• Title/Summary/Keyword: multiple shoot

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Plant Regeneration and Mutagenesis from Organogenic Callus of Dianthus Distributed in Gangwon Province (강원지역 패랭이꽃속의 캘러스로부터 식물체 재분화와 돌연변이체 유발)

  • Chang, Mi-Young;Hong, Sung-Won;Kim, Joon-Chul
    • Journal of Plant Biotechnology
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    • v.30 no.1
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    • pp.73-80
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    • 2003
  • Useful Dianthus species were collected and selected from two native and seven foreign species distributed in Gangwon province. For in vitro breeding,. callus was induced from the explants of apical meristem, leaf, stem and the in vitro adventitious shoots on MS basal medium with 2.0 mg/L 2,4-D and 0.5 mg/L BA at 27$^{\circ}C$ under continuous light. After 3 weeks of culture, calli initiated the most highly from the leaf explants of D. chinensis Organogenic calli were able to be selected from the adventitious shoot-derived calli. For shoot regeneration, these organogenic calli were cultured on MS medium with the combination of 0.1 mg/L NAA+1.0 mg/L BA under continuous light. Multiple shoots were proliferated with low frequency (about 30%) from those adventitious shootderived calli. Also, shoots initiated directly from the adventitious shoot explants without callus formation at high frequency of 52% when cultured on N6 medium containing 0.1 mg/L NAA and 1.0 mg/L BA in D. gratianopol. Multiple shoots and plantlets grew well and rooted on MS medium supplemented with 0.1 mg/L NAA. Regenerants with well-developed roots were transferred to 8-cm pots containing vermiculite at 85% relative humidity and 27$^{\circ}C$ These plantlets were acclimatized in artificial soil mixture and transferred to the greenhouse for flowering with normal phenotypes. M28 Mutant line was selected with white flowers from 0.03M EMS-treated organogenic calli derived from in vitro adventitious shoot explants of D. chinensis and set seeds.

In Vitro Propagation by Shoot-tip and Node-bud Culture of Rehmannia glutinosa (정단 및 마디조직 배양을 통한 지황의 기내 증식)

  • 백기엽;유광진;박상일
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.1
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    • pp.63-68
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    • 1998
  • Multiple shoots obtained in MS medium suppler with 5.0 mg/L BA though shoot-tip culture. The frequency of vitrified shoot was lower on Bacto-agar medium than on Gelrite as gelling agent. Addition of activated charcoal at concentrations of 0.1~0.3% reduced vitrification and markedly increased shoot growth, and formation and growth of roots, but significantly reduced the number of shoots formed. The ratio of fresh weight to dry weight was decreased by increasing light intensity and agar concentration. Eight-tenths times of macroelement of MS medium was observed to be effective for shoot formation. Addition of IAA effectively promoted shoot formation in both shoot tip and node-bud explants. Supplement of 5.0 mg/L BA, 0.3 mg/L IAA to MS medium was most effective in shoot proliferation on shoot tip and node-bud explants.Multiple shoots obtained in MS medium suppler with 5.0 mg/L BA though shoot-tip culture. The frequency of vitrified shoot was lower on Bacto-agar medium than on Gelrite as gelling agent. Addition of activated charcoal at concentrations of 0.1~0.3% reduced vitrification and markedly increased shoot growth, and formation and growth of roots, but significantly reduced the number of shoots formed. The ratio of fresh weight to dry weight was decreased by increasing light intensity and agar concentration. Eight-tenths times of macroelement of MS medium was observed to be effective for shoot formation. Addition of IAA effectively promoted shoot formation in both shoot tip and node-bud explants. Supplement of 5.0 mg/L BA, 0.3 mg/L IAA to MS medium was most effective in shoot proliferation on shoot tip and node-bud explants.

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Effect of TDZ (Thidiazuron) on Shoot Proliferation of Peace Poplar (Peace 포플러(Populus koreana X P. trichocarpa)의 줄기형성에 미치는 Thidiazuron 효과)

  • Kang, Ho-Duck;Moon, Heung-Kyu;Park, In-Sun;Lee, Min-Soon
    • Journal of Plant Biotechnology
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    • v.31 no.1
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    • pp.49-53
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    • 2004
  • Shoot formation was investigated from in vitro cultivation of exotic hybrid poplar (Populus koreana ${\times}$ P. trichocarpa) with a specific stomatal character occurring both upper and lower surface of leaves. Two different explants (stem and leaf segment) of Peace poplar were cultured on half strength Murashige and Skoog (MS) basal medium supplemented with the various concentrations of thidiazuron as a plant growth regulator. Most adventitious shoots were produced from excised ends of stem or mid-veins of leaf segments. The highest average numbers of shoots were 7.1 and 5.3 with the treatments of 0.02mg/L TDZ in both explants of stem and leaf segment. The highest shooting rates were achieved to 83.3% and 47.6% with the concentrations of 0.01mg/L and 0.02mg/L TDZ by axillary bud and leaf cultures, respectively.

Multiple Shoot Formation of Gentiana axillariflora Leveille by in Vitro Culture (큰용담의 기내증식에서 multiple shoot의 유기)

  • Lim, Jung-Dae;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.8 no.1
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    • pp.41-48
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    • 2000
  • This study was aimed to proliferate Gentiana axillariflora Leveille which is one of the important medicinal and ornamental plants, by establishment of multiple shoot formation and embryogenesis through tissue culture technique. Callus was formed on MS (Murashige and Skoog) medium supplemented with 2,4-D, CPA, but not formed with BAP. The addition of 2,4-D 2 mg/ l into the medium was effective for callus formation and the rate of callus formation was about 90%. Somatic embryos were obtained on MS medium for two months. When callus was cultured on MS medium with combination treatment of 2,4-D 0.5 mg/ l and BAP 0.5 mg/ l, the number of embryo formed was better than that of other single or combination treatments and the total numbers of embryo a were 18.8 (number of total embryo/number of explants incubated = 753/40) at mean. Callus induction from stem and node explants was increased by addition of TDZ 2 mg/ l in the presence of 2,4-D 2 mg/ l, respectively. The best result about the differentiation of shoots was obtained on MS medium added BAP 2 mg/ l from node culture. Multiple shoots from shoot apex were induced on MS medium containing BAP 1 mg/ I and TDZ 1 mg/ l , BAP 2 mg/ l and TDZ 1 mg/ l. The number of multiple shoots per one explant was above seventy plants. It was the most effective regeneration system for rapid multiplication of Gentiana axillariflora Leveille.

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Growth Characteristics of Teratoma Shoot derived from Crown Gall Callus of Nicotiana tabacum cv. NC2326 (연초 Crown Gall Callus 유래 Teratoma Shoot의 생장특성)

  • 양덕춘;최광태
    • Journal of the Korean Society of Tobacco Science
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    • v.13 no.1
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    • pp.36-42
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    • 1991
  • The present study was conducted to obtain some basic information on the shoot formation from crown gall callus and the characteristics of teratoma shoot derived from crown gall induced by inoculation of Agrobacterium tumefaciens C58. Crown gall callus could be continuously cultured on the phyohormone free basic medium. The growth of crown gall callus was inhibited when BA were added to the cultural media. Shoot formation from crown gall callus fail to be initiated except teratoma shoot which induced on the phytohormone free medium after several subculture on rare occasions. Teratoma shoot could not form root and grow as normal shoot. Addition of BA to cultural media was not effective for shoot elongation, reduction in multiple shoot formation, but IBA was somewhat effective for shoot elongation of teratoma shoot, never for root formation.

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Development of Plant Regeneration and Genetic Transformation System from Shoot Apices of Sorghum bicolor (L.) Moench

  • Syamala, D.;Devi, Prathibha
    • Journal of Plant Biotechnology
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    • v.6 no.2
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    • pp.77-85
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    • 2004
  • Development of efficient plant regeneration and genetic transformation protocols (using the Particle Inflow micro-projectile Gun and the shoot-tips as target tissue) of Sorghum bicolor (L.) Moench in terms of expression of the reporter gene, $\beta$-glucuronidase(uidA) is reported here. Two Indian cultivars of sorghum were used in the study, viz. M-35-1 and CSV-15. Plant regeneration was achieved from one-week-old seedling shoot-tip explants via multiple-shoot-clumps and also somatic embryos. The multiple-shoot-clumps were produced on MS medium containing BA (0.5, 1.0 or 2.0 mg/$L^{-1}$), with biweekly subculture. Somatic embryos were directly produced on the enlarged dome shaped expansive structures that developed from shoot-tip explants (without any callus formation) when cultured on MS medium supplemented both with BA (0.5, 1.0 or 2.0 mg/$L^{-1}$) and 2,4-D (0.5 mg/$L^{-1}$). Whereas each multiple-shoot-clump was capable of regenerating more than 80 shoots via an intensive differentiation of both axillary and adventitious shoot buds, the somatic embryos were capable of 90% germination, plant conversion and regeneration. The regenerated shoots could be efficiently rooted on MS medium containing 1.0mg/$L^{-1}$ IBA and successfully transplanted to the glasshouse and grown to maturity with a survival rate of 92%. The plant regeneration efficiency of both the genotypes were similar. After the micro-projectile bombardment, expression of uidA gene was determined by scoring blue transformed cell sectors in the bombarded tissue by an in situ enzyme assay. The optimal conditions comprising a helium pressure of 2200 K Pa, the target distance of 11 cm with helium inlet fully opened and the use of osmoticum have been defined to aid our future strategies of genetic engineering in sorghum with genes for tolerance to biotic and abiotic stresses.

Effect of Phytohormones on Multiple Shoot Bud Induction in cv. NARI-6 of Safflower (Carthamus tinctorius L.)

  • Kumar Jeya-Vijaya;Kumari B.D.Ranjitha
    • Journal of Plant Biotechnology
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    • v.7 no.3
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    • pp.149-153
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    • 2005
  • In the present study, in vitro multiple shoot induction was achieved from cotyledonary node and stem nodal explants of cv. NARI-6 of safflower (Carthamus tinctorius L.). Among various growth regulators tested, MS salts and B5 vitamins supplemented with BA (6-Benzy-laminopurine) $17.76\;{\mu}M$ and KN (Kinetin) $6.96\;{\mu}M$ phytohormonal combination was found to be the most effective in initiating numerous shoot buds after 30 days of culture than BA ($4.44-44.39\;{\mu}M$) or KN ($2.32-46.40\;{\mu}M$) alone in the medium. In addition, 0.8% (w/v) agar (Hi-media) and 3.0% sucrose (w/v) was the optimum level for the formation of adventitious shoots. Further results showed the maximum shoot elongation occurred on MS medium with BA ($8.88\;{\mu}M$) and $GA_3$($11.56\;{\mu}M$) combinations. Efficient rooting occurred on quarter strength MS medium with NAA $10.74\;{\mu}M$. The regenerated plantlets were acclimatized and successfully transferred to the field.

High Frequency Induction of Multiple Shoots from Nodal Explants of Vitex negundo L. Using Sodium Sulphate

  • Chandramu C.;Rao D. Manohar;Reddy V. Dashavantha
    • Journal of Plant Biotechnology
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    • v.5 no.2
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    • pp.107-113
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    • 2003
  • The effect of sodium sulphate on shoot induction and multiple shoot formation from nodal explants of Vitex negundo L. was tested on Murashige and Skoog's (MS) medium fortified with different auxins, cytokinins and sucrose. Highest percentage $(97.78\%)$ of explants for shoot induction and multiple shoot (20.68/explant) production were observed in the combination treatment of $N^6-Benzyl$ adenine (BA) $(17.80\;{\mu}M/L)$, ${\alpha}-Naphthalene$ acetic acid (NAA) $(2.15\;{\mu}M/L)$ and $5\%$ sucrose supplemented with 100 mg/L sodium sulphate. In vitro raised shoots were rooted on the half-strength MS medium fortified with different concentrations of NAA, Indole-3-acetic acid (IAA), and Indole-3-butyric acid (IBA) alone and in combinations. Among the treatments, $4.90\;{\mu}M/L$ of IBA was found most effective $(95.56\%)$ in inducing roots. The rooted plantlets were shifted to glasshouse for acclimatization and later transferred to the field with cent percent survival. Furthermore, in vitro flowering was observed in the shoots cultured on MS medium supplemented with BA $(8.90\;{mu}M/L)$ and NAA $(1.61\;{\mu}M/L)$.

Morphological Development and Histology of Multiple Shoots and Microbulbs of Garlic Cultured in Bioreactors (생물반응기에서 배양된 마늘 신초와 Microbulb의 형태발달 및 조직관찰)

  • Kim, Eun-Kyung;Hahn, Eun-Joo;Paek, Kee-Yoeup
    • Journal of Plant Biotechnology
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    • v.31 no.4
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    • pp.301-306
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    • 2004
  • Multiple shoots of garlic (Allium sativum L.) were propagated in bioreactors containing MS medium supplemented with 2% sucrose for 3 weeks. Microbulbs were induced on MS medium supplemented with 0.1mg/L NAA and 11% sucrose for 9 weeks. For multiple shoot proliferation, leaves in the shoot must be removed before cultures. When the multiple shoots were cultured without removal of leaves, more than 90% of hyperhydricity and no microbulb formation were observed. Histological observation also indicated irregular size and shape of the cells in hyperhydricity of the shoot. Microbulbs were strarted to form from the shoot after 7 weeks of culture by protuberance of adventitious shoot buds followed by inner periclinal divisions and simultaneous anticlinical division in the epidermis of meristematic bulge. Analysis of ploidy level indicated no phenotypic variations in both multiple shoots and microbulbs induced from the mother plant, suggesting genetic homogeneity among the regenerants.

Factors Affecting Efficiency of Shoot Induction in Citrus junos Sieb.

  • Kim, Mi-Hye;Lee, Hyoshin;Chung, Min-Sup;Jinki Jo
    • Journal of Plant Biotechnology
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    • v.3 no.3
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    • pp.141-144
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    • 2001
  • To enhance the shoot induction efficiency from nodal stem of yooza, the culture conditions such as basal medium, carbohydrate source, solidifying agent and the optimum concentration of plant growth regulators for shoot induction were investigated. The nodal explants were cultured better on MS medium than on MT, SH, B5 or W media in considering of shoot induction rate and mean shoot length. Solidifying agent in medium was better with 0.8% agar than with 0.3% agar, 1.2% agarose or 0.2% gelrite. Carbohydrate source in shoot induction medium was efficient with 30 g/L sucrose. The optimum concentrations of plant growth regulators were determined that 0.1 mg/L NAA as auxin was effective on the shoot induction, and 1.0 mg/L BAP as cytokinin induced multiple shoots efficiently. Shoot induction was the most effective on MS medium supplemented with 4 mg/L $GA_3$ in yooza.

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