• Title/Summary/Keyword: shoot culture

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Genetic Transformation and Plant Regeneration of Codonopsis lanceolata Using Agrobacterium (Agrobacterium에 의한 더덕의 형질전환과 식물체 재분화)

  • 최필선;김윤성;유장렬;소웅영
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.5
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    • pp.315-318
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    • 1994
  • To obtain transformed plants, we cocultured cotyledonary explants of Codonopsis lanceolata with Agrobacterium tumefaciens LBA4404, a disamed strain harboring a binary vector pBI121 carrying the CaMV35S promoter-$\beta$-glucuronidase (GUS) gene fusion used as a reporter gene and NOS promoter-neomycin phosphotransferase gene as a positive selection marker in MS liquid medium with 1mg/L BA. After 48 h of culture, explants were transferred onto MS solid medium with Img/L BA, 250mg/L carbenicillin, and 100mg/L kanamycin sulfate and cultured in the dark. Numerous adventitious buds formed on the cut edges of the explants after 2 weeks of culture. When subjected to GUS histochemical assay buds showed a positive response at a frequency of 15%. Explants formed adventitious shoot at a frequency of 56.7%, after 6 weeks of culture. Upon transfer onto the basal medium, most of the shoots were rooted and subsequently the regenerants were transplanted to potting soil. Southern blot analysis confirmed that the GUS gene was incorporated into the genomic DNA of the GUS-positive regenerants.

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Micropropagation of Heloniopsis orientalis (Thunb.) C. Tanska in vitro (조직배양을 이용한 처녀치마[Heloniopsis orientalis (Thunb) C. Tanaka] 대량 증식)

  • 윤세영;이명선;임상철;신중두
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.3
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    • pp.197-202
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    • 2000
  • The effect of cultural media and growth regulators on multiple plant regeneration from leaf explants of Heloniopsis orientalis (Thunb.) Tanaka was evaluated. The highest percentage of shoot and root formation were 20 and 30% on MS medium treated at 3.0 mg $l^{-1}$ of zeatin, respectively. Also 67 and 33% of high shoot formation appeared on 1/2 MS and 5 culture medium treated at zeatin 1.0 and 3.0 mg $l^{-1}$ respectively. With MS treated at 0.5 mg $l^{-1}$ of 2,4-D 1/2 MS and B5 culture media treated at each 1.0 and 3.0 mg $l^{-1}$ of zeatin the highest ratios of plant produced were 100, 280 and 310 % respectively relative to the other treatments. Generally, there was highest possibility for multiple propagation of Heloniopsis orientalis (Thunb.) C. Tanaka with B5 culture media supplemented 3.0 mg $l^{-1}$ of zeatin.

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Production of Virus-Free Stocks from Citrus Plant by the Shoot-Tip Grafting and Heat Treatment (열처리와 Shoot-Tip Grafting에 의한 감귤 바이러스 무독묘 생산)

  • Kim Daehyun;Shim Hyekyung;Kwon Hyeogmo;Hyun Jaewook;Kim Kwangsik;Lee Jinkyung;Lee Sukchan
    • Journal of Plant Biotechnology
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    • v.32 no.1
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    • pp.45-50
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    • 2005
  • Virus-free stocks was produced by the combination of the heat treatment of virus infected plant and shoot-tip grafting (STS). To produce virus-free stocks, the plants infected with citrus viruses were used for virus-free stock production using the modified method of STG in thermotherapy at $40^{\circ}C$ for 16 hours in the light, and at $30^{\circ}C$ for 8 hours of darkness for 4 weeks. Trifoliate orange (P. trifoliata) were used as rootstock seedling for STG. Percentages of virus-free stocks against citrus tristeza virus (CTV), satsuma dwarf virus (SDV) and citrus tatter leaf virus (CTLV) were $75.7\%,\;100.0,\%\;82.6\%$ respectively. Shoot tip size for successful STG were as small as possible. Less than $0.3\;\cal{mm}$ of shoot tips gave the hight efficiency of virus free plants but survival rates were low. And, survival rate after shoot-tip culture was analyzed and the rates were dependant on the cultivars; Yuzu cultivar showed the hight survival rate ($74.6\%$) and early satsuma mandarin (Iwasagi) was $13.3\%$ as the lowest cultivar. But citrus trees were not succeed to grown, turned brown, and died.

In vitro Conservation of Coleus forskohlii- an Endangered Medicinal Plant

  • Rajasekharan P.E.;Ambika S.R.;Ganeshan S.
    • Journal of Plant Biotechnology
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    • v.7 no.2
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    • pp.135-141
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    • 2005
  • Protocols for in vitro conservation was developed for Coleus forskohlii. Plants maintained both in field served as explant source. Shoot tips and single node cuttings were used to optimize protocols for in vitro multiplication. MS basal medium supplemented with $0.54\;{\mu}M$ naphthalene acetic acid (NAA) and $8.87\;{\mu}M$ benzy-ladenine (BA) induced multiple shoots in shoot tips and nodes. Shoot multiplication was amplified with a gradual decrease of BA concentration, leading to its final omission after 4 months. Concomitant rooting on multiplication media enabled successful establishment extra vitrum. For in vitro conservation studies, experiments were carried out with 2-3 week maintained in vitro plants under standard and reduced culture conditions (SCC, RCC). In vitro plants could be successfully conserved in full strength MS medium (FMS) under SCC for 6 months without subculture with full potential to regenerate, producing viable shoots and nodes. The root production remained unaffected due to conservation, showing high rooting activity in mannitol and low temperature treatments. Preset low temperature (15 and $10^{\circ}C$) and reduction in media constituents does not appear to favour conservation, although the former accomplished conservation levels equal to (FMS) under SCC.

Effects of Plant Growth Regulators on in vitro Propagation of Echinosophora koreensis Nakai

  • Yi, Jae-Seon;Lee, Hyunseok;An, Chanhoon
    • Journal of Forest and Environmental Science
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    • v.29 no.4
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    • pp.275-281
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    • 2013
  • To establish in vitro nodal culture conditions of Echinosophora koreensis Nakai, one of rare and endangered species famous for beautiful flowers in the Korean Peninsula, the influence of plant growth regulators (PGRs) on shooting and rooting from in vitro shoots was investigated. In shoot multiplication, addition of 6-benzylaminopurine (BA) to the half-strength Driver and Kuniyuki's media in the range of 2.22 to 8.88 ${\mu}M $induced 2.5 to 2.7 shoots per axillary bud; and addition of 2.27 ${\mu}M $ thidiazuron (TDZ) produced 3.2 shoots, during 4 weeks of culture, while zeatin and isopentenyl adenine (2ip) were not effective on shoot multiplication as observed from several combination treatments of BA with other PGRs. Shoots established were smaller than 2 cm in length, in most of the treatments. while in BA 8.88 ${\mu}M $ treatment more than 30% of shoots were longer than 2 cm and shorter than 4 cm. In rooting, naphthalene acetic acid (NAA) from 5.37 to 21.48 ${\mu}M $ showed the rooting rate from 40.0 to 62.5%. Indole butyric acid (IBA) addition had little effect on rooting (<10%), although some roots in IBA-containing media were longer than those in NAA. Micropropagation from axillary buds of nodular explants was applicable and promising to multiplication and conservation of Echinosophora koreensis Nakai.

Influence of Mineral Salts on Shoot Growth and Metabolite Biosynthesis in Tea Tree (Camellia sinensis L.)

  • Kim, Yong Duck;Yun, Jae Gill;Seo, Yeong Rong;Karigar, Chandrakant S.;Choi, Myung Suk
    • Horticultural Science & Technology
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    • v.33 no.1
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    • pp.106-113
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    • 2015
  • Effects of mineral salts (N, P, K, $Ca^{2+}$, $Mg^{2+}$, and $Fe^{3+}$) on the shoot growth and metabolite production of tea tree were studied using in vitro culture techniques. Among mineral s alts, ${H_2PO_4}^-$ was the most important for enhanced growth rate of tea tree, while $Mg^{2+}$ and $Ca^{2+}$ did not affect plant growth. Removal of ${NH_4}^+$ and $NO_3$ from the culture medium enhanced shoot multiplication compared to other treatments. Metabolite production was variable depending on mineral types and concentration. Removal of $Ca^{2+}$ decreased the production of caffeine; however, other treatments did not influence its production. $Ca^{2+}$, ${NH_4}^+$ and $Fe^{3+}$ were important factors for catechin production in tea tree. These results can be used as the basis for development of technical soil controls suitable for tea tree cultivation in the future.

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.

Thidiazuron-induced Shoot Formation of Hibiscus syriacus L. 'Honghwarang' by Suspension Culture (Thidiazuron이 무궁화 '홍화랑' 품종 액체 현탁 배양시 신초형성에 미치는 영향)

  • 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.525-527
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    • 1998
  • This study was conducted to determine the optimum cultural condition and method for in vitro mass production of Hibiscus syriacus L. 'Honghwarang'. When callus induced in MS solid medium supplemented with 0.01 mg/L TDZ was cultured in liquid medium containing 0.01mg/L TDZ, callus growth and shoot primordia formation was most effective. Formed shoot primordia were regenerated into shoot in MS or 1/2 MS medium of growth regulator-free condition. Effects of mesh size, shaking speed on callus and shoot primordia formation were examined after 5 weeks. Callus and shoot primordia formation was formed most effectively at 10 mesh and 80 rpm shaking speed in liquid medium.

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Salinity Tolerance of Progenies between Korean Cultivars and IRRI's New Plant Type Lines in Rice

  • Lee, Seung-Yeob;Dharmawansa Senadhira
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.43 no.4
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    • pp.234-238
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    • 1998
  • To select new germplasm for salinity tolerance from new plant type (NPT) breeding lines, the sixty F$_4$ lines selected from the crosses between Korean cultivars and IRRI's NPT lines were evaluated for salinity tolerance at the seedling stage with salinized culture solution (EC=12 dS/m) in the controlled conditions. Two NTP lines derived from a cross between 'Ilmibyeo' and 'IR66152-AC5-1', 'HR15258-7-1' and 'HR15258-27-1', were found to have good tolerance. The salinity tolerance of the lines was compared to their parents and the sensitive ('IR29') and tolerant ('Pokkali') checks in three salinity levels, no salinity (control) and an EC of 12 and 16 dS/m. Visual salinity score, shoot Na+ and Na-K ratio in two NPT lines was significantly low compared with the parents and IR29. Indicating that salinity tolerance of the lines might be derived from a transgressive segregation. The relative water content of the lines was higher than Pokkali, and the dry weight of shoot and root was proportionally decreased to salinity score and salinizing concentration. The visual salinity scores were significantly correlated with shoot Na concentration, Na-K ratio, relative water content, and reduction of dry weight (P<0.01). Their tolerance was attributed to root and shoot characteristics that led to high shoot water content, thus diluting the toxic effect of salts.

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Comparison of Adventitious Shoot Formation in Petiole Explant Cultures of 20 Cultivars of Catharanthus roseus

  • Lee, Soo-Young;Park, Pil-Son;Chung, Hwa-Jee;In, Dong-Soo;Park, Dong-Woog;Jang R. Liu
    • Journal of Plant Biotechnology
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    • v.5 no.1
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    • pp.59-61
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    • 2003
  • Petiole explants from 20 cultivars of Catharanthus roseus were cultured on various shoot-inducing media to assess their competence for adventitious shoot formation. After eight weeks of culture on Murashige and Skoog' s medium supplemented with 4.4 $\mu\textrm{m}$6-benzyladenine and 0.5 $\mu\textrm{m}$ $\alpha$-naphthaleneacetic acid, petiole explants from 'Cooler Icy Pink' exhibited the greatest frequency of adventitious shoot formation at 40%, which was followed by 'Little Bright Eye'. By comparing with a previous study on assessment of competence for adventitious shoot formation in hypocotyl explant cultures of various cultures, it is indicated that the relative degree of their competence among cultivars varies to the organ used for the source of explant. Excised adventitious shoots were readily rooted on half-strength MS basal medium. Regenerated plantlets were successfully transplanted to potting soil and grown to maturity in a greenhouse.