 순원기 배양)

  • 서호범;이수성
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.2
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    • pp.93-97
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    • 1999
  • Shoot tips with 2 leaf primordia were cultured to induce shoot primordia in MS liquid medium supplemented with several concentrations of BA and hIAA under the conditions of 10,000 lux illuminations for 24 h and of vertical shaking of 2 rpm in carrot. Two F$_1$ hybrids and two male sterility lines were used. Shoot primordia were only induced in the medium supplemented with 2.0 mg/L of BA and 0.2 mg/L of NAA. Genotypic specificity and seasonal effect of donor parents on shoot primordia induction were not observed and average 15-20% of the planted dornes developed to shoot primordia. The induced shoot primordia were successfully propagated by subculture in the same medium. However, they were grown into three different types during multiplication, that is, the type with multiple small shoots on the surface, the type of without any shoot, and the type of callus. Shoot primordia clusters with small shoots on the surface differentiated multiple shoots successfully in 1/2 MS solid medium supplemented with 0.2 to 1.0 mg/L of IAA and 0.2 to 1.0 mg/L of kinetin. New shoot primordia with small shoots were well formed when pieces bigger than 2 mm in diameter of the out layer of the shoot primordia cluster with small shoots were subcultured. No differences of multiplication and shooting ability and chromosomal variation of shoot primordia were observed until the 13th sub-culture.

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The simple assay of phosphinothricin acetyltransferase gene on the transgenic potato (형질전환 감자에서 제초제 저항성 유전자인 PAT gene의 간편한 확인)

  • 정재훈;양덕춘;방극수;최경화;한성수
    • Korean Journal of Plant Resources
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    • v.12 no.4
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    • pp.253-259
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    • 1999
  • In this study, three simple methods were established to confirm the transgenic potato plants. The leaf disc was used in the first method. After leaf discs of transgenic and non-transgenic potato were transfered into the liquid MS medium with bialaphos 5mg/l, 25 days, the chlorosis occurred in the non-transgenic leaf discs while it could not find in the transgenic leaf discs, In the second method, shoot tips of potato were transferred into MS medium supplemented with 0.5mg/l bialaphos and 0.6% agar. After 7-10 days, a lot of roots developed from the transgenic shoot tip, but the non-transgenic shoot tip was dead. The third method was using chlorophyll contents. Leaf discs were transferred into the liquid MS medium with bialaphos 0.5 mg/l. After 15 days, the content of chlorophyll A in transgenic plant was at least 2.5 times higher than in non-transgenic plant. In addition, the PAT enzyme activity were detected in the transgenic potato, but not detected in normal potato.

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